Center for Chemistry and Biomedicine, Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria.
Center for Chemistry and Biomedicine, Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria; Department of Pharmacy, Universitas Mandala Waluya, A.H.Nasution, Kendari 93231, Southeast Sulawesi, Indonesia.
J Colloid Interface Sci. 2023 Sep 15;646:290-300. doi: 10.1016/j.jcis.2023.05.051. Epub 2023 May 12.
Polyphosphate nanoparticles as phosphatase-degradable carriers for Penicillium chrysogenum antifungal protein (PAF) can enhance the antifungal activity of the protein against Candida albicans biofilm.
PAF-polyphosphate (PP) nanoparticles (PAF-PP NPs) were obtained through ionic gelation. The resulting NPs were characterized in terms of their particle size, size distribution and zeta potential. Cell viability and hemolysis studies were carried out in vitro on human foreskin fibroblasts (Hs 68 cells) and human erythrocytes, respectively. Enzymatic degradation of NPs was investigated by monitoring release of free monophosphates in the presence of isolated as well as C. albicans-derived phosphatases. In parallel, shift in zeta potential of PAF-PP NPs as a response to phosphatase stimuli was determined. Diffusion of PAF and PAF-PP NPs through C. albicans biofilm matrix was analysed by fluorescence correlation spectroscopy (FCS). Antifungal synergy was evaluated on C. albicans biofilm by determining the colony forming units (CFU).
PAF-PP NPs were obtained with a mean size of 300.9 ± 4.6 nm and a zeta potential of -11.2 ± 2.8 mV. In vitro toxicity assessments revealed that PAF-PP NPs were highly tolerable by Hs 68 cells and human erythrocytes similar to PAF. Within 24 h, 21.9 ± 0.4 μM of monophosphate was released upon incubation of PAF-PP NPs having final PAF concentration of 156 μg/ml with isolated phosphatase (2 U/ml) leading to a shift in zeta potential up to -0.7 ± 0.3 mV. This monophosphate release from PAF-PP NPs was also observed in the presence of C. albicans-derived extracellular phosphatases. The diffusivity of PAF-PP NPs within 48 h old C. albicans biofilm matrix was similar to that of PAF. PAF-PP NPs enhanced antifungal activity of PAF against C. albicans biofilm decreasing the survival of the pathogen up to 7-fold in comparison to naked PAF. In conclusion, phosphatase-degradable PAF-PP NPs hold promise as nanocarriers to augment the antifungal activity of PAF and enable its efficient delivery to C. albicans cells for the potential treatment of Candida infections.
多聚磷酸盐纳米颗粒作为磷酸酶可降解载体，用于携带青霉素曲霉菌抗真菌蛋白（PAF），可以增强该蛋白对白色念珠菌生物膜的抗真菌活性。
通过离子凝胶化获得 PAF-多聚磷酸盐（PP）纳米颗粒（PAF-PP NPs）。通过粒径、粒径分布和 Zeta 电位对所得纳米颗粒进行了表征。分别在人包皮成纤维细胞（Hs 68 细胞）和人红细胞上进行体外细胞活力和溶血研究。通过监测游离单磷酸盐的释放，研究了纳米颗粒在分离的和白色念珠菌衍生的磷酸酶存在下的酶促降解。同时，确定了 PAF-PP NPs 对磷酸酶刺激的 Zeta 电位变化。通过荧光相关光谱法（FCS）分析 PAF 和 PAF-PP NPs 通过白色念珠菌生物膜基质的扩散。通过测定集落形成单位（CFU）评估 PAF 和 PAF-PP NPs 对白色念珠菌生物膜的抗真菌协同作用。
获得了平均粒径为 300.9±4.6nm 和 Zeta 电位为-11.2±2.8mV 的 PAF-PP NPs。体外毒性评估显示，PAF-PP NPs 对 Hs 68 细胞和人红细胞的耐受性与 PAF 相似。在 24 小时内，当终浓度为 156μg/ml 的 PAF 与分离的磷酸酶（2U/ml）孵育时，PAF-PP NPs 释放出 21.9±0.4μM 的单磷酸盐，导致 Zeta 电位上升至-0.7±0.3mV。在存在白色念珠菌衍生的细胞外磷酸酶的情况下，也观察到 PAF-PP NPs 从 PAF-PP NPs 中释放出单磷酸盐。PAF-PP NPs 在 48 小时龄的白色念珠菌生物膜基质中的扩散率与 PAF 相似。与裸 PAF 相比，PAF-PP NPs 增强了 PAF 对白色念珠菌生物膜的抗真菌活性，使病原体的存活率降低了 7 倍。总之，磷酸酶可降解的 PAF-PP NPs 有望成为纳米载体，增强 PAF 的抗真菌活性，并使其能够有效递送至白色念珠菌细胞，从而为治疗白色念珠菌感染提供潜在的治疗方法。
