Author :
Jayagopal, A. ; Sussman, E.M. ; Shastri, V.P.
Abstract :
The objectives of this study were to synthesize and characterize functionalized solid lipid nanoparticles (fSLN) to investigate their interaction with endothelial cell monolayers and to evaluate their transendothelial transport capabilities. fSLN bearing tetramethylrhodamine-isothiocyanate-labeled bovine serum albumin (TRITC-BSA) and Coumarin 6 were prepared using a single-step phase-inversion process that afforded concurrent surface modification with a variety of macromolecules such as polystyrene sulfonate (PSS), poly-L-lysine (PLL), heparin (Hep), polyacrylic acid (PAA), polyvinyl alcohol, and polyethylene glygol (PEG). TRITC-BSA/Coumarin 6 encapsulated in fSLN with composite surface functionality (PSS-PLL and PSS-PLL-Hep) were also investigated. Size and surface charge of fSLN were analyzed using dynamic light scattering and transmission electron microscopy. Transport across bovine aortic endothelial cell (BAEC) monolayers was assessed spectrophotometrically using a transwell assay, and fSLN localization at the level of the cell and permeable support was analyzed using fluorescence microscopy. fSLN with tunable size and surface functionality were successfully produced, and had significant effects on cell localization and transport. Specifically, fSLN with PSS-PLL-Hep composite surface functionalization was capable of translocating 53.2 plusmn 8.7 mug of TRITC-BSA within 4 h, with fSLN-PEG, fSLN-PAA, and fSLN-PSS exhibiting near-complete apical, paracellular, and cytosolic localization, respectively. Coumarin 6 was released by fSLN as indicated by dye labeling of BAEC membranes. We have developed a rapid process for the production of fSLN bearing low- and high-molecular-weight payloads of varying physicochemical properties. These findings have implications for drug delivery and bioimaging applications, since due to tunable surface chemistry, fSLN internalization and/or translocation across intact endothelial cell monolayers is possible.
Keywords :
biomedical imaging; biomembranes; cellular biophysics; drug delivery systems; fluorescence; nanobiotechnology; nanoparticles; surface chemistry; transmission electron microscopy; BAEC membranes; Coumarin 6; bioimaging applications; bovine aortic endothelial cell; composite surface functionality; drug delivery; dynamic light scattering; endothelial cell monolayers; fluorescence microscopy; functionalized solid lipid nanoparticles; heparin; high-molecular-weight payloads; low-molecular-weight payloads; macromolecules; physicochemistry; poly-L-lysine; polyacrylic acid; polyethylene glycol; polystyrene sulfonate; polyvinyl alcohol; single-step phase-inversion process; surface charge; surface modification; tetramethylrhodamine-isothiocyanate-labeled bovine serum albumin; transendothelial delivery; transmission electron microscopy; tunable surface chemistry; Bovine; Fluorescence; Labeling; Light scattering; Lipidomics; Nanoparticles; Phase locked loops; Polyethylene; Solids; Transmission electron microscopy; Drug delivery; imaging; lipid; nanoparticles; Animals; Cattle; Cells, Cultured; Endothelial Cells; Lipids; Materials Testing; Nanoparticles; Pharmaceutical Vehicles;