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首页> 外文期刊>Photodiagnosis and Photodynamic Therapy >Effects of antimicrobial photodynamic therapy on antibiotic-resistant Escherichia coli
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Effects of antimicrobial photodynamic therapy on antibiotic-resistant Escherichia coli

机译：抗菌光动力治疗对抗生素抗性大肠杆菌的影响

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This study used Electron Cryo-tomography (ECT) and fluorescent images to evaluate antimicrobial photodynamic therapy (aPDT) on the envelope architecture of a Gram-negative bacteria and the effects of combined therapy of aPDT and antibiotics. Standard and clinical suspension of Escherichia coli were submitted to photo dynamic treatment with methylene blue solution (100 mu M) and a 100 mW LED emitting at 660 nm with 3 and 18 J of energy. As a control group, a suspension of E. coli was submitted to penicillin V for 60 min at 30 degrees C, to compare the damage in cell wall structure. After treatment, ECT images were collected and E. coli biofilms were grown in glass-cover slides and stained with live/dead staining for fluorescence analysis before and after treatments. Bacteria were also submitted to disc diffusion and MIC50 tests with Ampicillin, Amoxicillin + Clavulanic acid, Clindamycin and Erythromycin. For in vivo experiment Galleria mellonella larvae were infected with E. coli and treated with antibiotics, aPDT or combined therapy. ECT images presented damage to cell walls and vesicles structures inside and outside the bacteria and fluorescent images showed dose dependent effect of aPDT. Antibiotic or aPDT alone did not improve the survival of caterpillars, but the combined therapy significantly increased survival curve. ECT and fluorescent images shows that aPDT seems to promote micro damages to cell envelope and causes the production of membrane vesicles permeabilizing cell membranes. The results showed that pre-treating bacterial cells with a photosensitizer and light make them more susceptible to antibiotics and could be an alternative to local infection treatment by resistant bacteria.
机译：本研究使用了电子丧光断层扫描（ECT）和荧光图像来评估革兰阴性细菌的封套架构上的抗微生物光动力治疗（APDT）以及APDT和抗生素组合治疗的影响。将大肠杆菌的标准和临床悬浮液提交给用亚甲基蓝溶液（100μm）的照片动态处理，100 mW LED在660nm处发射，具有3和18 j的能量。作为对照组，将E.COLI的悬浮液在30摄氏度下向青霉素v提交60分钟，以比较细胞壁结构的损伤。处理后，收集图像，并在玻璃盖载玻片中生长E.Coli生物膜，并在处理前后用活/死染色进行荧光分析。用氨苄青霉素，阿莫西林+克拉​​维酸，克林霉素和红霉素提交细菌和MIC50试验。对于体内实验，Galleria Mellonella幼虫被大肠杆菌感染并用抗生素，APDT或组合治疗治疗。图像对细胞壁和细菌内外的囊泡结构造成的损坏，并且荧光图像显示出APDT的剂量依赖性效果。单独抗生素或APDT并未改善毛虫的存活，但组合治疗显着增加存活曲线。 ECT和荧光图像表明APDT似乎促进对细胞包膜的微观损伤，并导致膜囊泡的产生透化细胞膜。结果表明，用光敏剂和光预处理细菌细胞使它们更容易受到抗生素的影响，并且可以是抗性细菌的局部感染治疗的替代方法。

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