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

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

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。

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 Cl2 p 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)Bacterialcell 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.