Item Details

title

ENGINEERING ADIPONECTIN-LOADED MICROPARTICLES TO STIMULATE STEM CELLS TO SECRETE EXOSOMES & SUBSEQUENT CHARACTERIZATION FOR TREATMENT OF METABOLIC DISEASE

author

Bowlby, Joshua

abstract

There is significant interest in the use of exosomes for cell-free treatment of disease because of the advantages over cell therapy. The diversity and distribution of uncontrolled inflammation makes it extremely difficult to diagnose and control. Adiponectin is a well-known protein that is secreted by adipose tissue, which can regulate inflammatory responses in patients. Alginate is a highly studied biomaterial that provides a superb immobilization matrix for encapsulating cells and drugs within chemically crosslinked hydrogel microparticles. Small extracellular vehicles (ECVs) are a group of secretory vesicles that contain multiple intracellular signaling molecules, playing a role in reducing inflammation and restoring tissue homeostasis. The goal of the present study is to determine the optimal procedure utilizing air extrusion methodology to encapsulate adiponectin in alginate microbeads crosslinking in 100 mM CaCl2 solution and use it to stimulate and isolate exosomes from cells. Additionally, this study characterized extracellular vesicles secreted by adipose derived mesenchymal stem cells (Ad-MSCs) after prolonged exposure (3 days and 7 days) to adiponectin. This characterization comprised of three assays including quantifying protein content by bicinchoninic acid assay (BCA); identification of exosome biomarkers CD63, ALIX and Syntenin-1 by Western Blot; and nanoparticle tracking analysis (NTA) to analyze particle concentrations and size distribution. It is hypothesized that due to adiponectin’s interaction with cells, encapsulating the protein in alginate beads will stimulate exosome production that can contain anti-inflammatory signals to be utilized for improving inflammatory conditions outcome. In summary, this research elucidated exosome characterization secreted by Ad-MSCs and PSCs after exposure to adiponectin released from alginate microparticles over multiple days to determine the utility of these exosomes for potential therapeutic application.

subject

Adiponectin

Alginate

Characterization

Exosomes

Extracellular Vesicles

Extrusion

contributor

Opara, Emmanuel C (advisor)

Deep, Gagan (committee member)

Kerr, Bethany (committee member)

Levi, Nicole (committee member)

date

2023-07-25T17:48:45Z (accessioned)

2023-12-06T09:30:06Z (available)

2023 (issued)

degree

Biomedical Engineering (discipline)

embargo

2023-12-06 (terms)

identifier

http://hdl.handle.net/10339/102276 (uri)

language

en (iso)

publisher

Wake Forest University

type

Thesis