Hannah Levin

Utilizing Vivotex Scaffolds to Promote Matrix-Bound Nanovesicle Production

Faculty PI: Marley Dewey

Bone grafts are commonly used to repair critical bone defects, however, this ‘gold standard’ is limited by donor tissue availability and unpredictable healing outcomes, creating a need for more effective treatments to promote bone tissue regeneration. Biomaterials offer a customizable, reproducible alternative, mimicking bone structure and supporting cell growth. My research looks into PCL scaffolds, a biomaterial, and Extracellular Vesicles (EV), lipid nanoparticles produced by all cells that play critical roles in cell-cell communication and cargo delivery. Recently, a novel subtype of extracellular vesicles embedded within the extracellular matrix (ECM), known as matrix-bound nanovesicles (MBVs), have emerged as promising players in tissue regeneration, however, maximizing their yield in in-vitro models remains a limitation. My goal is to optimize biomaterials to produce scalable and reproducible MBVs. We culture mesenchymal stem cells (MSC) on scaffolds with five different architectures, varying in fiber alignment, spacing, and thickness. Following ECM deposition, scaffolds were decellularized, digested with liberase, and MBVs were isolated by size exclusion chromatography. Nanoparticle tracking analysis quantified MBV concentration and size. Two scaffold architectures produced higher MBV particle counts than the remaining scaffold designs and control. Our findings suggest scaffold architecture influences MBV production. Future studies will focus on two scaffold designs that maximize MBV production. We will evaluate ECM contents through histological staining, and characterize MBV cargo to determine how scaffold architecture influences vesicle cargo. Optimizing biomaterials to maximize MBV production may enable scalable production of these vesicles for preclinical studies and, ultimately, the development of MBV-based therapies for bone regeneration.