International Journal of Nanomedicine (Jul 2018)
Uptake and transport of pullulan acetate nanoparticles in the BeWo b30 placental barrier cell model
Abstract
Hongbo Tang,1 Ziwen Jiang,2 Haibo He,3 Xiaoqin Li,3 Haipeng Hu,1 Ning Zhang,1 Yinmei Dai,2 Zhimin Zhou4 1Department of Pharmacy, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100006, People’s Republic of China; 2Department of Gynecology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100006, People’s Republic of China; 3College of Biological and Pharmaceutical Sciences, China Three Gorges University, Yichang 443002, People’s Republic of China; 4Biomedical Barriers Research Center, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin Key Laboratory of Biomedical Materials, Tianjin, 300192, People’s Republic of China Introduction: Nanomedicine has shown a great potential in perinatal medicine because of its characteristics of sustained, controlled release and targeting ability; on the other hand, it may also lead to unexpected toxicities such as embryotoxicity and even malformation after crossing the placental barrier, but data concerning transplacental transport are scarce. Pullulan acetate (PA) nanoparticles (NPs) are a promising nanocarrier derived from natural polysaccharide; however, their transplacental transport ability and mechanism are unknown. Materials and methods: In this study, fluorescein isothiocyanate (FITC) conjugated PA (PA-FITC) was synthesized. PA-FITC NPs were characterized by dynamic light scattering, transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The cytotoxicity of PA-FITC NPs at concentrations of 15, 30, 60, 125, 250, 500, 1,000 and 2,000 μg/mL was studied by cell counting kit-8. The human chorionic gonadotrophin (HCG) cytokine assay was conducted to evaluate the biological function of BeWo b30 cells. Endocytic mechanisms of PA-FITC NPs were investigated via fluorescence analysis. The monolayer properties were characterized by TEM, tight junction staining, transepithelial electrical resistance and fluorescein sodium transportation. The transport ability was measured in the cell based transwell model by confocal imaging and SEM. Results: PA-FITC NPs were almost spherical shape with a size range of 200–300 nm. Cell viability of BeWo b30 cells was up to 100% in all groups. The concentrations of HCG increased with increasing numbers of cells and culture time, which showed the good biological function of BeWo b30 cells. PA-FITC NPs were rapidly endocytosed through caveolae-mediated endocytosis and pinocytosis, with uptake inhibition rates with nystatin (NY) and colchicines (Col) of 55% and 51% respectively. BeWo b30 cell monolayer was formed over 5 days. PA-FITC NPs were found in the cytoplasm of cells on the transwell membranes; while some NPs were found in the basolateral (fetal) compartment over 24 h. Conclusion: In summary, PA-FITC NPs are nontoxic, can cross the blood-placental barrier, and show mainly internalization to BeWo b30 cells through caveolae-mediated endocytosis and pinocytosis pathways, major via the former pathway. The results could benefit the adjustment and control of the transplacental transport of nanomedicines. Keywords: transport, embryotoxicity, endocytosis, nystatin, caveolae
