Quorum Sensing Laboratory, Centre for Research in Infectious Diseases (CRID), School of Chemical and Biotechnology, SASTRA Deemed to be University, Thanjavur, 613401, India.
School of Chemical and Biotechnology, SASTRA Deemed to be University, Thanjavur, 613401, India.
Sci Rep. 2023 Jan 23;13(1):1250. doi: 10.1038/s41598-023-28033-0.
Vibrio cholerae causes cholera, an acute diarrhoeal disease. The virulence in V. cholerae is regulated by the quorum-sensing mechanism and response regulator LuxO positively regulates the expression of virulence determinants adhesion, biofilm formation, and cholera toxin production. Previous in-silico studies revealed that 2-methoxy-4-vinylphenol could bind to the ATP binding site of LuxO and the complex was compact and stable in pHs like intestinal pHs. Here, we have explored the polymeric nano-formulation of 2-methoxy-4-vinylphenol using cellulose acetate phthalate for controlled drug release and their effectiveness in attenuating the expression of V. cholerae virulence. Physico-chemical characterization of the formulation showed particles with a mean size of 91.8 ± 14 nm diameter and surface charge of - 14.7 ± 0.07 mV. The uniform round polymeric nanoparticles formed displayed about 51% burst release of the drug at pH 7 by 3rd h, followed by a controlled linear release in alkaline pH. The polymeric nanoparticles demonstrated a tenfold increase in intestinal membrane permeability ex-vivo. At lower concentrations, the 2-methoxy-4-vinylphenol polymeric nanoparticles were non-cytotoxic to Int 407 cells. In-vitro analysis at pH 6, pH 7, pH 8, and pH 9 revealed that cellulose acetate phthalate-2-methoxy-4-vinylphenol nanoparticles were non-bactericidal at concentrations up to 500 μg/mL. At 31.25 μg/mL, the nanoparticles inhibited about 50% of the biofilm formation of V. cholerae MTCC 3905 and HYR14 strains. At this concentration, the adherence of V. cholerae MTCC 3905 and HYR14 to Int 407 cell lines were also significantly affected. Gene expression analysis revealed that the expression of tcp, qrr, and ct at pH 6, 7, 8, and 9 has reduced. The CAP-2M4VP nanoparticles have demonstrated the potential to effectively reduce the virulence of V. cholerae in-vitro.
霍乱弧菌引起霍乱，一种急性腹泻病。霍乱弧菌的毒力受群体感应机制和响应调节剂 LuxO 调节，正向调节粘附、生物膜形成和霍乱毒素产生等毒力决定因素的表达。以前的计算机研究表明，2-甲氧基-4-乙烯基苯酚可以与 LuxO 的 ATP 结合位点结合，并且在类似肠道 pH 的 pH 值下，复合物是紧凑和稳定的。在这里，我们探索了使用醋酸纤维素邻苯二甲酸酯的 2-甲氧基-4-乙烯基苯酚的聚合纳米制剂，用于控制药物释放及其在减弱霍乱弧菌毒力方面的有效性。制剂的物理化学特性表明，粒径为 91.8 ± 14nm 直径，表面电荷为-14.7 ± 0.07mV 的颗粒。形成的均匀圆形聚合物纳米粒子在第 3 小时通过 pH7 时表现出约 51%的药物突释，随后在碱性 pH 下进行控制的线性释放。聚合物纳米粒子在体外显示出肠膜通透性增加十倍。在较低浓度下，2-甲氧基-4-乙烯基苯酚聚合物纳米粒子对 Int407 细胞无细胞毒性。在 pH6、pH7、pH8 和 pH9 的体外分析表明，在高达 500μg/mL 的浓度下，醋酸纤维素邻苯二甲酸酯-2-甲氧基-4-乙烯基苯酚纳米粒子对细菌没有杀菌作用。在 31.25μg/mL 时，纳米粒子抑制了霍乱弧菌 MTCC3905 和 HYR14 菌株约 50%的生物膜形成。在该浓度下，霍乱弧菌 MTCC3905 和 HYR14 对 Int407 细胞系的粘附也受到显著影响。基因表达分析表明，tcp、qrr 和 ct 在 pH6、7、8 和 9 下的表达减少。CAP-2M4VP 纳米粒子已证明具有在体外有效降低霍乱弧菌毒力的潜力。
