聚多巴胺修饰还原石墨烯负载纳米银复合Gap19多肽水凝胶的制备及抗菌评价  

Preparation and antibacterial evaluation of nanosilver-reduced graphene oxide/polydopamine/methacrylated gelatin@Gap19 hydrogel

作　　者：李若彤 吴悦宁 邓云艺 陈世超 兰小蓉 李适廷 李广文 Li Ruotong;Wu Yuening;Deng Yunyi;Chen Shichao;Lan Xiaorong;Li Shiting;Li Guangwen(Department of Endodontics,Affiliated Stomatological Hospital,Southwest Medical University,Luzhou 646000,Sichuan Province,China;Luzhou Key Laboratory of Oral&Maxillofacial Reconstruction and Regeneration,Luzhou 646000,Sichuan Province,China;Institute of Stomatology,Southwest Medical University,Luzhou 646000,Sichuan Province,China;Department of Implantology,Affiliated Stomatological Hospital,Southwest Medical University,Luzhou 646000,Sichuan Province,China)

机构地区：[1]西南医科大学附属口腔医院,牙体牙髓病科,四川省泸州市646000 [2]口颌面修复重建和再生泸州市重点实验室,四川省泸州市646000 [3]西南医科大学口腔医学研究所,四川省泸州市646000 [4]西南医科大学附属口腔医院,种植科,四川省泸州市646000

出　　处：《中国组织工程研究》2025年第34期7333-7343,共11页Chinese Journal of Tissue Engineering Research

基　　金：四川省自然科学基金面上项目(2024NSFSC0680),项目负责人:李广文;四川省科技厅重点研发项目(22YFS0634),项目负责人:兰小蓉;泸州市科技局重点研发计划(面上)项目(2022-SYF-33),项目负责人:李广文;西南医科大学自科重点项目(2022ZD015),项目负责人:李广文;西南医科大学附属口腔医院重点项目(2022Z01),项目负责人:李广文;四川省卫生健康委员会科技项目资助(24QNMP018),项目负责人:李广文。

摘　　要：背景:活髓保存疗法是治疗深龋等口腔常见疾病的主要疗法之一,盖髓材料的抗菌性能对疗效至关重要。目前常用的盖髓材料抗菌性较弱并会激起牙髓组织一定的炎症反应,从而导致治疗失败。目的:探讨纳米银-还原氧化石墨烯/聚多巴胺/甲基丙烯酰化明胶@Gap19水凝胶材料的抗菌作用。方法:制备纳米银-还原氧化石墨烯。使用含不同质量浓度纳米银-还原氧化石墨烯的完全培养基培养人牙髓干细胞,采用CCK-8实验检测细胞增殖。采用抑菌圈实验检测不同质量浓度纳米银-还原氧化石墨烯对金黄色葡萄球菌的抑制作用。通过CCK-8与抑菌圈实验结果筛选促细胞增殖与抑菌效果最明显的纳米银-还原氧化石墨烯质量浓度,负载于水凝胶中。使用含不同浓度Gap19(一种针对连接蛋白43半通道的特异性抑制剂)的完全培养基培养人牙髓干细胞,采用CCK-8实验检测细胞增殖,筛选促细胞增殖效果最明显的Gap19浓度,负载于水凝胶中。制备纳米银-还原氧化石墨烯/聚多巴胺/甲基丙烯酰化明胶@Gap19(AgNPs-rGO/PDA/GelMA@Gap19)水凝胶,表征该水凝胶材料的理化性质。将金黄色葡萄球菌(或大肠杆菌、变异链球菌、乳酸杆菌)悬液分别与三氧化矿物凝聚体、聚多巴胺/甲基丙烯酰化明胶水凝胶、纳米银-还原氧化石墨烯/聚多巴胺/甲基丙烯酰化明胶水凝胶与AgNPs-rGO/PDA/GelMA@Gap19水凝胶共培养,以单独培养的细菌悬液为空白对照组,检测各组水凝胶的抗菌率。结果与结论:①通过CCK-8与抑菌圈实验结果确定纳米银-还原氧化石墨烯的最佳质量浓度为12.5μg/mL,通过CCK-8实验确定Gap19的最佳浓度为20μmol/L;②扫描电镜下可见AgNPs-rGO/PDA/GelMA@Gap19水凝胶呈褶皱多孔状,纳米银在水凝胶表面及内部均有负载;能谱分析结果显示纳米银-还原氧化石墨烯与Gap19成功负载至水凝胶上;③4种水凝胶对金黄色葡萄球菌、大肠�BACKGROUND:Vital pulp therapy is one of the main treatments for common oral diseases such as deep caries.The antibacterial properties of pulp-capping materials are crucial to the efficacy of the treatment.Currently,commonly used pulp-capping material has weak antibacterial properties and may induce a certain degree of inflammatory response in the pulp tissue,leading to treatment failure.OBJECTIVE:To investigate the antibacterial effects of nanosilver-reduced graphene oxide/polydopamine/methacrylated gelatin@Gap19(AgNPs-rGO/PDA/GelMA@Gap19)hydrogel material.METHODS:Nanosilver-reduced graphene oxide was prepared.Human dental pulp stem cells were cultured in complete medium containing different mass concentrations of nanosilver-reduced graphene oxide.Cell proliferation was detected by CCK-8 assay.The inhibition zone assay was used to detect the inhibitory effect of different mass concentrations of nanosilver-reduced graphene oxide on Staphylococcus aureus.The nanosilver-reduced graphene oxide mass concentration with the most obvious cell proliferation and antibacterial effects was screened by the results of CCK-8 and inhibition zone assays,and loaded into hydrogels.Human dental pulp stem cells were cultured in complete medium containing different concentrations of Gap19(a specific inhibitor of connexin 43 hemichannels),and cell proliferation was detected by CCK-8 assay.The Gap19 concentration with the most obvious cell proliferation effect was screened and loaded into hydrogels.AgNPs-rGO/PDA/GelMA@Gap19 hydrogel was prepared,and the physicochemical properties of the hydrogel material were characterized.The suspension of Staphylococcus aureus(or Escherichia coli,Streptococcus mutans,Lactobacillus)was co-cultured with mineral trioxide aggregates,polydopamine/methacrylated gelatin hydrogel,nanosilver-reduced graphene oxide/polydopamine/methacrylated gelatin hydrogel and AgNPs-rGO/PDA/GelMA@Gap19 hydrogel.The bacterial suspension cultured alone was used as the blank control group to detect the antibacterial rate of each

关 键 词：纳米银 还原氧化石墨烯 Gap19 甲基丙烯酰化明胶 水凝胶 抗菌性 工程化口腔材料 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学] R781.1