Cellular Uptake of Functional Nanogels Prepared by Inverse Miniemulsion ATRP with Encapsulated Proteins, Carbohydrates, and Gold Nanoparticles

Abstract

Atom transfer radical polymerization (ATRP) was used to produce a versatile drug delivery system capable of encapsulating a range of molecules. Inverse miniemulsion ATRP permitted the synthesis of biocompatible and uniformly cross-linked poly(ethylene oxide)-based nanogels entrapping gold nanoparticles, bovine serum albumin, rhodamine B isothiocyanate-dextran, or fluoresceine isothiocyanate-dextran. These moieties were entrapped to validate several biological outcomes and to model delivery of range of molecules. Cellular uptake of nanogels was verified by transmission electron microscopy, gel electrophoresis, Western blotting, confocal microscopy, and flow cytometry. Fluorescent co-localization of nanogels with a fluorophore-conjugated antibody for clathrin indicated clathrin-mediated endocytosis. Further, internalization of nanogels either with or without GRGDS cell attachment-mediating peptides was quantified using flow cytometry. After 45 minutes of incubation, the uptake of unmodified FITC-Dx-loaded nanogels was 62%, whereas cellular uptake increased to >95% with the same concentration of GRGDS-modified FITC-Dx- nanogels. In addition, a co-culture of human umbilical vascular endothelial cells (HUVECs) and human mesenchymal stem cells (hMSCs) validated cell endocytosis. Application of ATRP enabled the synthesis of a functionalized drug delivery system with a uniform network that is capable of encapsulating and delivering inorganic, organic, and biological molecules.

Injectable Hyaluronic Acid-co-Gelatin Cryogels for Tissue-Engineering Applications

“…Unlike other minimally invasive delivery approaches that require polymer injection and in situ gelation, gels made using cryopolymerization are able to retain predefined geometries, architecture, and porosity posti-njection [62,63,65,75,76,77,78]. Similarly, physical properties of cryogels were dependent on the fabrication temperature.…”

Aerosol Delivery of siRNA to the Lungs. Part 2: Nanocarrier-based Delivery Systems

“…and Matyjaszewski K., 2007, Nagavarma B. et al, 2012). Among the available controlled/living radical polymerization methods successfully and extensively studied methods include nitroxide-mediated polymerization (NMP), atom transfer radical polymerization (ATRP), and reversible addition and fragmentation transfer chain polymerization (RAFT) (Dire C. et al, 2008, Min K. et al, 2009, Nicolas J. et al, 2007, Rieger J. et al, 2010, Siegwart D.J. et al, 2009, Zhou X. et al, 2007).…”

Polymeric Nanoparticles Preparation Techniques and Applications for Drug Delivery System- A Brief Review.

“…Implementation of C/LRP in the industrially important aqueous dispersed systems, resulting in the formation of polymeric nanoparticles with precise particle size and size distribution control, is crucial for future commercial success of C/LRP. [77] Among the available controlled/living radical polymerization methods successful and extensively studied methods are 1) nitroxide-mediated polymerization (NMP) [78][79][80][81] , 2) atom transfer radical polymerization (ATRP) [82][83][84][85][86] and 3) reversible addition and fragmentation transfer chain polymerization (RAFT). [87][88][89][90][91] Ionic gelation or coacervation of hydrophilic polymers Polymeric nanoparticles are prepared by using biodegradable hydrophilic polymers such as chitosan, gelatin and sodium alginate.…”