Digital Light Processing Bioprinting Full-Thickness Human Skin for Modelling Infected Chronic Wounds in Vitro

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dc.contributor.author Stefanek, Evan
dc.date.accessioned 2022-08-08T16:53:26Z
dc.date.copyright 2022 en_US
dc.date.issued 2022-08-08
dc.identifier.uri http://hdl.handle.net/1828/14089
dc.description.abstract Chronic wounds have a detrimental impact on patient quality of life, a significant economic cost, and often lead to severe outcomes such as amputation, sepsis or death. The elaborate interplay between bacteria, cutaneous cells, immune cells, growth factors, and proteases in chronic wounds has complicated the development of new therapies that could improve outcomes for chronic wound patients. Existing in vitro models of chronic wounds do not appreciably mimic the complexity of the wound environment. In this work, tissue-engineered skin was developed with the goal of creating an in vitro platform appropriate for testing potential clinical therapies for chronic wounds. The Lumen-X, a digital light processing bioprinter, was used to create tissue-engineered skin from a 7.5% (w/v) gelatin methacryloyl hydrogel laden with primary dermal fibroblasts. This dermal layer was developed with an emphasis on providing a favourable microenvironment for the fibroblasts in order to mimic their in vivo phenotype. An epidermal layer of human keratinocytes was formed on the hydrogel surface and stratified through culture at the air-liquid-interface. The maturation of the epidermis was thoroughly characterized with histology, immunohistochemistry, and trans-epithelial electrical resistance analyses which showed a degree of maturation suitable for wound healing studies. To verify the suitability of this tissue-engineered skin for studying healing in vitro, sharp tweezers were used to create physical wounds in the epidermis which were then infected with Pseudomonas aeruginosa. Reepithelialisation, the production of the pro- inflammatory cytokine TNF-α, and the presence of bacteria were monitored over time, showing healing in wounds without infection and those treated with antibiotics, and potential biofilm formation in infected wounds. The tissue-engineered skin developed here is suitable for use as an in vitro model of the infected chronic wound environment. Future work includes developing better methods for creating the physical wound and characterizing the bacterial biofilm in order to improve the reproducibility and clarity of results. Such a model will then be well-poised to begin testing potential chronic wound therapies in vitro. en_US
dc.language English eng
dc.language.iso en en_US
dc.rights Available to the World Wide Web en_US
dc.subject Bioprinting en_US
dc.subject Tissue engineering en_US
dc.subject Skin en_US
dc.subject Wound healing en_US
dc.subject Hydrogels en_US
dc.subject Photo-crosslinking en_US
dc.title Digital Light Processing Bioprinting Full-Thickness Human Skin for Modelling Infected Chronic Wounds in Vitro en_US
dc.type Thesis en_US
dc.contributor.supervisor Akbari, Mohsen
dc.degree.department Department of Mechanical Engineering en_US
dc.degree.level Master of Applied Science M.A.Sc. en_US
dc.description.scholarlevel Graduate en_US
dc.description.embargo 2023-07-26

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