Topological design, permeability and mechanical behavior of additively manufactured functionally graded porous metallic biomaterials Acta Biomater. (IF 6.383) Pub Date : 2018-12-08 Xiang-Yu Zhang, Gang Fang, Sander Leeflang, Amir A. Zadpoor, Jie Zhou
Recent advances in additive manufacturing (AM) have enabled the fabrication of functionally graded porous biomaterials (FGPBs) for application as orthopedic implants and bone substitutes. Here, we present a step-wise topological design of FGPB based on diamond unit cells to mimic the structure of the femoral diaphysis. The FGPB was manufactured from Ti-6Al-4V powder using the selective laser melting (SLM) technique. The morphological parameters, permeability and mechanical properties of FGPB samples were measured and compared with those of the biomaterials with uniform porous structures based on the same type of the unit cell. The FGPB exhibited a low density (1.9 g/cm3), a moderate Young’s modulus (10.44 GPa), a high yield stress (170.6 MPa), a high maximum stress (201 MPa) and favorable ductility, being superior to the biomaterials with uniform porous structures in comprehensive mechanical properties. In addition, digital image correlation (DIC) and finite element (FE) simulation were used to unravel the mechanisms governing the deformation and yielding behavior of these biomaterials particularly at the strut junctions. Both DIC and FE simulations confirmed that the deformation and yielding of the FGPB occurred largely in the load-bearing layers but not at the interfaces between layers. Defect-coupled FE models based on solid elements provided further insights into the mechanical responses of the FGPB to compressive loads at both macro- and micro-scales. With the defect-coupled representative volume element model for the FGPB, the Young’s modulus and yield stress of the FGPBs were predicted with less than 2% deviations from the experimental data. The study clearly demonstrated the capabilities of combined experimental and computational methods to resolve the uncertainties of the mechanical behavior of FGPBs, which would open up the possibilities of applying various porosity variation strategies for the design of biomimetic AM porous biomaterials. Statement of Significance Functionally graded bone scaffolds significantly promote the recovery of segmental bone defect. In the present study, we present a step-wise topological design of functionally graded porous biomaterial (FGPB) to mimic the structure of the femoral diaphysis. The Ti-6Al-4V FGPB exhibited a superior combination of low density, moderate Young’s modulus, high yield stress and maximum stress as well as favorable ductility. The biomechanical performance of FGPB was studied in both macro and micro perspectives. The defect-coupled model revealed the significant yielding in the load-bearing parts and the Young’s modulus and yield stress of the FGPBs were predicted with less than 2% deviations from the experimental data. The superiority of combined experimental and computational methods has been confirmed.
Mixed-charge pseudo-zwitterionic mesoporous silica nanoparticles with low-fouling and reduced cell uptake properties Acta Biomater. (IF 6.383) Pub Date : 2018-12-06 Noemí Encinas, Mercedes Angulo, Carlos Astorga, Montserrat Colilla, Isabel Izquierdo-Barba, María Vallet-Regí
The design of drug delivery systems needs to consider biocompatibility and host body recognition for an adequate actuation. In this work, mesoporous silica nanoparticles (MSNs) surfaces were successfully modified with two silane molecules to provide mixed-charge brushes (-NH3⊕/-PO3⊝) and well evaluated in terms of surface properties, low-fouling capability and cell uptake in comparison to PEGylated MSNs. The modification process consists in the simultaneous direct-grafting of hydrolysable short chain amino (aminopropyl silanetriol, APST) and phosphonate-based (trihydroxy-silyl-propyl-methyl-phosphonate, THSPMP) silane molecules able to provide a pseudo-zwitterionic nature under physiological pH conditions. Results confirmed that both mixed-charge pseudo-zwitterionic MSNs (ZMSN) and PEG-MSN display a significant reduction of serum protein adhesion and macrophages uptake with respect to pristine MSNs. In the case of ZMSNs, his reduction is up to a 70-90% for protein adsorption and c.a. 60% for cellular uptake. This pseudo-zwitterionic modification has been focused on the aim of local treatment of bacterial infections through the synergistic effect between the inherent antimicrobial effect of mixed-charge system and the levofloxacin antibiotic release profile. These findings open promising future expectations for the effective treatment of bacterial infections through the use mixed-charge pseudo-zwitterionic MSNs furtive to macrophages and with antimicrobial properties.Statement of significanceHerein a novel antimicrobial mixed-charge pseudo-zwitterionic MSNs based system with low-fouling and reduced cell uptake behavior has been developed. This chemical modification has been performed by the simultaneous grafting of short chain organosilanes, containing amino and phosphonate groups, respectively. This nanocarrier has been tested for local infection treatment thought the synergistic effect between the inherent antimicrobial effect of mixed-charge brushes and the levofloxacin antibiotic release profile.
Nanotopographical Regulation of Pancreatic Islet-like Cluster Formation from Human Pluripotent Stem Cells using a Gradient-Pattern Chip Acta Biomater. (IF 6.383) Pub Date : 2018-12-06 Jong hyun Kim, Bo Gi Park, Suel-Kee Kim, Dong-Hyun Lee, Gyung Gyu Lee, Deok-Ho Kim, Byung-Ok Choi, Kyu Back Lee, Jong-Hoon Kim
Bioengineering approaches to regulate stem cell fates aim to recapitulate the in vivo microenvironment. In recent years, manipulating the micro- and nano-scale topography of the stem cell niche has gained considerable interest for the purposes of controlling extrinsic mechanical cues to regulate stem cell fate and behavior in vitro. Here, we established an optimal nanotopographical system to improve 3-dimensional (3D) differentiation of pancreatic cells from human pluripotent stem cells (hPSCs) by testing gradient-pattern chips of nano-scale polystyrene surface structures with varying sizes and shapes. The optimal conditions for 3D differentiation of pancreatic cells were identified by assessing the expression of developmental regulators that are required for pancreatic islet development and maturation. Our results showed that the gradient chip of pore-part 2 (Po-2, 200∼300 nm diameter) pattern was the most efficient setting to generate clusters of pancreatic endocrine progenitors (PDX1+ and NGN3+) compared to those of other pore diameters (Po-1, 100∼200 or Po-3, 300∼400 nm) tested across a range of pillar patterns and flat surfaces. Furthermore, the Po-2 gradient pattern-derived clusters generated islet-like 3D spheroids and tested positive for the zinc-chelating dye dithizone. The spheroids consisted of more than 30% CD200+ endocrine cells and also expressed NKX6.1 and NKX2.2. In addition, pancreatic β- cells expressing insulin and polyhormonal cells expressing both insulin and glucagon were obtained at the final stage of pancreatic differentiation. In conclusion, our data suggest that an optimal topographical structure for differentiation to specific cell types from hPSCs can be tested efficiently by using gradient-pattern chips designed with varying sizes and surfaces.Statement of SignificanceOur study provides demonstrates of using gradient nanopatterned chips for differentiation of pancreatic islet-like clusters.Gradient nanopatterned chips are consisted of two different shapes (nanopillar and nanopore) in three different ranges of nano sizes (100∼200, 200∼300, 300∼400nm). We found that optimal nanostructures for differentiation of pancreatic islet-like clusters were 200∼300 nm nano pores.Cell transplantation is one of the major therapeutic option for type 1 diabetes mellitus (DM) using stem cell-derived β-like cells. We generated 50 um pancreatic islet-like clusters in size, which would be an optimal size for cell transplantation. Futuremore, the small clusters provide a powerful source for cell therapy.Our findings suggest gradient nanopatterned chip provides a powerful tool to generate specific functional cell types of a high purity for potential uses in cell therapy development.
Cartilage Penetrating Cationic Peptide Carriers for Applications in Drug Delivery to Avascular Negatively Charged Tissues Acta Biomater. (IF 6.383) Pub Date : 2018-12-06 Armin Vedadghavami, Erica K. Wagner, Shikhar Mehta, Tengfei He, Chenzhen Zhang, Ambika G. Bajpayee
Drug delivery to avascular, negatively charged tissues like cartilage remains a challenge. The constant turnover of synovial fluid results in short residence time of administered drugs in the joint space and the dense negatively charged matrix of cartilage hinders their diffusive transport. Drugs are, therefore, unable to reach their cell and matrix targets in sufficient doses, and fail to elicit relevant biological response, which has led to unsuccessful clinical trials. The high negative fixed charge density (FCD) of cartilage, however, can be used to convert cartilage from a barrier to drug entry into a depot by making drugs positively charged. Here we design cartilage penetrating and binding cationic peptide carriers (CPCs) with varying net charge, spatial distribution and hydrophobicity to deliver large-sized therapeutics and investigate their electro-diffusive transport in healthy and arthritic cartilage. We showed that CPC uptake increased with increasing net charge up to +14 but dropped as charge increased further due to stronger binding interactions that hindered CPC penetrability and uptake showing that weak-reversible binding is key to enable their penetration through full tissue thickness. Even after 90% GAG depletion, while CPC +14 uptake reduced by over 50% but still had a significantly high value of 148x showing that intra-tissue long-range charge-based binding is further stabilized by short-range H-bond and hydrophobic interactions. The work presents an approach for rational design of cationic carriers based on tissue FCD and properties of macromolecules to be delivered. These design rules can be extended to drug delivery for other avascular, negatively charged tissues.Statement of significance:Osteoarthritis (OA) remains an untreatable disease partly due to short joint residence time of drugs and a lack of delivery methods that can effectively target the dense, avascular, highly negatively charged cartilage tissue. In this study, we designed cartilage penetrating and binding cationic peptide carriers (CPCs) that, due to their optimal charge provide adequate electrical driving force to rapidly transport OA drugs into cartilage and reach their cell and matrix targets in therapeutic doses before drugs exit the joint space. This way cartilage is converted from being a barrier to drug entry into a drug depot that can provide sustained drug release for several weeks. This study also investigates synergistic effects of short-range H-bond and hydrophobic interactions in combination with long-range electrostatic interactions on intra-cartilage solute transport. The work provides rules for rational design of cartilage penetrating charge-based carriers depending on the net charge of tissue (normal versus arthritic), macromolecule to be delivered and whether the application is in drug delivery or tissue imaging.
Antiadhesion effect of the C17 glycerin ester of isoprenoid-type lipid forming a nonlamellar liquid crystal Acta Biomater. (IF 6.383) Pub Date : 2018-12-06 Takahide Murakami, Ichiro Hijikuro, Kota Yamashita, Shigeru Tsunoda, Kenjiro Hirai, Takahisa Suzuki, Yoshiharu Sakai, Yasuhiko Tabata
Postoperative adhesion is a relevant clinical problem that causes a variety of clinical complications after abdominal surgery. The objective of this study is to develop a liquid-type antiadhesion agent and evaluate its efficacy in preventing tissue adhesion in a rat peritoneal adhesion model. The liquid-type agent was prepared by submicron-sized emulsification of C17 glycerin ester (C17GE), squalene, pluronic F127, ethanol, and water with a high-pressure homogenizer. The primary component was C17GE, which is an amphiphilic lipid of one isoprenoid-type hydrophobic chain and can form two phases of self-assembly nonlamellar liquid crystals. The C17GE agent consisted of nanoparticles with an internal inverted hexagonal phase when evaluated by small-angle X-ray scattering (SAXS) and cryo-transmission electron microscopy (cryo-TEM). Upon contact with the biological tissue, this agent formed a thin membrane with a bioadhesive property. After this agent was applied to a sidewall injury of rats, it showed a percentage average of adhesion significantly less than that obtained with the Seprafilm® antiadhesion membrane in a rat model. Additionally, the retention of the agent prolonged at the applied site in the peritoneal cavity of rats. In conclusion, the C17GE agent is promising as an antiadhesion material.Statement of SignificancePostoperative adhesion remains a common adverse effect. Although various materials have been investigated, there are few products commercially available to prevent adhesion. For the sheet-type agent, it is inconvenient to be applied through small laparotomy especially in laparoscopic surgery. Also, the liquid-type agent currently used requires complicated procedure to spray the targeted site.Our liquid-type anti-adhesion agent can form liquid crystal and act as a thin membrane like physical barrier between the peritoneum and tissues to prevent adhesion. Indeed, our agent prevent adhesion significantly compared with the anti-adhesion membrane clinically most used. Moreover, our agent is highly stable by itself and easy to use in laparoscopic surgery, leading to a promising new candidate as an anti-adhesion material.
Repurposing suramin for the treatment of breast cancer lung metastasis with glycol chitosan-based nanoparticles Acta Biomater. (IF 6.383) Pub Date : 2018-12-05 Bei Cheng, Feng Gao, Erica Maissy, Peisheng Xu
Suramin (SM), a drug for African sleeping sickness and river blindness therapy, has been investigated in various clinical trials for cancer therapy. However, SM was eventually withdrawn from the market because of its narrow therapeutic window and the side effects associated with multiple targets. In this work, we developed a simple but effective system based on a nontoxic dose of SM combined with a chemotherapeutic agent for the treatment of metastatic triple-negative breast cancer (TNBC). SM and glycol chitosan (GCS) formed nanogels because of the electrostatic effect, whereas doxorubicin (DOX) was incorporated into the system through the hydrophilic and hydrophobic interactions between DOX and GCS as well as the ionic interactions between DOX and SM to yield GCS-SM/DOX nanoparticles (NPs). GCS-SM/DOX NPs have a size of approximately 186 nm and a spherical morphology. In vitro experiments showed that GCS-SM NPs could effectively inhibit cancer cell migration and invasion, as well as angiogenesis. Furthermore, in a TNBC lung metastasis animal model, GCS-SM/DOX NPs significantly reduced tumor burden and extended the lifespan of animals, while not inducing cardio and renal toxicities associated with the DOX and SM, respectively. As all the components used in this system are biocompatible and easy for large-scale fabrication, the GCS-SM/DOX system is highly translatable for the metastatic breast cancer treatment. Statement of Significance The doxorubicin-loaded glycol chitosan-suramin nanoparticle (GCS-SM/DOX) is novel in the following aspects: SM acts as not only a gelator for the first time in the preparation of the nanoparticle but also an active pharmaceutical agent in the dosage form. GCS-SM/DOX NP significantly reduced tumor burden and extended the lifespan of animals with triple-negative breast cancer lung metastasis. GCS-SM/DOX NPs attenuate cardio and renal toxicities associated with the DOX and SM. The GCS-SM/DOX system is highly translatable because of its simple, one-pot, and easy-to-scale-up preparation protocol.
Micro-scaled topographies direct differentiation of human epidermal stem cells Acta Biomater. (IF 6.383) Pub Date : 2018-12-05 Sebastiaan Zijl, Aliaksei S. Vasilevich, Priyalakshmi Viswanathan, Ayelen Luna Helling, Nick R.M. Beijer, Gernot Walko, Ciro Chiappini, Jan de Boer, Fiona M. Watt
Human epidermal stem cells initiate terminal differentiation when spreading is restricted on ECM-coated micropatterned islands, soft hydrogels or hydrogel-nanoparticle composites with high nanoparticle spacing. The effect of substrate topography, however, is incompletely understood. To explore this, primary human keratinocytes enriched for stem cells were seeded on a topographical library with over 2000 different topographies in the micrometre range. Twenty-four hours later the proportion of cells expressing the differentiation marker Transglutaminase-1 was determined by high content imaging. As predicted, topographies that prevented spreading promoted differentiation. However, we also identified topographies that supported differentiation of highly spread cells. Topographies supporting differentiation of spread cells were more irregular than those supporting differentiation of round cells. Low topography coverage promoted differentiation of spread cells, whereas high coverage promoted differentiation of round cells. Based on these observations we fabricated a topography in 6-well plate format that supported differentiation of spread cells, enabling us to examine cell responses at higher resolution. We found that differentiated spread cells did not assemble significant numbers of hemidesmosomes, focal adhesions, adherens junctions, desmosomes or tight junctions. They did, however, organise the actin cytoskeleton in response to the topographies. Rho kinase inhibition and blebbistatin treatment blocked the differentiation of spread cells, whereas SRF inhibition did not. These observations suggest a potential role for actin polymerization and actomyosin contraction in the topography-induced differentiation of spread cells. Statement of significance The epidermis is the outer covering of the skin. It is formed by layers of cells called keratinocytes. The basal cell layer contains stem cells, which divide to replace cells in the outermost layers that are lost through a process known as differentiation. In this manuscript we have developed surfaces that promote the differentiation of epidermal stem cells in order to understand the signals that control differentiation. The experimental tools we have developed have the potential to help us to devise new treatments that control diseases such as psoriasis and eczema in which epidermal stem cell proliferation and differentiation are disturbed.
Degradable conductive self-healing hydrogels based on dextran-graft-tetraaniline and N-carboxyethyl chitosan as injectable carrier for myoblast cell therapy and muscle regeneration Acta Biomater. (IF 6.383) Pub Date : 2018-12-05 Baolin Guo, Jin Qu, Xin Zhao, Mengyao Zhang
Injectable conductive hydrogels have great potential as tissue engineering scaffolds and delivery vehicles for electrical signal-sensitive cell therapy. In this work, we present the synthesis of a series of injectable electroactive degradable hydrogels with rapid self-healing ability and their potential application as cell delivery vehicles for skeletal muscle regeneration. Self-healable conductive injectable hydrogels based on dextran-graft-aniline tetramer-graft-4-formylbenzoic acid and N-carboxyethyl chitosan were synthesized under physiological conditions. The dynamic Schiff base bonds between the formylbenzoic acid and the amine group of N-carboxyethyl chitosan endowed the hydrogels with rapid self-healing ability, which was verified by the rheological test. Equilibrated swelling ratio, morphology, mechanical strength, electrochemistry, and conductivity of the injectable hydrogels were fully investigated. The self-healable conductive hydrogels showed an in vivo injectability and a linear-like degradation behavior. Two different kinds of cells (C2C12 myoblasts and human umbilical vein endothelial cells (HUVECs)) were encapsulated in the hydrogels by the self-healing effect. The L929 fibroblast cell culture results indicated the biocompatibility of the hydrogels. Moreover, the C2C12 myoblast cells were released from the conductive hydrogels with a linear-like profile. The in vivo skeletal muscle regeneration was also studied in a volumetric muscle loss injury model. All these data indicated that these biodegradable self-healing conductive hydrogels are potential candidates as cell delivery vehicles and scaffolds for skeletal muscle repair. Statement of Significance Injectable hydrogels with self-healing and electrical conductivity are excellent candidates for skeletal muscle tissue engineering scaffolds and myoblast cell therapy. Self-healing property of the hydrogels can prolong their life-time. However, most of the reported conductive hydrogels are not degradable or do not have self-healing ability. We synthesized anti-bacterial conductive self-healing hydrogels as delivery carrier for cardiac cell therapy based on chitosan-grafted-tetraaniline in our previous work. However, acid solution was used to dissolve the polymers that may induce toxicity to cells. In this work, we have synthesized a series of injectable electroactive biodegradable hydrogels with rapid self-healing ability composed of N-carboxyethyl chitosan (CECS) and dextran-graft-aniline oligomers which can dissolve in pH 7.4 PBS solution, and we further demonstrated their potential application as cell delivery vehicles for skeletal muscle regeneration.
Extracellular matrix derived from chondrocytes promotes rapid expansion of human primary chondrocytes in vitro with reduced dedifferentiation Acta Biomater. (IF 6.383) Pub Date : 2018-12-05 Yong Mao, Travis Block, Anya Singh-Varma, Anne Sheldrake, Rachel Leeth, Sy Griffey, Joachim Kohn
A significant expansion of autologous chondrocytes in vitro is required for cell-based cartilage repair. However, the in vitro expansion of chondrocytes under standard culture conditions inevitably leads to the dedifferentiation of chondrocytes and contributes to suboptimal clinical outcomes. To address this challenge, we focused our efforts on developing an improved in vitro expansion protocol, which shortens the expansion time with decreased dedifferentiation. It is known that the tissue microenvironment plays a critical role in regulating the cellular functions of resident cells and provides guidance in tissue-specific regeneration. We hypothesized that chondrocyte extracellular matrix (ECM) mimics a native microenvironment and that it may support chondrocyte expansion in vitro. To test this hypothesis, we prepared decellularized ECMs from allogeneic human articular chondrocytes (HAC) (AC-ECM) and bone marrow stromal cells (BM-ECM) and studied their effects on the in vitro expansion of primary HAC. The differential composition and physical properties of these two ECMs were revealed by mass spectrometry and atomic force microscopy. Compared with standard tissue culture polystyrene (TCP) or BM-ECM, HAC cultured on AC-ECM proliferated faster and maintained the highest ratio of COL2A1/COL1A1. Furthermore, a pellet culture study demonstrated that cells expanded on AC-ECM produced a more cartilage-like ECM than cells expanded on BM-ECM or TCP. This is the first report on modulating chondrocyte expansion and dedifferentiation using cell type-specific ECM and on identifying AC-ECM as a preferred substrate for in vitro expansion of HAC cell-based therapies. Statement of Significance To reduce the dedifferentiation of chondrocytes during in vitro expansion, cell-type specific extracellular matrix (ECM), which mimics a native microenvironment, was prepared from human articular chondrocytes (AC-ECM) or bone marrow stromal cells (BM-ECM). As demonstrated by mass spectrometry and atomic force microscopy, AC-ECM and BM-ECM have differential ECM compositions and physical characteristics. Human articular chondrocytes (HAC) expanded faster and maintained a better chondrocyte phenotype on AC-ECM than on BM-ECM or a standard culture surface. AC-ECM has potential to be developed for expanding HAC for cell-based therapies.
Behavior of valvular interstitial cells on trilayered nanofibrous substrate mimicking morphologies of heart valve leaflet Acta Biomater. (IF 6.383) Pub Date : 2018-12-05 Soumen Jana, Amir Lerman
Heart valve tissue engineering could be an alternative to the current bioprosthetic heart valves and their limitations, especially for pediatric patients. However, heart valve tissue engineering remains challenging because leaflets — the primary component of a heart valve — have three diversely oriented layers — circumferential, random and radial. In order to mimic the orientations, we first designed three novel collectors to fabricate three nanofibrous layers with those orientations from a polymeric biomaterial in an electrospinning system. Then, we devised a novel direct electrospinning technique to develop a unified trilayered nanofibrous (TN) substrate comprising those oriented layers. The TN substrate supported the growth and orientations of seeded porcine valvular interstitial cells (PVICs) and their deposited collagen fibrils. After one month culture, the obtained trilayered tissue construct (TC) exhibited increased tensile properties over its TN substrate. Most importantly, the developed TC did not show any sign of shrinkage. Gene expression pattern of the PVICs indicated the developing stage of the TC. Their protein expression pattern was quite similar to that of leaflets. Statement of Significance This manuscript mainly talks about development of a novel trilayered nanofibrous substrate mimicking the morphologies of a heart valve leaflet. Then it describes the culturing of valvular interstitial cells that reside in the leaflet, in the substrate and compares the behavior of the cultured cells with that in the native leaflets in terms cell morphology, protein deposition and its orientation, and molecular signature. This study builds the groundwork for our future trilayered, tissue-engineered leaflet development. This research article would be of great interest to investigators and researchers in cardiovascular tissue engineering especially in cardiac valve tissue engineering through biomaterial-based tissue engineering.
Guided wave elastography of layered soft tissues Acta Biomater. (IF 6.383) Pub Date : 2018-12-05 Guo-Yang Li, Yang Zheng, Yu-Xuan Jiang, Zhaoyi Zhang, Yanping Cao
In vivo mechanical characterization of soft biological tissues has broad applications ranging from disease diagnosis to tissue engineering. Shear wave elastography based on the bulk wave theory has been widely used to measure the mechanical properties of soft tissues. Given that most soft tissues basically have layered structures, the dispersive feature of elastic waves should be considered when the thickness of the interested layer is comparable to or smaller than the wavelength. Bearing this fundamental issue in mind, we propose an ultrasound-based guided wave elastography (GWE) method to characterize the mechanical properties of layered soft tissues. The dispersion relations of guided waves in layered structures were derived first, and its explicit expression was achieved. An inverse approach based on the dispersion relation to characterize the mechanical properties of layered soft tissues was then established. Both finite element analysis (FEA) and phantom experiments were carried out to validate the new method. In vivo experiments on forearm skin demonstrate the usefulness of the present method in characterizing layered soft tissues. Statement of Significance Layered soft tissues and artificial soft materials are ubiquitous in both nature and engineering. Imaging their in vivo/in situ mechanical properties finds important applications and remains a great challenge to date. Here, we propose an ultrasound-based guided wave elastography method to in vivo characterize the elastic properties of layered soft materials. We validate the method via finite element analysis and phantom experiments and further demonstrate its usefulness in practice by performing in vivo measurements on forearm skins. Given that the dispersive feature of elastic waves in layered soft media is considered in our method, it provides the opportunity to assess the intrinsic elastic properties of an individual layer in a non-destructive manner as shown in our experiments.
Antimicrobial coatings prepared from Dhvar-5-click-grafted chitosan powders Acta Biomater. (IF 6.383) Pub Date : 2018-12-05 Mariana Barbosa, Fabíola Costa, Cláudia Monteiro, Filipa Duarte, M. Cristina L. Martins, Paula Gomes
Antimicrobial peptides (AMP) are powerful components of the innate immune system, as they display wide activity spectrum and low tendency to induce pathogen resistance. Hence, the development of AMP-based coatings is a very promising strategy to prevent biomaterials-associated infections. This work aims to investigate if Dhvar-5-chitosan conjugates, previously synthesized by us via azide-alkyne “click” reaction, can be applied as antimicrobial coatings. Ultrathin coatings were prepared by spin coater after dissolving Dhvar-5-chitosan conjugate powder in aqueous acetic acid. Peptide orientation and exposure from the surface was confirmed by ellipsometry and contact angle measurements. Bactericidal activity was evaluated against Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa, the most prevalent pathogens in implant-associated infections. Results showed that Dhvar-5-chitosan coatings displayed bactericidal effect. Moreover, since Dhvar-5 has head-to-tail amphipathicity, it was clear that the bactericidal potency was dependent on which domain of the peptide (cationic or hydrophobic) was exposed. In this context, Dhvar-5 immobilized through its C-terminus (exposing its hydrophobic end) presented higher antimicrobial activity against Gram-positive bacteria and reduced adhesion of Gram-negative bacteria. This orientation-dependent antimicrobial activity was further corroborated by the anti-biofilm assay, as covalent immobilization of Dhvar-5 through its C-terminus provided anti-biofilm properties to the chitosan thin film. Immobilization of Dhvar-5 showed no cytotoxic effect against HFF-1 cells, as both metabolic activity and cell morphology were similar to control. In conclusion, Dhvar-5-chitosan coatings are promising antimicrobial surfaces without cytotoxic effects against human cells. Statement of significance AMP-tethering onto ground biomaterial is still a poorly explored strategy in research. In this work, AMP-tethered onto ground chitosan is used to produce highly antibacterial ultrathin films. Powdered AMP-tethered chitosan appears as an alternative solution for antimicrobial devices production, as it is suitable for large scale production, being easier to handle for fabrication of different coatings and materials with antimicrobial properties and without inducing toxicity.
Co-delivery of cisplatin and doxorubicin by covalently conjugating with polyamidoamine dendrimer for enhanced synergistic cancer therapy Acta Biomater. (IF 6.383) Pub Date : 2018-12-05 Xue-Ling Guo, Xiao-Xuan Kang, Yue-Qi Wang, Xiao-Jie Zhang, Chang-Jian Li, Yang Liu, Li-Bo Du
Because of the synergistic effects of drugs and minimal drug dose for cancer therapy, combination chemotherapy is frequently used in the clinic. In this study, hyaluronic acid-modified amine-terminated fourth-generation polyamidoamine dendrimer nanoparticles were synthesized for systemic co-delivery of cisplatin and doxorubicin (HA@PAMAM-Pt-Dox). In vitro data showed that HA@PAMAM-Pt-Dox can enter the cells through the lysosome mediated-pathway in a time-dependent manner. Cell viability studies indicated that HA@PAMAM-Pt-Dox exhibited a higher anticancer activity on MCF-7 and MDA-MB-231 breast cancer cells at a relative low concentration. HA@PAMAM-Pt-Dox not only efficiently inhibited tumor growth but also significantly reduced the toxicity of Dox. Moreover, intravenous administration of HA@PAMAM-Pt-Dox to MDA-MB-231 tumor-bearing BALB/c nude mice resulted in the accumulation of HA@PAMAM-Pt-Dox at the tumor site, thereby significantly inhibiting tumor growth without apparent toxicity. These results suggested that HA@PAMAM-Pt-Dox has great potential to improve the chemotherapeutic efficacy of cisplatin and doxorubicin in breast cancer. Statement of Significance One of the main problems in cancer treatment is the development of drug resistance. To date, it is believed that combination chemotherapy might be an effective strategy for the above problem. However, for two completely different drugs, combination chemotherapy faces huge difficulties including the antagonistic nature of drugs, variations in drugs in terms of solubility, and limited tumor targeting. Recent developments in nanoscience and nanotechnology provide an effective approach for such disadvantages. Considering the advantages of dendrimers such as control of size and molecular weight, bioavailability, and biosafety, we used fourth-generation dendrimers modified by HA as drug vectors by covalently conjugating them with anticancer drugs (cisplatin and doxorubicin) to form a nanodrug delivery system, named HA@PAMAM-Pt-Dox. We observed that the HA@PAMAM-Pt-Dox system can effectively kill breast cancer cells both in vitro and in vivo, which showed a favorable synergistic effect. This strategy can be extended to other drugs, thus providing a highly effective strategy for cancer treatment.
Osteoclasts in Bone Regeneration under Type 2 Diabetes Mellitus Acta Biomater. (IF 6.383) Pub Date : 2018-11-30 Zhiai Hu, Chi Ma, Yongxi Liang, Shujuan Zou, Xiaohua Liu
Diabetes mellitus (DM) affects hundreds of million people worldwide and the impaired bone healing is an important DM-related complication. Understanding how DM affects the activities of osteoclasts and the underlying mechanisms is crucial to the development of effective approaches for accelerating bone healing in DM condition. To date, however, the influence of DM on osteoclasts remains obscure and controversial. In this study, we established a type 2 DM (T2DM) alveolar bone defect model, which closely simulates the pathogenesis of human T2DM, to explore the diabetic osteoclast activity during bone regeneration. We found that a high glucose concentration diminished the formation of osteoclasts, and the differentiation and function of osteoclasts from T2DM rats were suppressed. The degradation of matrix by osteoclasts was significantly reduced at a high glucose concentration. In vivo experiments further indicated that T2DM inhibited osteoclastogenesis and osteoclast activity, and delayed the degradation of matrix during the alveolar bone regeneration in T2DM rats. Our work clarifies the influence of T2DM on osteoclasts, and provides valuable insights for the design of novel scaffolding materials that target on osteoclasts for T2DM bone regeneration. Statement of Significance Impaired bone healing is one of the diabetes mellitus (DM)-related complications. Understanding how DM affects osteoclast activity and scaffolding matrix degradation is pivotal to the development of effective approaches for accelerating bone healing in DM condition. Currently, the influences of DM on osteoclast activity and matrix degradation in bone defect areas, however, remain controversial and obscure. Herein, we established a type 2 DM (T2DM) alveolar bone defect model and our results show that T2DM inhibited osteoclastogenesis and osteoclast activity, and delayed the degradation of scaffolding matrix. Our work clarifies the influence of T2DM on osteoclasts and matrix degradation, and provides insights for the design of novel scaffolding materials that target on osteoclasts for T2DM bone regeneration.
Biodegradable amino acid-based poly(ester amine) with tunable immunomodulating properties and their in vitro and in vivo wound healing studies in diabetic rats’ wounds Acta Biomater. (IF 6.383) Pub Date : 2018-11-30 Mingyu He, Luyao Sun, Xiaoling Fu, Sean P. McDonough, Chih-Chang Chu
The objective of this study is to design a new family of biodegradable synthetic polymeric biomaterials for providing a tunable inhibition of macrophage’s nitric oxide synthase (NOS) pathway. L-Arginine (Arg) is the common substrate for NOS and arginase. Both two metabolic pathways participate in the wound healing process. An impaired wound healing, such as diabetic or other chronic wounds is usually associated with an overproduction of NO by macrophages via the NOS pathway. In this study, a new family of L-nitroarginine (NOArg) based polyester amide (NOArg-PEA) and NOArg-Arg PEA copolymers (co-PEA) were designed and synthesized with different composition ratios. The NOArg-PEA and NOArg-Arg co-PEAs are biodegradable (more than 50% degradation in vitro in 4 days at 37 °C), biocompatible and did not activate the resting macrophage immune response per se. When classically activated or alternatively activated macrophages (CAM/AAM) were incubated with NOArg-PEA and NOArg-Arg co-PEAs, the treatments decreased the NO production of CAM, increased the arginase activity in both CAM and AAM, increased TGF-β1 production of CAM to various degrees and had no significant effect on TNF-α production. Diabetic rat models were used to evaluate the efficacy of NOArg-PEA and NOArg-Arg co-PEAs on wound healing. Diabetic rats treated with 2-NOArg-4 PEA, 2-NOArg-4-Arg-4 20/80, and 2-NOArg-4-Arg-4 50/50 biomaterials achieved 40%∼80% faster-wound healing when compared with the control on day 7. The data from the histological and immunohistochemical analysis showed that the 2-NOArg-4-Arg-4 20/80 and 2-NOArg-4-Arg-4 50/50 treatments led to more AAM phenotypes (CD206) and arginase I production in wound tissue than the control during the first 7 days, i.e., suggesting pro-healing wound microenvironment with improved re-epithelialization of wound healing. A similar trend was retained until Day 14. The 2-NOArg-4-Arg-4 20/80 and 2-NOArg-4-Arg-4 50/50 treatments also increased the collagen deposition and angiogenesis in the healing wound between day 7 to day 14. Both in vitro and in vivo data of this study showed that this new family of NOArg-Arg co-PEA biomaterials have the potential as viable alternatives for treating impaired wound healing, such as diabetic or other types of chronic wounds. Statement of significance Diabetic or other chronic wounds is usually associated with an overproduction of NO and pro-inflammatory signals by macrophages. Arginine supplement or NOS inhibitors administration failed to achieve an expected improved wound healing because of the dynamic complexity of arginine catabolism, the difficulty in transition from pro-inflammatory to pro-healing, and the short-term efficacy. We designed and synthesized a new family of water-soluble and degradable nitroarginine-arginine polyester amides to rebalance NOS/arginase metabolism pathways of macrophages. They showed tunable immunomodulating properties in vitro. The in vivo studies were performed to evaluate their efficacy in accelerating the healing. These new biomaterials have the potential as viable alternatives for treating impaired wound healing. The general audience of Acta Biomaterialia should be interested in these findings.
Functionalization of hyaluronic acid hydrogels with ECM-derived peptides to control myoblast behavior Acta Biomater. (IF 6.383) Pub Date : 2018-12-01 Juan Martin Silva Garcia, Alyssa Panitch, Sarah Calve
Volumetric muscle loss (VML) occurs when skeletal muscle injury is too large for the body to fully self-repair. Typically, fibrotic tissue fills the void, which reduces muscle functionality and limb movement. Although a wide variety of natural and synthetic scaffolds have been studied with the purpose of providing the appropriate structural support, to date no scaffold has significantly restored muscle functionality after VML. Satellite cells, adult stem cells within the muscle capable of restoring smaller injuries, are sensitive to the stiffness and composition of the surrounding environment. Scaffolds that only address structural support are not sufficient to restore functionality and instead need to be designed to both promote satellite cell activation and prevent excessive fibroblast recruitment. The objective of this study was to design a scaffold that mimicked the regenerative environment and determine how the biomechanical properties differentially influence myogenic precursor and connective tissue cells. One of the main extracellular matrix (ECM) molecules upregulated during regeneration is hyaluronic acid (HA). Therefore, thiol-modified HA and poly(ethylene glycol) diacrylate hydrogels were generated and functionalized with peptides based on ECM known to influence regeneration, including fibronectin, laminin and tenascin-C. Scaffolds with different stiffness were created by varying HA content. The influence of HA stiffness and peptide functionalization on myogenic precursor and connective tissue cell proliferation, migration and gene expression was quantified. Our results indicated that HA hydrogels functionalized with the laminin peptide, IKVAV, show potential due to the enhanced promotion of myogenic cell behaviors including migration, proliferation and an increase in relevant transcription factors. Statement of Significance The goal of this study was to identify hyaluronic acid (HA) hydrogels with peptide and stiffness combinations that will direct muscle-derived cells towards regenerating phenotypes. While the interaction of skeletal muscle with RGD-functionalized HA hydrogels has been investigated, none of the other peptides described in this study had been used in the context of HA-based scaffolds and skeletal muscle-derived cells. Notably, the response of cells to variations in mechanics was dependent on ECM coating and lineage. The 3% HA functionalized with the laminin peptide, IKVAV, showed the most promise for future in vivo studies, as these hydrogels best promoted myoblast cell proliferation, attachment and spreading, enhanced migration over connective tissue cells and upregulated transcription factors associated with activated satellite cells.
Calcium phosphate nanoparticle-mediated transfection in 2D and 3D mono- and co-culture cell models Acta Biomater. (IF 6.383) Pub Date : 2018-11-29 Viktoriya Sokolova, Leonardo Rojas-Sánchez, Nataniel Białas, Nina Schulze, Matthias Epple
The transfer of nucleic acids into living cells, i.e. transfection, is a major technique in current molecular biology and medicine. As nucleic acids alone are not able to penetrate the cell membrane, an efficient carrier is needed. Calcium phosphate nanoparticles can serve as carrier due to their biocompatibility, biodegradability and high affinity to nucleic acids like DNA or RNA. Their application was extended here from two-dimensional (2D) to three-dimensional (3D) cell culture models, including co-cultures. Compared to 2D monolayer cell cultures, a 3D culture system represents a more realistic spatial, biochemical and cellular environment. The uptake of fluorescent calcium phosphate nanoparticles (diameter 40 to 70 nm; cationic) was studied in 2D and 3D cell culture models by confocal laser scanning microscopy. The transfection of eGFP by calcium phosphate nanoparticles was compared in 2D and 3D cell culture, including co-cultures of green fluorescing HeLa-eGFP cells and MG-63 cells in 2D and in 3D models with the red fluorescent protein mCherry. This permitted a cell-specific assessment of the local transfection efficiency. In general, the penetration of nanoparticles into the spheroids was significantly higher than that of a model oligonucleotide carried by Lipofectamine. The transfection efficiency was comparable in 3D cell cultures with 2D cell cultures, but it occurred preferentially at the surface of the spheroids, following the uptake pathway of the nanoparticles.Statement of significanceThree-dimensional cell culture models can serve as a bridge between the in-vitro cell cultures and the in-vivo situation, especially when mass transfer effects have to be considered. This is the case for nanoparticles where the incubation effect in a two-dimensional cell culture strongly differs from a three-dimensional cell culture or a living tissue. We have compared the uptake of nanoparticles and a subsequent transfection of fluorescent proteins in two-dimensional and three-dimensional cell culture models. An elegant model to investigate the transfection in co-cultures was developed using HeLa-eGFP cells (green fluorescent) together with MG-63 cells (non-fluorescent) that were transfected with the red-fluorescing protein mCherry. Thereby, the transfection of both cell types in the co-culture was easily distinguished.
Stimuli-Responsive Polymer-Doxorubicin Conjugate: Antitumor Mechanism and Potential as Nano-Prodrug Acta Biomater. (IF 6.383) Pub Date : 2018-11-29 Kai Chen, Hao Cai, Hu Zhang, Hongyan Zhu, Zhongwei Gu, Qiyong Gong, Kui Luo
Polymer-drug conjugates has significantly improved the anti-tumor efficacy of chemotherapeutic drugs and alleviated their side effects. N-(1,3-dihydroxypropan-2-yl) methacrylamide (DHPMA) copolymer was synthesized via RAFT polymerization and polymer-doxorubicin (DOX) (diblock pDHPMA-DOX) were formed by conjugation, resulting in a self-aggregation-induced nanoprodrug with a favorable size of 21 nm and great stability. The nanoprodrug with a molecular weight (MW) of 95 kDa released drugs in response to tumor microenvironmental pH variations and they were enzymatically hydrolyzed into low MW segments (45 kDa). The nanoprodrug was transported through the endolysosomal pathway, released the drug into the cytoplasm and some was localized in the mitochondria, resulting in disruption of the cellular actin cytoskeleton. Cellular apoptosis was also associated with reduction in the mitochondrial potential caused by the nanoprodrug. Notably, the nanoprodrug had a significantly prolonged blood circulation time with an elimination half time of 9.8 h, displayed high accumulation within tumors, and improved the in vivo therapeutic efficacy against 4T1 xenograft tumors compared to free DOX. The tumor xenograft immunohistochemistry study clearly indicated tumor inhibition was through the inhibition of cell proliferation and antiangiogenic effects. Our studies demonstrated that the diblock pDHPMA-DOX nanoprodrug with a controlled molecular structure is promising to alleviate adverse effects of free DOX and have a great potential as an efficient anticancer agent.Statement of significanceIn this work, we prepared a biodegradable diblock DHPMA polymer-doxorubicin conjugate via one-pot of RAFT polymerization and conjugate chemistry. The conjugate-based nanoprodrug was internalized by endocytosis to intracellularly release DOX and further induce disruption of mitochondrial functions, actin cytoskeleton alterations and cellular apoptosis. The nanoprodrug with a high molecular weight (MW) (95 kDa) showed a long blood circulation time and achieved high accumulation into tumors. The nanoprodrug was degraded into low MW (∼45 kDa) products below the renal threshold, which ensured its biosafety. Additionally, the multi-stimuli-responsive nanoprodrug demonstrated an enhanced antitumor efficacy against 4T1 breast tumors and alleviated side effects, showing a great potential as an efficient and safe anticancer agent.
A versatile strategy to create active tumor-targeted chemo-photothermal therapy nanoplatform: a case of IR-780 derivative co-assembled with camptothecin prodrug Acta Biomater. (IF 6.383) Pub Date : 2018-11-29 Wenxiu He, Yue Jiang, Qian Li, Di Zhang, Zhonghao Li, Yuxia Luan
Self-assembled nanovehicles of chemotherapy drug with photothermal agent are regarded as intriguing chemo-photothermal therapy nanoplatform. However, most of the drugs and photothermal agents have poor water solubility and poor interactions to drive the formation of self-assembled nanovehicles, which is a bottleneck of co-assembled drug/photothermal agent for cancer therapy. Here, we proposed a versatile strategy to create self-assembled chemo-photothermal therapy nanoplatform based on the chemical modification of photothermal agent and drug. The IR-780 and camptothecin (CPT) were chosen as the studied models since they are important photothermal agent and anticancer drug, both of which have such poor water solubility with strong itself molecular interactions that they cannot co-assemble together. IR-780 was modified with an active targeting ligand lactobionic acid (LA) to result in amphiphilic IR780-LA while CPT was modified into redox-sensitive prodrug CPT-ss-CPT through a disulfide linkage to realize its assembly. Well-defined nanoparticles (NPs) could be created through the co-assembling of IR780-LA and CPT-ss-CPT. The IR780-LA/CPT-ss-CPT nanoparticles were demonstrated to be an excellent fluorescence imaging-guided, redox-responsive and enhanced synergistic chemo-photothermal therapy nanoplatform against tumors. Specifically, our chemical modification strategy offers a universal way to create self-assembled chemo-photothermal therapy nanoplatform, which solves the bottleneck of co-assembled drug/photothermal agent for cancer therapy.Statement of SignificanceSelf-assembled nanoparticles of chemotherapeutics with photothermic drugs are regarded as intriguing chemo-photothermal therapy nanoplatform. However, most drugs have too poor solubility and interactions to form into self-assembled nanoparticles. We proposed a versatile strategy to create co-assembled chemo-photothermal therapy nanoparticles based on the chemical modification of common drugs. The IR-780 was modified with an active targeting ligand LA to result in amphiphilic IR780-LA molecules, while CPT was modified into redox-sensitive prodrug CPT-ss-CPT through disulfide linkage. Well-defined IR780-LA/CPT-ss-CPT nanoparticles were created through the co-assembling of IR780-LA and CPT-ss-CPT. The nanoparticles were demonstrated to be an excellent fluorescence imaging-guided, redox-responsive, active targeting chemo-photothermal therapy nanoplatform against tumors. Our strategy offers a versatile way to construct smart chemo-photothermal therapy nanoplatform from common drugs.
3D collagen microfibers stimulate the functionality of preadipocytes and maintain the phenotype of mature adipocytes for long term cultures Acta Biomater. (IF 6.383) Pub Date : 2018-11-28 Fiona Louis, Shiro Kitano, João F. Mano, Michiya Matsusaki
Although adipose tissue is one of the most abundant tissues of the human body, its reconstruction remains a competitive challenge. The conventional in vitro two or three-dimensional (2D or 3D) models of mature adipocytes unfortunately lead to their quick dedifferentiation after one week, and complete differentiation of adipose derived stem cells (ADSC) usually requires more than one month. In this context, we developed biomimetic 3D adipose tissues with high density collagen by mixing type I collagen microfibers with primary mouse mature adipocytes or human ADSC in transwells. These 3D-tissues ensured a better long-term maintained phenotype of unilocular mature adipocytes, compared to 2D, with a viability of 96 ± 2% at day 14 and a good perilipin immunostaining,- the protein necessary for stabilizing the fat vesicles. For comparison, in 2D culture, mature adipocytes released their fat until splitting their single adipose vesicle into several ones with significantly 4 times smaller size. Concerning ADSC, the adipogenic genes expression in 3D-tissues was found at least doubled throughout the differentiation (over 8 times higher for GLUT4 at day 21), along with it, almost 4 times larger fat vesicles were observed (10 ± 4 µm at day 14). Perilipin immunostaining and leptin secretion, the satiety protein, attested the significantly doubled better functionality of ADSC in 3D adipose tissues. These obtained long-term maintained phenotype and fast adipogenesis make this model relevant for either cosmetic/pharmaceutical assays or plastic surgery purposes.Statement of significanceAdipose tissue has important roles in our organism, providing energy from its lipids storage and secreting many vital proteins. However, its reconstruction in a functional in vitro adipose tissue is still a challenge. Mature adipocytes directly extracted from surgery liposuctions quickly lose their lipids after a week in vitro and the use of differentiated adipose stem cells is too time-consuming. We developed a new artificial fat tissue using collagen microfibers. These tissues allowed the maintenance of viable big unilocular mature adipocytes up to two weeks and the faster adipogenic differentiation of adipose stem cells. Moreover, the adipose functionality confirmed by perilipin and leptin assessments makes this model suitable for further applications in cosmetic/pharmaceutical drug assays or for tissue reconstruction.
Biomimetic porous Mg with tunable mechanical properties and biodegradation rates for bone regeneration Acta Biomater. (IF 6.383) Pub Date : 2018-11-27 Min-Ho Kang, Hyun Lee, Tae-Sik Jang, Yun-Jeong Seong, Hyoun-Ee Kim, Young-Hag Koh, Juha Song, Hyun-Do Jung
The medical applications of porous Mg scaffolds are limited owing to its rapid corrosion, which dramatically decreases the mechanical strength of the scaffold. Mimicking the bone structure and composition can improve the mechanical and biological properties of porous Mg scaffolds. The Mg structure can also be coated with HA by an aqueous precipitation coating method to enhance both the corrosion resistance and the biocompatibility. However, due to the brittleness of HA coating layer, cracks tend to form in the HA coating layer, which may influence the corrosion and biological functionality of the scaffold. Consequently, in this study, hybrid poly(ether imide) (PEI)–SiO2 layers were applied to the HA-coated biomimetic porous Mg to impart the structure with the high corrosion resistance associated with PEI and excellent bioactivity with SiO2. The porosity of the Mg was controlled by adjusting the concentration of the sodium chloride (NaCl) particles used in the fabrication via the space-holder method. The mechanical measurements showed that the compressive strength and stiffness of the biomimetic porous Mg increased as the portion of the dense region increased. In addition, following results show that HA/(PEI–SiO2) hybrid-coated biomimetic Mg is a promising biodegradable scaffold for orthopedic applications. In-vitro testing revealed that the proposed hybrid coating reduced the degradation rate and facilitated osteoblast spreading compared to HA- and HA/PEI-coating scaffolds. Moreover, in-vivo testing with a rabbit femoropatellar groove model showed improved tissue formation, reduced corrosion and degradation, and improved bone formation on the scaffold.Statement of significancePorous Mg is a promising biodegradable scaffold for orthopedic applications. However, there are limitations in applying porous Mg for an orthopedic biomaterial due to its poor mechanical properties and susceptibility to rapid corrosion. Here, we strategically designed the structure and coating layer of porous Mg to overcome these limitations. First, porous Mg was fabricated by mimicking the bone structure which has a combined structure of dense and porous regions, thus resulting in an enhancement of mechanical properties. Furthermore, the biomimetic porous Mg was coated with HA/(PEI-SiO2) hybrid layer to improve both corrosion resistance and biocompatibility. As the final outcome, with tunable mechanical and biodegradable properties, HA/(PEI-SiO2)-coated biomimetic porous Mg could be a promising candidate material for load-bearing orthopedic applications.
Multifunctional Nanotheranostic Gold Nanocages for Photoacoustic Imaging Guided Radio/Photodynamic/Photothermal Synergistic Therapy Acta Biomater. (IF 6.383) Pub Date : 2018-11-27 Xiaoyu Xu, Yu Chong, Xiaoyun Liu, Han Fu, Chenggong Yu, Jie Huang, Zhijun Zhang
In this work, we developed a novel multifunctional nanoplatform based on hyaluronic acid modified Au nanocages (AuNCs-HA). The rational design of AuNCs-HA renders the nanoplatform three functionalities: (1) AuNCs-HA with excellent LSPR peak in the NIR region act as contrast agent for enhanced photoacoustic (PA) imaging and photothermal therapy (PTT); (2) the nanoplatform with high-energy rays (X-ray) absorption and auger electrons generation acts as a radiosensitizer for radiotherapy; (3) good photocatalytic property and large surface area make AuNCs-HA a photosensitive agent for photodynamic therapy (PDT). In vivo results demonstrated that AuNCs-HA presented excellent PA imaging performance after intravenous injection, which provided contour, size, and location information of the tumor. Moreover, because AuNCs-HA could combine radiotherapy and phototherapy together, the tumors treated with AuNCs-HA showed complete growth inhibition, comparing to that with each therapy alone. Taken together, our present study demonstrates that AuNCs-HA is of great potential as a multifunctional nanoplatform for PA imaging-guided radio- and photo-therapy of tumor.Statement of significanceIn this study, a commendable theranostic nanoplatform based on hyaluronic acid modified AuNCs (AuNCs-HA) was developed. In our approach, the dilute solution of Gold(III) chloride is slowly dripped into Ag nanocubes solution, then the Au nanocages were obtained by redox reaction, and followed by HA modification. We explored them, simultaneously, as radiosensitizers for RT, photosensitizers for PDT, and therapeutic agents for PTT. Compared to that of each therapies alone, the combination of radio-therapy and photo-therapy results in a considerably improved tumor eliminating effect and efficiently inhibited tumor growth. In addition, AuNCs-HA exhibited remarkably strong PA signals for precise identification of the location, size, and boundary of the tumor, thereby facilitating imaging-guided therapy. In brief, our design of AuNCs-HA represents a general and versatile strategy for building up cancer-targeted nanotheranostics with desired synergistic imaging and therapy functionalities.
Meshes in a mess: mesenchymal stem cell-based therapies for soft tissue reinforcement Acta Biomater. (IF 6.383) Pub Date : 2018-11-27 F. Marinaro, F.M. Sánchez-Margallo, V. Álvarez, E. López, R. Tarazona, M.V. Brun, R. Blázquez, J.G. Casado
Surgical meshes are frequently used for the treatment of abdominal hernias, pelvic organ prolapse, and stress urinary incontinence. Though these meshes are designed for tissue reinforcement, many complications have been reported. Both differentiated cell- and mesenchymal stem cell-based therapies have become attractive tools to improve their biocompatibility and tissue integration, minimizing adverse inflammatory reactions. However, current studies are highly heterogeneous, making it difficult to establish comparisons between cell types or cell coating methodologies. Moreover, only a few studies have been performed in clinically relevant animal models, leading to contradictory results. Finally, a thorough understanding of the biological mechanisms of mesenchymal stem cells in the context of foreign body reaction is lacking. This review aims to summarize in vitro and in vivo studies involving the use of differentiated and mesenchymal stem cells in combination with surgical meshes. According to preclinical and clinical studies and considering the therapeutic potential of mesenchymal stem cells, it is expected that these cells will become valuable tools in the treatment of pathologies requiring tissue reinforcement.Statement Of SignificanceThe implantation of surgical meshes is the standard procedure to reinforce tissue defects such as hernias. However, an adverse inflammatory response secondary to this implantation is frequently observed, leading to a strong discomfort and chronic pain in the patients. In many cases, an additional surgical intervention is needed to remove the mesh.Both differentiated cell- and stem cell-based therapies have become attractive tools to improve biocompatibility and tissue integration, minimizing adverse inflammatory reactions. However, current studies are incredibly heterogeneous and it is difficult to establish a comparison between cell types or cell coating methodologies. This review aims to summarize in vitro and in vivo studies where differentiated and stem cells have been combined with surgical meshes.
Early stage mechanical remodeling of collagen surrounding head and neck squamous cell carcinoma spheroids correlates strongly with their invasion capability Acta Biomater. (IF 6.383) Pub Date : 2018-11-27 Yin-Quan Chen, Jean-Cheng Kuo, Ming-Tzo Wei, Yen-Chih Chen, Muh-Hwa Yang, Arthur Chiou
Mechanical remodeling of stromal collagen, such as reorientation and deformation of collagen matrix, generated by invading cancer cells, plays an important role in the progression of cancer invasion and metastasis. In this study, we applied time-lapse microscopy in conjunction with particle displacement mapping to analyze time-dependent contraction and expansion deformations of collagen surrounding individual spheroids of head and neck squamous cell carcinoma cells (HNSCC), OECM-1 & SAS, as the cancer cells detached from the spheroid and invaded into the surrounding 3D collagen matrix. Our results revealed that highly-invasive HNSCC spheroids, stimulated by epidermal growth factor (EGF), generated a strong contraction deformation of the surrounding collagen in the very early stage, and aligned the collagen fibers radially with respect to the center of the spheroid. This initial collagen contraction deformation generated by the HNSCC spheroid bears a strong positive correlation with the overall extent of subsequent cancer cells invasion; hence, it may serve as an early indicator of the invasion capability of the HNSCC spheroids.Statement of significanceMechanical remodeling of extracellular matrix (ECM) generated by cancer cells plays an important role in the progression of cancer invasion and metastasis. We observed that the extent of collagen initial contraction deformation surrounding a head and neck squamous cell carcinoma cell (HNSCC) spheroid play an indispensable role in early stage to promote cancer cells invasion into the surrounding ECM. Our results revealed that more invasive HNSCC spheroids generated a larger extent of initial collagen contraction to align the surrounding collagen and to promote cancer cells invasion. This initial collagen contraction deformation generated by the HNSCC spheroids bears a strong positive correlation with the overall extent of cancer cells invasion; hence, it may serve as an early indicator of the invasion capability of the HNSCC spheroids.
Compositionally Graded Doped Hydroxyapatite Coating using Laser and Plasma Spray Deposition Acta Biomater. (IF 6.383) Pub Date : 2018-11-27 Dongxu Ke, Ashley A. Vu, Amit Bandyopadhyay, Susmita Bose
Plasma sprayed hydroxyapatite (HA) coating is known to improve the osteoconductivity of metallic implants. However, the adhesive bond strength of the coating is affected due to a mismatch in coefficients of thermal expansion (CTE) between the metal and the HA ceramic. In this study, a gradient HA coating was prepared on Ti6Al4V by laser engineered net shaping (LENSTM) followed by plasma spray deposition. In addition, 1 wt.% MgO and 2 wt.% Ag2O were mixed with HA to improve the biological and antibacterial properties of the coated implant. Results showed that the presence of an interfacial layer by LENSTM enhanced adhesive bond strength from 26 ± 2 MPa for just plasma spray coating to 39 ± 4 MPa for LENSTM and plasma spray coatings. Presence of MgO and Ag2O did not influence the adhesive bond strength. Also, Ag+ ions release dropped by 70% less with a gradient HA LENSTM layer due to enhanced crystallization of the HA layer. In vitro human osteoblast cell culture revealed presence of Ag2O had no deleterious effect on proliferation and differentiation when compared to pure HA as control and provided antibacterial properties against E. coli and S. aureus bacterial strands. This study presents an innovative way to improve interfacial mechanical properties of plasma sprayed HA coating for load-bearing orthopedic implants.Statement of SignificanceImplants are commonly composed of metals that lack osteoconductivity. Osteoconductivity is a propriety where bone grows on the surface meaning the material is compatible with the surrounding bone tissue. Plasma sprayed hydroxyapatite (HA) coating improves the osteoconductivity of metallic implants however the adhesive bond strength can be weak. This study incorporates a gradient HA coating by using an additive manufacturing technique, laser engineered net shaping (LENSTM), followed by plasma spray deposition to enhance the adhesive bond strength by incorporating a thermal barrier. The proposed system has not been well studied in the current literature and the results presented bring forth an innovative way to improve the interfacial mechanical properties of plasma sprayed HA coating for load-bearing orthopedic implants.
Bone-like features in skate suggest a novel elasmobranch synapomorphy and deep homology of trabecular mineralization patterns Acta Biomater. (IF 6.383) Pub Date : 2018-11-27 Oghenevwogaga J. Atake, David M.L. Cooper, B. Frank Eames
Bone is a defining characteristic of the vertebrate skeleton, and while chondrichthyans (sharks, skates, and other cartilaginous fishes) are vertebrates, they are hypothesized to have lost the ability to make bone during their evolution. Multiple descriptions of a bone-like tissue in neural arches of vertebrae in various shark species (selachians), however, challenge this hypothesis. Here, we extend this argument by analyzing vertebrae of two members of the batoids (the little skate Leucoraja erinacea, and Eaton’s skate Bathyraja eatonii), the sister group to selachians within elasmobranchs. Micro-CT images showed a bone-like mineralization pattern in neural arches of each skate species, and histological analyses confirmed that this bone-like tissue surrounded a cartilage core, exactly as described in sharks. Another mineralization pattern identified in skate vertebrae was distinct from the polygonal tesseral and areolar patterns that classically are associated with the chondrichthyan endoskeleton. Many regions of the vertebrae, including the neural spine and transverse processes, showed this perichondral mineralization pattern, termed here trabecular tesseral. Other than the cartilage core of the neural arch, all mineralized tissues in skate vertebrae had flattened cells surrounded by matrix with bone-like histology. Analyses of quantitative microstructural parameters revealed that, compared to rat vertebrae, the bone-like mineralization pattern in the neural arches of skate vertebrae was more similar to compact bone than trabecular bone. In contrast, the thickness of the trabecular tesseral pattern was more similar to trabecular bone than compact bone of rat vertebrae. In conclusion, a bone-like tissue in neural arches of skate vertebrae appears to be a novel elasmobranch synapomorphy. We propose that the trabecular tesseral mineralization pattern in the skate might have deep homology to the mineralization pattern utilized in trabecular bone.Statement of significanceMineralization patterns of skeletal tissues have not been investigated thoroughly in all vertebrate clades. Despite their designation as ‘cartilaginous fish’, chondrichthyans clearly evolved from ancestral vertebrates that made bone. The consensus that chondrichthyans lost the ability to make bone during their evolution, however, is challenged by reports of bone and bone-like tissues in the neural arches of vertebrae in extant sharks (selachians). Here, we provide evidence from micro-CT imaging and histological analyses to support our hypothesis that a bone-like tissue is present in the neural arches of batoids (the sister group to selachians within elasmobranchs). These results argue strongly that the neural arch bone-like tissue is a previously unknown synapomorphy of elasmobranchs. In addition to the bone-like mineralization pattern identified in the neural arches, micro-CT images also showed a novel mineralization pattern which we described as trabecular tesseral. Quantitative microstructural features shared between trabecular tesseral pattern and trabecular bone (from homologous rat vertebrae) suggest that both patterns might derive from an ancestral gene network driving trabecular mineralization (i.e., deep homology).
Tissue engineered hydrogels supporting 3D neural networks Acta Biomater. (IF 6.383) Pub Date : 2018-11-27 Ulises A Aregueta-Robles, Penny J Martens, Laura A Poole-Warren, Rylie A Green
Promoting nerve regeneration requires engineering cellular carriers to physically and biochemically support neuronal growth into a long lasting functional tissue. This study systematically evaluated the capacity of a biosynthetic poly(vinyl alcohol) (PVA) hydrogel to support growth and differentiation of co-encapsulated neurons and glia. A significant challenge is to understand the role of the dynamic degradable hydrogel mechanical properties on expression of relevant cellular morphologies and function. It was hypothesised that a carrier with mechanical properties akin to neural tissue will provide glia with conditions to thrive, and that glia in turn will support neuronal survival and development. PVA co-polymerised with biological macromolecules sericin and gelatin (PVA-SG) and with tailored nerve tissue-like mechanical properties were used to encapsulate Schwann cells (SCs) alone and subsequently a co-culture of SCs and neural-like PC12s. SCs were encapsulated within two PVA-SG gel variants with initial compressive moduli of 16 kPa and 2 kPa, spanning a range of reported mechanical properties for neural tissues. Both hydrogels were shown to support cell viability and expression of extracellular matrix proteins, however, SCs grown within the PVA-SG with a higher initial modulus were observed to present with greater physiologically relevant morphologies and increased expression of extracellular matrix proteins. The higher modulus PVA-SG was subsequently shown to support development of neuronal networks when SCs were co-encapsulated with PC12s. The lower modulus hydrogel was unable to support effective development of neural networks. This study demonstrates the critical link between hydrogel properties and glial cell phenotype on development of functional neural tissues.Statement of significanceHydrogels as platforms for tissue regeneration must provide encapsulated cellular progenitors with physical and biochemical cues for initial survival and to support ongoing tissue formation as the artificial network degrades. While most research focuses on tailoring scaffold properties to suit neurons, this work aims to support glia SCs as the key cellular component that physically and biochemically supports the neuronal network. The challenge is to modify hydrogel properties to support growth and development of multiple cell types into a neuronal network. Given SCs ability to respond to substrate mechanical properties, the significance of this work lies in understanding the relationship between dynamic hydrogel mechanical properties and glia SCs development as the element that enables formation of mature, differentiated neural networks.
Cell Armor for Protection Against Environmental Stress: Advances, Challenges and Applications in Micro- and Nanoencapsulation of Mammalian Cells Acta Biomater. (IF 6.383) Pub Date : 2018-11-24 Onur Hasturk, David L. Kaplan
Unlike unicellular organisms and plant cells surrounded with a cell wall, naked plasma membranes of mammalian cells make them more susceptible to environmental stresses encountered during in vitro biofabrication and in vivo cell therapy applications. Recent advances in micro- and nanoencapsulation of single mammalian cells provide an effective strategy to isolate cells from their surroundings and protect them against harsh environmental conditions. Microemulsification and droplet-based microfluidics have enabled researchers to encapsulate single cells within a variety of microscale hydrogel materials with a range of biochemical and mechanical properties and functionalities including enhanced cell-matrix interactions or on-demand degradation. In addition to microcapsules, nanocoatings of various organic and inorganic substances on mammalian cells have allowed for the formation of protective shells. A wide range of synthetic and natural polymers, minerals and supramolecular metal-organic complexes have been deposited as nanolayers on the cells via electrostatic interactions, receptor-ligand binding, non-specific interactions, and in situ polymerization/crosslinking. Here, current strategies in encapsulation of single mammalian cells along with challenges and advances are reviewed. Protection of encapsulated stem cells, fibroblasts, red and white blood cells and cancer cells against harsh in vitro and in vivo conditions including anoikis, UV radiation, physical forces, proteolytic enzymes and immune clearance are discussed. Statement of Significance The mechanical fragility of the plasma membrane and susceptibility to extracellular biochemical factors due to the lack of a physical barrier like a tough cell wall or exoskeleton make mammalian cells extra sensitive to harsh environmental conditions. This sensitively, in turn, limits the ex vivo storage, handling and manipulation of mammalian cells, as well as their in vivo applications. Environmental stresses such as exposure to UV, reactive chemicals and mechanical stress during biofabrication processes like 3D bioprinting can often compromise cell viability and function. Micro- and nanoencapsulation of single mammalian cells in protective shells have emerged as promising approaches to isolate cells from their surroundings and enhance resistance against perturbations in conditions during regenerative medicine and tissue engineering applications. In this review, the current state of art of single cell encapsulation strategies and the challenges associated with these technologies are discussed in detail. This is followed by the review of the protection provided by cell armor against a range of harsh in vitro and in vivo conditions.
Three-dimensional (3D) Printed Scaffold and Material Selection for Bone Repair Acta Biomater. (IF 6.383) Pub Date : 2018-11-24 Lei Zhang, Guojing Yang, Blake N. Johnson, Xiaofeng Jia
Critical-sized bone defect remains a substantial challenge in clinical settings and requires bone grafts or bone substitute materials. However, existing biomaterials often do not meet the clinical requirements of structural support, osteoinductive property, and controllable biodegradability. To treat large-scale bone defects, the development of three-dimensional (3D) porous scaffolds has received considerable focus within bone engineering. A variety of biomaterials and manufacturing methods including 3D printing have emerged to fabricate patient-specific bioactive scaffolds that possess controlled micro-architectures for bridging bone defects in complex configurations. During the last decade, with the development of the 3D printing industry, a large number of tissue-engineered scaffolds have been created for preclinical or clinical applications using novel materials and innovated technologies. Thus, this review provides a brief overview of current advances in existing biomaterials and tissue engineering scaffolds prepared by 3D printing technologies, with an emphasis on the material selection, scaffold design optimization, and their preclinical or clinical applications in the repair of critical-sized bone defects. Furthermore, it will elaborate the current limitations and potential future prospects of 3D printing technology. Statement of Significance 3D printing has emerged as a critical fabrication process for bone engineering due to its ability to control bulk geometry and internal structure of tissue scaffolds. The advancement of bioprinting methods and compatible ink materials for bone engineering have been a major focus to develop optimal 3D scaffolds for bone defect repair. Achieving a successful balance between the properties of a scaffold favorable to cellular function, cellular viability, and mechanical integrity under load-bearing conditions is critical. Hybridization of natural and synthetic polymer-based materials is a promising approach to create novel tissue engineered scaffolds that combines the advantages of both materials and meets various requirements, including biological activity, mechanical strength, easy fabrication and controllable degradation. 3D printing is linked to the future of bone grafts to create on-demand patient-specific grafts.
Gentle cyclic straining of human fibroblasts on electrospun scaffolds enhances their regenerative potential Acta Biomater. (IF 6.383) Pub Date : 2018-11-22 Mahshid Vashaghian, Chantal M. Diedrich, B. Zandieh-Doulabi, A. Werner, Theodoor H. Smit, Jan-Paul Roovers
The extracellular matrix of fascia-like tissues is a resilient network of collagenous fibers that withstand the forces of daily life. When overstretched, the matrix may tear, with serious consequences like pelvic organ prolapse (POP). Synthetic implants can provide mechanical support and evoke a host response that induces new matrix production, thus reinforcing the fascia. However, there is considerable risk of scar formation and tissue contraction which result in severe complications. Matrix producing fibroblasts are both mechanosensitive and contractile; their behavior depends on the implant’s surface texture and mechanical straining. Here we investigate the effect of both in a newly-designed experimental setting. Electrospun scaffolds of Nylon and PLGA/PCL and a non-porous PLGA/PCL film were clamped like a drumhead and seeded with fibroblasts of POP patients. Upon confluency, scaffolds were cyclically strained for 24 or 72h at 10% and 0.2Hz, mimicking gentle breathing. Non-loading condition was control. Strained fibroblasts loosened their actin-fibers, thereby preventing myofibroblastic differentiation. Mechanical loading upregulated genes involved in matrix synthesis (collagen I, III, V and elastin), matrix remodeling (α-SMA, TGF-β1, MMP-2) and inflammation (COX-2, TNF-α, IL8, IL1-β). Collagen genes were expressed earlier under mechanical loading and the ratio of I/III collagen increased. Matrix synthesis and remodeling were stronger on the electrospun scaffolds, while inflammation was more prominent on the non-porous film. Our findings indicate that mechanical straining enhances the regenerative potential of fibroblasts for the regeneration of fascia-type tissues and limit the risk of scar tissue formation. These effects are stronger on an electrospun texture. Statement of significance Pelvic organ prolapsed is a dysfunctional disease in female pelvic floor that can reduce the quality of life women. Currently, trans-vaginal knitted meshes are used to anatomically correct the dysfunctional tissues. However, the meshes can create sever adverse complications in some patients (e.g. chronic pain) in longer-term. As an alternative, we developed nanofibrous matrices by electrospinning based on different materials. We designed an in-vitro culture system and subjected cell-seeded matrices to cyclic mechanical loading. Results revealed that gentle straining of POP-cells on electrospun matrices, advances their regenerative potential at morphological and gene expression levels. Our findings, provide a proof-of-concept for using electrospun matrices as an alternative implant for pelvic floor repair, given that the parameters are designed efficiently and safely.
Blood interactions with nano- and microfibers: recent advances, challenges and applications in nano- and microfibrous hemostatic agents Acta Biomater. (IF 6.383) Pub Date : 2018-11-22 Paweł Nakielski, Filippo Pierini
Nanofibrous materials find a wide range of applications, such as vascular grafts, tissue-engineered scaffolds, or drug delivery systems. This phenomenon can be attributed to almost arbitrary biomaterial modification opportunities created by a multitude of polymers used to form nanofibers, as well as by surface functionalization methods. Among these applications, the hemostatic activity of nanofibrous materials is gaining more and more interest in biomedical research. It is therefore crucial to find both materials and nanofiber structural properties that affect organism responses. The present review critically analyzes the response of blood elements to natural and synthetic polymers, and their blends and composites. Also assessed in this review is the incorporation of pro-coagulative substances or drugs that can decrease bleeding time. The review also discusses the main animal models that were used to assess hemostatic agent safety and effectiveness. Statement of significance The paper contains an in-depth review of the most representative studies recently published in the topic of nanofibrous hemostatic agents. The topic evolved from analysis of pristine polymeric nanofibers to multifunctional biomaterials. Furthermore, this study is important because it helps clarify the use of specific blood-biomaterial analysis techniques with emphasis on protein adsorption, thrombogenicity and blood coagulation. The paper should be of interest to the readers of Acta Biomaterialia who are curious about the strategies and materials used for the development of multifunctional polymer nanofibers for novel blood-contacting applications.
Blood Coagulation Response and Bacterial Adhesion to Biomimetic Polyurethane Biomaterials Prepared with Surface Texturing and Nitric Oxide Release Acta Biomater. (IF 6.383) Pub Date : 2018-11-22 Li-Chong Xu, Mark E. Meyerhoff, Christopher A. Siedlecki
A dual functional polyurethane (PU) film that mimics aspects of blood vessel inner surfaces by combining surface texturing and nitric oxide (NO) release was fabricated through a soft lithography two-stage replication process. The fabrication of submicron textures on the polymer surface was followed by solvent impregnation with the NO donor, S-nitroso-N-acetylpenicillamine (SNAP). An in vitro plasma coagulation assay showed that the biomimetic surface significantly increased the plasma coagulation time and also exhibited reduced platelet adhesion and activation, thereby reducing the risk of blood coagulation and thrombosis. A contact activation assay for coagulation factor XII (FXII) demonstrated that both NO release and surface texturing also reduced FXII contact activation, which contributes to the inhibition of plasma coagulation. The biomimetic surface was also evaluated for bacterial adhesion in plasma and results demonstrate that this combined strategy enables a synergistic effect to reduce bacterial adhesion of Staphylococcus epidermidis, Staphylococcus aureus, and Pseudomonas aeruginosa microorganisms. The results strongly suggest that the biomimetic modification with surface texturing and NO release provides an effective approach to improve the biocompatibility of polymeric materials in combating thrombosis and microbial infection. Statement of significance Developed a dual functional polyurethane (PU) film that mimics blood vessel inner surface by combining surface texturing and nitric oxide (NO) release for combatting biomaterial associated thrombosis and microbial infection. Studied the blood coagulation response and bacterial adhesion to such biomimetic PU surfaces, and demonstrated that the combination of surface texturing and NO release synergistically reduced the platelet adhesion and bacterial adhesion in plasma, providing an effective approach to improve the biocompatibility of biomaterials used in blood-contacting medical devices. The NO releasing surface significantly inhibits the plasma coagulation via the reduction of contact activation of FXII, indicating the multifunctional roles of NO in improving the biocompatibility of biomaterials in blood-contacting medical devices.
In vitro and in vivo studies of electroactive reduced graphene oxide-modified nanofiber scaffolds for peripheral nerve regeneration Acta Biomater. (IF 6.383) Pub Date : 2018-11-22 Juan Wang, Yuan Cheng, Liang Chen, Tonghe Zhu, Kaiqiang Ye, Chao Jia, Hongjun Wang, Meifang Zhu, Cunyi Fan, Xiumei Mo
Graphene, as a promising biomaterial, has received great attention in biomedical fields due to its intriguing properties, especially the conductivity and biocompatibility. Given limited studies on the effects of graphene-based scaffolds on peripheral nerve regeneration in vitro and in vivo under electrical stimulation (ES), the present study was intended to systematically investigate how conductive graphene-based nanofibrous scaffolds regulate Schwann cell (SC) behavior including migration, proliferation and myelination, and PC12 cell differentiation in vitro via ES, and whether these conductive scaffolds could guide SC migration and promote nerve regeneration in vivo. Briefly, the reduced graphene oxide (RGO) was coated onto ApF/PLCL nanofibrous scaffolds via in situ redox reaction of the graphene oxide (GO). In vitro, RGO-coated ApF/PLCL (AP/RGO) scaffolds significantly enhanced SC migration, proliferation, and myelination including myelin-specific gene expression and neurotrophic factor secretion. The conditioned media of SCs cultured on AP/RGO scaffolds under ES could induce the differentiation of PC12 cells in a separate culture. In addition, PC12 cells cultured on the conductive AP/RGO scaffolds also showed elevated differentiation upon ES. In vivo implantation of the conductive AP/RGO nerve guidance conduits into rat sciatic nerve defects exhibited a similar healing capacity to autograft, which is the current gold standard in peripheral nerve regeneration. In view of the performance of AP/RGO scaffolds in modulating cell functions in vitro and promoting nerve regeneration in vivo, it is expected that the graphene-based conductive nanofibrous scaffolds would exhibit their potential in peripheral nerve repair and regeneration. Statement of Significance Despite the demonstrated capability of bridging the distal and proximal peripheral nerves, it remains a significant challenge with current artificial nerve conduits to achieve the desired physiological functions, e.g., the transmission of electrical stimuli. Herein, we explored the possibility of combining the conductive properties of graphene with electrospun nanofiber to create the electroactive biomimetic scaffolds for nerve tissue regeneration. In vitro and in vivo studies were carried out: 1) In vitro, the conductive nanofibrous scaffolds significantly promoted SC migration, proliferation and myelination including myelin specific gene expression and neurotrophic factor secretion, and induced PC12 cell differentiation with electrical stimulation. 2) In vivo, the conductive nerve guidance conduit exhibited similar effects with the gold standard autograft. In view of the performance of this conductive scaffold in modulating the cell functions in vitro and promoting nerve regeneration in vivo, it is expected that the graphene-modified nanofibrous scaffolds will exhibit their potential in peripheral nerve repair and regeneration.
Enzymatically Triggered Shape Memory Polymers Acta Biomater. (IF 6.383) Pub Date : 2018-11-22 Shelby L. Buffington, Justine E. Paul, Matthew M. Ali, Mark M. Macios, Patrick T. Mather, James H. Henderson
Cytocompatible shape memory polymers activated by thermal or photothermal triggers have been developed and established as powerful “smart material” platforms for both basic and translational research. Shape memory polymers (SMPs) that could be triggered directly by biological activity have not, in contrast, been reported. The goal of this study was to develop an SMP that responds directly to enzymatic activity and can do so under isothermal cell culture conditions. To achieve this goal, we designed an SMP with a shape fixing component, poly(ε-caprolactone) (PCL), that is vulnerable to enzymatic degradation and a shape memory component, Pellethane, that is enzymatically stable—as the shape fixing component undergoes enzymatically-catalyzed degradation, the SMP returns to its original, programmed shape. We quantitatively and qualitatively analyzed material properties, shape memory performance, and cytocompatibility of the enzymatically-catalyzed shape memory response. The results demonstrate enzymatic recovery, as contraction of tensile specimens, using bulk enzymatic degradation experiments and show that shape recovery is achieved by degradation of the PCL shape-fixing phase. The results further showed that both the materials and the process of enzymatic shape recovery are cytocompatible. Thus, the SMP design reported here represents both an enzyme responsive material capable of applying a programmed shape change or direct mechanical force and an SMP that could respond directly to biological activity. Statement of Significance Cytocompatible shape memory polymers activated by thermal or photothermal triggers have become powerful “smart material” platforms for basic and translational research. Shape memory polymers that could be triggered directly by biological activity have not, in contrast, been reported. Here we report an enzymatically triggered shape memory polymer that changes its shape isothermally in response to enzymatic activity. We successfully demonstrate enzymatic recovery using bulk enzymatic degradation experiments and show that shape recovery is achieved by degradation of the shape-fixing phase. We further show that both the materials and the process of enzymatic shape recovery are cytocompatible. This new shape memory polymer design can be anticipated to enable new applications in basic and applied materials science as a stimulus responsive material.
Enhanced extracellular vesicle production and ethanol-mediated vascularization bioactivity via a 3D-printed scaffold-perfusion bioreactor system Acta Biomater. (IF 6.383) Pub Date : 2018-11-22 Divya B. Patel, Christopher R. Luthers, Max J. Lerman, John P. Fisher, Steven M. Jay
Extracellular vesicles (EVs) have garnered significant interest in the biotechnology field due to their intrinsic therapeutic properties as well as their ability to serve as vehicles for bioactive cargo. However, the lack of an established biomanufacturing platform and limited potency of EVs in vivo remain critical bottlenecks for clinical translation. In this study, we utilized a 3D-printed scaffold-perfusion bioreactor system to assess the response of dynamic culture on extracellular vesicle production from endothelial cells (ECs). We also investigated whether ethanol conditioning, which was previously shown to enhance vascularization bioactivity of EC-derived EVs produced in standard 2D culture conditions, could be employed successfully for the same purpose in a 3D production system. Our results indicate that dynamic culture in a perfusion bioreactor significantly enhances EV production from human ECs. Moreover, the use of ethanol conditioning in conjunction with dynamic culture induces pro-vascularization bioactivity of EC-derived EVs that is correlated with increased EV levels of pro-angiogenic lncRNAs HOTAIR and MALAT1. Thus, this study represents one of the first reports of rationally-designed EV potency enhancement that is conserved between static 2D and dynamic 3D EV production systems, increasing the potential for scalable biomanufacturing of therapeutic EC-derived EVs for a variety of applications. Statement of Significance Extracellular vesicles (EVs) have substantial therapeutic potential in a variety of applications. However, translation of EV-based therapies may be hindered by biomanufacturing challenges. EV production to date has predominantly involved the use of tissue culture flasks. Here, we report, for the first time, the use of a tubular perfusion bioreactor system with an integrated 3D-printed biomaterial scaffold for EV production from human endothelial cells. This system increases EV yield by over 100-fold compared to conventional tissue culture systems. Further, we show that an ethanol-conditioning approach that our group previously developed in 2D culture for enhancing EV potency is compatible with this new system. Thus, potency enhancement of EVs for vascularization applications is possible even with significantly increased production rate.
Porous Scaffolds from Droplet Microfluidics for Prevention of Intrauterine Adhesion Acta Biomater. (IF 6.383) Pub Date : 2018-11-23 Yunlang Cai, Fangyuan Wu, Yunru Yu, Yuxiao Liu, Changmin Shao, Hongcheng Gu, Minli Li, Yuanjin Zhao
Severe intrauterine adhesions (IUAs) have a great negative impact on women's psychological and reproductive health. It remains a significant challenge to prevent postoperative IUAs because of the complications of various clinical preventive measures and incompatibility of uterine cavity morphology. Herein, we present a new drug-loaded porous scaffold based on a microfluidic droplet template, which combines the characteristics of the artificial biocompatible material GelMA and the natural polysaccharide material Na-alginate. By changing the containers that collect the microfluidic droplets, the porous scaffold conforming to the shape of the uterine cavity could be obtained. The porous structure, mechanical property, and flexibility impart the scaffold with compressibility and send it to the uterus through the vagina. In addition, the external–internal connected open structures could load and control the release of drugs to repair the damaged region continuously in vivo. To verify the antiadhesion and repair of drug-loaded porous scaffolds, we tested the system in the rat model of IUAs, and it was demonstrated that the system had the ability to improve neovascularization, cellularize the damaged tissue, and repair the endometrium. These features provide the drug-loaded porous scaffolds with new options for the improvement of postoperative IUAs. Statement of Significance Intrauterine adhesions are caused by various causes of damage to the endometrial basal layer, thus leading to part or entire adhesions in the cervical or uterine cavity. Clinically, various preventive measures reach the barrier effect through the physical barrier, which are difficult to further promote the repair of the damaged endometrium, and most of them have apparent side effects. This study aims to prepare compressible and biodegradable three-dimensional porous drug-loading biological scaffolds. GelMA and Na-alginate have desirable biocompatibility. The interconnect porous scaffolds, which were prepared through the combination of biomaterials and single emulsion microfluidics, not only have compressibility but also provide space for drug delivery and release. This system can further promote the repair of the endometrium while preventing adhesion.
Characterization of a tissue-engineered choroid Acta Biomater. (IF 6.383) Pub Date : 2018-11-23 Aïcha Dede Djigo, Julie Bérubé, Solange Landreville, Stéphanie Proulx
The choroid of the eye is a vascularized and pigmented connective tissue lying between the retina and the sclera. Increasing evidence demonstrates that, beyond supplying nutrients to the outer retina, the different choroidal cells contribute to the retina’s homeostasis, especially by paracrine signaling. However, the precise role of each cell type is currently unclear. Here, we developed a choroidal substitute using the self-assembly approach of tissue engineering. Retinal pigment epithelial (RPE) cells, as well as choroidal stromal fibroblasts, vascular endothelial cells and melanocytes, were isolated from human eye bank donor eyes. Fibroblasts were cultured in a medium containing serum and ascorbic acid. After six weeks, cells formed sheets of extracellular matrix (ECM), which were stacked to produce a tissue-engineered choroidal stroma (TECS). These stromal substitutes were then characterized and compared to the native choroid. Their ECM composition (collagens and proteoglycans) and biomechanical properties (ultimate tensile strength, strain and elasticity) were similar. Furthermore, RPE cells, human umbilical vein endothelial cells and choroidal melanocytes successfully repopulated the stromas. Physiological structures were established, such as a confluent monolayer of RPE cells, vascular-like structures and a pigmentation of the stroma. Our TECS thus recaptured the biophysical environment of the native choroid, and can serve as study models to understand the normal interactions between the RPE and choroidal cells, as well as their reciprocal exchanges with the ECM. This will consequently pave the way to derive accurate insight in the pathophysiological mechanisms of diseases affecting the choroid. Statement of significance The choroid is traditionally known for supplying blood to the avascular outer retina. There has been a renewed attention directed towards the choroid partly due to its implication in the development of age-related macular degeneration (AMD), the leading cause of blindness in industrialized countries. Since AMD involves the dysfunction of the choroid/retinal pigment epithelium (RPE) complex, a three-dimensional (3D) model of RPE comprising the choroid layer is warranted. We used human choroidal cells to engineer a choroidal substitute. Our approach takes advantage of the ability of cells to recreate their own environment, without exogenous materials. Our model could help to better understand the role of each choroidal cell type as well as to advance the development of new therapeutics for AMD.
A dual-targeted hyaluronic acid-gold nanorod platform with triple-stimuli responsiveness for photodynamic/photothermal therapy of breast cancer Acta Biomater. (IF 6.383) Pub Date : 2018-11-19 Weijun Xu, Junmin Qian, Guanghui Hou, Yaping Wang, Jinlei Wang, Tiantian Sun, Lijie Ji, Aili Suo, Yu Yao
Multi-stimuli-responsive theranostic nanoplatform integrating functions of both imaging and multimodal therapeutics holds great promise for improving diagnosis and therapeutic efficacy. In this study, we reported a pH, glutathione (GSH) and hyaluronidase (HAase) triple-responsive nanoplatform for HER2 and CD44 dual-targeted and fluorescence imaging-guided PDT/PTT dual-therapy against HER2-overexpressed breast cancer. The nanoplatform was fabricated by functionalizing gold nanorods (GNRs) with hyaluronic acid (HA) bearing pendant hydrazide and thiol groups via Au-S bonds, and subsequently chemically conjugating 5-aminolevulinic acid (ALA), Cy7.5 and anti-HER2 antibody onto HA moiety for PDT, fluorescence imaging and active targeting, respectively. The resulting versatile nanoplatform GNR-HA-ALA/Cy7.5-HER2 had uniform sizes, favorable dispersibility, as well as pH, GSH and HAase triple-responsive drug release manner. In vitro studies demonstrated that HER2 and CD44 receptor-mediated dual-targeting strategy could significantly enhance the cellular uptake of GNR-HA-ALA/Cy7.5-HER2. Under near-infrared (NIR) irradiation, MCF-7 cells could efficiently generate reactive oxygen species (ROS) and heat, and be more efficiently killed by a combination of PDT and PTT as compared with individual therapy. Pharmacokinetic and biodistribution studies showed that the nanoplatform possessed a circulation half-life of 1.9 h and could be specifically delivered to tumor tissues with an accumulation ratio of 12.8%. Upon the fluorescence imaging-guided PDT/PTT treatments, the tumors were completely eliminated without obvious side effects. The results suggest that the GNR-HA-ALA/Cy7.5-HER2 hold great potential for breast cancer therapy.Statement of significanceA combination of photodynamic therapy (PDT) and photothermal therapy (PTT) is emerging as a promising cancer treatment strategy. However, its therapeutic efficacy is compromised by the nonspecific delivery and unintended release of photo-responsive agents. Herein, we developed a multifunctional theranostic nanoplatform GNR-HA-ALA/Cy7.5-HER2 with pH, glutathione and hyaluronidase triple-responsive drug release for HER2 and CD44 dual-targeted and fluorescence imaging-guided PDT/PTT therapy against breast cancer. We demonstrated that HER2 and CD44 receptors-mediated dual-targeting strategy significantly enhanced the cellular uptake of GNR-HA-ALA/Cy7.5-HER2. We also demonstrated that the combined PDT/PTT treatment had significantly superior antitumor effect than PDT or PTT alone both in vitro and in vivo. Therefore, GNRs-HA-ALA//Cy7.5-HER2 could serve as a promising nanoplatform for HER2-postive breast cancer therapy.
Dual-ion delivery for synergistic angiogenesis and bactericidal capacity with silica-based microsphere Acta Biomater. (IF 6.383) Pub Date : 2018-11-19 Khaliun Boldbaatar, Khandmaa Dashnyam, Jonathan C. Knowles, Hae-Hyoung Lee, Jung-Hwan Lee, Hae-Won Kim
Inhibition of bacterial growth with the simultaneous promotion of angiogenesis has been challenging in the repair and regeneration of infected tissues. Here, we aim to tackle this issue through the use of cobalt-doped silicate microspheres that can sustainably release dual ions (silicate and cobalt) at therapeutically-relevant doses. The cobalt was doped up to 2.5 wt% within a sol-gel silicate glass network, and microspheres with the size of ∼300 μm were generated by an emulsification method. The cobalt and silicate ions released were shown to synergistically upregulate key angiogenic genes, such as HIF1-α, VEGF and the receptor KDR. Moreover, the incorporation of ions promoted the polarization, migration, homing and sprouting angiogenesis of endothelial cells. Neo-vascular formation was significantly higher in the dual-ion delivered microspheres, as evidenced in a chicken chorioallantoic membrane model. When cultured with bacterial species, the cobalt-doped microspheres effectively inhibited bacteria growth in both indirect or direct contacts. Of note, the bacteria/endothelial cell coculture model proved the efficacy of dual-ion releasing microcarriers for maintaining the endothelial survivability against bacterial contamination and their cell-cell junction. The current study demonstrates the multiple actions (proangiogenic and antibacterial) of silicate and cobalt ions released from microspheres, and the concept provided here can be extensively applied to repair and regenerate infected tissues as a growth factor- or drug-free delivery system.Statement of significanceWhile several ions have been introduced to biomaterials for therapeutic purposes, relaying the effects of antibacterial into tissue regenerative (e.g., angiogenesis) has been a significant challenge. In this study, we aim to develop a biomaterial platform that has the capacity of both ‘antibacterial’ and ‘proangiogenic’ from a microsphere sustainably releasing multiple ions (herein cobalt and silicate). Here, dual-actions of the microspheres revealed the stimulated endothelial functions as well as the inhibited growth of different bacterial species. In particular, protecting endothelial survivability against bacterial contamination was reported using the bacterial/endothelial co-culture model. The current concept of drug-free yet multiple-ion delivery biomaterials can be applicable for the repair and regeneration of infected tissues with dual actions of angiogenesis and suppressing bacterial activity.
A new glioblastoma cell trap for implantation after surgical resection Acta Biomater. (IF 6.383) Pub Date : 2018-11-19 Lila Autier, Anne Clavreul, Maximiliano L. Cacicedo, Florence Franconi, Laurence Sindji, Audrey Rousseau, Rodolphe Perrot, Claudia N. Montero-Menei, Guillermo R. Castro, Philippe Menei
Glioblastoma (GB) is a highly infiltrative tumor, recurring, in 90% of cases, within a few centimeters of the surgical resection cavity, even with adjuvant chemo/radiotherapy. Residual GB cells left in the margins or infiltrating the brain parenchyma shelter behind the extremely fragile and sensitive brain tissue and may favor recurrence. Tools for eliminating these cells without damaging the brain microenvironment are urgently required. We propose a strategy involving the implantation, into the tumor bed after resection, of a scaffold to concentrate and trap these cells, to facilitate their destruction by targeted therapies, such as stereotactic radiosurgery. We used bacterial cellulose (BC), an easily synthesized and modifiable random nanofibrous biomaterial, to make the trap. We showed that the structure of BC membranes was ideal for trapping tumor cells and that BC implants were biocompatible with brain parenchyma. We also demonstrated the visibility of BC on magnetic resonance imaging, making it possible to follow its fate in clinical situations and to define the target volume for stereotactic radiosurgery more precisely. Furthermore, BC membranes can be loaded with chemoattractants, which were released and attracted tumor cells in vitro. This is of particular interest for trapping GB cells infiltrating tissues within a few centimeters of the resection cavity. Our data suggest that BC membranes could be a scaffold of choice for implantation after surgical resection to trap residual GB cells.Statement of significanceGlioblastoma is a highly infiltrative tumor, recurring, in 90% of cases, within a few centimeters of the surgical resection cavity, even with adjuvant chemo/radiotherapy. Residual tumor cells left in the margins or infiltrating the brain parenchyma shelter behind the extremely fragile and sensitive brain tissue and contribute to the risk of recurrence. Finding tools to eliminate these cells without damaging the brain microenvironment is a real challenge. We propose a strategy involving the implantation, into the walls of the surgical resection cavity, of a scaffold to concentrate and trap the residual tumor cells, to facilitate their destruction by targeted therapies, such as stereotactic radiosurgery.
Methods for Producing Microstructured Hydrogels for Targeted Applications in Biology Acta Biomater. (IF 6.383) Pub Date : 2018-11-20 Cristobal Garcia Garcia, Kristi L. Kiick
Hydrogels have been broadly studied for applications in clinically motivated fields such as tissue regeneration, drug delivery, and wound healing, as well as in a wide variety of consumer and industry uses. While the control of mechanical properties and network structures are important in all of these applications, for regenerative medicine applications in particular, matching the chemical, topographical and mechanical properties for the target use/tissue is critical. There have been multiple alternatives developed for fabricating materials with microstructures with goals of controlling the spatial location, phenotypic evolution, and signaling of cells. The commonly employed polymers such as poly(ethylene glycol) (PEG), polypeptides, and polysaccharides (as well as others) can be processed by various methods in order to control material heterogeneity and microscale structures. We review here the more commonly used polymers, chemistries, and methods for generating microstructures in biomaterials, highlighting the range of possible morphologies that can be produced, and the limitations of each method. With a focus in liquid-liquid phase separation, methods and chemistries well suited for stabilizing the interface and arresting the phase separation are covered. As the microstructures can affect cell behavior, examples of such effects are reviewed as well.Statement of SignificanceHeterogeneous hydrogels with enhanced matrix complexity have been studied for a variety of biomimetic materials. A range of materials based on poly(ethylene glycol), polypeptides, proteins, and/or polysaccharides, have been employed in the studies of materials that by virtue of their microstructure, can control the behaviors of cells. Methods including microfluidics, photolithography, gelation in the presence of porogens, and liquid-liquid phase separation, are presented as possible strategies for producing materials, and their relative advantages and disadvantages are discussed. We also describe in more detail the various processes involved in LLPS, and how they can be manipulated to alter the kinetics of phase separation and to yield different microstructured materials.
Enhancing Chondrogenesis and Mechanical Strength Retention in Physiologically Relevant Hydrogels with Incorporation of Hyaluronic Acid and Direct Loading of TGF-β Acta Biomater. (IF 6.383) Pub Date : 2018-11-17 Yuhao Deng, Aaron X. Sun, Kalon J. Overholt, Gary Z. Yu, Madalyn R. Fritch, Peter G. Alexander, He Shen, Rocky S. Tuan, Hang Lin
Cell-loaded hydrogels are frequently applied in cartilage tissue engineering for their biocompatibility, ease of application, and ability to conform to various defect sites. As a bioactive adjunct to the biomaterial, transforming growth factor beta (TGF-β) has been shown to be essential for cell differentiation into a chondrocyte phenotype and maintenance thereof, but the low amounts of endogenous TGF-β in the in vivo joint microenvironment necessitate a mechanism for controlled delivery and release of this growth factor. In this study, TGF-β3 was directly loaded with human bone marrow-derived mesenchymal stem cells (MSCs) into poly-D,L-lactic acid/polyethylene glycol/poly-D,L-lactic acid (PDLLA-PEG) hydrogel, or PDLLA-PEG with the addition of hyaluronic acid (PDLLA/HA), and cultured in vitro. We hypothesize that the inclusion of HA within PDLLA-PEG would result in a controlled release of the loaded TGF-β3 and lead to a robust cartilage formation without the use of TGF-β3 in the culture medium. ELISA analysis showed that TGF-β3 release was effectively slowed by HA incorporation, and retention of TGF-β3 in the PDLLA/HA scaffold was detected by immunohistochemistry for up to 3 weeks. By means of both in vitro culture and in vivo implantation, we found that sulfated glycosaminoglycan production was higher in PDLLA/HA groups with homogenous distribution throughout the scaffold than PDLLA groups. Finally, with an optimal loading of TGF-β3 at 10 μg/mL, as determined by RT-PCR and glycosaminoglycan production, an almost twofold increase in Young’s modulus of the construct was seen over a 4-week period compared to TGF-β3 delivery in the culture medium. Taken together, our results indicate that the direct loading of TGF-β3 and stem cells in PDLLA/HA has the potential to be a one-step point-of-care treatment for cartilage injury. Statement of Significance Stem cell-seeded hydrogels are commonly used in cell-based cartilage tissue engineering, but they generally fail to possess physiologically relevant mechanical properties suitable for loading. Moreover, degradation of the hydrogel in vivo with time further decreases mechanical suitability of the hydrogel due in part to the lack of TGF-β3 signaling. In this study, we demonstrated that incorporation of hyaluronic acid (HA) into a physiologically stiff PDLLA-PEG hydrogel allowed for slow release of one-time preloaded TGFβ3, and when loaded with adult mesenchymal stem cells and cultured in vitro, it resulted in higher chondrogenic gene expression and constructs of significantly higher mechanical strength than constructs cultured in conventional TGFβ3-supplemented medium. Similar effects were also observed in constructs implanted in vivo. Our results indicate that direct loading of TGF-β3 combined with HA in the physiologically stiff PDLLA-PEG hydrogel has the potential to be used for one-step point-of-care treatment of cartilage injury.
Bioinspired Hydrogels for Drug-eluting Contact Lenses Acta Biomater. (IF 6.383) Pub Date : 2018-11-16 Carmen Alvarez-Lorenzo, Soledad Anguiano-Igea, Angela Varela-García, María Vivero-Lopez, Angel Concheiro
Efficient ocular drug delivery that can overcome the challenges of topical application has been largely pursued. Contact lenses (CLs) may act as light-transparent cornea/sclera bandages for prolonged drug release towards the post-lens tear fluid, if their composition and inner architecture are fitted to the features of the drug molecules. In this review, first the foundations and advantages of using CLs as ocular drug depots are revisited. Then, pros and cons of common strategies to prepare drug-loaded CLs are analyzed on the basis of recent examples, and finally the main section focuses on bioinspired strategies that can overcome some limitations of current designs. Most bioinspired strategies resemble a reverse engineering process to create artificial receptors for the drug inside the CL network by mimicking the human natural binding site of the drug. Related bioinspired strategies are being also tested for designing CLs that elute comfort ingredients mimicking the blinking-associated renewal of eye mucins. Other bioinspired approaches exploit the natural eye variables as stimuli to trigger drug release or take benefit of bio-glues to specifically bind active components to the CL surface. Overall, biomimicking approaches are being revealed as valuable tools to fit the amounts loaded and the release profiles to the therapeutic demands of each pathology.Statement of significanceBiomimetic and bioinspired strategies are remarkable tools for the optimization of drug delivery systems. Translation of the knowledge about how drugs interact with the natural pharmacological receptor and about components and dynamics of anterior eye segment may shed light on the design criteria for obtaining efficient drug-eluting CLs. Current strategies for endowing CLs with controlled drug release performance still require optimization regarding amount loaded, drug retained in the CL structure during storage, regulation of drug release once applied onto the eye, and maintenance of CL physical properties. All these limitations may be addressed through a variety of recently growing bioinspired approaches, which are expected to pave the way of medicated CLs towards the clinics.
The Multiscale Stiffness of Electrospun Substrates and Aspects of their Mechanical Biocompatibility Acta Biomater. (IF 6.383) Pub Date : 2018-11-15 Manuel Zündel, Alexander E. Ehret, Edoardo Mazza
In contrast to homogeneous materials, the mechanical properties of fibrous substrates depend on the probing lengthscale. This suggests that cells feel very different mechanical cues than expected from the macroscale characterisation of the substrate materials. By means of multiscale computational analyses we study here the mechanical environment of cells adhering to typical electrospun networks used in biomedical applications, with comparable macroscopic stiffness but different fibre diameters. The stiffness evaluated at the level of focal adhesions varies significantly, and the overall magnitude is strongly affected by the fibre diameter. The microscopic stiffness evaluated at cell scale depends substantially on the network topology and is about one order of magnitude lower than the macroscopic stiffness of the substrate, and two to three orders of magnitude below the fibres’ elastic modulus. Moreover, the translation of stiffness over the scales is modulated by global deformations of the scaffold. In particular, uniaxial or biaxial stretching of the substrate induces nonlinear microscopic stiffening. Finally, although electrospun networks allow long-range transmission of cell-induced deformations, the comparison between the range of forces measured in cell traction force microscopy and those required to markedly deform typical electrospun networks reveals an order of magnitude difference, suggesting that these scaffolds provide a rather rigid environment for cells. All these results underline that the achievement of mechanical biocompatibility at all relevant lengthscales, and over the whole range of physiological loading states is extremely challenging. At the same time, the study shows that the diameter, length and curvature of fibre segments might be tunable towards achieving this goal. Statement of Significance Electrospun fabrics have growing use as substrates and scaffolds in tissue engineering and other biomedical applications. Based on multiscale computational analyses, this study shows that substrates of comparable macroscopic stiffness can provide tremendously different mechanical micro-environments, and that cells adhering to fibrous substrates may thus experience by orders of magnitude different mechanical cues than it would be expected from macroscale material characterisation. The simulations further reveal that the transfer of stiffness over the length scales changes with macroscopic deformation, and identify some key parameters that govern the transfer ratio. We believe that such refined understanding of the multiscale aspects of mechanical biocompatibility is key to the development of successful scaffold materials.
Alum-functionalized graphene oxide nanocomplexes for effective anticancer vaccination Acta Biomater. (IF 6.383) Pub Date : 2018-11-15 Xiaoli Wang, Fengqiang Cao, Mengmeng Yan, Yijia Liu, Xianghui Zhu, Hongfan Sun, Guilei Ma
Aluminum-based adjuvant (e.g., aluminum oxyhydroxide (AlO(OH), known as the commercial Alhydrogel® (Alum)) is the first adjuvant to be used in human vaccines. Although Alum shows a robust induction of antibody-mediated immunity, its weak stimulation of cell-mediated immunity makes it a questionable adjuvant for cancer immunotherapy. Herein, we described a novel formulation of Alum-based adjuvant by preparing AlO(OH)-modified graphene oxide (GO) nanosheets (GO-AlO(OH)), which, in addition to maintaining the induction of humoral immune response by AlO(OH), could further elicit the cellular immune response by GO. Similar to Alum, GO-AlO(OH) vaccine formulation could be constructed by the incorporation of antigen using a facile mixing/adsorption approach. Antigen-loaded GO-AlO(OH) nanocomplexes facilitated cellular uptake and cytosolic release of antigens and promoted DC maturation, thereby eliciting higher antigen-specific IgG titers, inducing robust CD4+ and CD8+ T lymphocyte response, and inhibiting tumor growth in vivo. Furthermore, by employing tumor cell lysate-based cancer vaccines, GO-AlO(OH) nanocomplexes led to significant inhibition of tumor growth and can be implemented as a personalized treatment strategy for cancer vaccine development. Overall, GO-AlO(OH) nanocomplexes described herein may serve as a facile and efficient approach for effective anticancer vaccination. Statement of significance Herein, we described a novel formulation of aluminum-based adjuvant by preparing aluminum oxyhydroxide (AlO(OH)) (known as “Alum”)-modified graphene oxide (GO) nanocomplexes (GO-AlO(OH)), which, in addition to maintaining the induction of humoral immune response by AlO(OH), could further elicit the cellular immune response by GO. GO-AlO(OH) nanocomplexes can be prepared easily and in large scale by a chemical precipitation method. Similar to “Alum,” antigen-loaded GO-AlO(OH) vaccine formulation could be constructed by the incorporation of antigen using a facile mixing/adsorption approach. The very simple and reproductive preparation process of vaccines and the powerful ability to raise both humoral and cellular immune responses provide a novel approach for improving cancer immunotherapy efficacy.
Study on the effectiveness of ligand reversible shielding strategy in targeted delivery and tumor therapy Acta Biomater. (IF 6.383) Pub Date : 2018-11-15 Zhenpeng Hu, Xiaomin Li, Ming Yuan, Xinyu Wang, Yapei Zhang, Wei Wang, Zhi Yuan
We previously proved the superiority of the ligand reversible shielding strategy based on the pH-responsive self-assembly/disassembly of gold nanoparticles through computed tomography imaging in vivo. Herein, the practicality of this strategy in tumor therapy was investigated by a ligand reversible shielding system based on a temperature-responsive polymer. The ligand biotin, cisplatin-loaded chain poly(acrylic acid)-Pt, and the shielding segment thermo-sensitive poly(N-isopropylacrylamide-co-acrylamide) (P(NIPAAm-co-AAm)) were co-modified onto the surface of gold nanostars. In the blood circulation (37 °C), the ligand was shielded by the extension of P(NIPAAm-co-AAm), whose lower critical solution temperature (LCST) is approximately 39 °C. After the nanoparticles accumulate at the tumor site by the enhanced permeability and retention (EPR) effect, the heat generated from gold nanostars upon near-infrared light irradiation would trigger the contraction of P(NIPAAm-co-AAm), thus deshielding the ligand for enhanced tumor cellular uptake. Owing to the reversible extension–contraction transformation change of P(NIPAAm-co-AAm), the reversible shielding effect on the ligand could be accomplished even if the nanoparticles return to the blood circulation. The results indicated that the system could extend blood circulation (1.6-fold at 24 h), reduce immune system clearance (28% lower), and enhance tumor accumulation (37% higher) effectively compared with the irreversible ligand shielding system by analysis of platinum. This strategy showed significantly superior tumor inhibition (11% higher) than the irreversible system. All these results make clear that the ligand reversible shielding strategy is effective and offers important references for the design of nanomaterials for improving tumor accumulation. Statement of significance Herein, the practicality of the ligand reversible shielding strategy in tumor therapy was investigated. The ligand biotin, cisplatin loaded chain poly(acrylic acid)-Pt and the shielding segment thermo-sensitive poly(N-isopropylacrylamide-co-acrylamide) (P(NIPAAm-co-AAm) which LCST is about 39 °C) were co-modified onto the surface of gold nanostars. This well-designed NPs could shield target ligand in blood circulation (37 °C) and deshield it at tumor site (40 - 41 °C) reversibly. The results indicated that the system could extend blood circulation (1.6-fold at 24 h), reduce immune system clearance (28% lower) and enhance tumor accumulation (37% higher) effectively compared with the irreversible ligand shielding system by analysis of platinum. Significantly, the strategy showed superior tumor inhibition than the irreversible system (11% higher).
Osseointegration and current interpretations of the bone-implant interface Acta Biomater. (IF 6.383) Pub Date : 2018-11-13 Furqan A. Shah, Peter Thomsen, Anders Palmquist
Complex physical and chemical interactions take place in the interface between the implant surface and bone. Various descriptions of the ultrastructural arrangement to various implant design features, ranging from solid and macroporous geometries to surface modifications on the micron-, submicron-, and nano- levels, have been forwarded. The current knowledge regarding the structural organisation of the bone-implant interface is reviewed with a focus on solid devices, mainly metal (or alloy) intended for permanent anchorage in bone. Certain biomaterials that undergo surface and bulk degradation are also considered. The bone-implant interface is a heterogeneous zone consisting of mineralised, partially mineralised, and unmineralised areas. Within the meso-micro-nano-continuum, mineralised collagen fibrils form the structural basis of the bone-implant interface, in addition to accumulation of non-collagenous macromolecules such as osteopontin, bone sialoprotein, and osteocalcin. Based on the available literature, eight distinct interpretations of the ultrastructure of the bone-implant interface are revealed. The interpretation is influenced by the in vivo model and species-specific characteristics, healing time point(s), physico-chemical properties of the implant surface, implant geometry, sample preparation route(s) and associated artefacts, analytical technique(s) and their limitations, and non-compromised vs compromised local tissue conditions. The understanding of the ultrastructure of the interface during experimental conditions is rapidly evolving due to the introduction of novel techniques for the preparation and analysis of the interface. Nevertheless, the current understanding of the interface zone in humans in relation to clinical implant performance is still hampered by the shortcomings of clinical methods to determine the fine details of the bone-implant interface.Statement of SignificanceBeing a hierarchical material by design, the overall strength of bone is governed by composition and structure. Understanding the structure of the bone-implant interface is essential in the development of novel bone repair materials and strategies, and their long-term success. Here, the current knowledge regarding the eventual structural organisation of the bone-implant interface is reviewed, with a focus on solid devices intended for permanent anchorage in bone, and certain biomaterials that undergo surface and bulk degradation. The bone-implant interface is a heterogeneous zone consisting of mineralised, partially mineralised, and unmineralised areas. Within the meso-micro-nano-continuum, mineralised collagen fibrils form the structural basis of the bone-implant interface, in addition to accumulation of non-collagenous macromolecules such as osteopontin, bone sialoprotein, and osteocalcin.
Effect of stress on corrosion of high-purity magnesium in vitro and in vivo Acta Biomater. (IF 6.383) Pub Date : 2018-11-13 Yuanming Gao, Lizhen Wang, Linhao Li, Xuenan Gu, Kuo Zhang, Jie Xia, Yubo Fan
Magnesium-based implants are subjected to complicated stresses during implantation in the human body. The stress effects on corrosion of magnesium (Mg) in vitro were investigated in previous studies, whereas in this study, the corrosion behaviors of high-purity (HP) Mg under stress were comparatively studied in vitro in Hank’s solution and in vivo in the subcutaneous environment of rats. Loading devices were designed to apply compressive stress (15.1±0.5 MPa) and tensile stress (13.2±0.2 MPa) on HP Mg specimens both in vitro and in vivo. Corrosion rates of HP Mg were characterized by mass and volume losses. It was shown that the applied compressive stress had no effect on in vitro corrosion behaviors and the applied tensile stress accelerated the in vitro corrosion, thereby causing severe pitting corrosions and stress corrosion cracking (SCC). However, there was no significant change for corrosion behaviors in vivo under neither compressive stress nor tensile stress. Severe pitting corrosion and SCC did not occur in vivo. Histological evaluation revealed that a fibrotic capsule induced by foreign body reaction was formed on the corrosion surfaces of HP Mg in the subcutaneous environment. It was proposed that the fibrotic capsule suppressed the effects of stress in vivo by protecting the corrosion surfaces. These results provided new insights into understanding the stress effects on the corrosion of Mg both in vitro and in vivo.Statement of SignificanceMg and Mg alloys have shown potential as biodegradable metallic materials. During implantation, Mg is subjected to various mechanical environments in human body. It is necessary to clear different stress effect on Mg corrosion. However, few studies were performed in vivo. It is important to analyze the effect of quantitative stress on Mg corrosion in vivo. In this study, quantitative stresses were applied on Mg both in vitro and in vivo. The effects of stress on in vitro and in vivo corrosions of Mg were investigated and compared.
Lipid-hyaluronan synergy strongly reduces intrasynovial tissue boundary friction Acta Biomater. (IF 6.383) Pub Date : 2018-11-10 Weifeng Lin, Reut Mashiah, Jasmine Seror, Assaf Kadar, Oleg Dolkart, Tamir Pritsch, Ronit Goldberg, Jacob Klein
Hyaluronan (HA)-lipid layers on model (mica) surfaces massively reduce friction as the surfaces slide past each other, and have been proposed, together with lubricin, as the boundary layers accounting for the extreme lubrication of articular cartilage. The ability of such HA-lipid complexes to lubricate sliding biological tissues has not however been demonstrated. Here we show that HA-lipid layers on the surface of an intrasynovial tendon can strongly reduce the friction as the tendon slides within its sheath. We find a marked lubrication synergy when combining both HA and lipids at the tendon surface, relative to each component alone, further enhanced when the polysaccharide is functionalized to attach specifically to the tissue. Our results shed light on the lubricity of sliding biological tissues, and indicate a novel approach for lubricating surfaces such as tendons and, possibly, articular cartilage, important, respectively, for alleviating function impairment following tendon injury and repair, or in the context of osteoarthritis. Statement of Significance Lubrication breakdown between sliding biological tissues is responsible for pathologies ranging from dry eye syndrome to tendon-injury repair impairment and osteoarthtritis. These are increasing with human longevity and impose a huge economic and societal burden. Here we show that synergy of hyaluronan and lipids, molecules which are central components of synovial joints and of the tendon/sheath system, can strongly reduce friction between sliding biological tissues (the extrasynovial tendon sliding in its sheath), relative to untreated tissue or to either component on its own. Our results point to the molecular origins of the very low friction in healthy tendons and synovial joints, as well as to novel treatments of lubrication breakdown in these organs.
Hydrazone covalent adaptable networks modulate extracellular matrix deposition for cartilage tissue engineering Acta Biomater. (IF 6.383) Pub Date : 2018-11-10 Benjamin M. Richardson, Daniel G. Wilcox, Mark A. Randolph, Kristi S. Anseth
Dynamic control of hydrogel crosslinking via sortase-mediated reversible transpeptidation Acta Biomater. (IF 6.383) Pub Date : 2018-11-08 Matthew R. Arkenberg, Dustin M. Moore, Chien-Chi Lin
Glass-ceramics for cancer treatment: So close, or yet so far? Acta Biomater. (IF 6.383) Pub Date : 2018-11-09 Marta Miola, Yousef Pakzad, Sara Banijamali, Saeid Kargozar, Chiara Vitale-Brovarone, Abolfazl Yazdanpanah, Oana Bretcanu, Arash Ramedani, Enrica Vernè, Masoud Mozafari
Biomaterials and Glia: Progress on Designs to Modulate Neuroinflammation Acta Biomater. (IF 6.383) Pub Date : 2018-11-07 C. Tsui, K. Koss, M.A. Churchward, K.G. Todd
Construction of a biodegradable, versatile nanocarrier for optional combination cancer therapy Acta Biomater. (IF 6.383) Pub Date : 2018-11-07 Jia Wen, Yinghua Lv, Yongqian Xu, Pengfei Zhang, Hongjuan Li, Xiaoxu Chen, Xueliang Li, Lingkai Zhang, Fengyu Liu, Wenxian Zeng, Shiguo Sun
Isocyanate-terminated urethane-based dental adhesive bridges dentinal matrix collagen with adhesive resin Acta Biomater. (IF 6.383) Pub Date : 2018-11-07 Rongchen Xu, Fan Yu, Li Huang, Wei Zhou, Yan Wang, Fu Wang, Xiang Sun, Gang Chang, Ming Fang, Ling Zhang, Fang Li, Franklin Tay, Lina Niu, Jihua Chen
Freeze-Casting Porous Chitosan Ureteral Stents for Improved Drainage Acta Biomater. (IF 6.383) Pub Date : 2018-11-07 Kaiyang Yin, Prajan Divakar, Ulrike G.K. Wegst
Suicide-Gene Transfection of Tumor-tropic Placental Stem Cells employing Ultrasound-Responsive Nanoparticles Acta Biomater. (IF 6.383) Pub Date : 2018-11-07 Juan L. Paris, Paz de la Torre, M. Victoria Cabañas, Miguel Manzano, Ana I. Flores, María Vallet-Regí
Mammary Fibroblasts Remodel Fibrillar Collagen Microstructure in a Biomimetic Nanocomposite Hydrogel Acta Biomater. (IF 6.383) Pub Date : 2018-11-07 Chun Liu, Benjamin Chiang, Daniela Lewin Mejia, Kathryn E. Luker, Gary D. Luker, Andre Lee
Mineralization in Micropores of Calcium Phosphate Scaffolds Acta Biomater. (IF 6.383) Pub Date : 2018-11-05 Laurence E. Rustom, Michael J. Poellmann, Amy J. Wagoner Johnson
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