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Small extracellular vesicles as anti-scarring agents

Knight, Robert ORCID: 2019. Small extracellular vesicles as anti-scarring agents. PhD Thesis, Cardiff University.
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Introduction: Scar tissue formation during wound repair can be devastating for affected individuals and can lead to reduced tissue function or whole organ failure. Our group has previously isolated and characterised a novel, progenitor cell population from the buccal mucosa. These Oral Mucosa Lamina Propria-Progenitor Cells (OMLP-PCs) are multipotent,highly immuno-suppressive and anti-bacterial. The smallest of the Extracellular Vesicles(EVs), termed exosomes are of endosomal origin and typically range in size between 30-130nm and have recently demonstrated to play an important role in stem cell mediated repair. Methods: Three OMLP-PC cell strains were hTERT immortalised to produce three OMLPPC cell lines (OMLP-PCL). The line which demonstrated the greatest level of plasticity was used for EV isolation and EVs were isolated from the OMLP-PCL conditioned medium. EVs where characterised based upon the Journal of Extracellular Vesicles minimal essential criteria to characterise extracellular vesicles. The effects of OMLP-PCL, MSC and the two ReNeuron SEVs were assessed in a number of in vitro wound healing assays (proliferation,migration, myofibroblast formation) as well as a 3D ex-vivo corneal wound healing model. Results: Immortalised OMLP-PCLs demonstrated a normal fibroblast like morphology,expressed the expected progenitor cell surface markers but demonstrated some differences in respect to their multipotency and immunosuppressive properties. One OMLP-PCL completely mirrored the related cell strain and hence this was utilised to produce EVs. EVs isolated from this OMLP-PCL and EVs from MSCs and the two ReNeuron SEV products, were demonstrated to be SEVs as assessed by NTA, Cryo-EM, floatation density and Flow Cytometry. OMLP-PCL and MSC SEVs significantly increased both skin fibroblast proliferation and wound repopulation/migration in vitro whereas neither ReNeuron SEV effected cell proliferation. OMLP-PCL, MSC and one of the ReNeuron SEVs were also demonstrated to significantly inhibit myofibroblast formation in vitro. Fluorescently labelled SEVs demonstrated uptake into an ex vivo cornea culture model but demonstrated no significant increase in corneal re-epithelisation.

Item Type: Thesis (PhD)
Date Type: Completion
Status: Unpublished
Schools: Dentistry
Subjects: Q Science > Q Science (General)
Uncontrolled Keywords: wound healing; stem cell; EV; exosome; sEV; myofibroblast; progenitor cell; dental; scarless wound healing; hTERT,; oral progenitor
Date of First Compliant Deposit: 22 November 2019
Last Modified: 22 Nov 2022 02:30

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