Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-903, Brazil.
Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-900, Brazil.
Fungal Biol. 2020 May;124(5):297-303. doi: 10.1016/j.funbio.2019.08.004. Epub 2019 Aug 23.
The in vitro susceptibilities of Candida albicans and Candida tropicalis to Antimicrobial Photodynamic Treatment with aluminum phthalocyanine chloride in nanoemulsion (ClAlPc/NE) were investigated. PS concentration- and fluence-dependent cell survival after APDT were compared before and after unbound extracellular PS had been washed out. The PS uptake and its subcellular localization were also determined. Exposure to light in the absence of the PS and treatment with the PS in the absence of light did not kill the fungi. APDT with ClAlPc/NE resulted in a reduction of five orders of magnitude in viability for C. albicans and between four and five orders of magnitude for C. tropicalis. Washing the cells to remove unbound PS before light exposure did not impair fungal inactivation, suggesting that cell photosensitization was mainly carried out by cell bound ClAlPc. The degree of ClAlPc uptake was dependent on its concentration. Internalization of ClAlPc by C. albicans and C. tropicalis was confirmed by confocal fluorescence microscopy that showed the PS does not penetrate the nucleus and instead accumulates in specific regions of the cytoplasm. Our results show that incorporating the water-insoluble ClAlPc into a nanoemulsion leads to an efficient formulation capable of photoinactivating both Candida species.
研究了纳米乳状液中的铝酞菁氯(ClAlPc/NE)对白色念珠菌和热带念珠菌的抗微生物光动力治疗的体外敏感性。比较了 APDT 前后未结合的细胞外 PS 被冲洗掉后的 PS 浓度和剂量依赖性细胞存活率。还确定了 PS 的摄取及其亚细胞定位。在没有 PS 的情况下暴露于光下以及在没有光的情况下用 PS 处理均不会杀死真菌。用 ClAlPc/NE 进行 APDT 可使白色念珠菌的存活率降低五个数量级,而对热带念珠菌的存活率降低四个数量级至五个数量级。在暴露于光之前冲洗细胞以除去未结合的 PS 不会损害真菌失活,这表明细胞光敏化主要是由细胞结合的 ClAlPc 进行的。ClAlPc 的摄取程度取决于其浓度。共聚焦荧光显微镜证实了白色念珠菌和热带念珠菌对 ClAlPc 的内化,表明 PS 不会穿透细胞核,而是在细胞质的特定区域积累。我们的结果表明,将不溶于水的 ClAlPc 纳入纳米乳剂中可制成一种有效的配方,能够光灭活两种念珠菌。