Suzuki Luis Cláudio, Kato Ilka Tiemy, Prates Renato Araujo, Sabino Caetano Padial, Yoshimura Tania Mateus, Silva Tamires Oliveira, Ribeiro Martha Simões
Center for Lasers and Applications, IPEN-CNEN/SP, 05508-000, São Paulo, SP, Brazil.
The Engineering, Modelling and Applied Social Science Department, UFABC, 09210-580, Santo André, SP, Brazil.
Photodiagnosis Photodyn Ther. 2017 Mar;17:173-179. doi: 10.1016/j.pdpdt.2016.12.003. Epub 2016 Dec 18.
Candida albicans biofilm is a main cause of infections associated with medical devices such as catheters, contact lens and artificial joint prosthesis. The current treatment comprises antifungal chemotherapy that presents low success rates. Photodynamic inactivation (PDI) involves the combination of a photosensitizing compound (PS) and light to generate oxidative stress that has demonstrated effective antimicrobial activity against a broad-spectrum of pathogens, including C. albicans. This fungus senses glucose inducing an upregulation of membrane transporters that can facilitate PS uptake into the cell. The aim of this study was to evaluate the effects of glucose on methylene blue (MB) uptake and its influence on PDI efficiency when combined to a red LED with central wavelength at λ=660nm. C. albicans biofilms were grown on hydrogel disks. Prior to PDI assays, MB uptake tests were performed with and without glucose-sensitization. In this system, the optimum PS administration was determined as 500μM of MB in contact with the biofilm during 30min before irradiation. Irradiation was performed during 3, 6, 9, 12, 15 and 18min with irradiance of 127.3mW/cm. Our results showed that glucose was able to increase MB uptake in C. albicans cells. In addition, PDI without glucose showed a higher viability reduction until 6min; after 9min, glucose group demonstrated a significant decrease in cell viability when compared to glucose-free group. Taken together, our data suggest that glucose is capable to enhance MB uptake and modulate photodynamic inactivation of C. albicans biofilm.
白色念珠菌生物膜是与导管、隐形眼镜和人工关节假体等医疗设备相关感染的主要原因。目前的治疗方法包括抗真菌化疗,但成功率较低。光动力灭活(PDI)涉及将光敏化合物(PS)与光结合以产生氧化应激,已证明其对包括白色念珠菌在内的多种病原体具有有效的抗菌活性。这种真菌感知葡萄糖会诱导膜转运蛋白上调,从而促进PS进入细胞。本研究的目的是评估葡萄糖对亚甲蓝(MB)摄取的影响,以及当与中心波长为λ=660nm的红色发光二极管结合时对PDI效率的影响。白色念珠菌生物膜在水凝胶圆盘上生长。在进行PDI测定之前,进行了有无葡萄糖致敏的MB摄取试验。在该系统中,确定最佳的PS给药量为在照射前30分钟与生物膜接触的500μM MB。以127.3mW/cm的辐照度分别照射3、6、9、12、15和18分钟。我们的结果表明,葡萄糖能够增加白色念珠菌细胞对MB的摄取。此外,无葡萄糖的PDI在6分钟内显示出更高的活力降低;9分钟后,与无葡萄糖组相比,葡萄糖组的细胞活力显著下降。综上所述,我们的数据表明葡萄糖能够增强MB摄取并调节白色念珠菌生物膜的光动力灭活。