Effects of Cell Migration on Polyethylenimine/pDNA Polyplex Transfection Efficacy and Mechanism
Date
2023-05
Authors
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Journal ISSN
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Publisher
University of Delaware
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.