牙周膜干细胞在钛表面不同应答的实验研究

Response of Periodontal Ligament Stem Cells on Different treated Titanium Discs

目的:研究牙周膜干细胞在不同粗糙程度的钛板表面的增殖情况及成骨基因表达。方法:常规酶消化法培养牙周膜干细胞,按照接种面性质及钛盘表面不同的处理方法分为5组;将牙周膜干细胞按照相同的细胞浓度植于上述5组培养皿内,检测细胞的增值情况和成骨标志蛋白的表达情况。结果:SEM检测钛板表面处理情况:第1组常规机器处理组(G1);第两组75μm Al2O3喷砂组(G2);第3组125μm Al2O3喷砂组(G3);第4组阳极电压处理组(G4);第5组空白对照组(G5)。细胞增殖情况:第1天细胞增殖较慢,但是G2和G3细胞增殖明显高于其他组,统计学处理有显著性差异;第3天细胞增殖增快,G3和G4细胞增殖明显加快;第6天G2表现出最快的细胞增殖率,G5则表现出比其他各组明显缓慢的细胞增殖率。基因表达检测:培养后第1、3、6天GAPDH表达相对稳定,骨桥蛋白的表达在各组无显著性差异;骨钙蛋白的表达则有显著性,到第6天,骨钙蛋白在G5的表达明显降低,而在G2/3的表达明显增强。I型胶原的表达,在整个实验过程中在G5表达较高水平,而在G2,G3轻度降低,在G5明显降低。结论:牙周膜干细胞在粗糙面增殖优于光滑平面,与塑料表面相较,钛板表面能更利于牙周膜干细胞的骨向分化。

Objective.. To investigate the proliferation and differentiation of periodontal ligament stem cells in dif- ferent titanium surface. Methods： According to inoculated surface properties and different processing methods, five groups were divided. The same cell concentration are planted in the above 5 groups in petri dishes, RT--PCR was used to test the osteogenic differentiation surface marker. Results.. SEM observation of titanium plate surface treat- ment,Gl：conventional machined surface;G2：75 /xmA1203：G3： 125 μm A1203： G4： anode voltage treatment group; G5：blank control group. Cell proliferation： the first day cell proliferation is relatively slow, of which the G2 and G3 cell proliferation is much higher than in other groups. The third day cell proliferation obviously increased fast, G3 and G4 cell proliferation quickened. The sixth day G2 show the fastest rate of cell proliferation~ Cells in G5 show much slower proliferation. Gene expression detection： At 1st, 3rd, 6th day, expression of the house--keeping gene--GAPDH was almost unchanged. Osteopontin mRNA expression in each group has no significant difference. Osteocalcin expression is significant, on the sixth day, bone calcium protein in the expression of the G5 significantly reduced, and the expression of the G2/3 obviously increased. Conclusion： Periodontal ligament stem cells prolifera- tion in rough surface is better than the smooth plane. The titanium plate surface is more conducive to osteogenic dif- ferentiation of the periodontal ligament stem cells.