钛表面贻贝蛋白包被对人真皮成纤维细胞粘附和增殖的影响  

The influence of titanium surface coated with mussel adhesive protein on the adhesion and proliferation of human skin-derived fibroblasts

作　　者：王忠山[1] 秦海燕[2] 唐立辉[1] 董岩[1] 卢帅[1] 杜静[1] 赵铱民[1] 

机构地区：[1]第四军医大学口腔医学院,陕西西安710032 [2]解放军第454医院口腔科,江苏南京210002

出　　处：《牙体牙髓牙周病学杂志》2012年第12期699-704,736,共7页Chinese Journal of Conservative Dentistry

基　　金：国家自然科学基金资助项目(81200770);南京军区卫生科研基金(11MA055)

摘　　要：目的:观察人真皮成纤维细胞在两种贻贝足丝蛋白提取物Usun-afix和BD CELL-TAKTM试剂包被钛表面的粘附和增殖状况,研究贻贝蛋白包被法是否能够用于经皮种植体表面处理。方法:分别制备光滑钛片和两种贻贝蛋白包被的钛片样本,选取P3代人真皮成纤维细胞接种到3种培养基底上:①不做处理的光滑钛表面(对照组);②Usun-afix包被的钛表面;③BD CELL-TAKTM包被的钛表面。MTT法检测1、3、5 d细胞在3种钛表面的增殖数量;利用DiI-AcLDL荧光染色结合MTT法检测15、30 min和1、2、4 h时间点细胞的粘附状况。结果:培养1、3、5 d,成纤维细胞在3种表面活性均较好,3组之间无明显统计学差异(P>0.05);DiI-AcLDL荧光染色显示,在培养15、30 min和1 h各时间点,Usun-afix和BD CELL-TAKTM包被钛表面粘附细胞数目明显多于光滑钛表面,在1、2、4 h各时间点,两包被组与对照组相比有更好的细胞伸展形态;不同时间点MTT检测显示,两包被组在接种15、30 min和1、2 h各时间点的吸光度值均明显高于对照组(P<0.05),在4 h时,虽两包被组OD值比对照组略高,但无统计学差异(P>0.05);两包被组在各个时间点的促粘附效果相似,差异均无统计学意义(P>0.05)。结论:成纤维细胞接种早期(<4 h),可更容易粘附到两种贻贝足丝蛋白提取物Usun-afix和BD CELL-TAKTM试剂包被的钛表面,而贻贝蛋白对成纤维细胞的增殖能力没有明显的促进效果。AIM： To investigate the effect of titanium surfaces coated with Usun-afix and BD CELL-TAKTM on the proliferation and adhesion of human skin-derived fibroblasts. METHODS： Human skin-derived fibroblasts were seeded on three different titanium surfaces ： group 1, bare smooth titanium （ control group） ; group 2, titanium surface coated with Usun-afix; and group 3, titanium surface coated with BD CELL-TAKTM. The proliferation effects were evaluated by MTT assays in 5 days of cuhure . The adhesion effects were measured by DiI-AcLDL staining and MTY assays when cells were allowed to attach for 15 min,30 min, 1 h, 2 h and 4 h. RESULTS： Cell proliferation re- sults indicated that there was no significant difference among the samples at day 1,3,5 （P 〉 0.05）. Cell-adhesion an- alyses revealed that cell attachment rates of groups 2 and 3 were higher than those of control group （P 〈 0. 05 ） at 15 rain, 30 min, 1 h, 2 h time points, however there was no significant difference at 4 h time point （P 〉 0.05 ）. There was no significant difference between groups 2 and 3 at any time point （P 〉 0.05）. DiI-AcLDL staining results showed that the morphologies of ceils grown on protein-coated surfaces were better at 1 h,2 h,4 h time points compared with those of control group. CONCLUSION： Human skin-derived fibroblasts can adhere to titanium surface coated with mussel adhesive protein better than to normal surface within 4 hours after seeding. And these mussel adhesive proteins have no positive effect on cell proliferation.

关 键 词：贻贝足丝蛋白提取物 成纤维细胞 种植体经皮部 DiI-AcLDL荧光染色 MTT法 

分 类 号：R780.2[医药卫生—口腔医学]