转化生长因子β3对成骨细胞增殖和成骨能力的影响  

Effect of transforming growth factor-beta3 on the proliferation and osteogenic capability of osteoblasts

作　　者：艾力麦尔旦•艾尼瓦尔 王玲[1] 古丽[1] 迪丽达尔•塔西甫拉提 王珊 尹宏斌[1] Ailimaierdan·Ainiwaer;Wang Ling;Gu Li;Dilidaer•Taxifulati;Wang Shan;Yin Hongbin(Maxillofacial Surgery Clinic,the First Affiliated Hospital of Xinjiang Medical University(Affiliated Stomatological Hospital),Urumqi 830011,Xinjiang Uygur Autonomous Region,China)

机构地区：[1]新疆医科大学第一附属医院(附属口腔医院)颌面外科门诊,新疆维吾尔自治区乌鲁木齐市830011

出　　处：《中国组织工程研究》2021年第17期2664-2669,共6页Chinese Journal of Tissue Engineering Research

基　　金：新疆维吾尔自治区自然科学基金(2016D01C250),项目负责人:王玲。

摘　　要：背景:转化生长因子β3对成骨细胞增殖和成骨能力的影响相关研究甚少。目的:观察不同质量浓度的转化生长因子β3对成骨细胞生长增殖和成骨能力的影响。方法:采用课题组前期方法改良酶消化法分离获得新西兰幼兔颅盖骨成骨细胞,差速贴壁法纯化并鉴定。将成骨细胞分为对照组和实验组,对照组为常规培养的成骨细胞,实验组为分别含0.1,1,10,100μg/L的转化生长因子β3培养。各组每天观察细胞生长情况;培养1,3,5,7,9,11,13 d采用MTT法检测各组细胞增殖情况并绘制细胞生长曲线;培养1,3,5,7,9,11,13,15 d检测各组碱性磷酸酶水平;培养第7,14天用RT-qPCR法检测各组胶原蛋白1A1、Runx-2和骨钙素基因表达情况;培养21d茜素红染色法检测各组细胞矿化能力。实验方案经新疆医科大学第一附属医院动物实验伦理委员会批准。结果与结论:①成功分离培养、纯化并鉴定成骨细胞;②10μg/L转化生长因子β3组细胞增殖活性明显高于对照组(P<0.05);10,100μg/L转化生长因子β3组碱性磷酸酶水平高于对照组(P<0.05);③10μg/L转化生长因子β3组胶原蛋白1A1、Runx-2和骨钙素基因表达量始终高于对照组(P<0.05),100μg/L转化生长因子β3组胶原蛋白1A1,Runx-2和骨钙素基因表达量在第7天时高于对照组(P<0.05);④10,100μg/L转化生长因子β3组矿化结节较对照组大且多,其中10μg/L转化生长因子β3组的矿化结节数明显多于对照组(P<0.05);⑤结果说明,转化生长因子β3在10-100μg/L范围内可促进成骨细胞的增殖和成骨能力,其中10μg/L质量浓度的转化生长因子β3效果最佳。BACKGROUND:Less is reported on the effect of transforming growth factor-β3(TGF-β3)on the proliferation and osteogenic capability of osteoblasts till now.OBJECTIVE:To investigate the ability of TGF-β3 in different doses on the proliferation and osteogenic capability of osteoblasts.METHODS:We isolated osteoblasts from the calvarium of neonatal New Zealand white rabbit according to our previous method,purified and identified the cells using differential attachment method.The osteoblasts were divided into a control group and an experimental group.The osteoblasts in the control group were cultured with conventional culture medium,and those in the experimental group were cultured in the medium containing separately 0.1,1,10,and 100μg/L TGF-β3.The morphology and growth of osteoblasts were observed daily.Cell proliferation was detected by MTT assay and the growth curve was drawn at 1,3,5,7,9,11,and 13 days of culture.Alkaline phosphatase activity was measured at 1,3,5,7,9,11,13,and 15 days of culture.The expression levels of collagen 1A1,Runt related transcription factor 2(Runx-2)and osteocalcin genes were measured by RT-qPCR at 7 and 14 days of culture.Alizarin red S staining was carried out to testify mineralized matrix productivity of each group at 21 days of culture.The study was approved by the Animal Experimental Ethics Committee of the First Affiliated Hospital of Xinjiang Medical University.RESULTS AND CONCLUSION:The osteoblasts were isolated,purified and identified successfully.MTT assay results showed that the cell proliferation activity of 10μg/L TGF-β3 group was higher than that in the control group(P<0.05).The alkaline phosphatase activity of 10 and 100μg/L TGF-β3 groups was higher than that in the control group(P<0.05).RT-qPCR results revealed that the mRNA expressions of collagen 1A1,Runx-2 and osteocalcin in the 10μg/L TGF-β3 group were always higher than those in the control group(P<0.05).The mRNA expressions of collagen 1A1,Runx-2 and osteocalcin in the 100μg/L TGF-β3 group were higher than

关 键 词：骨 生长因子 成骨 成骨细胞 细胞增殖 碱性磷酸酶 基因 

分 类 号：R446[医药卫生—诊断学] R496[医药卫生—临床医学]