外源性Ⅵ型胶原对人后纵韧带细胞增殖和成骨转化的影响  

The influence of type VI collagen on the proliferation and osteogenic differentiation of human spinal ligament cells

作　　者：孙其志[1] 张竞 

机构地区：[1]山东省泰安市解放军88医院骨科中心,山东泰安271000 [2]上海市新华医院骨科,上海200092

出　　处：《生物骨科材料与临床研究》2014年第1期6-9,15,共5页Orthopaedic Biomechanics Materials and Clinical Study

基　　金：2010年度"上海市卫生局科研课题"经费支持(基金编号:2010138)

摘　　要：目的观察外源性VI型胶原蛋白（collagenVI,CⅥ）对人后纵韧带细胞增殖和成骨转化的影响.方法取颈椎后纵韧带骨化症（ossificationoftheposteriorlongitudinalligament,OPLL）患者手术切除的后纵韧带未骨化部分和非OPLL患者因外伤手术切除的正常后纵韧带,原代培养至第3代细胞,分别记作O细胞和N细胞;两种细胞均采用0、6.25、12.5、25、50、100g/mlCⅥ刺激;CCK8法测定刺激后两种细胞1～7天光密度（D）值,评价CⅥ对细胞增殖的影响;骨形态生成蛋白-2（BMP-2）对不同浓度CⅥ刺激的细胞进行诱导骨化,用反转录聚合酶链反应（RT-PCR）检测各组细胞成骨细胞转录因子-2（RunX2）、碱性磷酸酶（ALP）、骨钙素（OC）的基因表达情况.结果CⅥ对两种细胞的增殖均起促进作用,且随着CⅥ浓度增加,其对细胞增殖的促进作用随之增强,但达到一定浓度后这种作用会保持不变或减弱,同一时间点内50g/mlCⅥ刺激时两种细胞的D值最大;BMP-2诱导后的两组细胞有CⅥ刺激的ALP、RunX2和OC的基因表达均高于无CⅥ刺激的对照组（P〈0.05）;在无CⅥ刺激的情况下,O细胞的基因表达高于N细胞（P〈0.05）.结论CⅥ能够促进人后纵韧带细胞的增殖和成骨转化,而来自后纵韧带骨化患者的细胞则更易被促进增殖和诱导骨化,CⅥ可能具有促进后纵韧带细胞成骨的功能.Objective To investigate the influence of type VI collagen（CⅥ）on the proliferation and osteogenic differentiationof human spinal ligament cells. Methods Cells isolated fromthe patients with ossification of the posterior longitudinalligament（OPLL）（Ocells）and the patients with non-OPLL（Ncells）were stimulated by different concentrationsof CⅥ. The proliferation of the cell was measured by CCK8. After the cells were stimulated by different concentrationof CⅥin the presence of BMP-2, the gene expressions of alkaline phosphatase, Runx2 and osteocalcin were examinedby quantitative reverse transcriptase-polymerase chain reaction. Results CⅥpositively regulated the proliferation of bothO cells and N cells in a dose-dependent manner. The strongest effect was observed when the CⅥconcentration was 50g/ml. The gene expressions of alkaline phosphatase, Runx2 and osteocalcin of cells stimulated by CⅥwere higher thanthose of cells not be stimulated by CⅥ（ 〈0.05）. In the absence of CⅥ, the gene expressions of O cells were higherthan those ofNcells（ 〈0.05）.Conclusion CⅥpotentiated the proliferation and BMP-2 induced osteogenic differentiationof human spinal ligament. The cells fromOPLLwere more sensitive than the cells fromnon-OPLLto CⅥ. CⅥ may contributethe onset and progression of OPLL.

关 键 词：Ⅵ型胶原蛋白 后纵韧带骨化症 细胞增殖 成骨转化 

分 类 号：R318.08[医药卫生—生物医学工程]