Therapeutic Potential of Fibroblast-Derived Extracellular Vesicles and Matrix Bound Vesicles From Engineered Matrix

Presenter

Aoife Katherine Ruth

Campus

UMass Amherst

Sponsor

Cathal J. Kearney, Department of Biomedical Engineering, UMass Amherst

Schedule

Session 2, 11:30 AM - 12:15 PM [Schedule by Time][Poster Grid for Time/Location]

Location

Poster Board A73, Campus Center Auditorium, Row 4 (A61-A80) [Poster Location Map]

Abstract

Chronic wounds affect 6.5 million people in the United States and can take years to heal or simply never resolve. It has been discovered that extracellular vesicles (EVs) released by cells can assist in wound-healing by providing necessary signaling factors. Recently, researchers found a new type of vesicle, called matrix-bound vesicles (MBVs), which are EVs directly attached to the matrix. This project aims to isolate MBVs and EVs from fibroblast-seeded collagen glycosaminoglycan scaffolds (CG) to investigate their respective roles in wound-healing. To accomplish this, scaffolds were seeded with BJ fibroblasts for 3 weeks in an EV-depleted medium. Conditioned media was collected every 3 days for 21 days. At the end of the culture period, the CG scaffold was lyophilized and digested to release the MBVs. Both the digested scaffold solution and the conditioned media underwent differential centrifugation protocol to isolate the MBVs and EVs, respectively. MBV and EV size were characterized using dynamic light scattering (DLS), and a BCA assay was used to quantify protein content. Our lab has optimized this isolation protocol and through preliminary studies, successfully isolated EVs and MBVs. Functional assays such as migration, proliferation, and macrophage polarization assays will be used next to better understand the vesicles’ impact on cell regeneration and wound healing potential. This work will lay the groundwork for studying MBVs and EVs generated from scaffolds and for future manipulations to enhance their role in wound healing.

Keywords

Wound Healing, Matrix Bound Vesicles, Tissue Engineering, Extracellular Vesicles, Collagen

Research Area

Engineering

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