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Synthesis of hesperetin-loaded PLGA nanoparticles by two different experimental design methods and biological evaluation of optimized nanoparticles

机译：用两种不同的实验设计方法合成叶素酸素加载的PLGA纳米粒子及优化纳米粒子的生物学评价

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Hesperetin was effectively encapsulated into poly (d,l-lactic-co-glycolic acid) nanoparticles by using experimental design methods. A seven-factor Plackett-Burman design was used in order to determine the major process parameters. A significant linear equation, which shows the effect of each process parameter on encapsulation efficiency was developed, and then the most effective factors were determined. Further investigation and optimization was carried out by applying the three-factor three-level Box-Behnken design. Significant second-order mathematical models were developed by regression analysis of the experimental data for both responses: encapsulation efficiency and nanoparticle size. The two step experimental design allowed the synthesis of the desired nanoparticle formulations with maximum encapsulation efficiency (80.5 +/- 4.9%) and minimum particle size (260.2 +/- 16.5 nm) at optimum process conditions: 0.5% polyvinyl alcohol (PVA) concentration, 5.13 water: organic phase ratio, and 3.59 ml min(-1) flow rate of the emulsified solution into 0.1% PVA. Furthermore, the biological activity of these optimized nanoparticles were determined with antimicrobial activity and cytotoxicity studies; results were then compared to the free hesperetin. The cytotoxicity result revealed that hesperetin and hesperetin-loaded nanoparticles were biocompatible with normal cell line L929 fibroblast cells up to 184.83 and 190.88 mu g ml(-1) for 24 h, and up to 133.24 and 134.80 mu g ml(-1) for 48 h, respectively. In the antimicrobial study, the optimized nanoparticle showed inhibition activity (minimal inhibitory concentration (MIC) values were 125 mu g ml(-1) for Escherichia coli, and 200 mu g ml(-1) for Staphylococcus aureus), while the free hesperetin did not demonstrate activity in both strains (MIC value 200 mu g ml(-1)). These in vitro results may provide useful information for the investigation of hesperetin-loaded nanoparticles in diagnostic and therapeutic applications.
机译：橙皮素被有效封装到聚（d，1-乳酸 - 共 - 乙醇酸）通过使用实验设计方法的纳米颗粒。为了确定主要工艺参数被使用的七要素的Plackett-Burman设计。甲显著线性方程，其显示在包封效率每个过程参数的效果被开发，然后确定了最有效的因素。进一步的调查和优化是通过应用三因素三水平箱Behnken设计的设计进行。显著二阶数学模型，通过对两种反应的实验数据的回归分析，开发出：包封率和纳米粒子的大小。两个步骤的实验设计所允许的希望的纳米颗粒制剂与最佳工艺条件最大封装效率（80.5 +/- 4.9％）和最小粒径（260.2 +/- 16.5纳米）的合成：0.5％聚乙烯醇（PVA）浓度，5.13水：有机相比，3.59毫升分钟（-1）的乳化液的0.1％PVA的流速。此外，这些纳米颗粒的优化的生物活性的具有抗微生物活性和细胞毒性研究进行了测定;结果当时相比，免费的橙皮素。细胞毒性结果显示，橙皮素和橙皮素装载纳米颗粒的生物相容性与正常细胞系L929成纤维细胞高达184.83和190.88亩克毫升（-1）24小时，且至多133.24和134.80亩克毫升（-1） 48 h后，分别。在抗微生物的研究中，优化的纳米颗粒显示出抑制活性（最低抑菌浓度（MIC）值分别为125微米克毫升（-1）大肠杆菌，和200亩克毫升（-1）对金黄色葡萄球菌），而自由橙皮素没有证明在这两个菌株的活性（MIC值GT; 200亩克毫升（-1））。这些体外结果可为在诊断和治疗应用橙皮素纳米粒的调查的有用信息。

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