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

口腔生物医学 ›› 2025, Vol. 16 ›› Issue (2): 81-86.

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

韩兴怡¹,余洁¹,何杨天¹,阎茂伸¹,刘奕羚¹,索溪¹,李泽汉²,闫明³

1. 南京医科大学附属口腔医院牙体牙髓病科，口腔疾病研究与防治国家级重点实验室培育建设点，江苏省口腔转化医学工程研究中心
2. 南京医科大学附属口腔医院牙体牙髓科
3. 南京医科大学口腔医学研究所

收稿日期:2025-01-14 修回日期:2025-03-14 出版日期:2025-04-25 发布日期:2025-05-08
通讯作者: 闫明 E-mail:bean1212@163.com
基金资助:
国家自然科学基金

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

Received:2025-01-14 Revised:2025-03-14 Online:2025-04-25 Published:2025-05-08

摘要/Abstract

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（对照组）、1% NaClO组、MB-PDT组、1% NaClO+MB-PDT组、50 μg/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（SRRE）影响。结果：与0.9% NaCl对照组对比，MB-PDT组的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对照组（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。

关键词: 亚甲基蓝, 光动力疗法, 粪肠球菌, 生物膜

Abstract: 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 treatments: 0.9% NaCl; 1% NaClO; MB-PDT; 1% NaClO+MB-PDT; 50 μg/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 proteins (ACE), gelatinase (GELE), and serine protease (SPRE) to the mono-species biofilm of E. faecalis. Results: Compared with the control group of 0.9% NaCl, MB-PDT destroyed the biofilm of E. faecalis and significantly reduced the bacteria count (P＜0.05); the strength and activity of the regenerated biofilm were lower than those of the control 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). Conclusions: MB-PDT has a destructive effect on biofilms of E. faecalis and inhibits the ability of E. faecalis to reform biofilms.

Key words: Methylene Blue, Photodynamic Therapy, Enterococcus faecalis, Biofilm.

韩兴怡余洁何杨天阎茂伸刘奕羚索溪李泽汉闫明. 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[J]. 口腔生物医学, 2025, 16(2): 81-86.

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