Semaphorin7A intervention for titanium particles-induced apoptosis in mouse MC3T3-E1 osteoblasts / 中国组织工程研究

ABSTRACT

BACKGROUND:Semaphorin7A (Sema7A) is a kind of cel 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 cels were cultured alone; in standard control group, cel were cultured with titanium particles; in experimental groups 1 and 2, the cels were cultured with titanium particles+Sema7A overexpression plasmids and titanium particles+Sema7A siRNA, respectively. Apoptotic rate of MC3T3-E1 cels was detected by flow cytometry; the mRNA expression of bone sialoprotein, osteocalcin and type I colagen was detected by Q-PCR; western blot assay was adopted to detect the protein expression of bone sialoprotein, osteocalcin and type I colagen; 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 colagen 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.