中国实用口腔科杂志

中国实用口腔科杂志 ›› 2023, Vol. 16 ›› Issue (2): 194-199.DOI: 10.19538/j.kq.2023.02.013

• 论著 • 上一篇    下一篇

二氧化钛纳米管表面阿司匹林涂层的构建及其对骨形成的影响研究

李芯彦，李    毅，蔡东轩，宋    文，张玉梅   

  1. 军事口腔医学国家重点实验室，国家口腔疾病临床医学研究中心，陕西省口腔医学重点实验室，空军军医大学第三附属医院修复科，陕西  西安  710032 
  • 出版日期:2023-03-30 发布日期:2023-04-20
  • 基金资助:
    国家自然科学基金（81530051）

  • Online:2023-03-30 Published:2023-04-20

摘要： 目的    在二氧化钛纳米管（titania nanotubes，TNT）表面构建阿司匹林（acetylsalicylic acid，ASA）涂层，研究其对钛种植体骨结合的影响。方法    阳极氧化法制备TNT阵列，使用含ASA的正电性壳聚糖（chitosan，CS）溶液和负电性DNA溶液在TNT表面进行层层自组装（layer-by-layer self-assembly，LBL），最终得到含ASA的聚合物涂层（TNT-ASA）组，其间使用接触角测量仪检测表面亲水性的变化。另设单纯TNT组和无ASA的聚合物涂层（TNT-CS）组作为对照。通过扫描电镜、原子力显微镜、X射线光电子能谱仪表征试样表面，使用紫外分光光度计测量试样表面ASA在磷酸盐缓冲液中的释放曲线。在试样表面培养MC3T3-E1细胞，采用CCK-8检测细胞活力，使用扫描电镜观察细胞形态。将试样植入SD大鼠股骨远端4周后，采用Micro-CT、HE染色和Masson染色评价试样骨结合效果。结果    LBL过程中材料表面的接触角呈波浪形上下波动，最终于TNT表面制得的聚合物涂层厚度约400 nm。聚合物涂层加载ASA后C元素含量增加，O元素含量降低，其负载的ASA可在12 d内缓慢释放。MC3T3-E1细胞在TNT、TNT-CS和TNT-ASA表面均增殖良好，呈多边形伸展；各组间细胞活力的差异无统计学意义（P > 0.05）。Micro-CT结果显示，相较于TNT组、TNT-CS组，TNT-ASA组骨组织的骨体积分数更高（P < 0.05）。HE染色和Masson染色结果显示，TNT-ASA组植入试样周围具有更多的新骨形成。结论    通过LBL技术可实现ASA在TNT表面的有效加载，该涂层具备药物缓释、无细胞毒性和促进骨形成的优势。

关键词: 钛种植体, 二氧化钛纳米管, 阿司匹林, 骨结合

Abstract: Objective   To construct acetylsalicylic acid-loaded coating on the surface of titania nanotubes（TNT）and study its effect on the osseointegration of titanium implants. Methods   The TNT array was prepared by anodic oxidation. The positive chitosan（CS）solution containing aspirin（ASA）and negative DNA solution were used to carry out layer-by-layer self-assembly（LBL）on the surface of TNT，and finally the polymer coating（TNT-ASA）group containing ASA was obtained. The change of surface hydrophilicity was detected by contact angle measuring instrument. In addition，the TNT group and the polymer coating without ASA（TNT-CS）group were set as controls. The sample surface was characterized by scanning electron microscope，atomic force microscope and X-ray photoelectron spectroscopy，and the release curve of ASA on the sample surface in phosphate buffer salin（PBS）was measured by ultraviolet spectrophotometer. MC3T3-E1 cells were cultured on the surface of the sample，the cell viability was detected with CCK-8 kit，and the cell morphology was observed with scanning electron microscope. After the sample was implanted into the distal femur of SD rats for 4 weeks，the osseointegration of the sample was evaluated by Micro-CT，HE staining and Masson staining. Results   During the LBL process，the contact angle of the material surface fluctuated up and down in a wavy shape，and the thickness of the polymer coating on the TNT surface was about 400 nm. After loading ASA in the polymer coating，the content of C element was increased and the content of O element was decreased. The loaded ASA was released slowly within 12 days. MC3T3-E1 cells proliferated well on the surface of TNT，TNT-CS and TNT-ASA，with polygonal extension. There was no significant difference in cell viability among groups（P > 0.05）. Micro-CT results showed that the bone volume to total volume of bone tissue in TNT-ASA group was higher than that in TNT group and TNT-CS group（P < 0.05）. The results of HE staining and Masson staining showed that there were more new bone formation around the implants in TNT-ASA group. Conclusion    Effective loading of ASA on TNT surface can be achieved by LBL technology. The coating has the advantages of gradual drug release，no cytotoxicity and promoting bone formation.

Key words: titanium implant, titania nanotubes, acetylsalicylic acid, osseointegration