激素性股骨头坏死患者骨髓间充质干细胞体外增殖分化能力的研究

The Ability of Proliferation and Differentiation of Bone Marrow Mesenchymal Stem Cells In Vitro in Steroid-induced Osteonecrosis of Femoral Head

通过对激素性股骨头坏死患者股骨头部骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)增殖及成骨、成脂分化能力的研究,探讨激素性股骨头坏死的发病机理及自体BMSCs移植治疗股骨头坏死的可行性。选择激素性股骨头坏死患者10例,无股骨头坏死的股骨颈骨折患者10例作为对照组。两组患者在行全髋关节置换术中抽取股骨头部骨髓血,密度梯度离心法分离BMSCs并培养至第3代细胞,流式细胞仪检测细胞特异性表面抗原,成纤维细胞–集落形成实验以及成骨、成脂诱导培养后分析细胞的增殖及成骨、成脂分化能力。结果显示,股骨头坏死组细胞表达BMSCs特异性表面抗原CD44、CD73;股骨头坏死组BMSCs形成集落数明显低于对照组;股骨头坏死组碱性磷酸酶活性和钙结节数均低于对照组;股骨头坏死组的成脂细胞数明显高于对照组。该研究结果表明,激素性股骨头坏死患者股骨头BMSCs的增殖及成骨分化能力下降,但其成脂分化潜能增强,推测BMSCs增殖及分化能力的改变可能参与激素性股骨头坏死病理生理过程。

The aim of the study was to explore the pathogenesis of steroid-induced osteonecrosis of femoral head （ON） and the feasibility of autologous bone marrow mesenchymal stem cells （BMSCs） transplantation in the treatment of ON. We studied the ability of the proliferation, osteogenic and adipogenic differentiation of BMSCs which were isolated from femoral heads of patients with steroid-induced ON. 10 patients with steroid-induced ON were selected and 10 patients with femoral neck fractures without ON were treated as the control group. Bone marrow blood of the femoral heads were collected during the total hip arthroplasty （THA）. The BMSCs were isolated by density gradient centrifugation method and cultured to the 3rd passage. The specific antigens on cell surface were detected by flow cytometry. The ability of proliferation, osteogenic and adipogenic differentiation were analyzed by colony-forming unit fibroblast （CFU-F） assay, osteogenic and adipogenic induction. The results showed that specific surface antigen CD44 and CD73 of BMSCs were positive in the ON group. The CFU-F assay showed that the BMSCs obtained from the ON group reduced the amount of colonies compared with the control group. The alkaline phosphatase activity and the calcium nodules of ON group were lower than the control group. But the BMSCs of ON group had the higher ability to produce lipid droplets than the control group. These results showed that BMSCs from patients with steroid-induced ON possess reduced proliferation activity and less capability to differentiate into osteoblasts but more potential to differentiate into adipocytes. The changes of BMSCs proliferation and differentiation ability may be involved in the pathophysiological process of steroid-induced ON.