纯钛表面硅烷偶联剂复合氧化石墨烯涂层对施万细胞生物学行为的影响  

Effect of silane coupling agent composite graphene oxide coating on biological behavior of Schwann cells on pure titanium

作　　者：徐星星 文超举 李英 孟茂花 王勤英 陈镜桥 董强 Xu Xingxing;Wen Chaoju;Li Ying;Meng Maohua;Wang Qinying;Chen Jingqiao;Dong Qiang(School of Stomatology,Guizhou Medical University/Affiliated Stomatological Hospital of Guizhou Medical University,Guiyang 550004,Guizhou Province,China)

机构地区：[1]贵州医科大学口腔医学院/贵州医科大学附属口腔医院,贵州省贵阳市550004

出　　处：《中国组织工程研究》2023年第30期4783-4789,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81860192),项目负责人:董强;贵州医科大学附属口腔医院口腔医学院科研基金(GYKQKY-2020-05号),项目负责人:文超举。

摘　　要：背景:研究种植体表面处理时更看重是否能提高种植体骨结合的效率,而有关口腔种植体骨感知功能的研究则相对较少。目的:探讨纯钛表面硅烷偶联剂共价连接氧化石墨烯涂层对体外施万细胞生物学行为的影响。方法:在TA4钛片表面通过硅烷偶联剂的氨基基团共价连接制备不同质量浓度的氧化石墨烯涂层(0.25,0.5,0.75,1.0 mg/mL,分别命名为0.25-Ti-APTES-GO组、0.5-Ti-APTES-GO组、0.75-Ti-APTES-GO、1.0-Ti-APTES-GO组),通过扫描电镜、拉曼光谱和接触仪对涂层的微观结构、成分及接触角进行表征。分别在光滑钛片、硅烷偶联改性钛片及不同质量浓度氧化石墨烯涂层改性钛片上培养大鼠施万细胞,采用CCK-8法检测各组施万细胞的增殖活性,RT-qPCR和Western blot检测钛片上施万细胞分泌神经生长因子、胶质细胞源性神经营养因子的能力。结果与结论:①扫描电镜下可见,不同质量浓度的氧化石墨烯涂层呈云雾状,高质量浓度氧化石墨烯涂层呈现的云雾状突起更加致密;②与光滑钛片组相比,硅烷偶联改性组钛片表面接触角较小(P<0.05);随氧化石墨烯质量浓度的增加,钛片表面接触角逐渐变小,其中0.75-Ti-APTES-GO组、1.0-Ti-APTES-GO组组接触角小于光滑钛片组(P<0.05);③CCK-8实验显示,与光滑钛片组比较,硅烷偶联剂改性组、0.25-Ti-APTES-GO组、0.5-Ti-APTES-GO组细胞增殖无明显变化(P>0.05),0.75-Ti-APTES-GO组、1.0-Ti-APTES-GO组细胞增殖受到抑制(P<0.05),所以后续实验选择0.25-Ti-APTES-GO组、0.5-Ti-APTES-GO组;④RT-qPCR检测显示,0.5-Ti-APTES-GO组神经生长因子mRNA表达量高于其他3组(P<0.05);0.25-Ti-APTES-GO组胶质细胞源性神经营养因子mRNA表达量低于其他3组(P<0.05),0.5-Ti-APTES-GO组胶质细胞源性神经营养因子mRNA表达量高于光滑钛片组、硅烷偶联剂改性组(P<0.05);⑤Western blot检测显示,0.5-Ti-APTES-GO组神经生长因子蛋白表达量高于光滑钛片�BACKGROUND:The study of implant surface treatment pays more attention to whether it can improve the efficiency of implant osseointegration,while the study of oral implant bone perception function is relatively less.OBJECTIVE:To investigate the effect of silane coupling agent covalently bonded graphene oxide coating on the biological behavior of Schwann cells in vitro.METHODS:Different mass concentrations of graphene oxide coating(0.25,0.5,0.75,and 1.0 mg/mL,named as 0.25-Ti-APTES-GO group,0.5-Ti-APTES-GO group,0.75-Ti-APTES-GO,and 1.0-Ti-APTES-GO group)were prepared on the surface of TA4 titanium sheet by covalent bonding of amino groups of silane coupling agent.The microstructure,composition and contact angle of the coating were characterized by scanning electron microscopy,Raman spectroscopy and contact instrument.Rat Schwann cells were cultured on smooth titanium sheet,silane coupling modified titanium sheet and graphene oxide coating modified titanium sheet with different mass concentrations.The proliferation activity of Schwann cells in each group was detected by CCK-8 cell proliferation assay.RT-qPCR and western blot assay were used to detect the ability of Schwann cells to secrete nerve growth factor and glial cell-derived neurotrophic factor on titanium sheets.RESULTS AND CONCLUSION:(1)Scanning electron microscopy showed that silane coupling agent composite with different concentrations of graphene oxide coatings were cloudy,and high concentration of graphene oxide coating appeared more dense cloudy protrusions.(2)Compared with the smooth titanium sheet group,the contact angle of the silane coupling agent film titanium group was smaller(P<0.05).The contact angle of silane coupling agent composite graphene oxide coating became small with the increase of graphene oxide concentration.Among them,the contact angle of 0.75-Ti-APTES-GO group and 1.0-Ti-APTES-GO group was smaller than that of smooth titanium sheet group(P<0.05).(3)In the CCK-8 cell proliferation assay,compared with the smooth titanium sheet group

关 键 词：纯钛表面 氧化石墨烯 硅烷偶联剂 施万细胞 骨感知 神经营养因子 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学] R783.6