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为进一步探讨I型胶原及其受体系统是否为成骨细胞功能活动所必需, 使用Ⅰ型胶原和整合素α2β1的一抗阻断Ⅰ型胶原整合素α2β1系统,以细胞计数法比较成骨细胞的增殖能力,流式细胞仪分析成骨细胞的凋亡情况,RTPCR技术研究I型胶原、整合素α2β1 及骨钙素的mRNA表达情况。结果发现阻断I型胶原整合素α2β1系统后,成骨细胞的增殖能力减弱,凋亡率增高,Ⅰ型胶原、整合素α2β1 及骨钙素的mRNA表达减少,其中I型胶原一抗的阻断作用大于整合素α2β1的一抗,这种阻断作用具有可复性,随抗体的去除可部分恢复,表明Ⅰ型胶原整合素α2β1系统为成骨细胞功能活动所必需,构建骨组织工程的支架材料时要考虑到骨组织的基质成份,应为成骨细胞提供相对正常的胞外基质环境。
To further explore whether type I collagen and its receptor system are essential for osteoblast functional activity, type Ⅰ collagen and integrin α2β1 primary antibodies were used to block the type Ⅰ collagen-integrin α2β1 system and to compare osteogenic Cell proliferation, flow cytometry analysis of apoptosis of osteoblasts, RT PCR technique to study the type I collagen, integrin α2β1 and osteocalcin mRNA expression. The results showed that after blocking the type I collagen-integrin α2β1 system, the proliferation of osteoblasts decreased and the apoptosis rate increased. The mRNA expression of type Ⅰ collagen, integrin α2β1 and osteocalcin decreased. The type I collagen primary antibody Blocking effect than integrin α2β1 primary antibody, the blocking effect of a refolding, with the removal of antibodies can be partially restored, indicating that type Ⅰ collagen integrin α2β1 system is necessary for osteoblast functional activity, the construction of bone tissue Engineering scaffolds should take into account the matrix composition of bone tissue and should provide osteoblasts a relatively normal extracellular matrix environment.