Cell-based assay for characterizing cell adhesion properties of active targeted nanoparticles under static and flow condition using an integrated flow chamber

Abstract

Nanoparticle targeting of cancer cell surface markers to allow the specific delivery of chemotherapeutic agents is an attractive concept in cancer treatment. Prior to in vivo studies, newly developed nanomedicines must be tested in preclinical in vitro models fully representative of physiological conditions. The aim of this study was to develop an in vitro method for characterizing the cell binding properties of active targeting nanoparticles, using an integrated flow chamber. Anti-vascular endothelial growth factor (VEGF) antibody conjugated liposome/PLGA hybrid nanoparticles were generated, characterized, and utilized in binding studies with three different VEGF-positive cancer cell lines, in addition to VEGF-negative fibroblasts. Binding, and subsequent VEGF expression was examined through immunostaining, confocal microscopy and Western blot analysis. Our results indicated that targeted and non-targeted nanoparticles cannot be distinguished under static in vitro cell binding conditions, but that selective binding of targeted nanoparticles to VEGF-positive cells was more prominent under conditions of laminar flow. The flow chamber described herein thus provides a platform for cell-based assays, allowing effective evaluation of cell adhesion using active targeted nanoparticles.