生物活性玻璃对大鼠成骨细胞增殖及YAP/TAZ表达的影响  

Effects of bioactive glass on proliferation and YAP/TAZ expression of rat osteoblasts

作　　者：张高龙 王青宁 赵柏松 ZHANG Gaolong;WANG Qingning;ZHAO Baisong(Department of Anesthesiology,Guangzhou Women and Children's Medical Center,Guangzhou 510623,China)

机构地区：[1]广州市妇女儿童医疗中心麻醉科,广东广州510623

出　　处：《口腔生物医学》2024年第6期335-340,共6页Oral Biomedicine

基　　金：国家自然科学基金面上项目(82171219);广州市科学技术局基础研究项目(SL2022A03J01119)。

摘　　要：目的:探讨生物活性玻璃(BG)对大鼠成骨细胞增殖及Yes相关蛋白(YAP)/PDZ结合基序的转录共激活因子(TAZ)表达的影响。方法:构建高密度聚乙烯(HDPE)、羟基磷灰石(HA)+HDPE复合物和BG+HDPE复合物3种生物材料,通过扫描电子显微镜(SEM)和宽角X射线衍射观察3种材料高分子相的晶体形态和结构;3种材料紫外消毒后固定于细胞培养皿底部,将大鼠成骨细胞接种至培养皿中培养7 d,利用相差显微镜和激光共聚焦显微镜观察细胞结构,CCK-8与流式细胞仪分别检测各组细胞增殖和凋亡情况,采用Western blot检测各组成骨细胞碱性磷酸酶(ALP)、Ⅰ型胶原蛋白(COL-1)、骨桥蛋白(OPN)、Runt相关转录因子2(Runx2)、YAP和TAZ蛋白表达。结果:3种材料中HDPE基体相为典型取向结构,HA和BG分散于基体相内。3组成骨细胞为长梭状,细胞形态完整,细胞骨架连续完整无断裂无缺失,细胞核圆润立体细胞呈现有丝分裂状;BG+HDPE组成骨细胞增殖活性高于其他两组细胞(P<0.05),3组细胞凋亡率均低于5%;BG+HDPE组成骨细胞ALP、COL-1、OPN、Runx2、YAP、TAZ蛋白表达高于其他两组(P<0.05)。结论:BG可促进大鼠成骨细胞增殖,其机制可能与上调YAP/TAZ及Runx2表达有关。Objective:To investigate the effects of bioactive glass(BG)on the proliferation of osteoblasts and the expression of YAP/PDZ-binding motif of transcriptional coactivator(TAZ).Methods:High density polyethylene(HDPE),hydroxyapatite(HA)+HDPE complex and BG+HDPE complex were constructed.The crystal morphology and structure of polymer phase in the three biomaterials were observed by scanning electron microscopy(SEM)and wide-angle X-ray diffraction.The three materials were fixed in the bottom of the cell culture dish after ultraviolet disinfection,and the rat osteoblasts were inoculated and cultured in the dish for 7 days.The cell structure was observed by contrast microscope and laser confocal microscope,and the proliferation and apoptosis of the cells in each group were detected by CCK-8 and flow cytometry.Western blot was used to detect the expressions of alkaline phosphatase(ALP),collagen typeⅠ(COL-1),osteopontin(OPN),Runt-associated transcription factor 2(Runx2),YAP and TAZ proteins in osteoblasts in each group.Results:The HDPE matrix phase of the three materials showed a typical orientation structure,and HA and BG were dispersed in the matrix phase.The constituent bone cells were long fusiform,the cell morphology was complete,the cytoskeleton was continuous and complete without fracture or loss,and the cell nuclei were rounded and three-dimensional cells showed mitosis.The proliferation activity of bone cells composed of BG+HDPE was higher than that of the other two groups(P<0.05),and the apoptosis rate was lower than 5%in all three groups.The protein expressions of ALP,COL-1,OPN,Runx2,YAP and TAZ in bone cells composed of BG+HDPE were higher than those in the other two groups(P<0.05).Conclusions:BG can promote the proliferation of osteoblasts in rats,and its mechanism may be related to the up-regulation of YAP/TAZ and Runx2 expression.

关 键 词：生物活性玻璃 成骨细胞 颌骨缺损 增殖 YAP/TAZ 

分 类 号：R783.1[医药卫生—口腔医学]