Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmacy, The British University in Egypt, Cairo, Egypt.
Department of Pharmacology, Faculty of Pharmacy, The British University in Egypt, Cairo, Egypt.
Int J Nanomedicine. 2022 Feb 18;17:733-750. doi: 10.2147/IJN.S345870. eCollection 2022.
To design and optimize -cinnamic acid-loaded PLGA nanoparticles (CIN-PLGA-NPs) and assess its inhibitory effect on epithelial-mesenchymal transition (EMT) in triple-negative breast cancer.
The quality by design approach was used to correlate the formulation parameters (PLGA amount and Poloxamer188 concentration) and critical quality attributes (entrapment efficiency percent, particle size and zeta potential). Design of CIN-PLGA-NPs formulations was done based on central composite response surface design and formulated by nanoprecipitation method. In addition, the optimized CIN-PLGA-NPs formulation was further evaluated for morphology using transmission electron microscopy and in vitro dissolution test. The cytotoxicity of CIN-PLGA-NPs optimized formula in comparison to the free -cinnamic acid (CIN-Free) was investigated in vitro using MDA-MB-231, triple-negative breast cancer cells, followed by scratch wound assay for evaluating the impact on the migratory potential of MDA-MB-231 cells. In vivo antitumor activity was evaluated using Ehrlich ascites carcinoma solid tumor animal model where tumor volumes were measured at different time points and necrotic/apoptotic indices were estimated in tumor sections. EMT markers, E- and N-cadherin, were assessed in solid tumors as well.
The optimized formulation showed entrapment efficiency of 76.98%, particle size of 186.3 nm with a smooth spherical surface and zeta potential of -28.47 mV indicating its stability. Furthermore, CIN-PLGA-NPs optimized formula released 60.8±1.89% of the total CIN-Free within 24 hours compared to 29±1.25% of the raw CIN-Free indicating improved dissolution rate. The optimized formula showed superior cytotoxicity on MDA-MB-231 cells compared to its free counterpart as well as increased wound closure percentage along with reduced tumor size in mice and increased necrotic and apoptotic indices. Tumor levels of E-cadherin and N-cadherin were indicative of EMT inhibition.
Our findings proved the capability of PLGA nanoparticles in loading -cinnamic acid in addition to enhancing its antitumor efficacy in triple-negative breast cancer possibly via inhibiting EMT.
设计并优化 -肉桂酸-负载 PLGA 纳米粒（CIN-PLGA-NPs），并评估其对三阴性乳腺癌上皮-间充质转化（EMT）的抑制作用。
采用质量源于设计方法将制剂参数（PLGA 用量和泊洛沙姆 188 浓度）与关键质量属性（包封效率百分比、粒径和 Zeta 电位）相关联。基于中心复合响应面设计，采用纳米沉淀法制备 CIN-PLGA-NPs 制剂。此外，进一步通过透射电子显微镜和体外溶出试验对优化的 CIN-PLGA-NPs 制剂进行形态学评价。采用 MDA-MB-231，三阴性乳腺癌细胞，体外研究 CIN-PLGA-NPs 优化配方与游离 -肉桂酸（CIN-Free）的细胞毒性，然后通过划痕试验评估对 MDA-MB-231 细胞迁移潜力的影响。采用艾氏腹水癌实体瘤动物模型评价体内抗肿瘤活性，不同时间点测量肿瘤体积，在肿瘤切片中估计坏死/凋亡指数。还评估了 EMT 标志物 E-和 N-钙粘蛋白在实体瘤中的表达。
优化的配方显示包封效率为 76.98%，粒径为 186.3nm，表面光滑呈球形，Zeta 电位为-28.47mV，表明其稳定性。此外，与原始 CIN-Free 相比，CIN-PLGA-NPs 优化配方在 24 小时内释放了 60.8±1.89%的总 CIN-Free，表明其溶解速率提高。与游离 CIN-Free 相比，优化配方在 MDA-MB-231 细胞中的细胞毒性更高，伤口闭合百分比增加，小鼠肿瘤体积减小，坏死和凋亡指数增加。肿瘤组织中 E-钙粘蛋白和 N-钙粘蛋白的水平表明 EMT 抑制。
我们的研究结果证明了 PLGA 纳米粒能够负载 -肉桂酸，并且能够增强三阴性乳腺癌的抗肿瘤疗效，可能是通过抑制 EMT 实现的。
