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采用二极管激光 635nm 和甲苯胺蓝评估选定念珠菌菌株对光动力抗菌疗法的敏感性 - 体外研究。

Assessment of sensitivity of selected Candida strains on antimicrobial photodynamic therapy using diode laser 635 nm and toluidine blue - In vitro research.

机构信息

School of Medicine with the Division of Dentistry in Zabrze, Medical University of Silesia in Katowice, Department of Periodontal Diseases and Oral Mucosa Diseases, Pl. Traugutta 2, 41-800, Zabrze, Poland.

School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia in Katowice, Department of Microbiology and Virology, ul. Jagiellońska 4, 41-200, Sosnowiec, Poland.

出版信息

Photodiagnosis Photodyn Ther. 2019 Sep;27:241-247. doi: 10.1016/j.pdpdt.2019.06.007. Epub 2019 Jun 11.

Abstract

BACKGROUND

Photodynamic therapy is believed to be a promising treatment for Candida infections. This study evaluated the efficacy of antimicrobial photodynamic therapy (aPDT) using the 635 nm diode laser light and toluidine blue (TB) in the elimination of selected Candida species cultured on acrylic surface.

METHODS

108 acrylic plates (Methyl Methacrylate Polymer, routinely used for the production of prosthetic dentures) were placed in three sterile Petri dishes and poured with prepared suspensions of Candida strains: C. albicans, C. glabrata, and C. krusei. After all procedures of fungi incubation, fungal biofilm was visible on the plates' surfaces. The acrylic plates were divided into nine study groups (B) and nine control groups (K) for further experiments. In the study groups, the acrylic plates with fungal biofilm were immersed in TB and afterwards laser irradiation was applicated with different exposure parameters (groups: B1 - 400 mW, 24 J/cm, 30 s; B2 - 300 mW, 18 J/cm, 30 s; B3 - 200 mW, 12 J/cm, 30 s) separately for each Candida species. The control groups contained following parameters: no exposure to laser light or TB, treatment only with TB without laser irradiation, or only laser irradiation without previous immersion in TB. Calculations of colony forming units (CFUs) were conducted by using aCOlyte (Synbiosis). Differences in CFUs were analyzed by the Wilcoxon test.

RESULTS

In all study groups, the reduction in CFUs was statistically significant. The differences in CFUs before and after intervention were insignificant. The K3 control group showed a statistical reduction of Candida albicans after laser irradiation.

CONCLUSION

Our study confirmed the efficacy of aPDT against C. albicans, C. glabrata and C. krusei being dependent on the laser parameters and the type of fungus. The advantage of this study is the validation of aPDT effectiveness in in vitro studies to transpose this data into future clinical trials using photodynamic therapy in the treatment of oral candidiasis.

摘要

背景

光动力疗法被认为是治疗念珠菌感染的一种很有前途的治疗方法。本研究评估了使用 635nm 二极管激光和甲苯胺蓝(TB)消除培养在丙烯酸表面上的选定念珠菌物种的抗菌光动力疗法(aPDT)的疗效。

方法

将 108 块丙烯酸板(甲基丙烯酸甲酯聚合物,常规用于制作义齿)放置在三个无菌培养皿中,并倒入已制备好的念珠菌菌株悬浮液:白色念珠菌、光滑念珠菌和克柔念珠菌。在所有真菌孵育程序完成后,在平板表面可见真菌生物膜。将丙烯酸板分为 9 个实验组(B)和 9 个对照组(K)进行进一步实验。在实验组中,将有真菌生物膜的丙烯酸板浸入 TB 中,然后应用不同的暴露参数进行激光照射(组:B1-400mW,24J/cm,30s;B2-300mW,18J/cm,30s;B3-200mW,12J/cm,30s)分别针对每种念珠菌。对照组包含以下参数:不暴露于激光或 TB,仅用 TB 处理而不进行激光照射,或仅进行激光照射而不事先浸入 TB。使用 aCOlyte(Synbiosis)进行菌落形成单位(CFU)的计算。使用 Wilcoxon 检验分析 CFU 差异。

结果

在所有实验组中,CFU 的减少均具有统计学意义。干预前后 CFU 的差异无统计学意义。K3 对照组在激光照射后,白色念珠菌的 CFU 有统计学意义的减少。

结论

我们的研究证实了 aPDT 对白色念珠菌、光滑念珠菌和克柔念珠菌的疗效取决于激光参数和真菌类型。本研究的优势在于验证了 aPDT 在体外研究中的有效性,以便将这些数据转化为未来使用光动力疗法治疗口腔念珠菌病的临床试验。

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