中国组织工程研究

中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (2): 189-195.doi: 10.3969/j.issn.2095-4344.0005

• 组织工程口腔材料 tissue-engineered oral materials • 上一篇    下一篇

纯钛表面TiN/Ag多层复合结构制备及生物学性能的评价

马  明1，万荣欣2，吕晓飞1，褚珊珊1，李立军1，顾汉卿2，彭  诚1
  

  1. 1天津医科大学第二医院，天津市  300211；2天津市泌尿研究所，天津医科大学，天津市  300211
  • 收稿日期:2018-09-28 出版日期:2018-01-18 发布日期:2018-01-18
  • 通讯作者: 彭诚，主任医师，硕士生导师，天津医科大学第二医院口腔科，天津市 300211
  • 作者简介:马明，男，1991年生，辽宁省锦州市人，天津医科大学在读硕士，主要从事口腔种植体表面改性及抑菌成骨方面的研究。
  • 基金资助:
    国家高技术研究发展计划(863计划)项目(2015AA034702)；国家重点研发计划课题(2016YFC1100302)表面改性对生物材料综合性能的影响；天津市应用基础与前沿技术研究计划(重点项目)课题(15JCZDJC38200)；天津市卫计委科技基金课题(2015KZ094)

Preparation and biological properties of TiN/Ag composite coating on pure titanium surface 

Ma Ming1, Wan Rong-xin2, Lv Xiao-fei1, Chu Shan-shan1, Li Li-jun1, Gu Han-qing2, Peng Cheng1 
  

  1. 1The Second Hospital of Tianjin Medical University, Tianjin 300211, China; 2Tianjin Institute of Urological Surgery, Tianjin Medical University, Tianjin 300211, China
  • Received:2018-09-28 Online:2018-01-18 Published:2018-01-18
  • Contact: Peng Cheng, Chief physician, Master’s supervisor, the Second Hospital of Tianjin Medical University, Tianjin 300211, China
  • About author:Ma Ming, Studying for master’s degree, the Second Hospital of Tianjin Medical University, Tianjin 300211, China
  • Supported by:
     the National High-Tech Research and Development Program of China (863 Program), No. 2015AA034702; the National Key Research and Development Program of China, No. 2016YFC1100302; the Applied Basic and Cutting-Edge Technology Research Program of Tianjin, No. 15JCZDJC38200; the Scientific Research Foundation of the Health and Family Planning Commission of Tianjin, No. 2015KZ094

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文题释义：
多弧离子镀技术：是制备硬质薄膜的一项常用技术，原理为在真空条件下利用弧光放电原理，使阴极靶材料蒸发、离化，飞向阳极的机体表面，沉积成膜，其优点有沉积速率较高、操作简单、镀膜速度快、效率高等。
银的细胞毒性机制：尚不完全清楚，但其推测有以下两种说法：一种是银颗粒在溶液中释放银离子，银离子产生细胞毒性，其机制与纳米银产生细胞毒性的原理相似；另一种观点认为银离子可以胞吞的方式进入细胞，并与细胞的细胞器产生效应，干扰细胞的功能，影响细胞的正常生理功能，最终可能导致细胞死亡。
 
背景：在钛种植体表面建立一种能促进细胞早期黏附增殖、分化且无细胞毒性抑菌效果明显的界面至关重要。
目的：在纯钛种植体材料表面制备TiN/Ag多层复合结构涂层，分析其抗菌性能及对MC3T3-E1细胞生物学行为的影响。
方法：采用酸蚀喷砂技术及多弧离子镀技术在光滑纯钛表面制备TiN/Ag多层复合结构。取生长状态良好的MC3T3-E1细胞，分别接种于光滑钛片、酸蚀喷砂钛片及载TiN/Ag多层复合结构钛片表面，接种后24 h，激光共聚焦显微镜观察细胞黏附及细胞活化情况，扫描电镜观察细胞生长形态；接种后24，48，72 h，CCK-8法检测细胞增殖及毒性分级。将金黄色葡萄球菌菌液分别滴加于光滑钛片、酸蚀喷砂钛片及载TiN/Ag多层复合结构钛片表面，16 h后，激光共聚焦显微镜观察细菌生长情况。
结果与结论：①激光共聚焦显微镜显示，光滑组细胞呈两极或三极的纺锤状结构，F-肌动蛋白和丝状伪足表达量较少；酸蚀喷砂组与载银涂层组细胞分散附着排布，方向不同，并形成大量丝状伪足，相互交织融合在一起，细胞铺展较充分，F-肌动蛋白明显高表达，肌动蛋白纤维较粗；②扫描电镜显示，光滑组细胞黏附与铺展不够充分；酸蚀喷砂组、载银涂层组细胞铺展黏附良好，铺展大，细胞状态较好；③CCK-8实验显示接种72 h后，载银涂层组细胞生长良好，增殖明显，细胞毒性为1级；④激光共聚焦显微镜显示，光滑组和酸蚀喷砂组钛片表面金黄色葡萄球菌生长良好，载银涂层组钛片表面细菌出现大面积死亡；⑤结果表明，TiN/Ag多层复合结构涂层具有良好的细胞相容性及抗菌性能。

关键词: 生物材料, 口腔材料, 载银钛, 抑菌, 细胞增殖, 细胞毒性, MC3T3-E1细胞, 多弧离子镀, 涂层

Abstract:

BACKGROUND: It is of vital importance to fabricate an interface on the titanium implant surface which can promote early cell adhesion, proliferation, and differentiation, and exert better antibacterial effects with no cytotoxicity.
OBJECTIVE: To prepare a TiN/Ag composite coating on the surface of pure titanium implant, and to explore its antibacterial properties and effects on MC3T3-E1 biobehaviors.
METHODS: Acid etching blasting and multi-arc ion plating were adopted to prepare TiN/Ag composite coating on the smooth surface of pure titanium. Then, MC3T3-E1 cells that grew well were inoculated onto pure titanium plate, sandblasted and acid-etched titanium plate, and TiN/Ag-coated titanium plate. Twenty-four hours later, cell adhesion and viability were observed under confocal laser scanning microscope, and cell morphology was observed under scanning electron microscope. Cell counting kit-8 was used to detect cell proliferation and cytotoxicity at 24, 48, 72 hours after inoculation. In addition, Staphylococus aureus solution was dropped onto the smooth titanium plated, acid-etched and sandblasted titanium and TiN/Ag-coated titanium plate, and the growth of bacteria was observed by the laser confocal scanning microscope at 16 hours.
RESULTS AND CONCLUSION: Under the confocal laser scanning microscope, spindle cells with bipolar or three poles were observed on the smooth titanium surface, and there was less F-actin and filopodia expression; cells on the TiN/Ag-coated titanium surface and sandblasted and acid-etched titanium surface were scattered with a large amount of interconnected filopodia that were fully stretched and adhered to the titanium surface, highly expressed F-actin was detected, and actin fibers were thickened. Under the scanning electron microscope, the cells on the smooth titanium surface were not fully adhered and stretched, and those on the TiN/Ag-coated titanium surface or the sandblasted and acid-etched titanium surface exhibited better adhesion and extension. Findings from the cell counting kit-8 showed that after 72 hours of inoculation, the cells on the smooth titanium surface grew well, with cytotoxicity level 1. In addition, Staphylococus aureus grew well on the smooth titanium surface under the confocal laser scanning microscope, while a large amount of Staphylococus aureus died on the TiN/Ag-coated titanium surface or on the sandblasted and acid-etched titanium surface. These findings indicated that TiN/Ag coating has good biocompatibility and antibacterial properties. 

Key words: Dental Implants, Titanium, Silver, Cell Biology, Tissue Engineering

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