Macromolecular Crowding: Enhancing Cell Metabolism, Enhancing Regeneration

• Autor(a) principal: Coutinho, André Vieira Trigo dos Santos
• Data de Publicação: 2022
• Tipo de documento: Dissertação
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10362/145555
• topic: Tissue Engineering; macromolecular crowding; polyvinylpyrrolidone; Ficoll; extracellular matrix; fibroblast; Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias

Deep and extensive skin injuries represent a major worldwide healthcare problem without an efficient treatment option, with autologous skin grafts constituting the treatment gold standard. Tissue engineered skin constructs emerge as alternatives to this method, however, "cost-to-heal" factor or the need for long culture times are important drawbacks. The iSkin2 project aims to overcome these limitations, namely by incorporating the macromolecular crowding (MMC) effect, which shows potential in significantly reducing the prolonged healing times. This effect consists in emulating in vitro the heavily macromolecular-crowded in vivo environment, allowing to increase the rate of several biological reactions, namely cellular proliferation and extracellular matrix production. This dissertation assessed if the MMC effect is beneficial for the increase of collagen type I and fibronectin deposition by dermal fibroblasts seeded on polycaprolactone (PCL) scaffolds, and, if it is, which studied crowding agent (Ficoll cocktail or polyvinylpyrrolidone 55 kDa) had the best results. For this, fibroblast viabilitywas assessed in the matrices produced in the presence of the crowders, and immunocytochemistry procedures were performed to visualize and quantify the deposition of collagen type I and fibronectin in the constructs studied. We showed that the use of Ficoll cocktail, although not reaching as good results as the ones in literature, outperformed the use of polyvinylpyrrolidone 55 kDa, offering an interesting alternative to the uncrowded cell medium. We also present evidences that show the potential of using the MMC effect in 3D environments, specifically in PCL matrices.