Effects of Cell Migration on Polyethylenimine/pDNA Polyplex Transfection Efficacy and Mechanism
| Author(s) | Hewes, Jacob | |
| Date Accessioned | 2024-06-25T15:33:14Z | |
| Date Available | 2024-06-25T15:33:14Z | |
| Publication Date | 2023-05 | |
| Abstract | Nonhealing wounds have been the subject of decades of basic and clinical research. Despite new knowledge about the biology of impaired wound healing, little progress has been made in treating chronic wounds, leaving patients with few therapeutic options. The lack of treatments has created a major global burden on the healthcare system and resulted in high health care costs. In the past few decades, the field of nonviral gene delivery has garnered significant interest as one of the most promising strategies for the treatment of chronic wounds. Nonviral vectors have the advantage to deliver genes to target cells without the immunogenic or toxic responses associated with viral vectors, however, limited gene transfer efficacy remains a challenge. This work aims to address these challenge by optimizing the formulation of poly(ethylenimine) (PEI) nanocarriers for effective plasmid DNA (pDNA) delivery. PEI-pDNA polyplexes were synthesized with different N/P ratios to optimize polyplex properties, including size and zeta potential, and used to transfect NIH/3T3 cells in vitro. It was found that an N/P ratio of 10 produced the smallest and most positively charged polyplexes, resulting in the highest transgene expression. Furthermore, a simple in vitro wound model was developed and employed to study PEI-pDNA polyplex transfection during wound healing. Cells migrating to close the wound were found to be transfected more readily than cells that did not migrate. Additionally, the transfection efficiency in migrating cells was shown to decrease when less growth factors were present in the extracellular environment. Thus, this work demonstrates that PEI can be used to delivery therapeutic DNA and targeted delivery of transgenes to migrating cells during wound healing could serve as a viable strategy to enhance tissue repair and regeneration. | |
| Advisor | enter | |
| Program | enter | |
| URL | https://udspace.udel.edu/handle/19716/34518 | |
| Language | en_US | |
| Publisher | University of Delaware | |
| Title | Effects of Cell Migration on Polyethylenimine/pDNA Polyplex Transfection Efficacy and Mechanism | |
| Type | Thesis |