目的 研究紫外线照射后不同保存环境对钛表面成骨细胞黏附、增殖及分化的影响,探讨提高钛表面活性,延缓钛老化的方法.方法 钛试件经抛光、酸蚀后常温常压中储存4周,经紫外线照射后分别进行以下处理:即刻实验(光照组),密闭空气(空气组)或氮气(氮气组)环境中储存4周,以未经紫外线照射的钛试件为对照组；每组34个试件.扫描电镜观察各组钛表面形貌,接触角测试仪检测各组表面接触角.小鼠颅顶前骨细胞亚克隆14接种2、4和24 h后检测各组细胞黏附情况,扫描电镜观察黏附细胞形态；接种3、5及7d后检测各组细胞增殖情况；接种后第3天成骨诱导,7和14 d后检测碱性磷酸酶(alkaline phosphatase,ALP)活性.结果 各组钛试件表面形貌无明显差别.氮气组和光照组表面接触角[分别为(67.70±3.59)°和(0.70±0.28)°]显著小于空气组和对照组[分别为(85.09±1.52)°和(85.23±1.01)°](P＜0.05).接种2和4h后氮气组细胞黏附A值(0.237±0.006和0.578±0.039)均显著高于空气组(0.158±0.036和0.400±0.010)和对照组(0.161±0.024和0.390±0.011),显著低于光照组(0.268±0.015和0.844±0.040)(P＜0.05)；扫描电镜显示接种2h后氮气组和光照组细胞伸展良好,空气组和对照组接种4h后细胞仍未完全展开.培养3和5d后氮气组细胞增殖A值(0.743±0.026和1.548±0.046)均高于空气组(0.499±0.019和1.174±0.062)和对照组(0.508±0.015和1.209±0.025),显著低于光照组(1.250±0.041和1.714±0.096)(P＜0.05).相同时间点各组ALP差异无统计学意义(P＞0.05).结论 紫外线照射可提高钛表面的生物活性,氮气环境可延缓钛表面生物活性的降低.%Objective To evaluate the adhesion,proliferation and differentiation of osteoblast-like cells on the ultraviolet(UV)-treated titanium in different storing environment,and to find a way to enhance the bioactivity of titanium and to prevent its age-related degradation.Methods Acid-etched titanium disks stored under ambient conditions for 4 weeks and treated with UV light for 48 h.Then disks were divided into three groups and placed in a sealed container for 0 h (no-stored,NO group),4 weeks (air-stored,AS group) or in a sealed container filled with nitrogen for 4 weeks (nitrogen-stored,NS group) respectively.A group of UV-untreated titanium served as negative control (NC group).The surface morphology was evaluated using scanning electron microscopy (SEM),and hydrophilicity of disks were measured using contact angle measuring device.Cell counting kit-8 was used to measure the cell adhesion and proliferation.Cell differentiation was evaluated by testing alkaline phosphatase (ALP) activity using ALP reagent kit.Results There was no difference in surface topography among groups.Contact angels in NS group [(67.70 ±3.59)°] and NO group [(0.70 ±0.28)°]were smaller than the others (P ＜0.05).Cell adhesion in NS group at 2 h and 4 h point was (0.237 ±0.006) and (0.578 ±0.039),respectively,and proliferation at 3 d and 5 d point was (0.743 ±0.026) and (1.548 ±0.046) respectively,which were significantly higher than those in AS group [(0.158 ±0.036),(0.400 ±0.010),(0.499 ±0.019) and (1.174 ±0.062)] and inNCgroup [(0.161±0.024),(0.390±0.011),(0.508 ±0.015) and (1.209±0.025)] at the same time point (P ＜0.05).How ever the results mention above in the NS group were lower than those in the NO group (P ＜ 0.05).No difference were found between data from the AS group and NS group (P ＞ 0.05).Osteoblast-like cells had an abundant spread on NS and NO group during 2 h incubation,but did not exactly spread on AS and NC group after incubation for 4 h.No difference were found in ALP among groups.Conclusions UV treatmentcan enhance bioactivity of titanium,and nitrogen storage can slow down its biological aging.
展开▼
