In vitro bone formation by mesenchymal stem cells with 3D collagen/β-TCP composite scaffold

Recent years, various kinds of natural polymers and bioceramics has been used to develop porous scaffolds for bone tissue engineering. Among of them, collagen guarantees good biological conditions, and β-tricalcium phosphate (β-TCP) possesses good oseteoconductivity, cellular adhesion, accelerated differentiation and mechanical property. In this study, rat bone marrow mesenchymal stem cells (rMSC) were cultured in β-TCP/collagen composite scaffolds up to 28 days in order to assess the time-dependent behavior of the extracellular matrix formation and the mechanical performance of the scaffold-cell sysytem. The cell number and ALP activity were evaluated using a spectrophotometric plate reader. Gene expression of osteogenesis was analyzed using the real-time PCR reactions. Compression tests were also conducted periodically by using a conventional testing machine to evaluate the elastic modulus. The increasing behaviors of cell number and ALP activity in the composite scaffold were much better than in the collagen scaffold. The gene expression of osteocalcin and collagen type-I in collagen/β-TCP scaffold was higher than that of the collagen scaffold. The compressive modulus also increased up to 28 days. These results clearly showed that the distribution of micro β-TCP particles is very effective to increase the elastic modulus and promote cell growth.