Sema7A干预钛微粒诱导鼠MC3T3-E1成骨细胞的凋亡 |
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作者姓名: | 丛 宇 茹江英 赵云龙 俞 磊 包倪荣 许 斌 赵建宁 |
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作者单位: | 解放军南京军区南京总医院(解放军第二军医大学南京临床学院)骨科,江苏省南京市 210002 |
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基金项目: | 南京军区医药卫生科研基金面上项目(14MS112);江苏省临床医学科技专项资助(BL2012002) |
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摘 要: | BACKGROUND: Semaphorin7A (Sema7A) is a kind of cell surface protein, which can promote the fusion of osteoclasts and the migration of osteoblasts at the same time, affecting the dynamic balance of the bone. It is speculated that Sema7A siRNA may inhibit osteoblast apoptosis induced by titanium particles.OBJECTIVE: To study the effect of Sema7A on the preosteoblast activity inhibited by titanium particles.METHODS:Mouse MC3T3-E1 preosteoblasts at passages 6 and 7 were divided into four groups: in blank control group, MC3T3-E1 cells were cultured alone; in standard control group, cell were cultured with titanium particles; in experimental groups 1 and 2, the cells were cultured with titanium particles+Sema7A overexpression plasmids and titanium particles+Sema7A siRNA, respectively. Apoptotic rate of MC3T3-E1 cells was detected by flow cytometry; the mRNA expression of bone sialoprotein, osteocalcin and type I collagen was detected by Q-PCR; western blot assay was adopted to detect the protein expression of bone sialoprotein, osteocalcin and type I collagen; alizarin red calcium nodule staining was taken to detect the degree of osteoblast mineralization.RESULTS AND CONCLUSION: The expressions of bone sialoprotein, osteocalcin and type I collagen were decreased in the standard control group and experimental group 1, but these expression were significantly increased in the experimental group 2 compared with the standard control group (P < 0.05). Flow cytometry results suggested that the apoptotic rate of osteoblasts in the experimental group 1 was significantly higher than that in the other groups (P < 0.05), and the apoptotic rate in the experimental group 2 was lower than that in the standard control group (P < 0.05). Alizarin red staining showed that there were no obvious mineralized nodules in the experimental group 1, but mineralized nodules formed in the experimental group 2. In brief, the genetic interference technique that inhibits the activity of Sema7A can interfere the process of mouse MC3T3-E1 preosteoblast differentiation inhibited by titanium particles, and thus provide a feasible way for the clinical treatment of wear particles-induced osteolysis using biotechnology.
中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程
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关 键 词: | 组织构建 成骨细胞 Sema7A MC3T3-E1 骨涎蛋白 骨钙素 Ⅰ型胶原蛋白 人工关节 无菌性松动 骨溶解 |
收稿时间: | 2015-10-17 |
Semaphorin7A intervention for titanium particles-induced apoptosis in mouse MC3T3-E1 osteoblasts |
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Authors: | Cong Yu Ru Jiang-ying Zhao Yun-long Yu Lei Bao Ni-rong Xu Bin Zhao Jian-ning |
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Institution: | Department of Orthopedics, Nanjing General Hospital of Nanjing Military Region of Chinese PLA (Nanjing Clinical Medical School Affiliated to the Second Military University of Chinese PLA), Nanjing 210002, Jiangsu Province, China |
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