PDF(Pubmed)
Abstract：
UNASSIGNED: Proliferative vitreoretinal diseases (PVDs) represent a heterogeneous group of pathologies characterized by the presence of retinal proliferative membranes, in whose development retinal pigment epithelium (RPE) is deeply involved. As the only effective treatment for PVDs at present is surgery, we aimed to investigate the potential therapeutic activity of Nutlin-3a, a small non-genotoxic inhibitor of the MDM2/p53 interaction, on ARPE-19 cell line and on human RPE primary cells, as in vitro models of RPE and, more importantly, to formulate and evaluate Nutlin-3a loaded liposomes designed for ophthalmic administration.
UNASSIGNED: Liposomes were produced using an innovative approach by a microfluidic device under selection of different conditions. Liposome size distribution was evaluated by photon correlation spectroscopy and centrifugal field flow fractionation, while the liposome structure was studied by transmission electron microscopy and Fourier-transform infrared spectroscopy. The Nutlin-3a entrapment capacity was evaluated by ultrafiltration and HPLC. Nutlin-3a biological effectiveness as a solution or loaded in liposomes was evaluated by viability, proliferation, apoptosis and migration assays and by morphological analysis.
UNASSIGNED: The microfluidic formulative study enabled the selection of liposomes composed of phosphatidylcholine (PC) 5.4 or 8.2 mg/mL and 10% ethanol, characterized by roundish vesicular structures with 150-250 nm mean diameters. Particularly, liposomes based on the lower PC concentration were characterized by higher stability. Nutlin-3a was effectively encapsulated in liposomes and was able to induce a significant reduction of viability and migration in RPE cell models.
UNASSIGNED: Our results lay the basis for a possible use of liposomes for the ocular delivery of Nutlin-3a.
UNASSIGNED: Liposomes were produced using an innovative approach by a microfluidic device under selection of different conditions. Liposome size distribution was evaluated by photon correlation spectroscopy and centrifugal field flow fractionation, while the liposome structure was studied by transmission electron microscopy and Fourier-transform infrared spectroscopy. The Nutlin-3a entrapment capacity was evaluated by ultrafiltration and HPLC. Nutlin-3a biological effectiveness as a solution or loaded in liposomes was evaluated by viability, proliferation, apoptosis and migration assays and by morphological analysis.
UNASSIGNED: The microfluidic formulative study enabled the selection of liposomes composed of phosphatidylcholine (PC) 5.4 or 8.2 mg/mL and 10% ethanol, characterized by roundish vesicular structures with 150-250 nm mean diameters. Particularly, liposomes based on the lower PC concentration were characterized by higher stability. Nutlin-3a was effectively encapsulated in liposomes and was able to induce a significant reduction of viability and migration in RPE cell models.
UNASSIGNED: Our results lay the basis for a possible use of liposomes for the ocular delivery of Nutlin-3a.
摘要：
■增生性玻璃体视网膜疾病（PVDs）代表一组异质性的病理，其特征是存在视网膜增生膜，视网膜色素上皮（RPE）的发育密切相关。由于目前唯一有效的治疗方法是手术，我们旨在研究Nutlin-3a的潜在治疗活性，MDM2/p53相互作用的一种小型非基因毒性抑制剂，在ARPE-19细胞系和人RPE原代细胞上，作为RPE的体外模型，更重要的是,配制和评估设计用于眼科给药的装载Nutlin-3a的脂质体。
■脂质体是在选择不同条件下通过微流体装置使用创新方法生产的。通过光子相关光谱和离心场流分馏评估脂质体的尺寸分布，同时通过透射电子显微镜和傅里叶变换红外光谱研究了脂质体的结构。通过超滤和HPLC评估Nutlin-3a截留能力。Nutlin-3a作为溶液或装载在脂质体中的生物有效性通过活力进行评估，扩散,细胞凋亡和迁移测定以及形态学分析。
■微流体配方研究能够选择由磷脂酰胆碱（PC）5.4或8.2mg/mL和10％乙醇组成的脂质体，以平均直径为150-250nm的圆形囊泡结构为特征。特别是,基于较低PC浓度的脂质体具有较高的稳定性。Nutlin-3a被有效地包封在脂质体中并且能够在RPE细胞模型中诱导活力和迁移的显著降低。
■我们的结果为脂质体用于眼部递送Nutlin-3a的可能用途奠定了基础。
■脂质体是在选择不同条件下通过微流体装置使用创新方法生产的。通过光子相关光谱和离心场流分馏评估脂质体的尺寸分布，同时通过透射电子显微镜和傅里叶变换红外光谱研究了脂质体的结构。通过超滤和HPLC评估Nutlin-3a截留能力。Nutlin-3a作为溶液或装载在脂质体中的生物有效性通过活力进行评估，扩散,细胞凋亡和迁移测定以及形态学分析。
■微流体配方研究能够选择由磷脂酰胆碱（PC）5.4或8.2mg/mL和10％乙醇组成的脂质体，以平均直径为150-250nm的圆形囊泡结构为特征。特别是,基于较低PC浓度的脂质体具有较高的稳定性。Nutlin-3a被有效地包封在脂质体中并且能够在RPE细胞模型中诱导活力和迁移的显著降低。
■我们的结果为脂质体用于眼部递送Nutlin-3a的可能用途奠定了基础。
