Synthesis of bio-stabilized silver nanoparticles using Roccella montagnei, their anticandidal capacities & potential to inhibit the virulence factors in fluconazole-resistant Candida albicans

Shweta Bharti, Balwant Singh, Sanket Kumar, Rajesh Kumar, Jatinder Kumar
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Abstract

Candida species is the causative agent in approximately 80% of invasive mycoses and drug-resistant Candida albicans is among the four strains of ‘critical priority group’ framed by WHO. Lichens are endowed with some rare phytochemicals and a plethora of therapeutics viz. antifungal capacities of Roccella montagnei. Biosynthesis of silver nanoparticles (AgNPs) using lichen could offer an eco-friendly, and cost-effective alternative against emerging ‘microbial resistance.’ Therefore, the objective was to biosynthesize silver nanoparticles (Rm-AgNPs) using a Hydro-alcoholic (1:1) extract of R. montagnei to develop a potent anticandidal agent against Fluconazole-resistant C. albicans NBC099. UV-Spectroscopy identified AgNPs specific-peak of Rm-AgNPs at 420–440 nm and FTIR revealed the presence of amines, alcohol, aromatic compounds, and acids. SEM and TEM analysis indicated that Rm-AgNPs are spherical shaped with a size range of 10–50 nm. Zetasizer analysis indicated that particles are highly stable and have a mean hydrodynamic diameter of 116 nm with a zeta potential charge of − 41 mV. XRD analysis suggested face centered cubic crystal lattice structure. Results indicated that Rm-AgNPs strongly inhibited the growth of NBC099 at a minimum inhibitory concentration (IC50) of ≤ 15 µg. C. albicans culture treated with Rm-AgNPs at concentrations below IC50, down-regulates the production of different virulence factors in NBC099, viz. hyphal formation (> 85%), biofilms production (> 80%), phospholipase, esterase, proteinase activity. The apoptosis assay demonstrated the Rm-AgNPs induced apoptosis in NBC099 cells via oxidative stress. Interestingly, Rm-AgNPs showed negligible cytotoxicity (< 6%) in murine RAW 246.7 macrophage cells at a concentration above 15 µg/mL. Therefore, Rm-AgNPs have been offered as an anti-candida alternative that can be utilized to improve the efficacy of already available medications.

Graphical abstract

Abstract Image

利用蒙大拿洛可菌合成生物稳定银纳米粒子及其抗念珠菌能力和抑制耐氟康唑白色念珠菌毒力因子的潜力
白色念珠菌是大约 80% 侵袭性真菌病的病原体,耐药性白色念珠菌是世界卫生组织确定的 "关键优先群体 "中的四种菌株之一。地衣中含有一些稀有的植物化学物质和大量治疗药物,如 Roccella montagnei 的抗真菌能力。利用地衣进行银纳米粒子(AgNPs)的生物合成,可以提供一种生态友好型和具有成本效益的替代方法来对抗新出现的 "微生物抗药性"。因此,该研究的目的是利用蒙大拿地衣的水醇(1:1)提取物生物合成银纳米粒子(Rm-AgNPs),以开发出一种有效的抗白僵菌剂,对抗耐氟康唑的白僵菌 NBC099。紫外光谱鉴定出 Rm-AgNPs 在 420-440 纳米波长处的 AgNPs 特异峰,傅立叶变换红外光谱揭示了胺类、醇类、芳香族化合物和酸的存在。SEM 和 TEM 分析表明,Rm-AgNPs 呈球形,尺寸范围为 10-50 nm。Zetasizer 分析表明,颗粒高度稳定,平均流体力学直径为 116 nm,zeta 电位电荷为 - 41 mV。XRD 分析表明其为面心立方晶格结构。结果表明,Rm-AgNPs 能强烈抑制 NBC099 的生长,最低抑制浓度(IC50)≤ 15 µg。用浓度低于 IC50 的 Rm-AgNPs 处理白僵菌培养物,可下调 NBC099 中不同毒力因子的产生,即菌丝形成(85%)、生物膜产生(80%)、磷脂酶、酯酶和蛋白酶活性。细胞凋亡试验表明,Rm-AgNPs 可通过氧化应激诱导 NBC099 细胞凋亡。有趣的是,当 Rm-AgNPs 浓度超过 15 µg/mL 时,它对小鼠 RAW 246.7 巨噬细胞的细胞毒性(< 6%)可忽略不计。因此,Rm-AgNPs 被认为是一种抗念珠菌的替代品,可用于提高现有药物的疗效。
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