基于重水拉曼技术评价口腔抑菌剂对粪肠球菌抑菌效能的研究

doi:10.13701/j.cnki.kqyxyj.2021.11.007

口腔医学研究 ›› 2021, Vol. 37 ›› Issue (11): 989-993.DOI: 10.13701/j.cnki.kqyxyj.2021.11.007

基于重水拉曼技术评价口腔抑菌剂对粪肠球菌抑菌效能的研究

刘育含^1,2, 马玉莹^2,4, 张利娟³, 李修珍^1,2, 杨加震^2,4, 牛玉芬^2,4, 孙雁斐^1,2, 曾飞^2,4, 杨芳^1*

1.青岛大学口腔医学院山东青岛 266000;
2.青岛市市立医院口腔医学中心山东青岛 266071;
3.临沂市妇幼保健院口腔科山东临沂 276002;
4.大连医科大学口腔医学院辽宁大连 116044

收稿日期:2021-06-03 出版日期:2021-11-28 发布日期:2021-11-22
通讯作者: *杨芳,E-mail:yangf82@sina.com
作者简介:刘育含(1995~ ),女,山东人,硕士在读,医师,研究方向:口腔微生物。
基金资助:
国家自然科学基金(编号:81670979,31300424)

Efficacy Assessment of Clinical Disinfectants for Enterococcus Faecalis via Single-cell Raman Microspectroscopy

LIU Yuhan^1,2, MA Yuying^2,4, ZHANG Lijuan³, LI Xiuzhen^1,2, YANG Jiazhen^2,4, NIU Yufen^2,4, SUN Yanfei^1,2, ZENG Fei^2,4; YANG Fang^1*

1. School of Stomatology, Qingdao University, Qingdao 266000, China;
2. Stomatology Center, Qingdao Municipal Hospital, Qingdao 266071, China;
3. Department of Stomatology, Women & Children's Health Care Hospital of Linyi, Linyi 276002, China;
4. School of Stomatology, Dalian Medical University, Dalian 116044, China

Received:2021-06-03 Online:2021-11-28 Published:2021-11-22

摘要/Abstract

摘要： 目的: 开发有效且对患者副作用小的口腔抑菌剂配方,系统评价过氧化氢(hydrogen peroxide,H₂O₂)、次氯酸钠 (Sodium hypochlorite, NaClO)对粪肠球菌(Enterococcus faecalis ATCC29212)的抑菌效能。方法: 分别采用肉汤稀释法和重水拉曼技术(D₂O-labeled single-cell Raman micro-spectroscopy),通过最小抑菌浓度(minimum inhibitory concentration,MIC)和最低抑制代谢浓度(minimum inhibitory concentration based on metabolic activity, MIC-MA)定量评价了抑菌剂对粪肠球菌的生长和代谢抑制作用。结果: 对于过氧化氢和次氯酸钠,MIC分别为110 mg/L和0.45 g/L,尽管生长完全停止,但细菌细胞的代谢活性在8 h平均降低了71%、70%,显示出一种“非生长但代谢活跃” (NGMA)的状态,这种状态可能导致潜在的难治性反复感染。而MIC-MA分别为220 mg/L和0.9 g/L时,所有细胞的代谢活动在暴露8 h后完全停止。此外,NaClO+H₂O₂的组合使用优于单独使用次氯酸钠、过氧化氢。结论: MIC-MA有利于严格评估抗细菌疗效, NaClO+H₂O₂可作为细菌病原体更有效的抑菌剂方案。

关键词: 拉曼显微光谱, 重水, 粪肠球菌, 单细胞技术, 临床抑菌剂

Abstract: Objective: To evaluate the susceptibility of enterococcus faecalis (Ef) to clinical disinfectants for the development of effective yet patient-friendly disinfectant formulations. Methods: The growth and metabolism-inhibiting effects of intracanal disinfectants via minimum inhibitory concentration (MIC) and minimum inhibitory concentration were quantitatively assessed based on metabolic activity (MIC-MA), using broth dilution test and D2O-probed Single-Cell Raman Spectra (SCRS) respectively. Results: For hydrogen peroxide (H₂O₂) and sodium hypochlorite (NaClO), at their MIC of 110 mg/L and 0.45 g/L respectively, despite complete halt of growth, metabolic activity of bacterial cells was reduced averagely for 71% and 70% at 8 hours, revealing a "nongrowing but metabolically active" (NGMA) state that may underlie potential refractory infections. In contrast, at their MIC-MA of 220 mg/L and 0.9 g/L respectively, metabolic activity of all cells was completely halted throughout 8 hours' exposure. Moreover, the combination usage of NaClO+H₂O₂ outperformed the solo usage of NaClO or H₂O₂. Conclusion: MIC-MA is advantageous in critically assessing antibacterial efficacy, and NaClO+H₂O₂ can potentially serve as a more efficient disinfectant formula for bacterial pathogens.

Key words: raman micro-spectroscopy, heavy water, Enterococcus faecalis, single-cell technology, clinical disinfectants

刘育含, 马玉莹, 张利娟, 李修珍, 杨加震, 牛玉芬, 孙雁斐, 曾飞, 杨芳. 基于重水拉曼技术评价口腔抑菌剂对粪肠球菌抑菌效能的研究[J]. 口腔医学研究, 2021, 37(11): 989-993.

LIU Yuhan, MA Yuying, ZHANG Lijuan, LI Xiuzhen, YANG Jiazhen, NIU Yufen, SUN Yanfei, ZENG Fei; YANG Fang. Efficacy Assessment of Clinical Disinfectants for Enterococcus Faecalis via Single-cell Raman Microspectroscopy[J]. Journal of Oral Science Research, 2021, 37(11): 989-993.

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基于重水拉曼技术评价口腔抑菌剂对粪肠球菌抑菌效能的研究

Efficacy Assessment of Clinical Disinfectants for Enterococcus Faecalis via Single-cell Raman Microspectroscopy

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