微沟槽形貌对人牙龈成纤维细胞整联蛋白intergrin α5、β1表达的影响

摘要

目的：研究微沟槽形貌对人牙龈成纤维细胞（HGF）整联蛋白intergrin α5、integrn β1表达的影响。方法：用光刻技术制作沟槽宽度分别为15、30、60μm，沟槽深度分别为5、10μm（沟槽间隔分别等于各相应宽度）的微沟槽形貌，分别命名为T15／5、T15／10、T30／5、T30／10、T60／5、T60／10，光滑钛（T0）表面作为对照组；分别测量各组材料表面的微沟槽形貌及亲水性，用Real time－PCR和Western blotting 检测HGF在各组材料表面培养3 d时的intergrin α5和β1 mRNA及蛋白的表达水平。结果：T60／5表面亲水性最好，T15／10疏水性最大并明显大于光滑组T0（P＜0．05）；intergrin α5 mRNA表达水平以T60／10组最高，而T15／10时最低，各组间两两相比P＜0．05；沟槽宽度较小时（T15、T30），若深度增加则可减少intergrin α5 mRNA的表达，而沟槽宽度较大时（T60），若深度增加则可促进intergrin α5 mRNA的表达；intergrin β1 mRNA的表达水平以T60／10组最高，与其他各组相比P＜0．05；其余3组间比较均P＞0．05，intergrin α5、intergrin β1蛋白的表达趋势与其mRNA的表达基本一致。结论：intergrin α5和β1表达随着沟槽宽度增大而增加，深度对intergrin α5表达的影响与沟槽的宽度大小相关，沟槽宽度和深度对intergrin β1表达的影响程度低于intergrin α5。%AIM:To investigate the effects of microgroove surface morphology on the expression of intergrinα5 and intergrin β1 in human gingival fibroblasts(HGFs).METHODS:Microgroove titanium surface was fabricated by photolithography with parallel grooves:15 μm,30 μm or 60 μm in width and 5 μm or 10 μm in depth.The combi-nations of groove width and depth were hereafter denoted as T15/5,T15/10,T30/5,T30/10,T60/5 and T60/10. Smooth titanium surface(T0)was used as the control.Surface topography and hydrophilicity were detected.HGFs were cultured on the microgroove surfaces for 3 days.Intergrin α5 and intergrin β1 expression was examined by Real-time PCR and western blotting.RESULTS:T60/5 microgroove showed the highest surface hydrophilicity.However, T15/10 had the highest surface hydrophobicity,even higher than T0 (P<0.05 ).T60/10 group showed the highest expression of intergrin α5 and intergrin β1 mRNA,and in T15/10 group the lowest (P<0.05).In groups with lower n microgroove depth (T15,T30),expression of intergrin α5 decreased with the increase of microgroove depth;while in groups with higher microgroove depth (T60 ),expression of intergrin α5 mRNA increased with the increase of micro-groove depth.T60/10 group showed the highest expression of intergrin β1 mRNA (P <0.05 ),among the other 3 groups,P>0.05.The expression of intergrin α5 and β1 protein showed similar pattern with mRNA expression. CONCLUSION:The effects of microgroove depth on intergrinα5 and interginβ1 depended on micro-groove width.Mi-crogroove width and depth showed less effect on intergrin β1 mRNA and protein when compared to that on intergrin α5 .