钛种植体表面聚电解质多层膜刚度对变形链球菌黏附的影响  

Effect of stiffness of polyelectrolyte multilayer on titanium surface on bacterium adhesion

作　　者：王琼[1] 滕伟[2] 王琴梅[3] 黄洪章[1] 

机构地区：[1]中山大学光华口腔医学院·附属口腔医院口腔颌面外科·广东省口腔医学重点实验室,广州510055 [2]中山大学光华口腔医学院·附属口腔医院口腔修复科·广东省口腔医学重点实验室,广州510055 [3]中山大学附属第一医院卫生部辅助循环重点实验室

出　　处：《中华口腔医学杂志》2016年第3期166-171,共6页Chinese Journal of Stomatology

基　　金：国家自然科学基金（81371665、81571015）;广东省自然科学基金（2015A030313087）

摘　　要：目的 探讨钛种植体表面聚电解质多层膜改性后膜刚度对细菌黏附的影响,为种植体表面改性提供参考.方法 利用层层自组装技术,用儿茶酚化透明质酸（catechol functionalized hyaluronic acid,cHA）和脂多糖胺纳米囊泡（lipopolysaccharide-amine nanopolymersomes,NP）,在钛表面构建儿茶酚化聚电解质多层膜（catechol functionalized polyelectrolyte multilayers,cPEM）,通过调节cHA的儿茶酚摩尔比（5％、10％、20％、30％、40％、50％、60％、70％）调节膜刚度,用压痕划痕仪测量刚度,选择刚度最小、居中及最大的钛试件作为cPEM-L、cPEM-M和cPEM-H组,并以抛光钛为空白对照组.扫描电镜观察各组钛表面形貌.将4组试件分别与变形链球菌共培养,1和24 h后检测细菌黏附量和菌落形成单位（每组每个时间点样本量为5）,扫描电镜和激光扫描共聚焦显微镜观察试件表面细菌黏附形貌.结果 刚度结果显示,cHA中儿茶酚摩尔比分别为5％、30％、70％时对应钛试件cPEM刚度分别最小、居中和最大.空白对照组、cPEM-L、cPEM-M和cPEM-H组刚度分别为（107.1±8.7）、（10.7±4.5）、（21.0±5.8）和（32.6±6.9） GPa （F=16.773,P＜0.05）.扫描电镜可见组装cPEM后,钛片表面更光滑.细菌黏附1和24 h后,各cPEM组细菌黏附量和菌落形成单位均显著大于空白对照组（P＜0.05）.扫描电镜显示随刚度增加,1h细菌黏附量呈递减趋势;24h各组差别不明显.激光扫描共聚焦显微镜可见细菌黏附1h时各组以活菌为主,黏附量随刚度增加而逐渐减少,24 h时cPEM-L和空白对照组表面活菌比例较大,而cPEM-M、cPEM-H组表面死菌比例增大.结论 钛表面聚电解质多层膜刚度增加可抑制变形链球菌的黏附,刚度作为独立的因素可影响细菌的黏附.Objective To provide a theoretical basis for surface modification of titanium implants,the effects of the stiffness of polyelectrolyte multilayer films on titanium surface on bacterium adhesion was explored.Methods Via layer-by-layer technique,catechol functionalized polyelectrolyte multilayer film （cPEM） was constructed on titanium surface by using catechol functionalized hyaluronic acid（cHA） and lipopolysaccharide-amine nanopolymersomes（NP）.The stiffness of cPEM was controlled by adjusting the catechol substitution degree of cHA（5%,10%,20%,30%,40%,50%,60%,70%）.Titanium samples covered with cPEM were selected as test group,and the cPEM was constructed with the lowest,medium and highest stiffness.The polished titanium was used as a control.The surface topography of titanium before and after film construction was observed by scanning electron microscopy（SEM）.At 1 and 24 h after incubation,the adhesion and clonal formation of Streptococcus mutans（S.mutans） on different titanium surfaces were quantified,and their morphology and survival status were observed by SEM and laser scanning confocal microscope（LSCM）.Results When the catechol grafting ratio was 5%,30% and 70%,the lowest,medium and highest cPEM stiffness were obtained,and the cPEM stiffness were （10.69±4.54） GPa（cPEM-L）,（20.99± 5.81） GPa（cPEM-M） and （32.57±6.93） GPa（cPEM-H） respectively,and the stiffness of polished titanium was （107.12±8.68） GPa（P〈0.05）.SEM observation showed that after cPEM coating,the titanium surface became smoother.After incubation for 1 and 24 h,the amount of adhesion and clonal formation of S.mutans on cPEM were higher than those on control titanium,and the difference was statistically significant（P〈0.05）.SEM images showed that for 1 h incubation,softer surfaces were beneficial for S.mutans adhering and agglomerating,while this difference nearly disappeared at 24 h.Observation under LSCM revealed that most of bacteria were alive on titanium disks at 1 h,and their a

关 键 词：钛 牙种植体 细菌粘附 聚电解质多层膜 

分 类 号：R783.6[医药卫生—口腔医学]