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光动力对NIH 3T3细胞系和细菌的作用研究。

Study of photodynamic effects on NIH 3T3 cell line and bacteria.

作者信息

Hanakova Adela, Bogdanova Katerina, Tomankova Katerina, Binder Svatopluk, Bajgar Robert, Langova Katerina, Kolar Milan, Mosinger Jiri, Kolarova Hana

机构信息

Department of Medical Biophysics, Faculty of Medicine and Dentistry and Institute of Molecular and Translational Medicine, Palacky University Olomouc, Czech Republic.

出版信息

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2014 Jun;158(2):201-7. doi: 10.5507/bp.2012.057. Epub 2012 Jun 15.

Abstract

BACKGROUND

Bacterial resistance to antibiotics is a constantly growing challenge. Photodynamic therapy (PDT) offers a new approach to the treatment of bacterial and viral diseases. The aim of this study was to compare the efficacy of photosensitizers used in PDT applied to cell lines and bacterial strains.

METHODS

We tested the cytotoxicity and phototoxicity of 3 photosensitizers: TPPS4, ZnTPPS4 and TMPyP applied to the NIH3T3 cell line using two established methods for measuring ROS production and, MTT viability assay. Bacterial viability was determined spectrophotometrically over 24 h following PDT.

RESULTS

The most efficient photosensitiser was TMPyP as it reduced the viability of the NIH3T3 cell line by more than 85%. In general, the photosensitisers were more phototoxic to the two Gram-positive bacterial strains, Enterococcus faecalis and Staphylococcus aureus. The viability of E. faecalis was reduced to 78 % by a dose radiation 0.5 J/cm(2) and concentration of TMPyP 1.562 µmol/L. The viability of bacterium S. aureus was reduced to 23 % when exposed to a radiation dose 0.5 J/cm(2) and 100 µmol/L concentration of ZnTPPS4. The highest viability decrease (15 %) for Pseudomonas aeruginosa was caused by 0.5 J/cm(2) radiation dose and 50 µmol/L TMPyP concentration. Escherichia coli proved to be PDT resistant as the bacterial viability was higher than 90%.

CONCLUSIONS

The goal of the present study was to test the efficiency of photosensitizers on the NIH 3T3 cell line and bacterial cells. Subsequently we would like to study effectiveness of photosensitizers bound to carriers (for example cyclodextrins) on other cell line and bacterial strain.

摘要

背景

细菌对抗生素的耐药性是一个不断增加的挑战。光动力疗法(PDT)为细菌和病毒疾病的治疗提供了一种新方法。本研究的目的是比较用于光动力疗法的光敏剂对细胞系和细菌菌株的疗效。

方法

我们使用两种已确立的测量活性氧生成的方法以及MTT活力测定法,测试了3种光敏剂(四磺基苯基卟啉(TPPS4)、四磺酸锌卟啉(ZnTPPS4)和四甲基吡啶基卟啉(TMPyP))对NIH3T3细胞系的细胞毒性和光毒性。在光动力疗法后24小时内通过分光光度法测定细菌活力。

结果

最有效的光敏剂是TMPyP,因为它使NIH3T3细胞系的活力降低了85%以上。总体而言,光敏剂对两种革兰氏阳性细菌菌株粪肠球菌和金黄色葡萄球菌的光毒性更强。在辐射剂量为0.5 J/cm²和TMPyP浓度为1.562 μmol/L时,粪肠球菌的活力降低到78%。当暴露于辐射剂量为0.5 J/cm²和ZnTPPS4浓度为100 μmol/L时,金黄色葡萄球菌的活力降低到23%。对于铜绿假单胞菌,0.5 J/cm²的辐射剂量和50 μmol/L的TMPyP浓度导致其活力下降幅度最大(15%)。事实证明,大肠杆菌对光动力疗法具有抗性,因为其细菌活力高于90%。

结论

本研究的目的是测试光敏剂对NIH 3T3细胞系和细菌细胞的效率。随后,我们希望研究与载体(例如环糊精)结合的光敏剂对其他细胞系和细菌菌株的有效性。

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