Preliminary study of influence of bone tissue from osteonecrosis of femoral head on the proliferation and differentiation of canine bone marrow mesenchymal stem cells

OBJECTIVE: To examine the effects of bone tissue from osteonecrosis of femoral head on the proliferation and differentiation of canine bone marrow mesenchymal stem cells in vitro culture. METHODS: A canine model of femoral head osteonecrosis was induced by liquid nitrogen freezing. BMSC were isolated from dog ilium bone marrow by a combination of gradient centrifugation and adherent wall culture. Different bone tissues and BMSC were cultivated indirectly in vitro by co-cultured in Transwell plate. According to the culture media, 3 groups were established: blank group (10%FBS/DMEM), control group (10%FBS/DMEM+ bone tissue from natural femoral head) and experimental group (10%FBS/DMEM+ bone tissue from osteonecrosis of femoral head). Cell proliferation was measured by methyl-thiazol tetrazolium (MTT) method. Cell differentiation was examined by alkaline phosphatase (ALP) staining and its concentration examined. Alizarin red staining method was used to study the calcification effects and Oil red O staining method was used to detect if there was fat emergence. RESULTS: As compared with the blank group, the proliferation in the control and experiment groups were significantly promoted after culturing for Days 1, 3 and 5 (P experiment group > blank group (P < 0.05). Alizarin red staining show control group had the most calcium nodules (12.17 ± 2.48, P < 0.05) and the number of calcium nodules in the experiment group was more than the blank group (P < 0.05). Oil red O staining show there was no fat emergence after 21 days in every group. CONCLUSION: Both natural and osteonecrotic bone tissue of femoral head could promoted the proliferation of canine BMSC and induces them osteogenic differentiation but not lipo differentiation, bone tissue from osteonecrosis of the femoral head had more competence in promoting proliferation but less capability in inducing osteogenic differentiation than natural bone tissue of the femoral head.