10％纳米羟基磷灰石聚醚醚酮浸提液培养成骨细胞（MG63）Wnt3a／β－catenin信号通路的表达

目的：研究10％纳米羟基磷灰石聚醚醚酮（10％ HA／SPEEK／PEEK）复合材料浸提液培养MG63细胞后Wnt3a／β－catenin信号通路中特征性蛋白的变化，评价复合材料的生物相容性。方法：Wnt3a持续作用于10％HA／SPEEK／PEEK浸提液培养的MG63细胞。应用免疫荧光染色技术，定性检测Wnt信号通路中β－catenin蛋白的存在。应用Western－Blotting技术相对定量检测β－catenin蛋白的多少。应用实时定量PCR技术相对定量检测信号通路下游基因c－myc、cyclinD1的表达情况。结果：免疫荧光染色和Western－Blotting实验结果显示，Wnt3a作用于10％HA／SPEEK／PEEK浸提液中培养的MG63细胞，β－catenin蛋白绿色深染，位于细胞核中，表达量显著增加。实时定量RT－PCR实验结果显示Wnt3 a作用于10％HA／SPEEK／PEEK浸提液中培养的MG63细胞，信号通路的下游靶向基因c－myc、cyclinD1表达显著增加，Wnt通路开放。结论：10％HA／SPEEK／PEEK对MG63细胞有良好的成骨作用，可以指导临床实验研究。

Effects of PEEK/HA composite materials on MG63 cells

Objective:To study the variety of Wnt3a /β-catenin signaling pathway’s characteristic proteins after MG63 cell cultured with 10%nano-hydroxyapatite PEEK composites extracting solution,and evaluate the biocompatibility of composites.Methods:Wnt3a constantly have taken effect in MG63 cells cultured by 10% HA /SPEEK /PEEK extracting solution.Immunofluorescence staining technique qualitatively detected the presence of β-catenin protein in wnt signaling pathway.Western-Blotting detected relatively quantity of β-catenin protein.Real-time PCR technology detected downstream
genes c-myc, cyclinD1 expression of signal pathway.Results:Immunofluorescence staining and Western-Blotting results showed that Wnt3a taked effect in MG63 cells cultured by 10% HA /SPEEK /PEEK extracting solution.And β-catenin protein stained green, located in the nucleus with a significant increase in expression.Real-time quantitative PCR results showed that Wnt3a taked effect in MG63 cells cultured by 10%HA /SPEEK /PEEK extracting solution.The expression of downstream target genes in c-myc,cyclinD1 of signaling pathway was significantly increased,and Wnt signaling pathway was opened.Conclusion:10%HA /SPEEK /PEEK for MG63 cells have good osteogenesis,and could been safely used in clini-cal studies.