光动力疗法对粪肠球菌生物膜清除及抗成膜作用的体外研究  

In vitro study on the clearing and anti-biofilm formation effects of photodynamic therapy against Enterococcus faecalis biofilms

作　　者：韩兴怡 余洁 何杨天 阎茂伸 刘奕羚 索溪 李泽汉 闫明[1,2,3] HAN Xingyi;YU Jie;HE Yangtian;YAN Maoshen;LIU Yiling;SUO Xi;LI Zehan;YAN Ming(Department of Endodontics,The Affiliated Stomatological Hospital of Nanjing Medical University,Nanjing 210029,China;State Key Laboratory Cultivation Base of Research,Prevention and Treatment for Oral Diseases,Nanjing 210029,China;Jiangsu Province Engineering Research Center of Stomatological Translational Medicine,Nanjing 210029,China)

机构地区：[1]南京医科大学附属口腔医院牙体牙髓科,江苏南京210029 [2]口腔疾病研究与防治国家级重点实验室培育建设点,江苏南京210029 [3]江苏省口腔转化医学工程研究中心,江苏南京210029

出　　处：《口腔生物医学》2025年第2期81-86,共6页Oral Biomedicine

基　　金：国家自然科学基金(82201024);江苏省科教能力提升工程——江苏省研究型医院(YJXYYJSDW4),江苏省医学创新中心(CXZX202227)。

摘　　要：目的:研究亚甲基蓝介导的光动力疗法(MB-PDT)对粪肠球菌(E.faecalis)生物膜即时杀灭作用及其再成膜能力的影响。方法:体外建立E.faecalis 24 h细菌生物膜后分为0.9%NaCl组、1%NaClO组、MB-PDT组、1%NaClO+MB-PDT组、50μg/mL亚甲基蓝(MB)组,细菌涂布计数法检测各组菌落数,扫描电镜和激光共聚焦显微镜检测生物膜的即时破坏清除效果,MTT和结晶紫染色检测再生生物膜代谢活力及强度,实时荧光定量PCR分析MB-PDT对E.faecalis单菌种生物膜的胶原蛋白黏附素(ACE)、明胶酶(GELE)、丝氨酸蛋白酶(SPRE)影响。结果:与0.9%NaCl组对比,MB-PDT组E.faecalis生物膜且细菌数量明显减少(P<0.05),再生生物膜强度及活性均低于0.9%NaCl组(P<0.05),E.faecalis成膜的ACE、GELE、SPRE基因表达均显著下调(P<0.05)。结论:MB-PDT对E.faecalis生物膜有破坏作用,且有抑制E.faecalis再成膜的能力。Objective:To investigate the effect of methylene-blue-mediated photodynamic therapy(MB-PDT)on the clearance and reformation of biofilms of Enterococcus faecalis(E.faecalis).Methods:After establishing 24-hour bacterial biofilms of E.faecalis in vitro,the samples were divided into different groups for treatment:0.9%NaCl,1%NaClO,MB-PDT,1%NaClO+MB-PDT,50μg/mL MB.Bacteria coating counting method was used to detect the sterility of each group.Scanning electron microscopy and confocal laser microscopy were used to assess the immediate destruction and removal effects of the first four groups on the biofilm.These groups were then allowed to continue treatment to form regenerated biofilms,and MTT and crystal violet staining were employed to detect the metabolic activity and intensity of regenerated biofilm.qRT-PCR was used to analyze the effect of MB-PDT on the expression of genes encoding collagen-binding adhesin(ACE),gelatinase(GELE),and serine protease(SPRE)to the mono-species biofilm of E.faeca-lis.Results:Compared with the group of 0.9%NaCl,MB-PDT destroyed the biofilm of E.faecalis and significantly reduced the bacte-ria count(P<0.05);the strength and activity of the regenerated biofilm were lower than those of 0.9%NaCl group(P<0.05).qRT-PCR showed that the expression of genes related to biofilm formation in E.faecalis was significantly downregulated(P<0.05).Conclu-sions:MB-PDT has a destructive effect on biofilms of E.faecalis and inhibits the ability of E.faecalis to reform biofilms.

关 键 词：亚甲基蓝 光动力疗法 粪肠球菌 生物膜 

分 类 号：R780.2[医药卫生—口腔医学]