Publicação
Human plasma derived hydrogels for myocardial tissue engineering
| Resumo: | The development of novel strategies for cardiac tissue engineering (TE) has been deeply investigated. However, the lack of human-based alternatives that can be the basis to create completely xeno-free systems is a concern. Human blood-derived materials have been explored not only as substitutes for animal-derived serum supplementation for cell culture media but also during cell culture platforms and TE strategies development. Among human-blood materials, human platelet lysates (hPL) have been widely explored due to their richness in bioactive proteins and growth factors essential for cell maintenance and growth. Recently reported methacryloyl hPL (PLMA) have shown great potential as material to develop platforms for cell culture. PLMA-based hydrogels showed remarkable mechanical and stability properties and great ability to support cell culture. Driven by the lack of human-based platforms to create fully xeno-free environments for TE purposes, this thesis proposes the development of humanized platforms as completely xeno-free systems to ultimately be implemented in cardiac TE. Throughout this thesis, PLMA-based materials will be explored for the development of novel platforms for human cell culture capable of creating xeno-free environments. The versatility of PLMA was shown by using this human-derived material to fabricate 3D constructs employing various fabrication techniques. In an era where 3D bioprinting has been widely explored, the application of PLMA-based materials for extrusion printing was herein studied. A pre-gel with shear-thinning properties resulting from a coupling reaction between hPL and PLMA was successfully printed, and 3D fabricated constructs showed remarkable stability in vitro and potential for cell culture as well. PLMA-based porous scaffolds were also obtained by a simple freeze-drying methodology, showcasing the ability of PLMA-derived matrices to support stem cell culture avoiding the use of animal-derived serum supplements. Following this xeno-free approach, PLMA microparticles (MPs) produced by microfluidics allowed cardiac and endothelial cell attachment and maintenance over time when no animal-derived supplements were used with potential to be used as injectable systems for cardiac tissue regeneration. Moreover, PLMA were explored to culture human induced pluripotent stem cells-derived cardiomyocytes (hiPSC-CMs), showcasing the potential of PLMA hydrogels to support the culture of hiPSC-CMs with fast recovery of their beating function. Overall, these studies show the versatility of PLMA to be applied during cell culture and TE platforms development, opening new possibilities not only to culture cells in xeno-free 3D environments but also for application on the development of more reliable TE strategies. |
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| Autores principais: | Santos, Sara Catarina Nunes da Silva |
| Assunto: | Human platelet lysates Methacryloyl platelet lysates Hydrogels Inks Porous scaffolds Microparticles Cardiac tissue engineering Xeno free cell culture |
| Ano: | 2024 |
| País: | Portugal |
| Tipo de documento: | tese de doutoramento |
| Tipo de acesso: | acesso embargado |
| Instituição associada: | Universidade de Aveiro |
| Idioma: | inglês |
| Origem: | RIA - Repositório Institucional da Universidade de Aveiro |
| Resumo: | The development of novel strategies for cardiac tissue engineering (TE) has been deeply investigated. However, the lack of human-based alternatives that can be the basis to create completely xeno-free systems is a concern. Human blood-derived materials have been explored not only as substitutes for animal-derived serum supplementation for cell culture media but also during cell culture platforms and TE strategies development. Among human-blood materials, human platelet lysates (hPL) have been widely explored due to their richness in bioactive proteins and growth factors essential for cell maintenance and growth. Recently reported methacryloyl hPL (PLMA) have shown great potential as material to develop platforms for cell culture. PLMA-based hydrogels showed remarkable mechanical and stability properties and great ability to support cell culture. Driven by the lack of human-based platforms to create fully xeno-free environments for TE purposes, this thesis proposes the development of humanized platforms as completely xeno-free systems to ultimately be implemented in cardiac TE. Throughout this thesis, PLMA-based materials will be explored for the development of novel platforms for human cell culture capable of creating xeno-free environments. The versatility of PLMA was shown by using this human-derived material to fabricate 3D constructs employing various fabrication techniques. In an era where 3D bioprinting has been widely explored, the application of PLMA-based materials for extrusion printing was herein studied. A pre-gel with shear-thinning properties resulting from a coupling reaction between hPL and PLMA was successfully printed, and 3D fabricated constructs showed remarkable stability in vitro and potential for cell culture as well. PLMA-based porous scaffolds were also obtained by a simple freeze-drying methodology, showcasing the ability of PLMA-derived matrices to support stem cell culture avoiding the use of animal-derived serum supplements. Following this xeno-free approach, PLMA microparticles (MPs) produced by microfluidics allowed cardiac and endothelial cell attachment and maintenance over time when no animal-derived supplements were used with potential to be used as injectable systems for cardiac tissue regeneration. Moreover, PLMA were explored to culture human induced pluripotent stem cells-derived cardiomyocytes (hiPSC-CMs), showcasing the potential of PLMA hydrogels to support the culture of hiPSC-CMs with fast recovery of their beating function. Overall, these studies show the versatility of PLMA to be applied during cell culture and TE platforms development, opening new possibilities not only to culture cells in xeno-free 3D environments but also for application on the development of more reliable TE strategies. |
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