Electronic Thesis and Dissertation Repository

Thesis Format

Monograph

Degree

Master of Science

Program

Anatomy and Cell Biology

Collaborative Specialization

Musculoskeletal Health Research

Supervisor

Flynn, Lauren E.

Abstract

There is a need for new treatments that promote the healing of chronic wounds, which are associated with high costs to the Canadian healthcare system and high mortality rates as current treatments are frequently inefficacious. To address this need, this thesis focused on the development of a “cell-assembled” bioscaffolding technology platform that incorporates pro-regenerative adipose-derived stromal cells (ASCs) dispersed throughout a cell-supportive extracellular matrix derived from decellularized adipose tissue (DAT). To advance towards pre-clinical testing in an impaired wound healing model in genetically-diabetic db/db mice, scaffolds generated with mouse ASCs sourced from C57BL/6 versus db/db mice were compared. Stable scaffolds were generated with both sources, and scaffold contraction, biochemical composition, cell viability, and levels of secreted VEGF-A, HGF and IL-6 were similar between the groups. Next, the cell-assembly methods were successfully adapted using porcine ASCs to generate scaled up scaffolds sized for future testing in a porcine wound healing model.

Summary for Lay Audience

Chronic wounds are wounds that do not heal properly within 3 months and their causes are poorly understood. However, these wounds are likely to affect those with pre-existing health conditions. In particular, type 2 diabetics have a 25% lifetime risk of developing chronic wounds, compared to a ∼2 % rate in the general population. Chronic wound patients experience pain and a high 5-year mortality rate, due to the fact that current treatments are often ineffective, as well as being costly and requiring long treatment times.

Cell therapies involving the delivery of pro-regenerative cells including adipose-derived stromal cells (ASCs) sourced from fat have emerged as promising alternatives to promote wound healing. The wound environment is hostile, and a challenge is that ASC survival and localized retention following delivery can be very limited. As a solution, ASCs can be seeded into 3-D scaffolds that can be implanted into chronic wounds. In particular, this thesis investigated scaffolds derived from pro-regenerative proteins isolated from fat tissue discarded as surgical waste, termed decellularized adipose tissue (DAT). DAT can be applied off-the-shelf without stimulating a negative immune response, supports ASC attachment, and may promote ASC survival and wound healing activity. Recently, a technique to generate scaffolds with an enhanced ASC density was developed. However, key questions related to scaffold development remain, including: 1) whether ASCs sourced from diabetic donors will respond similarly within the scaffolds to those sourced from healthy donors, and 2) whether the methods can be scaled-up to generate scaffolds sized for use in humans or for pre-clinical testing in a pig model as a next-step towards clinical translation.

First, scaffolds were generated using mouse ASCs sourced from healthy versus genetically diabetic mice and were shown to have similar properties in terms of handling, cell viability, tissue structure and composition. Next, the fabrication methods were successfully adapted to generate large scaffolds that contained a high density of viable porcine ASCs. Overall, this thesis highlights the versatility of the cell assembly approach for generating robust scaffolds and represents key progress towards future pre-clinical testing of a patented cell delivery platform for wound healing applications.

Creative Commons License

Creative Commons Attribution 4.0 License
This work is licensed under a Creative Commons Attribution 4.0 License.

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