das Neves José, Sarmento Bruno
INEB - Instituto de Engenharia Biomédica, Universidade do Porto, Porto, Portugal; CESPU, Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da Saúde, Gandra PRD, Portugal; Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal.
INEB - Instituto de Engenharia Biomédica, Universidade do Porto, Porto, Portugal; CESPU, Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da Saúde, Gandra PRD, Portugal; Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal.
Acta Biomater. 2015 May;18:77-87. doi: 10.1016/j.actbio.2015.02.007. Epub 2015 Feb 17.
Polymeric nanoparticles (NPs) have the potential to provide effective and safe delivery of antiretroviral drugs in the context of prophylactic anti-HIV vaginal microbicides. Dapivirine-loaded poly(d,l-lactic-co-glycolic acid) (PLGA) NPs were produced by an emulsion-solvent evaporation method, optimized for colloidal properties using a 3-factor, 3-level Box-Behnken experimental design, and characterized for drug loading, production yield, morphology, thermal behavior, drug release, in vitro cellular uptake, cytotoxicity and pro-inflammatory potential. Also, drug permeability/membrane retention in well-established HEC-1-A and CaSki cell monolayer models as mediated by NPs was assessed in the absence or presence of mucin. Box-Behnken design allowed optimizing monodisperse 170nm drug-loaded NPs. Drug release experiments showed an initial burst effect up to 4h, followed by sustained 24h release at pH 4.2 and 7.4. NPs were readily taken up by different genital and macrophage cell lines as assessed by fluorescence microscopy. Drug-loaded NPs presented lower or at least similar cytotoxicity as compared to the free drug, with up to around one-log increase in half-maximal cytotoxic concentration values. In all cases, no relevant changes in cell pro-inflammatory cytokine/chemokine production were observed. Dapivirine transport across cell monolayers was significantly decreased when mucin was present at the donor side with either NPs or the free drug, thus evidencing the influence of this natural glycoprotein in membrane permeability. Moreover, drug retention in cell monolayers was significantly higher for NPs in comparison with the free drug. Overall, obtained dapivirine-loaded PLGA NPs possess interesting technological and biological features that may contribute to their use as novel safe and effective vaginal microbicides.
在预防性抗HIV阴道杀菌剂的背景下，聚合物纳米颗粒（NPs）有潜力实现抗逆转录病毒药物的有效和安全递送。通过乳液-溶剂蒸发法制备了载有达匹韦林的聚（d，l-乳酸-共-乙醇酸）（PLGA）纳米颗粒，采用三因素、三水平的Box-Behnken实验设计对其胶体性质进行了优化，并对药物负载量、产率、形态、热行为、药物释放、体外细胞摄取、细胞毒性和促炎潜力进行了表征。此外，在不存在或存在粘蛋白的情况下，评估了由纳米颗粒介导的达匹韦林在成熟的HEC-1-A和CaSki细胞单层模型中的药物渗透性/膜保留情况。Box-Behnken设计使得能够优化单分散的170nm载药纳米颗粒。药物释放实验显示，在4小时内有初始突释效应，随后在pH 4.2和7.4条件下持续释放24小时。通过荧光显微镜评估，纳米颗粒很容易被不同的生殖道和巨噬细胞系摄取。与游离药物相比，载药纳米颗粒的细胞毒性更低或至少相似，半数最大细胞毒性浓度值最多增加约一个对数。在所有情况下，均未观察到细胞促炎细胞因子/趋化因子产生的相关变化。当在供体侧存在粘蛋白时，无论是纳米颗粒还是游离药物，达匹韦林跨细胞单层的转运均显著降低，从而证明了这种天然糖蛋白对膜通透性的影响。此外，与游离药物相比，纳米颗粒在细胞单层中的药物保留率显著更高。总体而言，所获得的载有达匹韦林的PLGA纳米颗粒具有有趣的技术和生物学特性，这可能有助于它们作为新型安全有效的阴道杀菌剂的应用。
