不同微结构磷酸三钙材料对Wnt非经典信号通路相关标志物蛋白的表达影响  

Effects of tricalcium phosphate materials with different microstructures on the expression of non-canonical Wnt signaling pathway-related markers

作　　者：陈凯瑞 包崇云[3] 刘显[3] 肖宇 谭艳林 刘秀飘 熊博凯 张强[2] Chen Kairui;Bao Chongyun;Liu Xian;Xiao Yu;Tan Yanlin;Liu Xiupiao;Xiong Bokai;Zhang Qiang(Department of Oral and Maxillofacial Surgery,Guangzhou Women and Children's Medical Center,Guangzhou 510000,Guangdong Province,China;Department of Oral and Maxillofacial Surgery,the First Affiliated Hospital of Nanchang University,Nanchang 330006,Jiangxi Province,China;State Key Laboratory of Oral Diseases,West China School/Hospital of Stomatology,Sichuan University,Chengdu 400030,Sichuan Province,China)

机构地区：[1]广州市妇女儿童医疗中心口腔外科,广东省广州市510000 [2]南昌大学第一附属医院口腔颌面外科,江西省南昌市330006 [3]四川大学华西口腔医院口腔疾病国家重点实验室,四川省成都市400030

出　　处：《中国组织工程研究》2019年第34期5430-5435,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81560189),项目负责人：张强~~

摘　　要：背景:骨诱导的发生机制目前还不明确,骨诱导过程中支架材料微结构通过调控相关骨向分化信号通路从而发挥关键性作用。目的:探讨不同微结构磷酸三钙材料对MG63细胞中Wnt非经典信号通路相关标志物表达水平的影响。方法:利用H2O2发泡及控制烧结温度合成具有不同微结构的磷酸三钙材料(TCP-compact,TCP-middle,TCP-big),通过压汞实验、扫描电镜扫描确证材料微结构;同时将已确证的3组具有不同微结构磷酸三钙材料与MG63细胞共培养,检测Wnt非经典信号通路的标志物(Wnt5a,FZD1,ROR2,DKK1)的表达。结果与结论:与具有微孔结构材料共培养的细胞中Wnt5a,FZD1,ROR2,DKK1表达水平显著提高(P<0.05),不同孔径的材料之间Wnt5a,ROR2,DKK1表达存在差异(P<0.05);在共培养末期材料组之间4组标记物表达无显著差异(P>0.05)。材料微结构影响Wnt非经典信号通路的表达水平,其相关标志蛋白表达水平的差异为材料微孔结构所调控,在一定范围内材料孔径越大Wnt非经典信号通路被明显促进。BACKGROUND:The mechanism of osteoinduction is still unclear.The scaffold material microstructure plays a key role in the process of osteoinduction by regulating the relevant bone differentiation signaling pathway.OBJECTIVE:To investigate the effects of tricalcium phosphate with different microstructures on the expression levels of related markers in non-canonical Wnt signaling pathway in MG63 cells.METHODS:Tricalcium phosphate materials with different microstructures(TCP-compact,TCP-middle,TCP-big)were synthesized by controlling H2O2 foaming and sintering temperature.The microstructure of the material was confirmed by mercury intrusion porosimetry and scanning electron microscopy.At the same time,three types of tricalcium phosphate materials with different microstructures were co-cultured with MG63 cells to detect the expression of non-canonical Wnt signaling pathway-related markers(Wnt5a,FZD1,ROR2,DKK1).RESULTS AND CONCLUSION:The expression levels of Wnt5a,FZD1,ROR2 and DKK1 were significantly increased in MG63 cells co-cultured with microporous materials(P<0.05).The expressions of Wnt5a,ROR2 and DKK1 were significantly different between materials with different pore sizes(P<0.05).There was no significant difference in the expression of Wnt5a,FZD1,ROR2 and DKK1 between three material groups(P>0.05).The material microstructure affected the expression level of non-canonical Wnt signaling pathway.The difference of the expression level of related markers was regulated by material microstructure.The larger the pore size of the material,the greater the Wnt non-canonical signaling pathway was promoted.

关 键 词：骨缺损 骨诱导 骨向分化 磷酸三钙 微结构 孔隙率 材料孔径 Wnt非经典信号通路 Wnt5a ROR2 

分 类 号：R459.9[医药卫生—治疗学] R782.13[医药卫生—临床医学]