关键词: 动态培养；骨髓间充质干细胞；β磷酸三钙支架；成骨分化

Abstract:

Objective  To study the osteogenic differentiation of hBMSCs seeded in the three dimensional porous scaffold and cultured in a dynamic environment. Methods   Human bone marrow-derived mesenchymal stem cells (BMSCs) were isolated from human bone marrow and cultured. After passage and proliferation,  human BMSCs were seeded on the porous β-tricalcium phosphate (β-TCP) scaffold and cultured in a dynamic perfusion environment. After 28 days of culture,  cell proliferation, alkaline phosphatase activity, and the secretion of osteopontin and osteocalcin were measured. The calcification of the extracellular matrix was observed through SEM, histological assay and histomorphometry. A static culture was used as a control. Results  After 28 days of culture, the proliferation of cells and expression of alkaline phosphatase activity were higher in the dynamic culture group than that in the static culture group. In the culture period, the secretion of osteopontin and osteocalcin were higher in the dynamic culture group than that in the static culture group. In the static culture group, human BMSCs only survived and proliferated on the periphery of the β-TCP scaffold and the new bone volume was lower than in the dynamic culture group. However, in the dynamic culture  condition,cells survived and proliferated on the whole scaffold, and the new bone volume was more than that in the static culture group. Conclusion  The dynamic culture environment benefits the proliferation and osteogenic differentiation of human BMSCs in the three dimensional porous scaffold.

Key words: Dynamic culture; Bone marrow-derived mesenchymal stem cells; β-tricalcium phosphate scaffold; Osteogenic differentiation