Towards injectable cell-based tissue-engineered bone: the effect of different calcium phosphate microparticles and pre-culturing.
Bone tissue engineering by combining bone marrow stromal cells (BMSCs) with a porous scaffold is a promising technology. Current major challenges are to upscale the technique for clinical application and to improve the handling characteristics. With respect to minimal invasive surgery, moldable and/or injectable formulations are highly preferable. Ceramic microparticles of different HA/TCP formulations (100/0, 70/30, 60/40, 40/60, and 0/100) with varying surface roughness were sieved to select 200 microg aliquots of the 212-300 microm fraction. Goat BMSCs were seeded on different aliquots one week prior to in vivo implantation. These constructs and remaining cells were cultured for one week. By then, the remaining cells were harvested and resuspended in a specific binder: hyaluronic acid, alginate, or blood plasma, combined with aliquots of 60/40 microparticles peroperatively. All constructs were implanted in nude rats (n = 10) and analyzed for their bone yield histomorphometrically after 6 weeks. All precultured constructs showed consistent bone formation of comparable quantity. No significant differences were observed between the different material compositions. Peroperatively prepared constructs hardly showed any bone formation. The present study demonstrated the osteogenic potential of a tissue- engineered bone substitute made of microparticles of various HA/TCP compositions. There was an obvious advantage when the constructs were pre-cultured.
Department of Orthopaedics, University Medical Center, Utrecht, The Netherlands. mkruyt@studenten.net
Moyo C
MC
Kruyt
Charlotte
C
Persson
Gunilla
G
Johansson
Wouter J A
WJ
Dhert
Joost D
JD
de Bruijn
D000464:Alginates / Q000378:metabolism; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D018786:Bone Substitutes; D002130:Calcium Phosphates / Q000737:chemistry*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000378:metabolism; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006041:Goats; D006820:Hyaluronic Acid / Q000378:metabolism; D006882:Hydroxyapatites / Q000737:chemistry; D020007:Implants, Experimental; D010006:Osteoblasts / Q000166:cytology / Q000502:physiology / Q000637:transplantation*; D010316:Particle Size; D010636:Phenols; D010949:Plasma / Q000378:metabolism; D016062:Porosity; D011897:Random Allocation; D051381:Rats; D011923:Rats, Nude; D017154:Stromal Cells / Q000166:cytology; D013454:Sulfoxides; D023822:Tissue Engineering / Q000379:methods; D016042:Transplantation, Heterotopic; D014961:X-Ray Diffraction; D014992:Xylenes
D000464:Alginates; D018786:Bone Substitutes; D002130:Calcium Phosphates; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006882:Hydroxyapatites; D010636:Phenols; D013454:Sulfoxides; D014992:Xylenes; C083098:calcein green; D006820:Hyaluronic Acid; C016833:xylenol orange