Kunihiro, Joshua2025-01-202025-01-202025-01-202025-01-14https://hdl.handle.net/10012/21392Corneal diseases such as recurrent corneal erosion and dry eye disease are common diseases which can significantly interfere with quality of life. The lack of effective treatments for these diseases necessitates the development of novel treatment methods. Skin wounds are another disease area which presents opportunities for improvement of clinical solutions. Gene delivery solutions have gathered interest for both corneal disease treatment and skin wound healing applications. Matrix metalloproteinase 9 (MMP9) enzymes are highly upregulated in both corneal diseases and in chronic skin wounds. Gelatin methacrylate (GelMA) is a biocompatible hydrogel which is degraded in the presence of MMP9 enzyme. It was thus hypothesized that GelMA could serve as an enzyme-responsive controlled release scaffold for polyplexes. The objective of this project was to demonstrate the potential of GelMA hydrogel as an enzyme- responsive controlled release scaffold for chitosan-graft-polyethyleneimine (CS-PEI) polyplexes as a non-viral gene delivery platform to treat corneal diseases or skin wounds. Key criteria of a controlled-release system are tunable release kinetics and maintained bioactivity of the therapeutic molecule after loading and release. The aims of this work were to characterize the release kinetics of the polyplexes and assess the bioactivity of polyplexes released from the GelMA hydrogel system. Methods were developed to quantitate the concentration of the released CS-PEI polyplexes in solution, and the release profile of polyplexes from GelMA in different MMP9 concentrations was measured. The release was found to be sustained over a 5-day period in the presence of physiologically relevant MMP9 concentrations. The in vitro transfection efficiency of released CS-PEI polyplexes was also explored to demonstrate the bioactivity of the polyplexes. The released polyplexes successfully transfected COS-7 cells in an enzyme-responsive and dose- dependent manner.enhydrogelpolyplexnon-viral gene deliverycontrolled-releasegelatin methcrylatechitosan-graft-polyethylenimineMaster Thesis