A Protective Extracellular Matrix-Based Gene Delivery Reservoir Fabricated by Electrostatic Charge Manipulation

Gene therapy is a field that offers hope and promise for the treatment of diseases and traumas of a genetic nature and otherwise. However, progress toward the clinic has been delayed because of concerns over the safety of viral vectors and the efficacy of safer nonviral systems, which have low transfection efficacy and short transgene expression. This study describes the fabrication and characterization of a safe gene delivery reservoir system that has the potential to overcome issues associated with nonviral systems. Harnessing the electrostatic charges of collagen and polystyrene, microspheres were fabricated using a template-based method and characterized by microscopy techniques such as scanning electron microscopy, transmission electron microscopy, and atomic force microscopy, with the removal of the polystyrene template confirmed by Fourier transform infrared spectroscopy analysis. Loading and release of polyplexes confirmed the ability of the system to prolong polyplex delivery, with minimal cytotoxicity observed from viability studies on 3T3 fibroblasts. Finally, biological activity of released polyplexes was confirmed by reporter gene expression. Taken together, these properties indicate the potential of this system as a reservoir for gene delivery.