The Comparison between Energy Density of Blue and Red Light which Activation Silver Nanoparticles to Inhibition Candida albicans Biofilms

Pryandi M. Tabaika, S. Astuty, S. Dewang, Nur Umriani Permatasari, Wahiduddin Wahiduddin
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Abstract

Photodynamic inactivation (PDI) is a technique to inhibit microbial biofilm growth through the toxicity of Reactive Oxygen Species (ROS) compounds. ROS can be attack membrane, lipids, DNA and nucleic acid then initiate cell necrosis. This study aims to analyze the potential of red and blue LEDs to activating silver nanoparticles (AgNPs) to produce significant amounts of ROS that are believed to be toxic and lethal to Candida albicans biofilm cells. The effectiveness of the treatment in this study was evaluated through cell viability represented by Optical Density values and malondialdehyde levels. There were 4 treatment groups used as samples, namely the control group, the photosensitizer group, the light group, and the combination group of light with photosensitizer. The duration of light exposure ranged from 2 to 10 min with a power of 100 MW. The biofilm staining done to detection some indicator as an impact of photodynamic against mortality and survive cell with 2 dyes are XTT assay as cell viability values and the Thiobarbituric Acid Reactive Substances assay for malondialdehyde levels. The results showed that photoinactivation of Candida albicans biofilm with the lowest viability occurred in the treatment group of the combination of blue light with AgNPs with an irradiation duration of 10 min, namely 0.076 ± 0.005 and the treatment group of the combination of red light with AgNPs with an irradiation duration of 10 min, namely 0.131 ± 0.021. The data resulted in an inactivation rate of 94.68 ± 0.55 % for blue light and 90.98 ± 0.02 % for red light. The malondialdehyde levels were 1.563 nmol/mL for blue light and 1.514 nmol/mL for red light. The comparison of blue light treatment with red light is based on penetration in the cell, where blue light has low penetration but high energy which gives more opportunities to produce ROS at the triplet level. The combination of blue LED spectrum with AgNPs is highly effective in inactivating the metabolic activity of pathogenic microbial cells. HIGHLIGHTS Candida albicans biofilm is very rigid and has strong potential as a chronic infection. The research focuses on the application of photodynamic inactivation with LED light and antimicrobial AgNPs. Identification of the results with XTT assay 94.65 % inhibition and TBARS assay at MDA level of 1,864 nmol/mL. GRAPHICAL ABSTRACT
蓝光和红光激活银纳米粒子抑制白色念珠菌生物膜的能量密度比较
光动力灭活(PDI)是一种通过活性氧(ROS)化合物的毒性抑制微生物生物膜生长的技术。ROS 可攻击细胞膜、脂质、DNA 和核酸,进而导致细胞坏死。本研究旨在分析红色和蓝色 LED 活化银纳米粒子(AgNPs)产生大量 ROS 的潜力,ROS 被认为对白色念珠菌生物膜细胞具有毒性和致命性。本研究通过光学密度值和丙二醛水平来评估细胞存活率。共有 4 个处理组作为样本,即对照组、光敏剂组、光照组和光照与光敏剂组合组。光照时间为 2 至 10 分钟,功率为 100 兆瓦。用两种染料对生物膜进行染色,以检测光动力对细胞死亡率和存活率的影响:XTT 法检测细胞存活率,硫代巴比妥酸活性物质法检测丙二醛含量。结果表明,在蓝光与 AgNPs 组合(照射时间为 10 分钟)的处理组中,白色念珠菌生物膜的光灭活率最低,为 0.076 ± 0.005;在红光与 AgNPs 组合(照射时间为 10 分钟)的处理组中,白色念珠菌生物膜的光灭活率最低,为 0.131 ± 0.021。数据显示,蓝光的灭活率为 94.68 ± 0.55 %,红光的灭活率为 90.98 ± 0.02 %。蓝光下的丙二醛水平为 1.563 nmol/mL,红光下为 1.514 nmol/mL。蓝光处理与红光处理的比较是基于对细胞的穿透力,蓝光穿透力低,但能量高,因此有更多机会产生三重态的 ROS。蓝光 LED 光谱与 AgNPs 的结合在灭活病原微生物细胞的代谢活性方面非常有效。本研究的重点是应用 LED 光和抗菌 AgNPs 进行光动力灭活。用 XTT 检测法鉴定抑制率为 94.65%,TBARS 检测法鉴定 MDA 水平为 1,864 nmol/mL。
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