Liquified capsules containing nanogrooved microdiscs and co-encapsulated endothelial and mesenchymal stem cells for bone tissue engineering

Detalhes bibliográficos
Autor(a) principal: Carreira, Mariana da Silva
Data de Publicação: 2019
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
Texto Completo: http://hdl.handle.net/10773/30068
Resumo: The emerging of the Tissue Engineering field led to the substitution of natural bone grafts by engineered ones, termed as scaffolds. Scaffolds are temporary structures that can be combined with cells, and include biochemical or biophysical cues to promote the regeneration of tissues. Surface topography has been shown to influence cell behavior and direct the differentiation of mesenchymal stem cells (MSCs) into distinct lineages. Whereas this has been verified in a bi-dimensional (2D) context, the role of topography in tri-dimensional (3D), which better mimics the natural cell environment, needs to be explored. Therefore, the main goal of this work is to incorporate topographical featured microparticles in a 3D system, and to assess their ability to guide MSCs osteoblastic differentiation in the absence of osteogenic differentiation factors. A co-culture of Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) and human umbilical vein endothelial cells (HUVECs) is encapsulated in a 3D system. Such system consists in a permselective liquified environment containing freely dispersed spherical microparticles (spheres) or nanogrooved microdiscs (microdiscs). Microdiscs presenting 358±23 nm grooves and 944±49 nm ridges are produced via nanoimprinting of spherical polycaprolactone microparticles between water-soluble polyvinyl alcohol counter-moulds of optical media (CDs). Spheres and microdiscs are cultured in vitro in basal or osteogenic media for 21 days. It is hypothesized that, without additional osteogenic differentiation factors, the topographical features present in the microdiscs would induce the osteoblastic differentiation of adhered WJ-MSCs through contact guidance. Results show an enhanced osteoblastic differentiation in microdiscs compared to spheres, even in basal medium, evidenced by ALP activity and extracellular matrix deposition. The developed 3D bioencapsulation system comprising nontopographical features might be suitable as smart and autonomous bone tissue engineered approaches requiring minimum in vitro manipulation.
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spelling Liquified capsules containing nanogrooved microdiscs and co-encapsulated endothelial and mesenchymal stem cells for bone tissue engineering3D-cultureBioencapsulationBone regenerationCo-cultureEndothelial cellsIn vitro vascularizationNanotopographyStem cellsUmbilical cordThe emerging of the Tissue Engineering field led to the substitution of natural bone grafts by engineered ones, termed as scaffolds. Scaffolds are temporary structures that can be combined with cells, and include biochemical or biophysical cues to promote the regeneration of tissues. Surface topography has been shown to influence cell behavior and direct the differentiation of mesenchymal stem cells (MSCs) into distinct lineages. Whereas this has been verified in a bi-dimensional (2D) context, the role of topography in tri-dimensional (3D), which better mimics the natural cell environment, needs to be explored. Therefore, the main goal of this work is to incorporate topographical featured microparticles in a 3D system, and to assess their ability to guide MSCs osteoblastic differentiation in the absence of osteogenic differentiation factors. A co-culture of Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) and human umbilical vein endothelial cells (HUVECs) is encapsulated in a 3D system. Such system consists in a permselective liquified environment containing freely dispersed spherical microparticles (spheres) or nanogrooved microdiscs (microdiscs). Microdiscs presenting 358±23 nm grooves and 944±49 nm ridges are produced via nanoimprinting of spherical polycaprolactone microparticles between water-soluble polyvinyl alcohol counter-moulds of optical media (CDs). Spheres and microdiscs are cultured in vitro in basal or osteogenic media for 21 days. It is hypothesized that, without additional osteogenic differentiation factors, the topographical features present in the microdiscs would induce the osteoblastic differentiation of adhered WJ-MSCs through contact guidance. Results show an enhanced osteoblastic differentiation in microdiscs compared to spheres, even in basal medium, evidenced by ALP activity and extracellular matrix deposition. The developed 3D bioencapsulation system comprising nontopographical features might be suitable as smart and autonomous bone tissue engineered approaches requiring minimum in vitro manipulation.Com o desenvolvimento da Engenharia de Tecidos, diferentes tipos de scaffolds têm sido propostos como alternativa aos enxertos ósseos. Estes scaffolds podem incluir células e fatores físico-químicos para promover a regeneração do tecido. A diferenciação de células mesenquimais estaminais (MSCs) em diferentes linhagens pode ser induzida por superfícies topográficas. Apesar de se constatar esta diferenciação celular mediada por características topográficas em sistemas bidimensionais (2D), é necessário explorar se o mesmo se verifica para sistemas tridimensionais (3D), visto que estes recapituam melhor o ambiente celular nativo. O objetivo principal deste trabalho é investigar se num contexto 3D, micropartículas com topografia conseguem guiar a diferenciação de MSCs para osteoblastos, mesmo sem adição de fatores de diferenciação ao meio de cultura. MSCs isoladas da geleia de Wharton (WJ-MSCs) são encapsuladas num sistema 3D em co-cultura com células endoteliais isoladas da veia do cordão umbilical, e com micropartículas esféricas (esferas) ou microdiscos com nanotopografia (microdiscos). Os microdiscos são produzidos através da nanoimpressão do padrão presente em discos óticos (CDs) com contra-moldes de álcool polivinílico, apresentando sulcos com 358±23 nm e cumes de 944±49 nm. As esferas e os microdiscos são cultivados in vitro durante 21 dias em meio basal ou osteogénico. A hipótese é que, na ausência de fatores de diferenciação osteogénica, a topografia dos microdiscos induza a diferenciação osteoblástica das WJ-MSCs. A quantificação da atividade enzimática da fosfatase alcalina revela um aumento na diferenciação osteoblástica induzida pelos microdiscos, comparando com as esferas, mesmo em meio basal. Este resultado é corroborado com a deposição de matriz celular e pela deteção de osteopontina. Espera-se que o sistema proposto seja adequado para o desenvolvimento de aplicações autónomas para regeneração óssea através de engenharia de tecidos, com o mínimo de manipulação in vitro.2020-12-15T14:59:51Z2019-12-01T00:00:00Z2019-12info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10773/30068engCarreira, Mariana da Silvainfo:eu-repo/semantics/openAccessreponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiainstacron:RCAAP2024-05-06T04:29:10Zoai:ria.ua.pt:10773/30068Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireinfo@rcaap.ptopendoar:https://opendoar.ac.uk/repository/71602025-05-28T14:10:08.823994Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiafalse
dc.title.none.fl_str_mv Liquified capsules containing nanogrooved microdiscs and co-encapsulated endothelial and mesenchymal stem cells for bone tissue engineering
title Liquified capsules containing nanogrooved microdiscs and co-encapsulated endothelial and mesenchymal stem cells for bone tissue engineering
spellingShingle Liquified capsules containing nanogrooved microdiscs and co-encapsulated endothelial and mesenchymal stem cells for bone tissue engineering
Carreira, Mariana da Silva
3D-culture
Bioencapsulation
Bone regeneration
Co-culture
Endothelial cells
In vitro vascularization
Nanotopography
Stem cells
Umbilical cord
title_short Liquified capsules containing nanogrooved microdiscs and co-encapsulated endothelial and mesenchymal stem cells for bone tissue engineering
title_full Liquified capsules containing nanogrooved microdiscs and co-encapsulated endothelial and mesenchymal stem cells for bone tissue engineering
title_fullStr Liquified capsules containing nanogrooved microdiscs and co-encapsulated endothelial and mesenchymal stem cells for bone tissue engineering
title_full_unstemmed Liquified capsules containing nanogrooved microdiscs and co-encapsulated endothelial and mesenchymal stem cells for bone tissue engineering
title_sort Liquified capsules containing nanogrooved microdiscs and co-encapsulated endothelial and mesenchymal stem cells for bone tissue engineering
author Carreira, Mariana da Silva
author_facet Carreira, Mariana da Silva
author_role author
dc.contributor.author.fl_str_mv Carreira, Mariana da Silva
dc.subject.por.fl_str_mv 3D-culture
Bioencapsulation
Bone regeneration
Co-culture
Endothelial cells
In vitro vascularization
Nanotopography
Stem cells
Umbilical cord
topic 3D-culture
Bioencapsulation
Bone regeneration
Co-culture
Endothelial cells
In vitro vascularization
Nanotopography
Stem cells
Umbilical cord
description The emerging of the Tissue Engineering field led to the substitution of natural bone grafts by engineered ones, termed as scaffolds. Scaffolds are temporary structures that can be combined with cells, and include biochemical or biophysical cues to promote the regeneration of tissues. Surface topography has been shown to influence cell behavior and direct the differentiation of mesenchymal stem cells (MSCs) into distinct lineages. Whereas this has been verified in a bi-dimensional (2D) context, the role of topography in tri-dimensional (3D), which better mimics the natural cell environment, needs to be explored. Therefore, the main goal of this work is to incorporate topographical featured microparticles in a 3D system, and to assess their ability to guide MSCs osteoblastic differentiation in the absence of osteogenic differentiation factors. A co-culture of Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) and human umbilical vein endothelial cells (HUVECs) is encapsulated in a 3D system. Such system consists in a permselective liquified environment containing freely dispersed spherical microparticles (spheres) or nanogrooved microdiscs (microdiscs). Microdiscs presenting 358±23 nm grooves and 944±49 nm ridges are produced via nanoimprinting of spherical polycaprolactone microparticles between water-soluble polyvinyl alcohol counter-moulds of optical media (CDs). Spheres and microdiscs are cultured in vitro in basal or osteogenic media for 21 days. It is hypothesized that, without additional osteogenic differentiation factors, the topographical features present in the microdiscs would induce the osteoblastic differentiation of adhered WJ-MSCs through contact guidance. Results show an enhanced osteoblastic differentiation in microdiscs compared to spheres, even in basal medium, evidenced by ALP activity and extracellular matrix deposition. The developed 3D bioencapsulation system comprising nontopographical features might be suitable as smart and autonomous bone tissue engineered approaches requiring minimum in vitro manipulation.
publishDate 2019
dc.date.none.fl_str_mv 2019-12-01T00:00:00Z
2019-12
2020-12-15T14:59:51Z
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reponame_str Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
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