镍钛—不锈钢激光焊复合弓丝细胞毒性的检测

发布时间：2018-07-11 14:17

本文选题：正畸金属丝 + 焊接　；参考：《吉林大学》2014年硕士论文

【摘要】：随着材料加工工艺的发展，镍钛-不锈钢异种金属弓丝的激光焊接方法不断革新，目前中间加层焊接方法较普遍，其中间加层金属有纯镍、钴、铜。与以往激光焊接金属弓丝相比加铜中间层激光焊接的镍钛-不锈钢金属弓丝具有焊接处光滑平整、抗拉强度大、镍钛弓丝侧变形恢复率高、接头弯曲角大等优点。该项发明已申请国家专利，相关研究已检测该复合弓丝的机械性能、腐蚀性能及离子析出，然而针对该复合弓丝的生物安全性检测尚未报道，本文首先检测其细胞安全性。为该复合弓丝的临床应用提供生物安全依据。目的：检测镍钛-不锈钢激光焊复合弓丝的体外细胞毒性，以及其细胞毒性与时间和浓度的相关性，并且与临床常用不锈钢弓丝、镍钛弓丝比较，探讨复合弓丝的细胞安全性。方法：实验分为两部分。实验一：将镍钛-不锈钢复合弓丝经95%酒精浸泡、超声荡洗、去离子水冲洗、烘干、高温高压灭菌后浸泡于高糖细胞培养液（Dulbecco’s modified Eagle’s medium，DMEM），含10%血清与1%双抗（青霉素100000U/L和链霉素100000μg/L），浸泡42h。将浸提液配置成体积分数为50%、40%、30%、20%、10%的稀释液（n=6），与铺板48h的小鼠成纤维细胞系L929细胞共培养，采用甲基噻唑基四唑（methyl thiazolyl tetrazolium，MTT）检测培养24h、48h后各稀释液对小鼠成纤维细胞L929的细胞毒性。实验二：将复合弓丝组与不锈钢弓丝组、镍钛弓丝组（二者处理方式同实验一中复合弓丝）浸泡于高糖细胞培养液（含10%血清与1%双抗）42h。将浸提液配置成体积分数为100%、75%、50%、25%的稀释液(n=6)，与铺板48h的小鼠成纤维细胞系L929细胞共培养，采用MTT检测并比较48h后各稀释液对L929的细胞毒性。阴性对照组加入高密度聚乙烯浸提液（100%体积分数），阳性对照组加入二甲基亚砜。结果：各实验组浸提液细胞毒性均为0～1级，阴性对照组细胞毒性为0级，，阳性对照组细胞毒性为4级。实验一：24h与48h共培养结果显示随着50%至10%复合弓丝浸提液体积分数的降低，光密度值（optical density，OD）呈显著增高趋势；除20%复合弓丝浸提液外，相同体积分数复合弓丝浸提液与L929共培养48h后光密度值均显著低于共培养24h。实验二：100%、75%、50%、25%复合弓丝组浸提液OD值（1.964±0.122、2.084±0.056、2.056±0.071、2.096±0.050）均显著低于与不锈钢组（2.168±0.091、2.227±0.160、2.302±0.052、2.301±0.060）、镍钛弓丝组浸提液（2.138±0.105、2.262±0.050、2.271±0.082、2.294±0.056）（P0.05）。而相同体积分数的不锈钢组与镍钛弓丝组的吸光度值无统计学差异。结论：镍钛-不锈钢激光焊复合弓丝的细胞毒性作用随着浸提液体积分数降低呈现降低；复合弓丝的细胞毒性作用随着培养时间的延长呈略增高趋势；复合弓丝的细胞毒性作用较不锈钢弓丝、镍钛弓丝明显。综上所述，该复合弓丝的细胞毒性作用较小，在材料毒性反应安全级别范围内。
[Abstract]:With the development of material processing technology, the laser welding method of nickel-titanium stainless steel dissimilar metal arch wire is innovated continuously. At present, the welding method of interlayer welding is relatively common, and the interlayer metals include pure nickel, cobalt and copper. Compared with the previous laser welding of metal bow wire, the Ni-Ti stainless steel arc wire with copper interlayer laser welding has the advantages of smooth welding place, high tensile strength, high recovery rate of deformation on the side of Ni-Ti bow wire, and large bending angle of joint, etc. The invention has been applied for a national patent. The mechanical properties, corrosion performance and ion precipitation of the composite bow wire have been examined by relevant research. However, the biosafety detection of the composite bow wire has not been reported. In this paper, the cell safety of the composite bow wire has been first tested. To provide the biosafety basis for the clinical application of the composite arch wire. Objective: to detect the cytotoxicity of Ni-Ti stainless steel laser welded composite arch wire in vitro and its correlation with time and concentration, and to investigate the cell safety of composite wire compared with stainless steel arch wire and nickel titanium arch wire commonly used in clinic. Methods: the experiment was divided into two parts. Experiment 1: the Ni-Ti stainless steel composite bow wire was soaked in 95% alcohol, washed by ultrasonic swing, washed with deionized water, and dried. After sterilizing under high temperature and high pressure, the cells were immersed in high glucose cell culture medium (Dulbeccos modified Eagleus medium DMEM), containing 10% serum and 1% double antibody (penicillin 100000U / L and streptomycin 100000 渭 g / L) for 42 h. The extractant was co-cultured with the mouse fibroblast cell line L929 for 48 hours. The cytotoxicity of each diluent to L929 was detected by (methyl thiazolyl tetrazoliumMTT for 24 h or 48 h. Experiment 2: the composite wire group, stainless steel arch wire group and nickel titanium arch wire group (both treated in the same way as in experiment 1) were immersed in high glucose cell culture medium (containing 10% serum and 1% double antibody) for 42 h. The extract was co-cultured with the mouse fibroblast cell line L929 for 48 hours. The cytotoxicity of the diluents to L929 was measured by MTT assay and compared after 48 hours. The negative control group added high density polyethylene extract (100% volume fraction), the positive control group added dimethyl sulfoxide. Results: the cytotoxicity of the extract in each experimental group was 0 grade 1, that in the negative control group was 0 grade, and that in the positive control group was grade 4. The results of experiment 1: 24 h and 48 h coculture showed that the optical density OD increased significantly with the decrease of 50% ~ 10% composite wire extraction liquid integral, except 20% compound bow wire extraction solution. The optical density was significantly lower than that of co-cultured for 24 h after co-culture of the same volume fraction of the compound arch wire extract and L929 for 48 h. In experiment 2, the OD value of the extract in the 25% composite wire group (1.964 卤0.122) was significantly lower than that in the stainless steel group (2.168 卤0.091 卤2.227 卤0.160 卤2.302 卤0.0522.301 卤0.060) and the nickel titanium wire group (2.138 卤0.105 卤2.262 卤0.0502.271 卤0.0822.294 卤0.056) (P 0.05), and was significantly lower than that in the stainless steel group (2.168 卤0.091) and the nickel titanium wire group (2.138 卤0.105) 2.262 卤0.0502.271 卤0.0822.294 卤0.056 (P0.05). However, there was no significant difference in absorbance between stainless steel group and nickel titanium arch wire group. Conclusion: the cytotoxicity of Ni-Ti stainless steel laser welded composite wire decreased with the decrease of the number of extracted liquid, and the cytotoxicity of composite wire increased slightly with the prolongation of culture time. The cytotoxicity of composite arch wire was more obvious than that of stainless steel arch wire and nickel titanium arch wire. To sum up, the cytotoxicity of the composite arch wire is relatively small, and the safety level of the material toxicity reaction is within the range.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R783.1

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