四种瓷烤冠基底金属对人牙龈成纤维细胞P70S6k表达的影响

背景:研究已证实,部分烤瓷冠基底金属在体外具有抑制人牙龈成纤维细胞增殖作用,并诱导其凋亡,但烤瓷冠基底金属抑制细胞的具体机制尚不清楚.目的:观察4种烤瓷冠基底合金在体外对人牙龈成纤维细胞P70S6k表达的影响.设计、时间及地点:以人牙龈成纤维细胞P70S6k的表达量为观察对象,对比观察实验.于2009-07/10在辽宁医学院科技楼免疫组化及免疫印迹实验室完成.材料:镍铬合金、纯钛、金合金、钴铬合金由深圳美冠达牙科有限公司提供.选择年龄12-18岁,因正畸需拔除的健康第一前磨牙的牙颈部牙龈缘组织进行原代培养,取其5代对数期的细胞进行指标测定.方法:将镍铬合金、纯钛、金合金、钴铬合金加入DMEM培养液中制备出合金浸提液,以10%浓度加入培养的人牙龈成纤维细胞培养液中.阴性对照组为含体积分数为10%胎牛血清的DMEM培养液.主要观察指标:各组合金浸提液干预72 h后采用免疫印迹分析检测人牙龈成纤维细胞P70s6k表达量.结果:免疫印记结果显示,P70S6k表达的平均灰度值在阴性对照组、金合金组、纯钛组之间差异无显著性意义(P>0.05),金合金组与纯钛组、镍铬合金与钴铬合金组之间差异也无显著性意义(P>0.05).而镍铬合金、钴铬合金与阴性对照组、金合金、纯钛组之间差异均具有显著性意义(P<0.01).结论:金合金、纯钛对人牙龈成纤维细胞的增殖活性影响不明显,而镍铬、钴铬合金组对人牙龈成纤维细胞的增殖活性有抑制作用.

Effects of four kinds of ceramic casting alloys on P70S6k expression of human gingival fibroblasts

BACKGROUND: It has been confirmed that some casting alloy can inhibit the activity of human gingival fibroblasts in vitro, which can be used for inducing apoptosis of human gingival fibroblasts (HGFs). However, the mechanism of this effect is poorly understood.OBJECTIVE: To investigate the effect of Ni-Cr, Co-Cr, Au and pure Ti ceramic alloys on the expression of P70S6k by HGFs in vitro.DESIGN, TIME AND SETTING: Grouping controlled experiments with the expression of P70s6k by HGFs in each group as observation objects. Experiments were performed at the Institute of Immunohistochemistry and Western blotting, Technology Building of Liaoning Medical University from July to October 2009.MATERIALS: Ni-Cr, Co-Cr, Au and pure Ti ceramic alloys were processed by Shenzhen Mecodent Dental Laboratory Co., Ltd. The gingival from gingival neck of the first premolar (which goes to be extracted due to the need of orthodontic treatment) of teenager who was15 years old without any clinical disease was obtained, followed by the primary culture of HGFs. The fifth passage of cells in logarithmic phase were used to measure indexes.METHODS: Ni-Cr, Co-Cr, Au and pure Ti ceramic alloys were incubated in DMEM to prepare alloy leaching liquor, which were then added in HGF medium at 10% concentration. DMEM containing 10% fetal bovine serum was used in negative controls. MAIN OUTCOME MEASURES: The expression of P70S6k in each group was measured by Western Blotting following 72 hours of intervention with alloy leaching liquor.RESULTS: Western-blot results showed that there was no significant difference in the average gray value of P70S6k expression in Au, pure Ti ceramic alloys and control groups (P > 0.05). There was also no significant difference between Au ceramic alloys and pure Ti ceramic alloys groups or between Ni-Cr ceramic alloys and Co-Cr ceramic alloys groups. (P > 0.05), but Ni-Cr ceramic alloys group or Co-Cr ceramic alloys group showed significant difference compared with the Au, pure Ti ceramic alloys and control groups (P < 0.01).CONCLUSION: Au ceramic alloys and pure Ti ceramic alloys show that they have no obvious effect on the proliferation activity of HGFs. Ni-Cr and Co-Cr ceramic alloys showed an inhibitory effect on the proliferation activity of HGFs.