中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (21): 3332-3337.doi: 10.12307/2022.640

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

表没食子儿茶素没食子酸酯/己二胺涂层接枝氯乙定改善种植体的抗菌性能

颜楷泉，刘惠珊，王晓玮，吴晓琴，张维波，李向阳，王银龙，陈佳龙   

  1. 安徽医科大学口腔医学院/附属口腔医院，口腔疾病研究安徽省重点实验室，安徽省合肥市  230032
  • 收稿日期:2021-08-25 接受日期:2021-10-11 出版日期:2022-07-28 发布日期:2022-01-27
  • 通讯作者: 王银龙，教授，安徽医科大学口腔医学院/附属口腔医院，口腔疾病研究安徽省重点实验室，安徽省合肥市 230032 陈佳龙，教授，安徽医科大学口腔医学院/附属口腔医院，口腔疾病研究安徽省重点实验室，安徽省合肥市 230032
  • 作者简介:颜楷泉，男，1997年生，江西省抚州市人，汉族，安徽医科大学在读硕士，医师，主要从事口腔材料学研究。
  • 基金资助:
    安徽省重点研究与开发计划(202104j07020039)，课题名称：“釉质靶向/抗菌/促矿化”的新型纳米颗粒防治龋病的效果评价，项目负责人：陈佳龙；国家自然科学基金(31670967)，课题名称：构建具有抗菌、促矿化和釉质靶向的纳米复合材料并评价其在龋齿防治中的效果，项目负责人：陈佳龙； 安徽省高等学校自然科学研究项目(KJ2019A0251)，课题名称：抗菌、促矿化和促成骨的生物活性表面并评价其对种植体感染和骨整合不良的防治效果，项目负责人：张维波；国家自然科学基金(32000932)，课题名称：原位构建糖皮质激素负载和一氧化氮释放双功能涂层用于血管支架表面改性研究、项目负责人：李向阳

Epigallocatechin gallate/hexanediamine coating grafted chlorhexidine improves the antibacterial properties of implants

Yan Kaiquan, Liu Huishan, Wang Xiaowei, Wu Xiaoqin, Zhang Weibo, Li Xiangyang, Wang Yinlong, Chen Jialong   

  1. Department of Stomatology/Affiliated Stomatological Hospital of Anhui Medical University, Anhui Provincial Key Laboratory of Oral Disease Research, Hefei 230032, Anhui Province, China
  • Received:2021-08-25 Accepted:2021-10-11 Online:2022-07-28 Published:2022-01-27
  • Contact: Wang Yinlong, Professor, Department of Stomatology/Affiliated Stomatological Hospital of Anhui Medical University, Anhui Provincial Key Laboratory of Oral Disease Research, Hefei 230032, Anhui Province, China Chen Jialong, Professor, Department of Stomatology/Affiliated Stomatological Hospital of Anhui Medical University, Anhui Provincial Key Laboratory of Oral Disease Research, Hefei 230032, Anhui Province, China
  • About author:Yan Kaiquan, Master candidate, Physician, Department of Stomatology/Affiliated Stomatological Hospital of Anhui Medical University, Anhui Provincial Key Laboratory of Oral Disease Research, Hefei 230032, Anhui Province, China
  • Supported by:
    Key Research and Development Program of Anhui Province, No. 202104j07020039 (to CJL); National Natural Science Foundation of China, No. 31670967 (to CJL); Natural Science Research Project of Anhui Province Colleges and Universities, No. KJ2019A0251 (to ZWB); National Natural Science Foundation of China, No. 32000932 (to LXY)

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摘要：

文题释义：
表没食子儿茶素没食子酸酯：是一种在绿茶、乌龙茶和红茶叶中广泛存在的多酚类物质，大量研究表明其具有抗癌、抗突变、预防和治疗心脑血管系统疾病以及调理内分泌、免疫系统等生物活性，同时在机体的解毒功能及药物联合应用方面有着重要应用价值。
氯己定：作为一种阳离子双胍类抗菌剂，不仅对革兰阳性菌、革兰阴性菌、酵母菌、病毒(包括人类免疫缺陷病毒和乙型肝炎病毒)有抑制能力，对细菌生物膜的形成也有抑制作用，已经作为局部抗菌剂、擦洗剂和冲洗剂被广泛运用于临床治疗。

背景：细菌感染引起的并发症严重影响种植体的长期疗效，因此抗菌性能在种植体表面设计中具有重要意义。
目的：在多孔钛表面构建载氯己定的酚胺交联涂层，评价涂层的抗菌性能及细胞相容性。
方法：将钛片打磨清洗，干燥后浸泡于NaOH溶液中，置于60 ℃干燥箱24 h，沸水清洗干燥后记为对照组；将对照组钛片浸泡于含表没食子儿茶素没食子酸酯与己二胺的Tris溶液中24 h，清洗干燥后记为酚胺组；将酚胺组钛片浸泡在氯己定溶液中24 h，清洗干燥后记为氯己定组。通过扫描电镜、X射线光电子能谱、水接触角测量仪、NH基团定量对3组样品进行材料学表征；采用活/死菌染色、抑菌环法及浊度法评价3组样品的抗菌能力，MTT法和细胞荧光染色评价3组样品的细胞相容性，细菌-细胞共培养评价有菌环境中3组样品的细胞黏附能力。
结果与结论：①氯己定组钛片表面被大量球状颗粒覆盖，出现Cl2p峰，水接触角上升至(24.6±3.3)°，表面NH密度增至(46.14±7.63) nmol/cm2，证明涂层构建成功；②抗菌结果显示，氯己定组表面细菌黏附量最低且均为死细菌，周围具有透亮的抑菌环，培养体系中细菌未增殖，证明涂层对材料表面和周围环境的细菌均具有抑制能力；对照组与酚胺组钛片表面细菌黏附较多且以活细菌多见，周围无明显的抑菌环，培养体系中细菌增殖明显；③细胞培养结果显示，氯己定组钛片轻微抑制细胞黏附但不影响细胞的增殖；④细菌-细胞共培养结果显示，仅氯己定组黏附大量细胞且细胞形态良好；⑤结果表明，氯己定接枝表面构建成功且具有良好的抗菌能力，其在有菌环境中有助于成骨细胞的黏附和增殖。
缩略语：表没食子儿茶素没食子酸酯：epigallocatechin gallate，EGCG

https://orcid.org/0000-0002-4466-0479 (颜楷泉) 

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

关键词: 种植体, 钛片, 表面改性, 表没食子儿茶素没食子酸酯, 氯己定, 抗菌

Abstract: BACKGROUND: The complications caused by bacterial infection seriously affect the long-term efficacy of implants, so the antibacterial performance is of great significance in the design of implant surface. 
OBJECTIVE: To construct chlorhexidine-loaded phenolic amine cross-linked coating on the surface of porous titanium, and to evaluate the antibacterial properties and osteoblast compatibility of the coating.
METHODS: The titanium sheet was polished, cleaned, dried, and soaked in NaOH solution, placed in a 60℃ drying box for 24 hours, washed and dried in boiling water, and then recorded as the control group. The titanium sheet of the control group was immersed in the Tris solution containing epigallocatechin gallate and hexanediamine for 24 hours, washed and dried, and recorded as the phenolamine group. The titanium sheet of the phenolamine group was soaked in the chlorhexidine solution for 24 hours, washed and dried, and recorded as the chlorhexidine group. The samples were characterized by scanning electron microscopy, X-ray photoelectron spectroscopy, water contact angle meter, and NH group quantification. The antibacterial ability of each group was evaluated by live/dead bacteria staining, bacteriostatic ring method, and turbidity method. The biocompatibility of each group was evaluated by methyl thiazolyl tetrazolium method and fluorescent staining. The cell adhesion ability of each group in the bacterial environment was evaluated by bacterial-cell coculture. 
RESULTS AND CONCLUSION: (1) The surface of titanium sheet in the chlorhexidine group was covered by a large number of spherical particles, and the Cl2p peak appeared. The water contact angle rose to (24.6±3.3)°, and the NH content was (46.14±7.63) nmol/cm2, which proved that the coating was successfully constructed. (2) The antibacterial results showed that the bacterial adhesion on the surface of chlorhexidine group was the lowest and all of them were dead bacteria. There was a transparent bacteriostatic ring around the coating, and the bacteria did not proliferate in the culture system. It is proved that the coating can inhibit bacteria on and around the material. There were more bacteria on the surface of titanium sheets in the control group and the phenolamine group, and live bacteria were more common. There was no obvious antibacterial ring around, and the bacterial proliferation in the culture system was obvious. (3) Cell results showed that the titanium sheet in the chlorhexidine group slightly inhibited cell adhesion but did not affect cell proliferation. (4) Bacterial-cell coculture results showed that only chlorhexidine group adhered to a large number of cells, and the cell morphology was good. (5) The results confirmed that chlorhexidine grafted surface was successfully constructed and had good antibacterial ability, which was beneficial to the adhesion and proliferation of osteoblasts in the bacterial environment. 

Key words: implants, titanium sheet, surface modification, epigallocatechin gallate, chlorhexidine, antibacterial

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