In vitro and In vivo Osteogenic Induction of Mesenchymal Stem Cells from Rabbit Bone Marrow / 대한정형외과연구학회지

ABSTRACT

PURPOSE: The aim of this study was to analyze the osteogenic effect of cultured rabbit mesenchymal stem cells(MSCs). MATERIALS AND METHODS: MSCs were obtained from rabbit femur and were cultured in a Dulbecco's Modified Eagle's Medium(DMEM) with beta-glycerophosphatate, L-ascorbic acid, and dexamethasone to proliferate and differentiate into osteoprogenitor cells until 12 weeks. The expression of osteogenic markers was detected by reverse transcription-polymerase chain reaction(RT-PCR) and the release of osteocalcin was measured by Enzyme-linked Immunosorbent Assay(ELISA). MSCs that were cultured on the porous poly-L-lactic-co-glycolic acid(PLGA) scaffold were implanted into athymic nude mouse to observe the osteogenic activity. RESULTS: As the time, we observed osteoblastic-like cells on the culture flask. Mineralized nodules were observed at 3-4 weeks. Osteogenic markers such as osteopontin, osteonectin, type I collagen, and alkaline phophatase were all identified at 2 weeks. But, expression of osteocalcin was only detected after cells differentiation. The amount of osteocalcin which is a specific protein in osteoblast, increased gradually from 2 weeks until 7 weeks. Scanning electron microscopy revealed that the MSCs were well adhered and proliferated within the PLGA scaffold. Immature bone was identified after 10 weeks in the histological examination of transplanted cell-scaffold composit. CONCLUSION: Our results demonstrate gradual differentiation of MSCs into osteoblastic cells. The adhesion and proliferation of the cells within the biodegradable scaffold represents the possibility of bone formation using cell-scaffold composites.