Hydrogels derived from human chorionic membrane as an innovative platform for 3D cell culture

Detalhes bibliográficos
Autor(a) principal: Martins, Elisa Alexandra Gonçalves
Data de Publicação: 2021
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10773/30888
Resumo: Regenerative medicine and tissue engineering have emerged as alternatives to therapies currently used in the treatment and replacement of damaged tissues or organs, due to the very limited capacity of the human body to regenerate. This area combines the culture of cells in biomaterials and the presence of signals. In native tissue, cells are surrounded by an extracellular matrix (ECM) composed of several proteins, glycosaminoglycans and soluble factors. ECM is very important in the cellular response, as it influences processes such as migration, proliferation and differentiation. Thus, an effort has been made to create materials that mimic this function. The human placenta is a virtually unlimited source of ECM, without associated ethical issues, immunoprivileged, biocompatible and capable of healing. In addition, the successful use of fetal membranes as an allograft has been reported in the literature and more recently the chorionic membrane has been described as effective in periodontal regeneration. In addition to the base material, the used morphology is also important. Hydrogels are a unique class of materials in terms of their ability to mimic ECM. They are 3D hydrophilic polymeric networks capable of capturing large amounts of water, allowing cell fixation and migration, exchange of nutrients, oxygen and cellular waste. However, most hydrogels produced from decellularized ECM have low mechanical properties, since these must mimic the native tissue. Thus, the objective of this work is to produce, as far as we know, for the first time, hydrogels derived from photocrosslinkable human chorionic membrane (CM) with adjustable mechanical properties, for the 3D culture of cells. In this work, CM prepolymers were produced by the addition of methacrylic anhydride, with two degrees of modification: one with a lower degree (CMMA100) and another with a higher degree of modification (CMMA250). By adding a photoinitiator to these prepolymers and in the presence of light with a specific wavelength it is possible to produce hydrogels. Quantifications of some of the most important ECM proteins allowed to verify their retention in the obtained material. The evaluation of the mechanical properties allowed to realize that it was possible to produce robust hydrogels and whose mechanical properties and water absorption capacity vary with their concentration and which are in the range of most human tissue elastic modulus. Moreover, it was possible to verify that CMMA hydrogels allowed the culture of cells that proliferated and remained viable for at least 7 days. The results obtained so far suggest that these CMMA based hydrogels have potential for 3D cell culture.
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spelling Hydrogels derived from human chorionic membrane as an innovative platform for 3D cell cultureHorionic membraneExtracellular matrixCardiac tissue engineeringThree-dimensionalHydrogelPlacentaRegenerative medicine and tissue engineering have emerged as alternatives to therapies currently used in the treatment and replacement of damaged tissues or organs, due to the very limited capacity of the human body to regenerate. This area combines the culture of cells in biomaterials and the presence of signals. In native tissue, cells are surrounded by an extracellular matrix (ECM) composed of several proteins, glycosaminoglycans and soluble factors. ECM is very important in the cellular response, as it influences processes such as migration, proliferation and differentiation. Thus, an effort has been made to create materials that mimic this function. The human placenta is a virtually unlimited source of ECM, without associated ethical issues, immunoprivileged, biocompatible and capable of healing. In addition, the successful use of fetal membranes as an allograft has been reported in the literature and more recently the chorionic membrane has been described as effective in periodontal regeneration. In addition to the base material, the used morphology is also important. Hydrogels are a unique class of materials in terms of their ability to mimic ECM. They are 3D hydrophilic polymeric networks capable of capturing large amounts of water, allowing cell fixation and migration, exchange of nutrients, oxygen and cellular waste. However, most hydrogels produced from decellularized ECM have low mechanical properties, since these must mimic the native tissue. Thus, the objective of this work is to produce, as far as we know, for the first time, hydrogels derived from photocrosslinkable human chorionic membrane (CM) with adjustable mechanical properties, for the 3D culture of cells. In this work, CM prepolymers were produced by the addition of methacrylic anhydride, with two degrees of modification: one with a lower degree (CMMA100) and another with a higher degree of modification (CMMA250). By adding a photoinitiator to these prepolymers and in the presence of light with a specific wavelength it is possible to produce hydrogels. Quantifications of some of the most important ECM proteins allowed to verify their retention in the obtained material. The evaluation of the mechanical properties allowed to realize that it was possible to produce robust hydrogels and whose mechanical properties and water absorption capacity vary with their concentration and which are in the range of most human tissue elastic modulus. Moreover, it was possible to verify that CMMA hydrogels allowed the culture of cells that proliferated and remained viable for at least 7 days. The results obtained so far suggest that these CMMA based hydrogels have potential for 3D cell culture.A medicina regenerativa e a engenharia de tecidos surgiram como alternativas às terapias atualmente usadas no tratamento e substituição de tecidos ou órgãos danificados, pois o corpo humano tem capacidade muito limitada de regeneração. Esta área combina a cultura de células em biomateriais com biomoléculas sinalizadoras. No tecido nativo, as células estão envolvidas por matriz extracelular (ECM) composta por diversas proteínas, glicosaminoglicanos e fatores solúveis. A ECM é muito importante na resposta celular, pois tem influência em processos com a migração, proliferação e diferenciação. Assim, tem sido feito um esforço para criar materiais que mimetizem essa função. A placenta humana trata-se uma fonte de ECM virtualmente ilimitada, sem questões éticas associadas, imunoprivilegiada devido à falta de expressão de antigénios, possui elevada biocompatibilidade, propriedades imunossupressoras, anti-inflamatórias e regenerativas. Para além disso, está reportado na literatura o sucesso do uso de membranas fetais como aloenxerto e mais recentemente a membrana coriónica foi descrita como eficaz na regeneração periodontal. Para além do material base, também é importante a morfologia usada. Os hidrogéis são uma classe única de materiais relativamente à capacidade de mimetizar a ECM. São redes poliméricas hidrofílicas 3D capazes de captar grandes quantidades de água, permitindo a fixação e migração celular, trocas de nutrientes, oxigénio e resíduos celulares. No entanto, a maioria dos hidrogéis produzidos a partir de ECM descelularizada, tem reduzidas propriedades mecânicas e pouca estabilidade in vitro. Assim, o objetivo deste trabalho é produzir, pelo que sabemos, pela primeira vez, hidrogéis derivados de membrana coriónica (CM) humana foto-reticuláveis com propriedades mecânicas ajustáveis, para a cultura de células 3D. Neste trabalho foram produzidos pré-polímeros de CM através da reação com metacrílico anidrido, com dois graus de modificação: um de menor grau (CMMA100) e outro com maior grau de modificação (CMMA250). Ao adicionar um fotoiniciador a estes pré-polímeros e na presença de luz com comprimento de onda específico é possível produzir hidrogéis. A quantificação de DNA permitiu verificar uma descelularização eficiente da CM, enquanto que as quantificações de algumas das mais importantes proteínas da ECM permitiram verificar a sua retenção no material obtido. A avaliação das propriedades mecânicas permitiu perceber que foi possível produzir hidrogéis robustos e cuja rigidez e capacidade de absorção de água são ajustáveis. Para além disso foi possível verificar que os hidrogéis permitem a cultura de células, que proliferaram e mantiveram-se viáveis por pelo menos 7 dias. Os resultados obtidos até agora sugerem que estes hidrogéis baseados em CMMA têm potencial para cultura de células 3D.2023-03-01T00:00:00Z2021-02-26T00:00:00Z2021-02-26info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10773/30888engMartins, Elisa Alexandra Gonçalvesinfo:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-02-22T11:59:43Zoai:ria.ua.pt:10773/30888Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:02:55.079690Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Hydrogels derived from human chorionic membrane as an innovative platform for 3D cell culture
title Hydrogels derived from human chorionic membrane as an innovative platform for 3D cell culture
spellingShingle Hydrogels derived from human chorionic membrane as an innovative platform for 3D cell culture
Martins, Elisa Alexandra Gonçalves
Horionic membrane
Extracellular matrix
Cardiac tissue engineering
Three-dimensional
Hydrogel
Placenta
title_short Hydrogels derived from human chorionic membrane as an innovative platform for 3D cell culture
title_full Hydrogels derived from human chorionic membrane as an innovative platform for 3D cell culture
title_fullStr Hydrogels derived from human chorionic membrane as an innovative platform for 3D cell culture
title_full_unstemmed Hydrogels derived from human chorionic membrane as an innovative platform for 3D cell culture
title_sort Hydrogels derived from human chorionic membrane as an innovative platform for 3D cell culture
author Martins, Elisa Alexandra Gonçalves
author_facet Martins, Elisa Alexandra Gonçalves
author_role author
dc.contributor.author.fl_str_mv Martins, Elisa Alexandra Gonçalves
dc.subject.por.fl_str_mv Horionic membrane
Extracellular matrix
Cardiac tissue engineering
Three-dimensional
Hydrogel
Placenta
topic Horionic membrane
Extracellular matrix
Cardiac tissue engineering
Three-dimensional
Hydrogel
Placenta
description Regenerative medicine and tissue engineering have emerged as alternatives to therapies currently used in the treatment and replacement of damaged tissues or organs, due to the very limited capacity of the human body to regenerate. This area combines the culture of cells in biomaterials and the presence of signals. In native tissue, cells are surrounded by an extracellular matrix (ECM) composed of several proteins, glycosaminoglycans and soluble factors. ECM is very important in the cellular response, as it influences processes such as migration, proliferation and differentiation. Thus, an effort has been made to create materials that mimic this function. The human placenta is a virtually unlimited source of ECM, without associated ethical issues, immunoprivileged, biocompatible and capable of healing. In addition, the successful use of fetal membranes as an allograft has been reported in the literature and more recently the chorionic membrane has been described as effective in periodontal regeneration. In addition to the base material, the used morphology is also important. Hydrogels are a unique class of materials in terms of their ability to mimic ECM. They are 3D hydrophilic polymeric networks capable of capturing large amounts of water, allowing cell fixation and migration, exchange of nutrients, oxygen and cellular waste. However, most hydrogels produced from decellularized ECM have low mechanical properties, since these must mimic the native tissue. Thus, the objective of this work is to produce, as far as we know, for the first time, hydrogels derived from photocrosslinkable human chorionic membrane (CM) with adjustable mechanical properties, for the 3D culture of cells. In this work, CM prepolymers were produced by the addition of methacrylic anhydride, with two degrees of modification: one with a lower degree (CMMA100) and another with a higher degree of modification (CMMA250). By adding a photoinitiator to these prepolymers and in the presence of light with a specific wavelength it is possible to produce hydrogels. Quantifications of some of the most important ECM proteins allowed to verify their retention in the obtained material. The evaluation of the mechanical properties allowed to realize that it was possible to produce robust hydrogels and whose mechanical properties and water absorption capacity vary with their concentration and which are in the range of most human tissue elastic modulus. Moreover, it was possible to verify that CMMA hydrogels allowed the culture of cells that proliferated and remained viable for at least 7 days. The results obtained so far suggest that these CMMA based hydrogels have potential for 3D cell culture.
publishDate 2021
dc.date.none.fl_str_mv 2021-02-26T00:00:00Z
2021-02-26
2023-03-01T00:00:00Z
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