Department of Microbiology, Faculty of Biological Sciences, Falavarjan Branch, Islamic Azad University, University Boulevard, Falavarjan 84515/155, Isfahan, Iran.
Department of Biology, Faculty of Biological Sciences, Falavarjan Branch, Islamic Azad University, University Boulevard, Falavarjan 84515/155, Isfahan, Iran.
FEMS Microbiol Lett. 2021 Nov 30;368(20). doi: 10.1093/femsle/fnab143.
The prevalence of multidrug-resistant (MDR) strains has caused serious problems in the treatment of burn infections. MDR Enterobactercloacae and Enterobacterhormaechei have been defined as the causative agents of nosocomial infections in burn patients. In this situation, examination of phages side effects on human cell lines before any investigation on human or animal that can provide beneficial information about the safety of isolated phages. The aim of this study was to isolate and identify the specific bacteriophages on MDR E. cloacae and E. hormaechei isolated from burn wounds and to analyze the efficacy, cell viability and cell cytotoxicity of phages on A-375 and HFSF-PI cell lines by MTT (3-(4, 5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide) colorimetric assay and lactate dehydrogenase (LDH) release assay. Phages were isolated from urban sewage Isfahan, Iran. Enterobactercloacae strain Iau-EC100 (GenBank accession number: MZ314381) and E. hormaechei strain Iau-EHO100 (GenBank accession number: MZ348826) were sensitive to the isolated phages. Transmission electron microscopy (TEM) results revealed that PɸEn-CL and PɸEn-HO that were described had the morphologies of Myovirus and Inovirus, respectively. Overall, MTT and LDH assays showed moderate to excellent correlation in the evaluation of cytotoxicity of isolated phages. The results of MTT and LDH assays showed that, phages PɸEn-CL and PɸEn-HO had no significant toxicity effect on A375 and HFSF-PI 3 cells. Phage PɸEn-HO had a better efficacy on the two tested cell lines than other phage. Our results indicated that, there were significant differences between the two cytotoxicity assays in phage treatment compared to control.
多药耐药（MDR）菌株的流行给烧伤感染的治疗带来了严重问题。阴沟肠杆菌和海氏肠杆菌已被定义为烧伤患者医院感染的病原体。在这种情况下，在对人类或动物进行任何调查之前，检查噬菌体对人细胞系的副作用，可以提供有关分离噬菌体安全性的有益信息。本研究的目的是分离和鉴定从烧伤伤口中分离出的多药耐药阴沟肠杆菌和海氏肠杆菌的特异性噬菌体，并通过 MTT（3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四唑溴盐）比色法和乳酸脱氢酶（LDH）释放试验分析噬菌体对 A-375 和 HFSF-PI 细胞系的疗效、细胞活力和细胞细胞毒性。噬菌体从伊朗伊斯法罕市的城市污水中分离出来。肠杆菌属 cloacae 菌株 Iau-EC100（GenBank 登录号：MZ314381）和 E. hormaechei 菌株 Iau-EHO100（GenBank 登录号：MZ348826）对分离出的噬菌体敏感。透射电子显微镜（TEM）结果表明，描述的 PɸEn-CL 和 PɸEn-HO 具有肌病毒和肌病毒的形态。总体而言，MTT 和 LDH 测定在评估分离噬菌体的细胞毒性方面具有中度至极好的相关性。MTT 和 LDH 测定的结果表明，噬菌体 PɸEn-CL 和 PɸEn-HO 对 A375 和 HFSF-PI3 细胞没有明显的毒性作用。噬菌体 PɸEn-HO 对两种测试细胞系的疗效优于其他噬菌体。我们的结果表明，与对照相比，两种细胞毒性测定法在噬菌体处理方面存在显著差异。
