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Photoinactivation of Escherichia coli (SURE2) without intracellular uptake of the photosensitizer

机译:大肠杆菌(SURE2)的光灭活而细胞内不吸收光敏剂

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Aim This study was performed to investigate the possibility to photodynamically inactivate Gram-negative bacteria without intracellular uptake of the photosensitizer. The efficiency of the photodynamic growth inhibition of Escherichia coli (SURE2) was proved in a comparative study of a neutral and a cationic photosensitizer. Methods and Results We used confocal laser scanning microscopy (CLSM) to investigate the uptake of the photosensitizer by the bacteria to show that both chlorin e6 and TMPyP are not accumulated in the cells. Fluorescence lifetime imaging (FLIM) and phototoxicity experiments were used to investigate the photodynamic inactivation of the Gram-negative bacteria. The phototoxicity experiments were carried out using a white light LED-setup to irradiate the bacterial suspensions. The viability of the bacteria was obtained by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-assay. For the cationic TMPyP, photodynamic inactivation without intracellular uptake was observed, whereas for chlorin e6 such behaviour was not found. Conclusions It was proven that in general, it is possible to photodynamically inactivate Gram-negative bacteria without photosensitizer accumulation in the bacterial cells. This fact is especially interesting, considering that the development of resistances may be prevented, leaving the active components outside the bacterium. Significance and Impact of the Study In a world with bacteria that gain the ability to withstand the effects of antibiotics and are able to transmit on these resistances to the next generation, it becomes necessary to develop new approaches to inhibit the growth of multi-resistant bacteria. The photodynamic inactivation of bacteria is based on a three-component system by which the growth of the bacterial cells is inhibited. The well-directed oxidative damage is initiated by visible light of a certain wavelength, which excites a nontoxic photoactive molecule, called photosensitizer. Its reaction with oxygen causes the generation of cytotoxic species like singlet oxygen, which is highly reactive and causes the inactivation of the growth of bacteria.
机译:目的进行这项研究是为了研究光动力灭活革兰氏阴性细菌而细胞内不吸收光敏剂的可能性。在对中性光敏剂和阳离子光敏剂的比较研究中,证明了对大肠杆菌(SURE2)的光动力生长抑制的效率。方法和结果我们使用共聚焦激光扫描显微镜(CLSM)来研究细菌对光敏剂的吸收情况,结果表明细胞中没有积累二氢卟酚e6和TMPyP。荧光寿命成像(FLIM)和光毒性实验用于研究革兰氏阴性细菌的光动力灭活。使用白光LED设置进行光毒性实验,以照射细菌悬浮液。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四唑鎓(MTT)测定获得细菌的生存力。对于阳离子TMPyP,观察到没有细胞内摄取的光动力失活,而对于二氢卟酚e6,则未发现这种行为。结论事实证明,一般而言,光动力灭活革兰氏阴性细菌而不会使光敏剂在细菌细胞中积聚是可能的。考虑到可以防止耐药性的发展,使活性成分留在细菌外部,这一事实特别有趣。该研究的意义和影响在一个细菌能够承受抗生素作用并能够将这些抗药性传递给下一代的世界中,有必要开发新的方法来抑制多重耐药性细菌的生长。细菌的光动力失活是基于三组分系统,通过该系统可以抑制细菌细胞的生长。定向氧化损伤是由一定波长的可见光引起的,该波长激发一种无毒的光敏分子,称为光敏剂。它与氧气的反应会导致细胞毒性物质(如单线态氧)的产生,后者具有很高的反应性,会导致细菌生长失活。

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