酚胺交联涂层接枝氯己定改善钛表面抗菌性能研究  

Chlorhexidine-grafted phenolamine coating to improve antibacterial property of the titanium surface

作　　者：丁思勰 洪慧蕾 徐凌寒 王晓玮 张维波[1] 李向阳 王银龙[1] 陈佳龙[1] DING Sixie;HONG Huilei;XU Linghan;WANG Xiaowei;ZHANG Weibo;LI Xiangyang;WANG Yinlong;CHEN Jialong(Stomatologic Hospital and College,Key Laboratory of Oral Diseases Research of Anhui Province,Anhui Medical University,Hefei Anhui,230032,P.R.China)

机构地区：[1]安徽医科大学口腔医学院/附属口腔医院口腔材料教研室口腔疾病研究安徽省重点实验室,合肥230032

出　　处：《中国修复重建外科杂志》2022年第3期335-342,共8页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(31670967、32000932);安徽省重点研究与开发计划(202104j0702003);安徽省高等学校自然科学研究项目(KJ2019A0251);安徽省高等学校省级质量工程项目(2019jyxm0138)~。

摘　　要：目的探究以酚胺交联涂层接枝氯己定(chlorhexidine,CHX)制备纯钛表面的理化性能,并评价改性后材料体外抗菌性及成骨细胞相容性。方法纯钛经过碱热处理(对照组)后,浸泡于表没食子儿茶素没食子酸酯与己二胺混合液,表面沉积酚胺涂层(涂层组),再将其浸泡于CHX溶液,制得CHX接枝表面(接枝组)。扫描电镜观察样品表面形貌、X射线光电子能谱仪分析表面元素组成、水接触角测量分析表面亲水性;活/死细菌染色、浊度法、抑菌环实验评价抗菌性能;通过MTT法和细胞荧光染色评价细胞毒性;细菌-MC3T3-E1细胞共培养评价有菌环境中细胞黏附能力。结果扫描电镜观察示涂层组和接枝组材料表面沉积物依次增加,逐渐覆盖孔隙;X射线光电子能谱仪分析示接枝组N1s峰增强并出现Cl2p特征峰;水接触角测量显示对照组、涂层组、接枝组亲水角依次增加且组间差异有统计学意义(P<0.05)。活/死细菌染色示接枝组表面黏附细菌最少且死细菌比例高,仅接枝组周围有透亮的抑菌环,其培养体系菌液吸光度(A)值未显著增加,且显著低于对照组和涂层组(P<0.05)。MTT和细胞荧光染色结果示接枝组表面黏附细胞最少,但黏附细胞具有良好的增殖活性。细菌-细胞共培养显示仅接枝组表面无细菌且有形态良好的活细胞黏附。结论酚胺交联/CHX抗菌涂层具有抑制细菌黏附与增殖的能力,并在有菌环境中有效保障细胞黏附与增殖。Objective To investigate the physicochemical properties of pure titanium surface grafted with chlorhexidine(CHX)by phenolamine coating,and to evaluate its antibacterial activity and osteoblast-compatibility in vitro.Methods Control group was obtained by alkali and thermal treatment,and then immersed in the mixture of epigallocatechin-3-gallate/hexamethylene diamine(coating group).Phenolamine coating was deposited on the surface,and then it was immersed in CHX solution to obtain the grafted surface of CHX(grafting group).The surface morphology was observed by scanning electron microscope,the surface element composition was analyzed by X-ray photoelectron spectroscopy,and the surface hydrophilicity was measured by water contact angle test.Live/dead bacterial staining,nephelometery,and inhibition zone method were executed to evaluate the antibacterial property.Cytotoxicity was evaluated by MTT assay and cell fluorescence staining.Bacteria-MC3 T3-E1 cells co-culture was conducted to evaluate the cell viability on the samples under the circumstance with bacteria.Results Scanning electron microscope observation results showed that deposits of coating group and grafting group increased successively and gradually covered the porous structure.X-ray photoelectron spectroscopy results showed the peak of N1 s enhanced and the peak of Cl2 p appeared in grafting group.Water contact angle test results showed that the hydrophilic angle of three groups increased in turn,and there was significant difference between groups(P<0.05).Live/dead bacteria staining results showed that the grafting group had the least amount of bacteria adhered to the surface and the proportion of dead bacteria was high.The grafting group had a transparent inhibition zone around it and the absorbance(A)value did not increase,showing significant difference when compared with control group and coating group(P<0.05).MTT assay and cell fluorescence staining results showed that the number of adherent cells on the surface of the grafting group was the least,but

关 键 词：氯己定 钛 酚胺交联 涂层 抗菌性能 

分 类 号：R318.08[医药卫生—生物医学工程]