流体剪切力作用下喷砂酸碱处理钛表面对原代成骨细胞Wnt/β-catenin信号通路关键蛋白mRNA表达的影响

Effect of sandblasted and acid-alkali treated titanium surface on the expression of key molecule of primaryosteoblast Wnt/β-catenin signaling under fluid shear stress loading

目的探讨流体剪切力及喷砂酸碱处理钛表面在不同时相对原代成骨细胞Wnt/β-catenin信号通路关键蛋白表达的影响。方法制备抛光处理纯钛钛片(P组)、喷砂酸碱处理纯钛钛片(S组),取新生第一天SD大鼠颅骨进行原代成骨细胞培养并分别接种于玻片(G组)、P组钛片、S组钛片表面,培养72h后,置于流体加载系统中,在0dynes/cm2(静止组)和12dynes/cm2(受力组)大小的流体剪切力下分别作用0h、0.25h、0.5h、1h、2h、4h,通过实时荧光定量聚合酶链反应检测低密度脂蛋白受体相关蛋白5(low-density lipoprotein receptor-relat-ed protein5,LRP5)和β-连环蛋白(β-catenin)的mRNA表达变化。结果 3种表面受力组LRP5和β-catenin的mRNA表达水平均比静止组高(P<0.05)。LRP5的mRNA表达水平在S组最高(P<0.05);细胞受力时G组在1h、P组和S组在0.5h时LRP5的mRNA表达量达到最高峰。β-catenin的mRNA表达水平在S组最高(P<0.05);细胞受力时表达量最高峰在3种表面均出现在0.5h。结论适宜大小的流体剪切力以及喷砂酸碱处理钛表面均能促进成骨细胞Wnt/β-catenin信号通路的激活,并且喷砂-酸碱处理钛表面提高了成骨细胞Wnt/β-catenin信号通路对流体剪切力刺激的敏感性。

Objective To investigate the effect of sandblasted and acid-alkali treated titanium surface on the expression of key molecule of primary osteoblast Wnt/β-catenin signaling under fluid shear stress at different time pionts. Methods The primary osteoblasts of newborn SD rats skull were cultured and seeded on glass slides （group G）, po-lished titanium surface （group P） and sandblasted and acid-alkali treated titanium surface （group S） respectively for72 h, then loaded under 0 dynes/cm^2 （static group） and 12 dynes/cm^2 （loading group） fluid shear stress respectively for 0, 0.25, 0.5, 1, 2 and 4 h. The mRNA expression of low-density lipoprotein receptor-related protein 5 （Lrp5） andβ-catenin were detected by real-time fluorescence quantitative PCR. Results Lrp5 and β-catenin mRNA expression in all three surfaces in loading group were higher than that in static group （P〈0.05）. Lrp5 mRNA level was the highest in group S （P〈0.05）. The expression peaks were detected at 1 h in group G and 0.5 h both in group P and group S.β-catenin mRNA level was the highest in group S （P〈0.05）. The expression peaks were detected at 0.5 h in all three surfaces. Conclusion Appropriate fluid shear stress and sandblasted and acid-alkali treated titanium surface can pro-mote osteoblast Wnt/β-catenin signaling, and sandblasted and acid-alkali treated titanium surface may increase the sensitivity of osteoblast Wnt/β-catenin signal pathway to fluid shear stress.