| Objective：To investigate the effects of icariin on the differentiation andproliferation of the canine bone marrow mesenchyme stem cell under the conditionsof Osteonecrosis microenvironment.Methods：Several healthy12-month-old dogs of clean grade were chosen, whichsex were unlimited and weight were about15-20kg. We established the animal modelof osteonecrosis of canine bilateral femoral heads using liquid nitrogen freezingmethod．And then obtained10ml bone marrow sample in canine model after twoweeks， used the treatment of bone marrow cell kits to separate and collectmononuclear cells．The cells cultured in containing10％fetal bovine serum DMEM.Cut the femoral heads of model into small pieces at the same time, then cultured themin the same medium. Changed the medium every two days, then gathered medium inwhich necrotic bone have been cultured. The second generation of cells were culturedin DMEM which contain5%necrotic liquid and10%fetal bovine serum. The finalconcentration of icariin were2.58,4.18,6.77,10.95,17.72μg/ml respectively. Weused MTT to evaluate the proliferation of cells, the Alkaline phosphatase activitywere measured at the seventh and eighth day under the condition of osteogenicinduction culture, then alizarin red stain were proceeded and calcified tubercle werecounted at the fourteenth day．Result：The types of the primary culture cells were similar to the types offibroblast．Necrotic liquid could promote the proliferation of BMSCs, but had noeffect on the differentiation of BMSCs, however icariin could inhibited theproliferation of BMSCs, but the Alkaline phosphatase activity and the number ofcalcified tubercle were advanced steeply.Conclusion：It is a feasible way to mimic the osteonecrosis microenvironmentby using necrotic liquid method. The icariin could promote the differentiation ofBMSCs into osteogenesis under the osteonecrosis microenvironment, whichsuggested icariin may as a adjuvant drug to improve the curative effects of BMSCs therapy in necrosis of the femoral head. |
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