摘要
目的探讨松动假体周围的假膜组织中成纤维母细胞样基质细胞在假体周围骨溶解中的作用。方法从人工假体周围的假膜标本中分离成纤维母细胞样基质细胞并作体外培养,传代后与外周血单核细胞共培养,并向各组内分别加入骨水泥颗粒和破骨细胞形成抑制因子(OPG)等,检测是否有破骨细胞形成及其活性;并采用半定量RT-PCR法检测成纤维母细胞样基质细胞中核因子κB受体活化因子配基(RANKL)和OPG等破骨细胞调节因子mRNA表达量的变化。结果细胞共培养结束时观察到抗酒石酸磷酸酶(TRAP)和玻璃蛋白酶受体(VNR)阳性的多核细胞及骨吸收陷窝形成,这一过程可被OPG彻底阻断;骨水泥颗粒组骨吸收陷窝的数量较对照组增加。同时基质细胞表达RANKL和OPGmRNA,骨水泥颗粒组的表达量较对照组分别增加,并且RANKL/OPG的比率增大。结论人工假体周围假膜中成纤维母细胞样基质细胞表达膜蛋白型及可溶性RANKL,支持破骨细胞分化及其骨吸收活性,从而促进假体周围骨溶解。
[Objective] To examine whether fibroblastic stromal cells from failure prosthetic pseudomembrane can express solvable RANKL and support osteoclast formation,and to study the effect of bone cement particles on the expression of RANKL and OPG in these cells.[Methods] Fibroblastic stromal cells were isolated from the inner granulation tissue of fresh pseudomembrane from 7 patients undergoing revision arthroplasty for aseptic loosening of the hip.The stromal cells were cultured and passaged for two times.Part of these fibroblastic stromal cells were cocultured with peripheral blood monocytes,and transwells were used to keep these two sorts of cells from contacting with each other.OPG and bone cement particles were also added to the cultures respectively.At the end of the culture,histochemistry staining was performed to examine the expression of tartrate resistant acid phosphatase（TRAP） and vitronectin receptor（VNR）,and the number of lacuna resorption pits on dentine slices in each group was also compared.Another part of the fibroblastic stromal cells was cultured in the presence and absence of bone cement particles.Semiquantitive RT-PCR was performed to study the effect of bone cement particles on the expression of RANKL and OPG mRNA in the stromal cells.[Results] Multinucleated cells both positive for TRAP and VNR were observed both in the presence and absence of bone cement particles,while the number of lucuna resorption pits in the particles group was significantly increased [（20.67±4.28）vs（18.00±3.36）,P =0.03）].OPG completely blocked the formation of TRAP and VNR positive multinucleared cells and lacuna resorption pits.The expression of RANKL and OPG mRNA in particles group was increased 1.6 folds and 1.4 folds respectively（P =0.005,0.008 respectively） compared to the control group,and the ratio of RANKL/OPG was also significantly increased（P =0.01）.[Conclusion] Fibroblastic stromal cells from the inner granulation tissue of failure prosthetic pseudomembrane can express solvable RANKL and support osteoclast formation.Bone cement particles promote the expression of RANKL and OPG in these cells.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2011年第16期1864-1867,1872,共5页
China Journal of Modern Medicine
基金
陕西省科技厅社发公关基金资助项目(No:2009k17-01)
