Abstract
Molybdenum disulphide (MoS2) exhibits unique properties that are useful for various biomedical applications. Owing to its distinct characteristics and osteogenic differentiation promotion effect, this material has been studied extensively. However, the effect of cell density on osteogenic differentiation between MoS2-based biomaterials and cells is still unknown. In this study, we used MoS2/polyacrylonitrile (PAN) composite nanofibres as substrates to evaluate the effect of cellular density on the osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs). We created different experimental groups with increasing cell seeding densities and investigated cellular behaviours, biocompatibility, proliferation and osteogenic properties. The results show that MoS2/PAN composite nanofibres can positively regulate osteogenic differentiation. More importantly, in the presence of standard culture conditions, 1.0 × 104 cells/cm2 is the most efficient and suitable cell seeding density for BMSCs osteogenic differentiation. Our findings suggest that the optimal cell density for osteogenesis is vital for the osteogenic differentiation of BMSCs when these cells are cultured onto MoS2-based biomaterials.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ahmad T, Byun H, Lee J, Perikamana SKM, Shin YM, Kim EM, Shin H (2019) Stem cell spheroids incorporating fibers coated with adenosine and polydopamine as a modular building blocks for bone tissue engineering. Biomaterials 230:119652. https://doi.org/10.1016/j.biomaterials.2019.119652
Anderson-Baron M, Kunze M, Mulet-Sierra A, Adesida AB (2020) Effect of cell seeding density on matrix-forming capacity of meniscus fibrochondrocytes and nasal chondrocytes in meniscus tissue engineering. FASEB J 00:1–14. https://doi.org/10.1096/fj.201902559R
Barba A, Diez-Escudero A, Espanol M, Bonany M, Sadowska JM, Guillem-Marti J, Öhman-Mägi C, Persson C, Manzanares MC, Franch J, Ginebra MP (2019) Impact of biomimicry in the design of osteoinductive bone substitutes: Nanoscale matters. ACS Appl Mater Interfaces 11(9):8818–8830. https://doi.org/10.1021/acsami.8b20749
Berent ZT, Wagoner Johnson AJ (2020) Cell seeding simulation on micropatterned islands shows cell density depends on area to perimeter ratio, not on island size or shape. Acta Biomater 20:30123–30129. https://doi.org/10.1016/j.actbio.2020.02.035
Cámara-Torres M, Sinha R, Mota C, Moroni L (2019) Improving cell distribution on 3D additive manufactured scaffolds through engineered seeding media density and viscosity. Acta Biomater 101:183–195. https://doi.org/10.1016/j.actbio.2019.11.020
Camasão DB, Pezzoli D, Loy C, Kumra H, Levesque L, Reinhardt DP, Candiani G, Mantovani D (2019) Increasing cell seeding density improves elastin expression and mechanical properties in collagen gel-based scaffolds cellularized with smooth muscle cells. Biotechnol J 14(3):1700768. https://doi.org/10.1002/biot.201700768
Choi C, Choi MK, Liu S, Kim MS, Park OK, Im C, Kim J, Qin X, Lee GJ, Cho KW, Kim M, Joh E, Lee J, Son D, Kwon SH, Jeon NL, Song YM, Lu N, Kim DH (2017) Human eye-inspired soft optoelectronic device using high-density MoS2-graphene curved image sensor array. Nat Commun 8:1664. https://doi.org/10.1038/s41467-017-01824-6
Corin KA, Gibson LJ (2010) Cell contraction forces in scaffolds with varying pore size and cell density. Biomaterials 31(18):4835–4845. https://doi.org/10.1016/j.biomaterials.2010.01.149
Eivazzadeh-Keihan R, Maleki A, de la Guardia M, Bani MS, Chenab KK, Pashazadeh-Panahi P, Baradaran B, Mokhtarzadeh A, Hamblin MR (2019) Carbon based nanomaterials for tissue engineering of bone: Building new bone on small black scaffolds: a review. J Adv Res 18:185–201. https://doi.org/10.1016/j.jare.2019.03.011
Elango J, Zhang J, Bao B, Palaniyandi K, Wang S, Wenhui W, Robinson JS (2016) Rheological, biocompatibility and osteogenesis assessment of fish collagen scaffold for bone tissue engineering. Int J Biol Macromol 91:51–59. https://doi.org/10.1016/j.ijbiomac.2016.05.067
Erickson IE, Kestle SR, Zellars KH, Farrell MJ, Kim M, Burdick JA, Mauck RL (2012) High mesenchymal stem cell seeding densities in hyaluronic acid hydrogels produce engineered cartilage with native tissue properties. Acta Biomater 8(8):3027–3034. https://doi.org/10.1016/j.actbio.2012.04.033
Eyckmans J, Lin GL, Chen CS (2012) Adhesive and mechanical regulation of mesenchymal stem cell differentiation in human bone marrow and periosteum-derived progenitor cells. Biol Open 1:1058–1068. https://doi.org/10.1242/bio.20122162
Farrell MJ, Comeau ES, Mauck RL (2012) Mesenchymal stem cells produce functional cartilage matrix in three-dimensional culture in regions of optimal nutrient supply. Eur Cell Mater 23:425–440. https://doi.org/10.22203/eCM.v023a33
Harvestine JN, Saiz AM Jr, Leach JK (2019) Cell-secreted extracellular matrix influences cellular composition sequestered from unprocessed bone marrow aspirate for osteogenic grafts. Biomater Sci 7(5):2091–2101. https://doi.org/10.1039/C8BM01478G
Herrmann M, Hildebrand M, Menzel U, Fahy N, Alini M, Lang S, Benneker L, Verrier S, Stoddart MJ, Bara JJ (2019) Phenotypic characterization of bone marrow mononuclear cells and derived stromal cell populations from human iliac crest, vertebral body and femoral head. Int J Mol Sci 20(14):3454. https://doi.org/10.3390/ijms20143454
Hu L, Ren Y, Yang H, Xu Q (2014) Fabrication of 3D hierarchical MoS2/polyaniline and MoS2/C architectures for lithium-ion battery applications. ACS Appl Mater Interfaces 6(16):14644–14652. https://doi.org/10.1021/am503995s
Jin L, Yue D, Xu ZW, Liang GB, Zhang YL, Zhang JF, Zhang XC, Wang ZL (2014) Fabrication, mechanical properties, and biocompatibility of reduced graphene oxide-reinforced nanofiber. Mats RSC Adv 4:35035–35041. https://doi.org/10.1039/c4ra03987d
Khetan S, Guvendiren M, Legant WR, Cohen DM, Chen CS, Burdick JA (2013) Degradation-mediated cellular traction directs stem cell fate in covalently crosslinked three-dimensional hydrogels. Nat Mater 12(5):458–465. https://doi.org/10.1038/nmat3586
Kolanthai E, Sindu PA, Khajuria DK, Veerla SC, Kuppuswamy D, Catalani LH, Mahapatra DR (2018) Graphene oxide-A tool for the preparation of chemically crosslinking free alginate-chitosan-collagen scaffolds for bone tissue engineering. ACS Appl Mater Interfaces 10(15):12441–12452. https://doi.org/10.1021/acsami.8b00699
Lee WY, Wang B (2017) Cartilage repair by mesenchymal stem cells: Clinical trial update and perspectives. J Orthop Transl 9:76–88. https://doi.org/10.1016/j.jot.2017.03.005
Lee YH, Zhang XQ, Zhang W, Chang MT, Lin CT, Chang KD, Yu YC, Wang JT, Chang CS, Li LJ, Lin TW (2012) Synthesis of large-area MoS2 atomic layers with chemical vapor deposition. Adv Mater 24(17):2320–2325. https://doi.org/10.1002/adma.201104798
Li XL, Li YD (2004) MoS2 Nanostructures: Synthesis and electrochemical Mg2+ Intercalation. J Phys Chem B 108(37):13893–13900. https://doi.org/10.1021/jp0367575
Liu H, Li W, Wen W, Luo B, Liu M, Ding S, Zhou C (2017) Mechanical properties and osteogenic activity of poly(l-lactide) fibrous membrane synergistically enhanced by chitosan nanofibers and polydopamine layer. Mater Sci Eng C Mater Biol Appl 81:280–290. https://doi.org/10.1016/j.msec.2017.08.010
Lopes D, Martins-Cruz C, Oliveira MB, Mano JF (2018) Bone physiology as inspiration for tissue regenerative therapies. Biomaterials 185:240–275. https://doi.org/10.1016/j.biomaterials.2018.09.028
Luo Z, Ouyang Y, Zhang H, Xiao M, Ge J, Jiang Z, Wang J, Tang D, Cao X, Liu C, Xing W (2018) Chemically activating MoS2 via spontaneous atomic palladium interfacial doping towards efficient hydrogen evolution. Nat Commun 9:2120. https://doi.org/10.1038/s41467-018-04501-4
Mao AS, Shin JW, Mooney DJ (2016) Effects of substrate stiffness and cell-cell contact on mesenchymal stem cell differentiation. Biomaterials 98:184–191. https://doi.org/10.1016/j.biomaterials.2016.05.004
Mehrian M, Lambrechts T, Marechal M, Luyten FP, Papantoniou I, Geris L (2020) Predicting in vitro human mesenchymal stromal cell expansion based on individual donor characteristics using machine learning. Cytotherapy 22(2):82–90. https://doi.org/10.1016/j.jcyt.2019.12.006
Mei L, Zhang X, Yin W, Dong X, Guo Z, Fu W, Su C, Gu Z, Zhao Y (2019) Translocation, biotransformation-related degradation, and toxicity assessment of polyvinylpyrrolidone-modified 2H-phase nano-MoS2. Nanoscale 11(11):4767–4780. https://doi.org/10.1039/C8NR10319D
Mobasseri R, Tian L, Soleimani M, Ramakrishna S, Naderi-Manesh H (2017) Bio-active molecules modified surfaces enhanced mesenchymal stem cell adhesion and proliferation. Biochem Biophys Res Commun 483(1):312–317. https://doi.org/10.1016/j.bbrc.2016.12.146
Qian Y, Zhou X, Sun H, Yang J, Chen Y, Li C, Wang H, Xing T, Zhang F, Gu N (2018) Biomimetic domain-active electrospun scaffolds facilitating bone regeneration synergistically with antibacterial efficacy for bone defects. ACS Appl Mater Interfaces 10(4):3248–3259. https://doi.org/10.1021/acsami.7b14524
Qiu K, Chen B, Nie W, Zhou X, Feng W, Wang W, Chen L, Mo X, Wei Y, He C (2016) Electrophoretic deposition of dexamethasone-loaded mesoporous silica nanoparticles onto poly (l-lactic acid)/poly(epsilon-caprolactone) composite scaffold for bone tissue engineering. ACS Appl Mater Interfaces 8(6):4137–4148. https://doi.org/10.1021/acsami.5b11879
Rufaihah AJ, Cheyyatraivendran S, Mazlan MDM, Lim K, Chong MSK, Mattar CNZ, Chan JKY, Kofidis T, Seliktar D (2018) The effect of scaffold modulus on the morphology and remodeling of fetal mesenchymal stem cells. Front Physiol 9:1555. https://doi.org/10.3389/fphys.2018.01555
Singh SJ, Turner W, Glaser DE, McCloskey KE, Filipp FV (2017) Metabolic shift in density-dependent stem cell differentiation. Cell Commun Signal 15(1):44. https://doi.org/10.1186/s12964-017-0173-2
Stoker MG, Rubin H (1967) Density dependent inhibition of cell growth in culture. Nature 215(5097):171–172. https://doi.org/10.1038/215171a0
Suhito IR, Han Y, Kim DS, Son H, Kim TH (2017) Effects of two-dimensional materials on human mesenchymal stem cell behaviors. Biochem Biophys Res Commun 493(1):578–584. https://doi.org/10.1016/j.bbrc.2017.08.149
Sun AX, Lin H, Fritch MR, Shen H, Alexander PG, DeHart M, Tuan RS (2017) Chondrogenesis of human bone marrow mesenchymal stem cells in 3-dimensional, photocrosslinked hydrogel constructs: Effect of cell seeding density and material stiffness. Acta Biomater 58:302–311. https://doi.org/10.1016/j.actbio.2017.06.016
Tang J, Peng R, Ding J (2010) The regulation of stem cell differentiation by cell-cell contact on micropatterned material surfaces. Biomaterials 31(9):2470–2476. https://doi.org/10.1016/j.biomaterials.2009.12.006
Teo WZ, Chng EL, Sofer Z, Pumera M (2014) Cytotoxicity of exfoliated transition-metal dichalcogenides (MoS2, WS2, and WSe2) is lower than that of graphene and its analogues. Chemistry 20(31):9627–9632. https://doi.org/10.1002/chem.201402680
Venugopal B, Mogha P, Dhawan J, Majumder A (2018) Cell density overrides the effect of substrate stiffness on human mesenchymal stem cells' morphology and proliferation. Biomater Sci 6(5):1109–1119. https://doi.org/10.1039/C7BM00853H
Wang H, Li Y, Zuo Y, Li J, Ma S, Cheng L (2007) Biocompatibility and osteogenesis of biomimetic nano-hydroxyapatite/polyamide composite scaffolds for bone tissue engineering. Biomater 28(22):3338–3348. https://doi.org/10.1016/j.biomaterials.2007.04.014
Wang S, Li K, Chen Y, Chen H, Ma M, Feng J, Zhao Q, Shi J (2015) Biocompatible PEGylated MoS2 nanosheets: controllable bottom-up synthesis and highly efficient photothermal regression of tumor. Biomaterials 39:206–217. https://doi.org/10.1016/j.biomaterials.2014.11.009
Wang S, Qiu J, Guo W, Yu X, Nie J, Zhang J, Zhang X, Liu Z, Mou X, Li L, Liu H (2017) A nanostructured molybdenum disulfide film for promoting neural stem cell neuronal differentiation: toward a nerve tissue-engineered 3D scaffold. Adv Biosys 1(5):1600042. https://doi.org/10.1002/adbi.201600042
Wang H, Li C, Fang P, Zhang Z, Zhang JZ (2018) Synthesis, properties, and optoelectronic applications of two-dimensional MoS2 and MoS2-based heterostructures. Chem Soc Rev 47(16):6101–6127. https://doi.org/10.1039/C8CS00314A
Wang D, Li J, Zheng A, Ma H, Pan Z, Qu W, Wang L, Han J, Wang C, Tian Z (2019) Quasi-single-layer MoS2 on MoS2/TiO2 nanoparticles for anthracene hydrogenation. ACS Appl Nano Mater 2(8):5096–5107. https://doi.org/10.1021/acsanm.9b01001
Wiesner M, Berberich O, Hoefner C, Blunk T, Baue-Kreisel P (2018) Gap junctional intercellular communication in adipose-derived stromal/stem cells is cell density-dependent and positively impacts adipogenic differentiation. J Cell Physiol 233(4):3315–3329. https://doi.org/10.1002/jcp.26178
Wu S, Wang J, Jin L, Li Y, Wang Z (2018) Effects of polyacrylonitrile/MoS2 composite nanofibers on the growth behavior of bone marrow mesenchymal stem cells. ACS Appl Nano Mater 1(1):337–343. https://doi.org/10.1021/acsanm.7b00188
Wu YK, Tu YK, Yu J, Cheng NC (2020) The Influence Of Cell Culture Density On The Cytotoxicity Of Adipose-Derived Stem Cells Induced by l-ascorbic acid-2-phosphate. Sci Rep 10(1):104. https://doi.org/10.1038/s41598-019-56875-0
Xia Y, Sun J, Zhao L, Zhang F, Liang XJ, Guo Y, Weir MD, Reynolds MA, Gu N, Xu HHK (2018) Magnetic field and nano-scaffolds with stem cells to enhance bone regeneration. Biomaterials 183:151–170. https://doi.org/10.1016/j.biomaterials.2018.08.040
Yassin MA, Leknes KN, Pedersen TO, Xing Z, Sun Y, Lie SA, Finne-Wistrand A, Mustafa K (2015) Cell seeding density is a critical determinant for copolymer scaffolds-induced bone regeneration. J Biomed Mater Res A 103(11):3649–3658. https://doi.org/10.1002/jbm.a.35505
Yuan Z, Tao B, He Y, Liu J, Lin C, Shen X, Ding Y, Yu Y, Mu C, Liu P, Cai K (2019) Biocompatible MoS2/PDA-RGD coating on titanium implant with antibacterial property via intrinsic ROS-independent oxidative stress and NIR irradiation. Biomaterials 217:119290. https://doi.org/10.1016/j.biomaterials.2019.119290
Zhu H, Qin X, Cheng L, Azcatl A, Kim J, Wallace RM (2016) Remote plasma oxidation and atomic layer etching of MoS2. ACS Appl Mater Interfaces 8(29):19119–19126. https://doi.org/10.1021/acsami.6b04719
Acknowledgements
This work was supported by the National Natural Science Foundation of China Youth Fund [Grant Numbers 81900977, U1904176], the Guangdong Basic and Applied Basic Research Foundation [Grant Number 2019A1515010592], the Guangzhou Science and Technology Planning Project [Grant Number 201904010150], and the Guangdong Financial Fund for High-Calibre Hospital Construction (174–2018-XMZC-0001–03-0125/D-19).
Author information
Authors and Affiliations
Contributions
All authors contributed to the study conception and design. YL supervised the project. SW and LJ designed the research. SL and JX conducted experiments, analysed the datas and prepared the manuscript. XZ provided language help. LZ and RY provided writing assistance. All authors read and approved the final manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that there is no conflict of interests regarding the publication of this research.
Research involving animals
Several 28 to 3-week-old male Sprague Dawley rats from laboratory animal centre of Sun Yat-sen University were used to collect bone marrow-derived mesenchymal stem cells (BMSCs). All relevant ethical safeguards have been met in relation to animal experimentation.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Luo, S., Wu, S., Xu, J. et al. Osteogenic differentiation of BMSCs on MoS2 composite nanofibers with different cell seeding densities. Appl Nanosci 10, 3703–3716 (2020). https://doi.org/10.1007/s13204-020-01473-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13204-020-01473-0