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体外研究核黄素存在与否时蓝光对耐甲氧西林金黄色葡萄球菌(MRSA)和人角质形成细胞的不同光动力效应。

Different photodynamic effects of blue light with and without riboflavin on methicillin-resistant Staphylococcus aureus (MRSA) and human keratinocytes in vitro.

机构信息

Department of Ophthalmology, Faculty of Medicine and Health, Örebro University, Örebro, Sweden.

Department of Ophthalmology, Örebro University Hospital, SE-701 85, Örebro, Sweden.

出版信息

Lasers Med Sci. 2019 Dec;34(9):1799-1805. doi: 10.1007/s10103-019-02774-9. Epub 2019 Mar 30.

DOI:10.1007/s10103-019-02774-9
PMID:30929100
Abstract

Methicillin-resistant Staphylococcus aureus (MRSA) is an important cause of infections in humans. Photodynamic therapy using blue light (450 nm) could possibly be used to reduce MRSA on different human tissue surfaces without killing the human cells. It could be less harmful than 300-400 nm light or common disinfectants. We applied blue light ± riboflavin (RF) to MRSA and keratinocytes, in an in vitro liquid layer model, and compared the effect to elimination using common disinfection fluids. MRSA dilutions (8 × 10/mL) in wells were exposed to blue light (450 nm) ± RF at four separate doses (15, 30, 56, and 84 J/cm). Treated samples were cultivated on blood agar plates and the colony forming units (CFU) determined. Adherent human cells were cultivated (1 × 10/mL) and treated in the same way. The cell activity was then measured by Cell Titer Blue assay after 24- and 48-h growth. The tested disinfectants were chlorhexidine and hydrogen peroxide. Blue light alone (84 J/cm) eliminated 70% of MRSA. This dose and riboflavin eradicated 99-100% of MRSA. Keratinocytes were not affected by blue light alone at any dose. A dose of 30 J/cm in riboflavin solution inactivated keratinocytes completely. Disinfectants inactivated all cells. Blue light alone at 450 nm can eliminate MRSA without inactivation of human keratinocytes. Hence, a high dose of blue light could perhaps be used to treat bacterial infections without loss of human skin cells. Photodynamic therapy using riboflavin and blue light should be explored further as it may perhaps be possible to exploit in treatment of skin diseases associated with keratinocyte hyperproliferation.

摘要

耐甲氧西林金黄色葡萄球菌(MRSA)是人类感染的重要原因。使用蓝光(450nm)的光动力疗法可能用于减少不同人体组织表面的MRSA,而不会杀死人类细胞。它可能比 300-400nm 光或常用消毒剂的危害更小。我们将 MRSA 和角质形成细胞应用于体外液体层模型中,用常用的消毒液进行比较,以评估蓝光±核黄素(RF)的消除效果。MRSA 稀释液(8×10/mL)在孔中暴露于蓝光(450nm)±RF 下,剂量分别为 15、30、56 和 84J/cm。处理后的样品在血琼脂平板上培养并确定菌落形成单位(CFU)。以同样的方式培养附着的人类细胞。然后通过 Cell Titer Blue 测定法在 24 和 48 小时生长后测量细胞活性。测试的消毒剂是洗必泰(chlorhexidine)和过氧化氢。单独的蓝光(84J/cm)消除了 70%的 MRSA。该剂量和核黄素消除了 99-100%的 MRSA。单独的蓝光在任何剂量下都不会影响角质形成细胞。核黄素溶液中的 30J/cm 剂量可完全灭活角质形成细胞。消毒剂可使所有细胞失活。单独的 450nm 蓝光可以消除 MRSA,而不会使人类角质形成细胞失活。因此,高剂量的蓝光或许可以用于治疗细菌感染而不会损失人类皮肤细胞。使用核黄素和蓝光的光动力疗法应进一步探索,因为它可能可以用于治疗与角质形成细胞过度增殖相关的皮肤病。

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