氧化锆微米涂层对成骨细胞增殖和分化的影响

Effects of zirconia micron coating on the proliferation and differentiation of osteoblasts

目的探讨氧化锆微米涂层对其表面成骨细胞增殖和分化的影响，为氧化锆种植体表面改性提供参考。方法制备直径15.0 mm、厚度1.5 mm的氧化锆陶瓷试件40个，20个经水砂纸抛光作为对照组，20个制备氧化锆微米涂层后作为涂层组。各组取1个试件扫描电镜观察表面形貌。将成骨细胞MC3T3-E1接种于两组试件表面，分别于培养第1、3、5天用扫描电镜观察细胞形态；培养第1、3天用甲基噻唑基四唑法检测细胞增殖能力；分化培养第3、7天实时定量PCR法检测Runt相关转录因子2 （Runt-related transcription factor-2，RUNX2）、特异AT序列结合蛋白2 （special AT-rich sequence binding protein-2, SATB2）、碱性磷酸酶（alkaline phosphatase，ALP）、骨桥蛋白、骨钙蛋白mRNA相对表达量。使用独立样本t检验比较两组差异。结果涂层组试件表面涂覆氧化锆微米颗粒高温烧结后，试件表面可见1-20 μm大小的孔隙。两组试件表面成骨细胞的细胞形态相似。培养第1天两组细胞数量相似（P〉0.05）；培养第3天涂层组成骨细胞数量（1.763±0.165）显著大于对照组（1.067±0.077）（P〈0.05）。分化培养第3天涂层组ALP、骨桥蛋白、骨钙蛋白mRNA相对表达量（分别为1.63±0.28、1.99±0.41、1.60±0.30）均显著大于对照组（均为1.00±0.00）（P〈0.05 ）；分化培养第7天涂层组RUNX2、SATB2、ALP、骨桥蛋白、骨钙蛋白mRNA相对表达量（分别为1.33±0.19、1.64±0.36、1.78± 0.40、2.25±0.36、1.88±0.21）均显著大于对照组（均为1.00±0.00）（P〈0.05）。结论氧化锆微米涂层能促进其表面成骨细胞的增殖和分化。

ObjectiveTo investigate the effects of zirconia micro coating on the proliferation and differentiation of osteoblasts on the surface of zirconia ceramic, and to provide a strategy for zirconia implant surface treatment.MethodsForty tablets of zirconia ceramic, with the diameter of 15 mm and the thickness of 1.5 mm, were prepared. Then, twenty tablets polished by water sandpaper were taken as the control group, and 20 pieces of the zirconia coating after sintering micron were taken as the experimental group. The micromorphology of the surface of the two groups were observed by scanning electron microscope. The cell morphology after inoculation with MC3T3-E1 of osteoblasts on the surface of the material was investigated for 1, 3, and 5 days by scanning electron microscope. The cell proliferation was detected at 1 and 3 days by methyl thiazolyl tetrazolium. The cell differentiation ability was detected at 3 and 7 days by real-time quantitative PCR. Statistical analysis was conducted by independent sample t test.ResultsAfter coating with zirconia micron particles, pores with the diameter of 1-20 μm could be observed on the surface of the test group of tiles through high temperature sintering. The growth of osteoblasts on the surface of the ceramic chip in the test group and control group exhibited the similar cell morphology. As they were cultured for 1 day, the experimental group exhibited a similar quality of cells as those in the test group （P〉0.05）. After 3 days＇ incubation, comparing with the cell quality of the test group （1.067 ± 0.077） （P〈0.05）, the quality of osteoblasts on the surface of zirconia ceramics coating increased to 1.763±0.165, and the expression of mRNA in alkaline phosphatase （ALP）, osteopotin （OPN） and osteocalcin （OCN） also increased with the amount of 1.63±0.28, 1.99±0.41 and 1.60±0.30, respectively, compared with the test group （1.00± 0.00） （P〈0.05）. Seven days later, the expression of mRNA in Runt-related transcription factor-2 （RNUX2） （1.33±0.19）, special AT-rich sequence binding protein-2 （SATB2） （1.64 ± 0.36）, as well as alkaline phosphatase （ALP） （1.78±0.40）, OPN （2.25±0.36）, and OCN （1.88±0.21）, showed a remarkably increase compared with the test group （1.00±0.00） （P〈0.05）.ConclusionsZirconia micro coating on the surface of zirconia ceramics promoted the proliferation and differentiation of osteoblasts adhered.