Lambrechts Saskia A G, Schwartz Kevin R, Aalders Maurice C G, Dankert Jacob B
Laser Center K01-225-5, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands,
Lasers Med Sci. 2005 Sep;20(2):62-7. doi: 10.1007/s10103-005-0338-x. Epub 2005 Jun 7.
An important determinant of the clinical applicability and value of antimicrobial photodynamic inactivation (PDI) is the cytotoxicity of the treatment to human cells. We evaluated the in vitro cytotoxicity of PDI to human dermal fibroblasts using 5-phenyl-10,15,20-tris(N-methyl-4-pyridyl)porphyrin chloride (TriP[4]) as the photosensitiser. The fibroblasts were exposed to a PDI regime that is known to be sufficient for the inactivation of Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans. The PDI experiments were carried out in phosphate-buffered saline (PBS) and in 6.25%, 12.5%, 25% and 50% fetal calf serum (FCS)/PBS suspensions. Cell viability subsequent to exposure was evaluated after 0 h, 6 h and 18 h using the methylthiazoletetrazolium (MTT) assay and compared to pretreatment values. At a TriP[4] concentration previously demonstrated to induce a 5 log(10)-unit reduction in a viable count for S. aureus, 79% of the fibroblasts were photo-inactivated. Increasing the FCS concentration in the medium protected the fibroblasts against PDI. Based on our in vitro results, we propose that in vivo PDI of S. aureus holds potential; however, PDI of P. aeruginosa and C. albicans will probably require such a strong PDI regime that it will induce substantial damage to fibroblasts.
抗菌光动力灭活(PDI)临床适用性和价值的一个重要决定因素是该治疗方法对人体细胞的细胞毒性。我们使用氯化5-苯基-10,15,20-三(N-甲基-4-吡啶基)卟啉(TriP[4])作为光敏剂,评估了PDI对人皮肤成纤维细胞的体外细胞毒性。将成纤维细胞暴露于已知足以灭活金黄色葡萄球菌、铜绿假单胞菌和白色念珠菌的PDI方案下。PDI实验在磷酸盐缓冲盐水(PBS)以及6.25%、12.5%、25%和50%胎牛血清(FCS)/PBS悬浮液中进行。使用甲基噻唑基四氮唑(MTT)法在暴露后0小时、6小时和18小时评估细胞活力,并与预处理值进行比较。在先前证明能使金黄色葡萄球菌活菌数减少5个对数(10)单位的TriP[4]浓度下,79%的成纤维细胞被光灭活。增加培养基中FCS的浓度可保护成纤维细胞免受PDI的影响。基于我们的体外研究结果,我们认为金黄色葡萄球菌的体内PDI具有潜力;然而,铜绿假单胞菌和白色念珠菌的PDI可能需要如此强烈的PDI方案,以至于会对成纤维细胞造成实质性损伤。