Abstract
Three-dimensional (3D)-printed scaffolds have attracted considerable attention in recent years as they provide a suitable environment for bone cell tissue regeneration and can be customized in shape. Among many other challenges, the material composition and geometric structure have major impacts on the performance of scaffolds. Hydroxyapatite and tricalcium phosphate (HA/TCP), as the major constituents of natural bone and teeth, possess attractive biological properties and are widely used in bone scaffold fabrication. Many fabrication methods have been investigated in attempts to achieve HA/TCP scaffolds with microporous structure enabling cell growth and nutrient transport. However, current 3D printing methods can only achieve the fabrication of HA/TCP scaffolds with certain range of microporous structure. To overcome this challenge, we developed a slurry-based microscale mask image projection stereolithography, allowing us to form a HA/TCP-based photocurable suspension with complex geometry including biomimetic features and hierarchical porosity. Here, the curing performance and physical properties of the HA/TCP suspension were investigated, and a circular movement process for the fabrication of highly viscous HA/TCP suspension was developed. Based on these investigations, the scaffold composition was optimized. We determined that a 30 wt% HA/TCP scaffold with biomimetic hierarchical structure exhibited superior mechanical properties and porosity. Cell proliferation was investigated in vitro, and the surgery was conducted in a nude mouse in vivo model of long bone with cranial neural crest cells and bone marrow mesenchymal stem cells. The results showed our 3D-printed HA/TCP scaffold with biomimetic hierarchical structure is biocompatible and has sufficient mechanical strength for surgery.
Similar content being viewed by others
References
Basu B (2017) Natural bone and tooth: structure and properties. In: Basu B, Ghosh S (eds) Biomaterials for musculoskeletal regeneration. Springer, Singapore, pp 45–85
Wang Q et al (2017) Artificial periosteum in bone defect repair—a review. Chin Chem Lett 28(9):1801–1807
Yu JC et al (1996) An experimental study of the effects of craniofacial growth on the long-term positional stability of microfixation. J Craniofacial Surg 7(1):64–68
Stelnicki EJ, Hoffman W (1998) Intracranial migration of microplates versus wires in neonatal pigs after frontal advancement. J Craniofacial Surg 9(1):60–64
Macchetta A, Turner IG, Bowen CR (2009) Fabrication of HA/TCP scaffolds with a graded and porous structure using a camphene-based freeze-casting method. Acta Biomater 5(4):1319–1327
Miao X et al (2008) Mechanical and biological properties of hydroxyapatite/tricalcium phosphate scaffolds coated with poly (lactic-co-glycolic acid). Acta Biomater 4(3):638–645
Ho CMB, Ng SH, Yoon Y-J (2015) A review on 3D printed bioimplants. Int J Precis Eng Manuf 16(5):1035–1046
Bose S, Vahabzadeh S, Bandyopadhyay A (2013) Bone tissue engineering using 3D printing. Mater Today 16(12):496–504
Rengier F et al (2010) 3D printing based on imaging data: review of medical applications. Int J Comput Assist Radiol Surg 5(4):335–341
Kim S-S et al (2006) Poly (lactide-co-glycolide)/hydroxyapatite composite scaffolds for bone tissue engineering. Biomaterials 27(8):1399–1409
Sato M et al (2006) Increased osteoblast functions on undoped and yttrium-doped nanocrystalline hydroxyapatite coatings on titanium. Biomaterials 27(11):2358–2369
Shao H et al (2017) Bone regeneration in 3D printing bioactive ceramic scaffolds with improved tissue/material interface pore architecture in thin-wall bone defect. Biofabrication 9(2):025003
Xu S et al (2017) Effects of HAp and TCP in constructing tissue engineering scaffolds for bone repair. J Mater Chem B 5(30):6110–6118
Cao H, Kuboyama N (2010) A biodegradable porous composite scaffold of PGA/β-TCP for bone tissue engineering. Bone 46(2):386–395
Rakovsky A et al (2014) β-TCP-polylactide composite scaffolds with high strength and enhanced permeability prepared by a modified salt leaching method. J Mech Behav Biomed Mater 32:89–98
Nakahira A et al (2005) Fabrication of porous hydroxyapatite using hydrothermal hot pressing and post-sintering. J Am Ceram Soc 88(5):1334–1336
Zhou S et al (2011) Fabrication of hydroxyapatite/ethylene-vinyl acetate/polyamide 66 composite scaffolds by the injection-molding method. Polym Plast Technol Eng 50(10):1047–1054
Bose S et al (2003) Pore size and pore volume effects on alumina and TCP ceramic scaffolds. Mater Sci Eng, C 23(4):479–486
Tarafder S et al (2015) SrO-and MgO-doped microwave sintered 3D printed tricalcium phosphate scaffolds: mechanical properties and in vivo osteogenesis in a rabbit model. J Biomed Mater Res Part B Appl Biomater 103(3):679–690
Tarafder S et al (2013) Microwave-sintered 3D printed tricalcium phosphate scaffolds for bone tissue engineering. J Tissue Eng Regen Med 7(8):631–641
Zeng Y et al (2018) 3D printing of hydroxyapatite scaffolds with good mechanical and biocompatible properties by digital light processing. J Mater Sci 53(9):6291–6301
Huang W et al (2013) Fabrication of HA/β-TCP scaffolds based on micro-syringe extrusion system. Rapid Prototyp J 19(5):319–326
Nyberg E et al (2017) Comparison of 3D-printed poly-ɛ-caprolactone scaffolds functionalized with tricalcium phosphate, hydroxyapatite, bio-oss, or decellularized bone matrix. Tissue Eng Part A 23(11–12):503–514
Leukers B et al (2005) Hydroxyapatite scaffolds for bone tissue engineering made by 3D printing. J Mater Sci Mater Med 16(12):1121–1124
Cox SC et al (2015) 3D printing of porous hydroxyapatite scaffolds intended for use in bone tissue engineering applications. Mater Sci Eng, C 47:237–247
Wu C et al (2015) Graphene-oxide-modified β-tricalcium phosphate bioceramics stimulate in vitro and in vivo osteogenesis. Carbon 93:116–129
Kon E et al (2000) Autologous bone marrow stromal cells loaded onto porous hydroxyapatite ceramic accelerate bone repair in critical-size defects of sheep long bones. J Biomed Mater Res 49(3):328–337
Kim J et al (2012) Rapid-prototyped PLGA/β-TCP/hydroxyapatite nanocomposite scaffolds in a rabbit femoral defect model. Biofabrication 4(2):025003
Witek L (2015) Extrusion-based, three-dimensional printing of calcium-phosphate scaffolds. Dissertation of Oklahoma State University
Diogo GS et al (2014) Manufacture of β-TCP/alginate scaffolds through a Fab@ home model for application in bone tissue engineering. Biofabrication 6(2):025001
He F et al (2017) Fabrication of β-tricalcium phosphate composite ceramic sphere-based scaffolds with hierarchical pore structure for bone regeneration. Biofabrication 9(2):025005
Castilho M et al (2014) Direct 3D powder printing of biphasic calcium phosphate scaffolds for substitution of complex bone defects. Biofabrication 6(1):015006
Zhang Y et al (2017) 3D-printed bioceramic scaffolds with antibacterial and osteogenic activity. Biofabrication 9(2):025037
Trombetta R et al (2017) 3D printing of calcium phosphate ceramics for bone tissue engineering and drug delivery. Ann Biomed Eng 45(1):23–44
Castilho M et al (2015) Fabrication of individual alginate-TCP scaffolds for bone tissue engineering by means of powder printing. Biofabrication 7(1):015004
Nadeem D et al (2015) Three-dimensional CaP/gelatin lattice scaffolds with integrated osteoinductive surface topographies for bone tissue engineering. Biofabrication 7(1):015005
Di Luca A et al (2016) Toward mimicking the bone structure: design of novel hierarchical scaffolds with a tailored radial porosity gradient. Biofabrication 8(4):045007
Yang Y et al (2018) Recent progress in biomimetic additive manufacturing technology: from materials to functional structures. Adv Mater 30(36):1706539
Holmes B et al (2016) A synergistic approach to the design, fabrication and evaluation of 3D printed micro and nano featured scaffolds for vascularized bone tissue repair. Nanotechnology 27(6):064001
Salerno A et al (2017) Synthetic scaffolds with full pore interconnectivity for bone regeneration prepared by supercritical foaming using advanced biofunctional plasticizers. Biofabrication 9(3):035002
Yang J-Z et al (2015) Structure design and manufacturing of layered bioceramic scaffolds for load-bearing bone reconstruction. Biomed Mater 10(4):045006
Yang Y et al (2018) 3D-printed biomimetic super-hydrophobic structure for microdroplet manipulation and oil/water separation. Adv Mater 30(9):1704912
Li X, Chen Y (2017) Micro-scale feature fabrication using immersed surface accumulation. J Manuf Process 28:531–540
Li X et al (2018) Mask video projection based stereolithography with continuous resin flow to build digital models in minutes. In: ASME 2018 13th international manufacturing science and engineering conference. American Society of Mechanical Engineers
Zhou C, Chen Y, Waltz RA (2009) Optimized mask image projection for solid freeform fabrication. In: ASME 2009 international design engineering technical conferences and computers and information in engineering conference. American Society of Mechanical Engineers
Li X et al (2018) 3D printing temporary crown and bridge by temperature controlled mask image projection stereolithography. Procedia Manuf 26:1023–1033
Zhou C et al (2013) Digital material fabrication using mask-image-projection-based stereolithography. Rapid Prototyp J 19(3):153–165
Song X et al (2015) Ceramic fabrication using mask-image-projection-based stereolithography integrated with tape-casting. J Manuf Process 20:456–464
Song X et al (2017) Piezoelectric component fabrication using projection-based stereolithography of barium titanate ceramic suspensions. Rapid Prototyp J 23(1):44–53
Frisch U, Hasslacher B, Pomeau Y (1986) Lattice-gas automata for the Navier–Stokes equation. Phys Rev Lett 56(14):1505
Zissi S et al (1996) Stereolithography and microtechniques. Microsyst Technol 2(2):97–102
Jacobs PF (1992) Rapid prototyping & manufacturing: fundamentals of stereolithography. Society of Manufacturing Engineers, Dearborn
Griffith ML, Halloran JW (1993) Freeform fabrication of ceramics via stereolithography. J Am Ceram Soc 79(10):2601–2608
Song X et al (2017) Porous structure fabrication using a stereolithography-based sugar foaming method. J Manuf Sci Eng 139(3):031015
Hamad AJ (2017) Size and shape effect of specimen on the compressive strength of HPLWFC reinforced with glass fibres. J King Saud Univ Eng Sci 29(4):373–380
Miller RG Jr (1997) Beyond ANOVA: basics of applied statistics. Chapman and Hall, Boca Raton
Jordan MM et al (2008) Influence of firing temperature and mineralogical composition on bending strength and porosity of ceramic tile bodies. Appl Clay Sci 42(1–2):266–271
Deng XG et al (2016) Effects of firing temperature on the microstructures and properties of porous mullite ceramics prepared by foam-gelcasting. Adv Appl Ceram 115(4):204–209
Rice RW (1993) Comparison of stress concentration versus minimum solid area based mechanical property-porosity relations. J Mater Sci 28(8):2187–2190
Heunisch A, Dellert A, Roosen A (2010) Effect of powder, binder and process parameters on anisotropic shrinkage in tape cast ceramic products. J Eur Ceram Soc 30(16):3397–3406
Boccaccini AR, Trusty PA (1998) In situ characterization of the shrinkage behavior of ceramic powder compacts during sintering by using heating microscopy. Mater Charact 41(4):109–121
Hollister SJ (2009) Scaffold design and manufacturing: from concept to clinic. Adv Mater 21(32–33):3330–3342
Kwok T-H et al (2017) Mass customization: reuse of digital slicing for additive manufacturing. J Comput Inf Sci Eng 17(2):021009
Xu K, Kwok T-H, Chen Y (2016) A reverse compensation framework for shape deformation in additive manufacturing. In: ASME 2016 11th international manufacturing science and engineering conference. American Society of Mechanical Engineers
Leung Y-S et al (2019) Challenges and status on design and computation for emerging additive manufacturing technologies. J Comput Inf Sci Eng 19(2):021013
Chung I-H et al (2009) Stem cell property of postmigratory cranial neural crest cells and their utility in alveolar bone regeneration and tooth development. Stem Cells 27(4):866–877
Leucht P et al (2008) Embryonic origin and Hox status determine progenitor cell fate during adult bone regeneration. Development 135(17):2845–2854
Acknowledgements
The work was supported by the Alfred E. Mann Institute at University of Southern California as a grant to Yang Chai and Yong Chen. The authors also acknowledge the support of National Science Foundation (NSF) grants 1151191 and 1335476 and the Core Center of Excellence in Nano Imaging (CNI) at USC for the use of microscopic measuring equipment.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare there is no conflict of interest.
Ethical approval
The animal experiments in this study were approved by the Institutional Animal Care and Use Committee of the University of Southern California. All applicable guidelines and protocols for care and use of animals were followed.
Rights and permissions
About this article
Cite this article
Li, X., Yuan, Y., Liu, L. et al. 3D printing of hydroxyapatite/tricalcium phosphate scaffold with hierarchical porous structure for bone regeneration. Bio-des. Manuf. 3, 15–29 (2020). https://doi.org/10.1007/s42242-019-00056-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s42242-019-00056-5