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喷砂复合微弧氧化处理纯钛表面后对成骨细胞活性的影响

The inlfuences on activity of osteoblasts on pure titanium surface treated by sandblast-micro arc oxidation

摘要目的:研究喷砂复合微弧氧化处理纯钛材料后成骨细胞的附着、增殖及活性,从而探讨喷砂复合微弧氧化处理技术在钛种植体表面改性中的应用价值和可能性。方法首先将实验分为3组:A组为纯钛对照组、B组为喷砂酸蚀(SLA)组、C组为喷砂复合微弧氧化组,每组35片;取第4代成骨细胞接种于处理好的3组钛片上,培养到测定的时间点;在细胞对数生长期每组取1块钛片,采用扫描电镜观察成骨细胞在钛片表面生长的情况并拍片记录。用血球计数板记录每组钛片表面在1、2、3、6、7、8 d的细胞数,观察细胞的生长曲线。通过MTT比色法测定不同时间点各组细胞的毒性与存活、增殖情况。用碱性磷酸酶(ALP)ELISA检测试剂盒检测细胞的活性。结果MTT法细胞毒性与存活、增殖的测试中,培养1、2 d中测得的各组成骨细胞差异无统计学意义(P>0.05);培养3 d后细胞增殖率具有统计学意义(P<0.05),喷砂复合微弧氧化组>SLA组>纯钛对照组。ALP活性检测中,3、6 d两个检测时间点上,各组ALP活性都具有统计学意义(P<0.05),第6天的活性比3d前有明显增高,喷砂复合微弧氧化组>SLA组>纯钛对照组。结论喷砂复合微弧氧化处理方法比传统的表面改性处理方法更具优势。

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abstractsObjectiveTo investigate the attachment, proliferation and activity of osteoblasts on pure titanium surface treated by sandblast-acid etch micro arc oxidation and to provide experience base by comparing SLA group and control group; to discuss the application value and possibility of S-MAO technology about surface modification of titanium implants.MethodsFirst, this experiment was divided into three groups: 35 of them were not treated in group A, 35 of them were treated by SLA in group B, and 35 of them were treated by sandblast-acid etch micro arc oxidation in group C; second, the forth generation of osteoblasts of SD suckling mouse which were cultured in vitro were placed on the surfaces of titanium samples of three groups and then to culture them to the measured time; To observe the morphology of osteoblasts by hematoxylin- hematoxylin-eosin staining(HE staining). To identify osteoblasts by ALP histological stain and Alizarin red stain mineralization nodules cytology stain. To record the number of cells of surface of titanium in 1d, 2d, 3d, 6d, 7d, 8d by blood counts plate and observe growth curve of cells. To test toxicity, survival and proliferation at different time by MTT method. To test the activity of cells by ALP ELISA.ResultsThere was no obvious statistical significance of osteoblasts among three groups during (1d, 2d)(P>0.05) in the test of toxicity, survival and proliferation by MTT method; after 3 days' culturing, the rate of proliferation of cells had statistical significance (P<0.05), S-MAO group>SLA group>pure titanium group. In the test of ALP activity, the activity of each group had statistical significance at time of 3d and 6d(P<0.05), the activity of 6d was significantly higher than the activities before 3d, S-MAO group>SLA group>pure titanium group. Osteoblasts were well attached on the surface of titanium, the number of cells increased significantly but in 6~8 d which was the lag phrase, the number of cells decreased; all the cell counting had statistical meaning(P<0.05), according to the amount of cells the order was S-MAO group>SLA group>pure titanium group.Conclusion The comparing result of attachment, proliferation and ALP activity of titanium of three groups was S-MAO was better than two others. The S-MAO method not only obtained better microstructure of surface but also improved roughness to form bioactive film to enhance the attachment of osteoblasts. Thus S-MAO method were more advantageous than the traditional modification processing method of surface.

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DOI 10.3760/cma.j.issn.1673-8799.2014.06.012
发布时间 2015-01-16(万方平台首次上网日期,不代表论文的发表时间)
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