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临床分离牙龈卟啉单胞菌 fimA 基因表达谱对光动力消毒治疗的反应。

Gene expression profiling of fimA gene encoding fimbriae among clinical isolates of Porphyromonas gingivalis in response to photo-activated disinfection therapy.

机构信息

Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran; Dental Research Center, Dentistry Research Institute, Tehran University of Medical Sciences, Tehran, Iran.

Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran; Dental Research Center, Dentistry Research Institute, Tehran University of Medical Sciences, Tehran, Iran.

出版信息

Photodiagnosis Photodyn Ther. 2017 Dec;20:1-5. doi: 10.1016/j.pdpdt.2017.08.001. Epub 2017 Aug 7.

DOI:10.1016/j.pdpdt.2017.08.001
PMID:28797828
Abstract

BACKGROUND

The endodontic disinfection therapy majorly aims to eradicate microbial pathogens. Photo-activated disinfection (PAD), also known as antimicrobial photodynamic therapy, is an alternative antimicrobial modality used to control the microorganisms causing endodontic infections. Notably, microorganisms may be exposed to sub-lethal doses of PAD (sPAD), influencing microbial virulence. The present study assessed the effects of sPAD on expression profiling of the gene associated with the biofilm formation being the most essential virulence factor in Porphyromonas gingivalis strain cells that survive the photodynamic reatment in vivo.

MATERIALAS AND METHODS

Sixteen clinical strains of PAD resistant P. gingivalis that were isolated in vivo, were further photosensitized in vitro with toluidine blue O (TBO), methylene blue (MB), and indocyanine green (ICG) as the photosensitizing agents, which were excited with specific wavelength of light based on the photosensitizer. After evaluating sPAD, its effects on the fimA gene expression were assessed using real-time quantitative reverse transcription PCR (qRT-PCR).

RESULTS

In this study, maximum values of sPAD against P. gingivalis were 6.25μg/mL TBO at a fluency of 171.87J/cm, 15.6μg/mL ICG at fluency of 15.6J/cm, and 25μg/mL MB at fluency of 93.75J/cm. MB-, TBO-, and ICG-sPAD could cause about 4.6-, 14.4-, and 17.3-fold suppression of fimA expression, respectively. P. gingivalis strains expressed less virulence in cells surviving PAD.

CONCLUSIONS

In conclusion, the gene expression profiling was reduced in the bacterial cells, wherein greater reduction was observed for ICG-sPAD than TBO- and MB-sPAD.

摘要

背景

牙髓消毒治疗主要目的是消除微生物病原体。光激活消毒(PAD),也称为抗菌光动力疗法,是一种替代的抗菌方式,用于控制引起牙髓感染的微生物。值得注意的是,微生物可能会暴露于亚致死剂量的 PAD(sPAD)下,从而影响微生物的毒力。本研究评估了 sPAD 对与生物膜形成相关基因表达谱的影响,生物膜形成是牙龈卟啉单胞菌菌株细胞中最重要的毒力因子,这些细胞在体内光动力治疗后存活下来。

材料与方法

从体内分离出 16 株对 PAD 有耐药性的牙龈卟啉单胞菌临床株,然后在体外用甲苯胺蓝 O(TBO)、亚甲蓝(MB)和吲哚菁绿(ICG)作为光敏剂进行光敏化,根据光敏剂用特定波长的光激发。在评估 sPAD 后,使用实时定量逆转录 PCR(qRT-PCR)评估其对 fimA 基因表达的影响。

结果

在这项研究中,TBO 的 sPAD 对牙龈卟啉单胞菌的最大抑制浓度为 6.25μg/mL,在通量为 171.87J/cm 时;ICG 的最大抑制浓度为 15.6μg/mL,在通量为 15.6J/cm 时;MB 的最大抑制浓度为 25μg/mL,在通量为 93.75J/cm 时。MB-sPAD、TBO-sPAD 和 ICG-sPAD 分别可导致 fimA 表达抑制约 4.6 倍、14.4 倍和 17.3 倍。在 PAD 存活的细胞中,牙龈卟啉单胞菌菌株的毒力表达降低。

结论

总之,基因表达谱在细菌细胞中减少,其中 ICG-sPAD 比 TBO-sPAD 和 MB-sPAD 观察到更大的减少。

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