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芦丁-镓配合物介导的抗菌光动力疗法:抗变形链球菌生物膜的体外研究。

Rutin-gallium complex mediated antimicrobial photodynamic therapy: An in vitro studies against Streptococcus mutans biofilms.

机构信息

Department of Orthodontics, School of Dentistry, Shahed University, Tehran, Iran.

Department of Orthodontics, School of Dentistry, Shahed University, Tehran, Iran.

出版信息

Photodiagnosis Photodyn Ther. 2023 Jun;42:103367. doi: 10.1016/j.pdpdt.2023.103367. Epub 2023 Feb 24.

Abstract

BACKGROUND

Discoloration of teeth following antimicrobial photodynamic therapy (aPDT) is a serious concern. Common photosensitizers are colored, and access to a photosensitizer that does not leave color on the teeth or is the same color as the enamel and dentin is highly demanded. The physicochemical characterization, anti-virulence, and antimicrobial effects of a novel rutin-gallium (III) (Rt-Ga) complex as novel photosensitizer are presented herein.

MATERIALS AND METHODS

Photophysical properties and cytotoxicity of the Rt-Ga complex were evaluated in comparison with the parental rutin. Intracellular reactive oxygen species (ROS) generation following Rt-Ga complex-mediated aPDT was measured using the fluorescent 2',7'-dichlorofluorescein diacetate (H2DCF-DA) method. The anti-biofilm effects of Rt-Ga complex-mediated aPDT on Streptococcus mutans were assessed using a colorimetric assay. The virulence‑associated gtfB gene expression was assessed following Rt-Ga complex-mediated aPDT by quantitative real‑time PCR.

RESULTS

The photophysical properties of the Rt-Ga complex revealed a significant blue-shift in absorption (60 nm shift) and increased extinction coefficient (4100 M -1 cm -1; at λmax = 450 nm). Average (± SEM) DCF fluorescence intensities in an arbitrary unit (A.U.) were 7.1 ± 0.9, 4.1 ± 0.5, and 1.7 ± 0.3 for 10.0 μM of Rt-Ga complex-mediated aPDT, 7.5 μM of Rt-Ga complex-mediated aPDT, and 10.0 μM of Rt-Ga complex alone, respectively. The corresponding DCF fluorescence intensities were 710% (P = 0.001), 410% (P = 0.001), and 170% (P = 0.02) of the untreated S. mutans as the control group (1.0 ± 0.1 A.U.), respectively. The novel Rt-Ga complex-mediated aPDT exhibited no toxicity against primary human gingival fibroblast cells, a dose dependent decrease in S. mutans biofilm cell survival and virulence were observed (91.4% and 11.7-fold, respectively).

CONCLUSION

The Rt-Ga complex-mediated anti-virulence and antimicrobial photodynamic effects were superior to the one caused by rutin alone making the Rt-Ga complex a more promising photosensitizer than the parent material.

摘要

背景

抗菌光动力疗法(aPDT)后牙齿变色是一个严重的问题。常见的光敏剂是有色的,因此人们非常需要一种不会在牙齿上留下颜色或与牙釉质和牙本质颜色相同的光敏剂。本文介绍了新型芦丁-镓(III)(Rt-Ga)配合物作为新型光敏剂的物理化学特性、抗毒力和抗菌作用。

材料与方法

与芦丁相比,评价了 Rt-Ga 配合物的光物理性质和细胞毒性。使用荧光 2',7'-二氯荧光素二乙酸酯(H2DCF-DA)法测量 Rt-Ga 配合物介导的 aPDT 后细胞内活性氧(ROS)的产生。使用比色法评估 Rt-Ga 配合物介导的 aPDT 对变形链球菌生物膜的抗作用。通过定量实时 PCR 评估 Rt-Ga 配合物介导的 aPDT 后与毒力相关的 gtfB 基因表达。

结果

Rt-Ga 配合物的光物理性质显示吸收(60nm 位移)明显蓝移和消光系数增加(4100M -1 cm -1;在λmax = 450nm 时)。在任意单位(A.U.)中,10.0μM 的 Rt-Ga 配合物介导的 aPDT、7.5μM 的 Rt-Ga 配合物介导的 aPDT 和 10.0μM 的 Rt-Ga 配合物分别为 7.1 ± 0.9、4.1 ± 0.5 和 1.7 ± 0.3。相应的 DCF 荧光强度分别为未经处理的 S. mutans 的 710%(P = 0.001)、410%(P = 0.001)和 170%(P = 0.02),作为对照组(1.0 ± 0.1 A.U.)。新型 Rt-Ga 配合物介导的 aPDT 对原代人牙龈成纤维细胞没有毒性,观察到 S. mutans 生物膜细胞存活和毒力呈剂量依赖性下降(分别为 91.4%和 11.7 倍)。

结论

与芦丁单独作用相比,Rt-Ga 配合物介导的抗毒力和抗菌光动力作用更优,使 Rt-Ga 配合物成为比母体材料更有前途的光敏剂。

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