奥硝唑药物对牙髓再生根管内血管化的潜在影响  

Potential effects of ornidazole on intracanal vascularization in endodontic regeneration

作　　者：黎梓楷 张程程 熊嘉颖 杨曦瑞 阳晶 石海山 Li Zikai;Zhang Chengcheng;Xiong Jiaying;Yang Xirui;Yang Jing;Shi Haishan(School of Stomatology,Jinan University,Guangzhou 510632,Guangdong Province,China)

机构地区：[1]暨南大学口腔医学院,广东省广州市510632

出　　处：《中国组织工程研究》2025年第14期2892-2898,共7页Chinese Journal of Tissue Engineering Research

基　　金：广州市科技计划项目基础与应用基础研究项目(202102021127),项目负责人:石海山。

摘　　要：背景:在牙髓治疗中,组织的再生修复和根尖的进一步发育依赖于血运重建及细菌感染的有效控制。有研究报道,将奥硝唑载于盖髓材料或血管化支架材料中可控制牙髓感染,但其血管化影响有待研究。目的:探究奥硝唑在根管内的残留浓度规律,评价奥硝唑对内皮细胞增殖、迁移、分化的影响以及对血管的刺激性。方法:①将奥硝唑包封于离体牙髓腔后浸泡于Hanks缓冲液中,7 d后去除髓腔内的奥硝唑并重新包封无菌水,再次浸泡于Hanks缓冲液中,采用比色法定期检测髓腔液中奥硝唑质量浓度。②将人脐静脉内皮细胞接种于孔板内,观察细胞贴壁后加入脂多糖刺激24 h,然后分别加入0,1,2,5,8,10μg/mL奥硝唑共培养,检测细胞活性与迁移能力。将人脐静脉内皮细胞接种于孔板内,分别加入不同质量浓度(0,1,2,5,8,10μg/mL)奥硝唑共培养,或者经脂多糖刺激24 h后分别加入不同质量浓度(0,1,2,5,8,10μg/mL)奥硝唑共培养,检测血管内皮生长因子、碱性成纤维细胞生长因子的基因表达与血管内皮生长因子的蛋白表达。③采用鸡胚绒毛膜实验评估2,10μg/mL奥硝唑对血管的刺激性。结果与结论:①离体牙中残留的奥硝唑在最初6 d内迅速释放,随后释放速度减慢,8 d以后根尖奥硝唑质量浓度约为2μg/mL。②在脂多糖诱导的炎症条件下,CCK-8与细胞活死荧光染色显示奥硝唑(1-10μg/mL)对人脐静脉内皮细胞的活性无明显影响,细胞划痕实验显示奥硝唑(1-10μg/mL)对人脐静脉内皮细胞的迁移能力无明显影响。RT-qPCR检测显示,与单独的奥硝唑共培养后,人脐静脉内皮细胞中血管内皮生长因子、碱性成纤维细胞生长因子mRNA表达整体呈下降趋势;在脂多糖诱导的炎症条件下与奥硝唑共培养后,人脐静脉内皮细胞中两种因子的mRNA表达呈上升趋势。Western blot检测显示,在脂多糖诱导的炎症条件下与奥硝唑共培养后,人BACKGROUND:In endodontics,revascularization and effective control of bacterial infection are prerequisite for regenerative repair of tissues and further development of the root apex.Ornidazole,carried in pulp-capping materials or vascularized scaffolding materials may control pulpal infections,but its effect on vascularization need to be investigated.OBJECTIVE:To investigate the residual concentration pattern of ornidazole in root canals and to evaluate the effects of ornidazole on endothelial cell proliferation,migration,and differentiation,as well as on vascular irritation.METHODS:(1)Ornidazole was encapsulated in the isolated pulp cavity and then immersed in Hank’s balanced salt solution for 7 days.Ornidazole was then removed from the pulp cavity,reencapsulated in sterile water,and again immersed in Hank’s balanced salt solution.The mass concentration of ornidazole in the pulp cavity fluid was measured periodically by colorimetric method.(2)Human umbilical vein endothelial cells were inoculated into well plates.Adherent cells were stimulated by the addition of lipopolysaccharide for 24 hours,and then co-cultured by the addition of 0,1,2,5,8,10μg/mL ornidazole,to detect the cellular activity and migratory ability.Human umbilical vein endothelial cells were inoculated in well plates and co-cultured with different mass concentrations(0,1,2,5,8,10μg/mL)of ornidazole or stimulated by lipopolysaccharide for 24 hours followed by the addition of different mass concentrations(0,1,2,5,8,10μg/mL)of ornidazole.The gene expression of vascular endothelial growth factor and basic fibroblast growth factor as well as the protein expression of vascular endothelial growth factor was detected.(3)The chorioallantoic membrane assay was employed to assess the vascular irritation of 2 and 10μg/mL ornidazole.RESULTS AND CONCLUSION:Residual ornidazole in exfoliated teeth was rapidly released within the initial 6 days,with a subsequent decrease in release rate,maintaining a concentration of approximately 2μg/mL at the root apex

关 键 词：奥硝唑 内皮细胞 血管再生 药物残留 牙髓感染 根管内血管化 

分 类 号：R459.9[医药卫生—治疗学] R34[医药卫生—临床医学] R781.3