Laboratory of Molecular Biology of Fungi, University of Brasilia, Brasilia 70910-900, Brazil.
LAQV/REQUIMTE, Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal.
Int J Mol Sci. 2024 Jul 16;25(14):7769. doi: 10.3390/ijms25147769.
is one of the agents of invasive candidiasis, a life-threatening disease strongly associated with hospitalization, particularly among patients in intensive care units with central venous catheters. This study aimed to evaluate the synergistic activity of the antifungal peptide ToAP2 combined with fluconazole against biofilms grown on various materials. We tested combinations of different concentrations of the peptide ToAP2 with fluconazole on biofilms. These biofilms were generated on 96-well plates, intravenous catheters, and infusion tubes in RPMI medium at two maturation stages. Scanning electron microscopy and atomic force microscopy were employed to assess the biofilm structure. We also evaluated the expression of genes previously proven to be involved in biofilm formation in planktonic and biofilm cells after treatment with the peptide ToAP2 using qPCR. ToAP2 demonstrated a synergistic effect with fluconazole at concentrations up to 25 µM during both the early and mature stages of biofilm formation in 96-well plates and on medical devices. Combinations of 50, 25, and 12.5 µM of ToAP2 with 52 µM of fluconazole significantly reduced the biofilm viability compared to individual treatments and untreated controls. These results were supported by substantial structural changes in the biofilms observed through both scanning and atomic force microscopy. The gene expression analysis of cells treated with 25 µM of ToAP2 revealed a decrease in the expression of genes associated with membrane synthesis, along with an increase in the expression of genes involved in efflux pumps, adhesins, and filamentation. Our results highlight the efficacy of the combined ToAP2 and fluconazole treatment against biofilms. This combination not only shows therapeutic potential but also suggests its utility in developing preventive biofilm tools for intravenous catheters.
是侵袭性念珠菌病的病原体之一，这种危及生命的疾病与住院治疗密切相关，尤其是在重症监护病房中使用中心静脉导管的患者。本研究旨在评估抗真菌肽 ToAP2 与氟康唑联合使用对各种材料上生物膜的协同活性。我们测试了不同浓度的肽 ToAP2 与氟康唑联合使用对生物膜的效果。这些生物膜是在 RPMI 培养基中于 96 孔板、静脉导管和输液管上在两个成熟阶段生成的。我们使用扫描电子显微镜和原子力显微镜来评估生物膜结构。我们还通过 qPCR 评估了肽 ToAP2 处理后浮游细胞和生物膜细胞中先前证明与生物膜形成有关的基因的表达。在 96 孔板和医疗器械上，ToAP2 在生物膜形成的早期和成熟阶段与氟康唑联合使用时，在高达 25µM 的浓度下表现出协同作用。与单独治疗和未处理的对照组相比，浓度为 50、25 和 12.5µM 的 ToAP2 与 52µM 的氟康唑联合使用可显著降低生物膜的活力。通过扫描和原子力显微镜观察到生物膜的结构发生了实质性变化，这一结果得到了支持。用 25µM 的 ToAP2 处理后的细胞的基因表达分析显示，与膜合成相关的基因表达减少，而与外排泵、黏附素和丝状形成相关的基因表达增加。我们的研究结果突出了 ToAP2 和氟康唑联合治疗生物膜的有效性。这种组合不仅具有治疗潜力，还表明它在开发用于静脉导管的预防生物膜工具方面具有应用价值。
