钛种植体载抗菌肽涂层的抗菌性及其对成骨细胞活性的影响  

Study on antibacterial properties and osteoblast activity of antimicrobial peptide coatings on titanium implants

作　　者：孙丰权 李慕勤[3] 彭书浩 张慧明[4] 刘苗[1] 曲学延[2] Sun Fengquan;Li Muqin;Peng Shuhao;Zhang Huiming;Liu Miao;Qu Xueyan(Department of Implantology, Stomatological Hospital Affiliated to Jiamusi University, Jiamusi Heilongjiang 154007, China;Key Laboratory of Biomaterials, College of Materials Science and Engineering, Jiamusi University, Jiamusi Heilongjiang 154007, China;Life Science Center, Basic Medicine of Jiamusi University, Jiamusi Heilong]iang 154007, China;Department of Oral and MaxiUofacial Surgery, Stomatological Hospital Affiliated to Jiamnsi University, Jiamusi Heilongjiang 154007, China)

机构地区：[1]佳木斯大学附属口腔医院种植科,黑龙江154007 [2]佳木斯大学附属口腔医院口腔颌面外科,黑龙江154007 [3]佳木斯大学材料科学与工程学院生物材料重点实验室,黑龙江154007 [4]佳木斯大学基础医学院生命科学中心,黑龙江154007

出　　处：《中华口腔医学杂志》2018年第6期419-424,共6页Chinese Journal of Stomatology

基　　金：国家自然科学基金（31370979）

摘　　要：目的探讨钛种植体表面载抗菌肽涂层对牙龈卟啉单胞菌的抗菌性及其对成骨细胞黏附、增殖的影响，为构建具有抗菌功能及良好生物活性的口腔种植体功能性生物涂层提供依据。方法纯钛试件经超声微弧氧化、碱处理、硅烷化处理后，分别加入0.25、0.50、0.75 g/L抗菌肽Pac-525得到3组抗菌肽组试件，对照组钛试件仅进行超声微弧氧化、碱处理和硅烷化处理（每组样本量均为45）。用扫描电镜及能谱分析仪观察各组涂层形貌并分析元素变化，吖定橙-溴化乙锭双染色检测各组试件与牙龈卟啉单胞菌共培养72 h后的活菌平均百分比和生物膜厚度，细胞计数试剂盒检测成骨细胞增殖情况，免疫荧光染色观察成骨细胞黏附数量及生长形态。结果扫描电镜观察可见抗菌肽颗粒嵌入涂层表面的微孔中，各抗菌肽组C、N元素均明显多于对照组；对照组、0.25、0.50、0.75 g/L抗菌肽组的活菌平均百分比分别为0.58%、0.45%、0.34%和0.28%，各组间差异均有统计学意义（P〈0.05）。0.50和0.75 g/L抗菌肽组牙龈卟啉单胞菌的生物膜厚度[分别为（98.3±1.2）和（94.5±2.5）μm]均显著小于对照组和0.25 g/L抗菌肽组[分别为（117.6±1.5）和（118.0±1.3）μm]（P〈0.05）；各抗菌肽组成骨细胞的黏附和增殖数量均显著大于对照组（P〈0.05），且细胞铺展较好。结论钛种植体载抗菌肽涂层可阻碍细菌生物膜生成，有较好的抗菌性且具有促进成骨细胞黏附及增殖的作用。Objective To investigate the antibacterial property and biological activity of Ti dental implant with antimicrobial peptide Pac-525 coatings, and to study the effect of peptide Pac-525 coatings on Porphyromonas gingivalis＇s antibacterial performance and osteoblast proliferation and adhesion.Methods After ultrasonic micro arc oxidation, alkali treatment and silane treatment, forty-five pure titanium specimens were exposed to antibacterial peptide Pac-525 in different concentration （0.25, 0.50, 0.75 g/L）. The titanium specimens in the control group were only treated with ultrasonic micro arc oxidation, alkali treatment and silane treatment. The morphologies of coatings were observed by scanning electron microscope （SEM）, and the element changes were detected by energy spectrum analyzer. Orange acridine-ethidium bromide double staining was used to detect the average percentage of live bacteria and biofilm thickness, after the specimens in each group and Porphyromonas gingivalis were co-cultured for 72 hours. Cell counting Kit-8 method and immunofluorescence staining were used to test the proliferation of osteoblasts, the number and growth morphologies of adherent cells, respectively.Results SEM and energy spectrum analysis showed that the Pac-525 particles loaded on the surface of the coating, and the C and N elements in the Pac-525 coating group were significantly more than those in the control group. The average percentage of living bacteria in the control group, 0.25, 0.50 and 0.75 g/L antimicrobial peptides were 0.58%, 0.45%, 0.34% and 0.28%, respectively, and the difference between each group was statistically significant （P〈0.05）. The biofilm thickness of Porphyromonas gingivalis in 0.50 and 0.75 g/L antibacterial peptide group were （98.3±1.2） and （94.5±2.5） μm respectively, which were significantly less than those in control group and 0.25 g/L antibacterial peptide group [（117.6±1.5） and （118.0±1.3） μm] （P〈0.05）, respectively. The number of bone cell adhesion and pr

关 键 词：牙种植体 抗菌肽类 紫单胞菌 龈 微弧氧化 

分 类 号：R783.1[医药卫生—口腔医学]