骨桥蛋白及其受体在黄韧带骨化症黄韧带细胞中的表达及其意义  

Expression and role of osteopontin and its receptors in ligmentum flavum cells derived from patients with ossification of ligamentum flavum

作　　者：许政[1] 周盛源[1] 李学斌[1] 刘晓东[1] 陈雄生[1] 王智清[1] 

机构地区：[1]解放军第二军医大学附属上海长征医院脊柱三科,上海市200003

出　　处：《中国组织工程研究》2016年第2期179-184,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81171753)~~

摘　　要：背景:黄韧带骨化症的机制尚不明确,骨桥蛋白可能是其骨化过程中重要的影响因素。目的:观察骨桥蛋白及其受体CD44、整合素β3在人黄韧带细胞及人骨化黄韧带细胞中的表达及其意义。方法:胸/腰椎后路减压术中取人骨化节段黄韧带组织和非黄韧带骨化黄韧带组织各8例,均来自成人,两组组织进行体外细胞分离、培养及鉴定,分别行骨桥蛋白、CD44、整合素β3免疫组织化学染色,用倒置相差显微镜观察结果,提取总RNA行半定量PCR检测。结果与结论:免疫细胞化学染色结果显示,黄韧带组织组中骨化节段黄韧带细胞骨桥蛋白、CD44、整合素β3表达强度均大于非黄韧带骨化组(P<0.01)。PCR检测结果显示,黄韧带组织组中骨化节段黄韧带细胞骨桥蛋白、CD44、整合素β3 m RNA表达强度明显强于非黄韧带骨化组(P<0.05)。结果表明,黄韧带骨化时黄韧带细胞中骨桥蛋白及其受体CD44、整合素β3基因表达均升高。BACKGROUND： The exact pathogenesis of ossification of ligamentum flavum has not been elucidated yet. And osteopontin may be an important factor involved in the ossification of ligamentum flavum. OBJECTIVE： To clarify the expression and significance of osteopontin and its receptors, CD44 and integrin-β3, in ligamentum flavum cells between normal controls and patients with ossification of ligamentum flavum.METHODS： Ligamentum flavum tissues were obtained from normal adult controls and adult patients with ossification of ligamentum flavum（n=8 per group） who underwent thoracic/lumbar posterior decompression surgery. Ligmentum flavum cells were separated, cultured and identified in vitro, and osteopontin, CD44, integrin-β3 were stained using immunocytochemistry method and observed under inverted phase contrast microscope. And the m RNA expressions of osteopontin, CD44, integrin-β3 were measured by RT-PCR. RESULTS AND CONCLUSION： Immunocytochemistry results showed that the stronger positive staining for osteopontin, CD44, integrin-β3 was observed in the ossification of ligamentum flavum group than the control group（P〈0.01）. The m RNA expressions of osteopontin, CD44 and integrin-β3 were also higher in the ossification of ligamentum flavum group than the control group（P〈0.05）. These findings indicate that osteopontin and its receptors, CD44 and integrin-β3, in ligamentum flavum cells may play an important role in ossification of ligamentum flavum.

关 键 词：组织工程 骨桥蛋白 黄韧带 组织构建 肌肉肌腱韧带 CD44 整合素Β3 黄韧带细胞 黄韧带骨化症 原代培养 细胞鉴定 免疫荧光染色 PCR 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]