Elizabeth Wright
Pronouns: she/her/hers
Research Mentor(s): David Kohn
Co-Presenter:
Research Mentor School/College/Department: Departments of Biologic and Materials Sciences and Biomedical Engineering / Engineering
Presentation Date: April 20
Presentation Type: Poster
Session: Session 1 – 10am – 10:50am
Room: League Ballroom
Authors: Elizabeth Wright, Merjem Mededovic, David Kohn
Presenter: 45
Abstract
Of the 15 million fractures that occur annually in the US, 750,000 bone injuries do not heal properly due to a limited ability to regenerate. A potential treatment is using Mesenchymal Stem Cells (MSCs), which contribute to bone regeneration through differentiation into osteoblasts. Increased expression of Connexin 43 (Cx43), a gap junction protein, can lead to higher levels of differentiation and therefore increased bone healing. The expression of Connexin 43 was increased by gene delivery using a lentivirus to transduce GJA1 (the gene that encodes the protein) into both MSCs and HUVECs. For fracture healing to occur, MSCs must be able to migrate to the fracture site. This study investigates the effectiveness of using Gap Junction Intercellular Communication (GJIC) modification as a treatment for bone fractures. To determine effectiveness of this method, transfection efficiency, cell viability post transduction, and duration of gene incorporation were investigated. A Transwell assay in the presence of chemokines (SDF-1) was used to determine effects of increased GJIC on cell migration. A western blot for Cx43 showed that Cx expression was increased in transduced cells, indicating successful gene delivery. A cell viability assay showed a high proportion of the cells as viable post transfection. In addition, a Mono vs Coculture study displayed no decrease in transfection efficiency when HUVECs and MSCs are grown together. Finally, the transwell assay showed that transduction does not affect migration, as migration of the transfected cells were not inhibited in comparison to the control cells. Overall, we were able to show that gene delivery using a lentivirus is a potential tool to treat delayed bone healing. Investigating the effect of upregulated GJIC on MSC differentiation and migration will advance understanding of the use of gene delivery for treatment of delayed bone healing.
Engineering, Interdisciplinary
