{"id":{"repo_id":"minho-thes","oai_identifier":"oai:repositorium.uminho.pt:1822/96268"},"canonical_url":"https://search.dev.ndltd.org/etd/minho-thes/oai:repositorium.uminho.pt:1822/96268","repository":{"repo_id":"minho-thes","name":"Universidade do Minho","base_url":"http://repositorium.sdum.uminho.pt/oai/request"},"display":{"title":"Advancing bone tissue engineering research using bone-derived cells laden in biomimetic gellan gum-based biomaterials","abstract":"Bone health is critically challenged by issues such as slow fracture healing, osteoporosis, and bone infections. Osteoporosis, causing over 8.9 million fractures annually, burdens global healthcare system and is currently managed with bisphosphonates, bone grafts, and growth factors. However, these treatments frequently fall short due to the need for repeated interventions, limited effectiveness, and the disease’s complex underlying factors. This thesis tackles the challenge of developing ineffective treatments for bone disorders like osteoporosis by focusing on the development of three-dimensional (3D) bone-engineered tissue models. These highly realistic models can be further used to enhance our understanding of bone conditions and evaluate the efficacy of potential new therapies. Gellan Gum (GG) spongy-like hydrogels are recognized in tissue engineering for finely represent the extracellular matrix owed to their high-water content, microstructure, mechanical performance, and cell adhesiveness. In this thesis, bioactive and osteoconductive cues were introduced in spongy-like hydrogels polymeric by adding lactoferrin (Lf) and hydroxyapatite (HAp). Particularly, the addition of HAp stimulated osteoblasts to osteocyte differentiation and mineralization, being selected for further studies. Subsequently, GG/HAp spongy-like hydrogels were shaped into an outer ring and an inner disc and cultured with human bone marrow-derived stem cells (HBM-MSCs) (outer ring) and with both HBM-MSCs and endothelial cells (inner disc) to build a 3D tissue model that anatomically represents the cortical and spongy bone. Once the sole constructs achieved osteogenesis and capillary-like structures maturity, they were assembled and formed an integrated 3D vascular-bone-like tissue model, offering a suitable platform for studying bone physiology and evaluating therapies. Acute inflammation was induced in this model through TNF-α supplementation to mimic the initial trigger of inflammatory bone diseases like osteoporosis and further evaluate its impact on vascular assembly, osteogenesis, and inflammation. Findings showed that TNF-α did not impact cell viability or significantly change the expression of angiogenic and osteogenic markers. However, it increased pro-inflammatory cytokine levels in a dose-dependent manner and inversely proportional to the culture time, suggesting its resolution along the time. Lastly, the GG/HAp polymer(s) content was tailored to obtain a printable bioactive ink to meet the increasing need of on-demand bone tissue models. 3D bone tissue analogues were printed and supported SaOs-2 cell viability and osteogenesis in vitro, which highlights the osteoconductive and osteogenic properties of the composite inks.","abstract_html":"Bone health is critically challenged by issues such as slow fracture healing, osteoporosis, and bone infections. Osteoporosis, causing over 8.9 million fractures annually, burdens global healthcare system and is currently managed with bisphosphonates, bone grafts, and growth factors. However, these treatments frequently fall short due to the need for repeated interventions, limited effectiveness, and the disease’s complex underlying factors. This thesis tackles the challenge of developing ineffective treatments for bone disorders like osteoporosis by focusing on the development of three-dimensional (3D) bone-engineered tissue models. These highly realistic models can be further used to enhance our understanding of bone conditions and evaluate the efficacy of potential new therapies. Gellan Gum (GG) spongy-like hydrogels are recognized in tissue engineering for finely represent the extracellular matrix owed to their high-water content, microstructure, mechanical performance, and cell adhesiveness. In this thesis, bioactive and osteoconductive cues were introduced in spongy-like hydrogels polymeric by adding lactoferrin (Lf) and hydroxyapatite (HAp). Particularly, the addition of HAp stimulated osteoblasts to osteocyte differentiation and mineralization, being selected for further studies. Subsequently, GG/HAp spongy-like hydrogels were shaped into an outer ring and an inner disc and cultured with human bone marrow-derived stem cells (HBM-MSCs) (outer ring) and with both HBM-MSCs and endothelial cells (inner disc) to build a 3D tissue model that anatomically represents the cortical and spongy bone. Once the sole constructs achieved osteogenesis and capillary-like structures maturity, they were assembled and formed an integrated 3D vascular-bone-like tissue model, offering a suitable platform for studying bone physiology and evaluating therapies. Acute inflammation was induced in this model through TNF-α supplementation to mimic the initial trigger of inflammatory bone diseases like osteoporosis and further evaluate its impact on vascular assembly, osteogenesis, and inflammation. Findings showed that TNF-α did not impact cell viability or significantly change the expression of angiogenic and osteogenic markers. However, it increased pro-inflammatory cytokine levels in a dose-dependent manner and inversely proportional to the culture time, suggesting its resolution along the time. Lastly, the GG/HAp polymer(s) content was tailored to obtain a printable bioactive ink to meet the increasing need of on-demand bone tissue models. 3D bone tissue analogues were printed and supported SaOs-2 cell viability and osteogenesis in vitro, which highlights the osteoconductive and osteogenic properties of the composite inks.","abstract_has_math":false,"creators":["Bastos, Ana Raquel Fernandes"],"institution":"Universidade do Minho","degree_name":"Tese de doutoramento em Engenharia de Tecidos, Medicina Regenerativa e Células Estaminais","degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Correlo, V. M.","Reis, R. L."],"committee_chairs":[],"committee_members":[],"year":2025,"date_issued":"2025-04-15","date_published":"2025-04-15","updated_at":"2026-08-21T16:46:39Z","subjects":["Bone cells","Gellan Gum","Hydrogels","Hydroxyapatite","Three-dimensional models","Células ósseas","Hidrogéis","Hidroxiapatite","Modelos tridimensionais"],"languages":["eng"],"rights":["embargoedAccess (2 Years)"],"rights_urls":["http://creativecommons.org/licenses/by/4.0/"],"identifier_entries":[]},"links":{"outbound_url":"https://hdl.handle.net/1822/96268","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"source_record":{"url":"http://repositorium.sdum.uminho.pt/oai/request?verb=GetRecord&metadataPrefix=dim&identifier=oai%3Arepositorium.uminho.pt%3A1822%2F96268","prefix":"dim"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Correlo, V. M.","Reis, R. 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Osteoporosis, causing over 8.9 million fractures annually, burdens global healthcare system and is currently managed with bisphosphonates, bone grafts, and growth factors. However, these treatments frequently fall short due to the need for repeated interventions, limited effectiveness, and the disease’s complex underlying factors. This thesis tackles the challenge of developing ineffective treatments for bone disorders like osteoporosis by focusing on the development of three-dimensional (3D) bone-engineered tissue models. These highly realistic models can be further used to enhance our understanding of bone conditions and evaluate the efficacy of potential new therapies. Gellan Gum (GG) spongy-like hydrogels are recognized in tissue engineering for finely represent the extracellular matrix owed to their high-water content, microstructure, mechanical performance, and cell adhesiveness. In this thesis, bioactive and osteoconductive cues were introduced in spongy-like hydrogels polymeric by adding lactoferrin (Lf) and hydroxyapatite (HAp). Particularly, the addition of HAp stimulated osteoblasts to osteocyte differentiation and mineralization, being selected for further studies. Subsequently, GG/HAp spongy-like hydrogels were shaped into an outer ring and an inner disc and cultured with human bone marrow-derived stem cells (HBM-MSCs) (outer ring) and with both HBM-MSCs and endothelial cells (inner disc) to build a 3D tissue model that anatomically represents the cortical and spongy bone. Once the sole constructs achieved osteogenesis and capillary-like structures maturity, they were assembled and formed an integrated 3D vascular-bone-like tissue model, offering a suitable platform for studying bone physiology and evaluating therapies. Acute inflammation was induced in this model through TNF-α supplementation to mimic the initial trigger of inflammatory bone diseases like osteoporosis and further evaluate its impact on vascular assembly, osteogenesis, and inflammation. Findings showed that TNF-α did not impact cell viability or significantly change the expression of angiogenic and osteogenic markers. However, it increased pro-inflammatory cytokine levels in a dose-dependent manner and inversely proportional to the culture time, suggesting its resolution along the time. Lastly, the GG/HAp polymer(s) content was tailored to obtain a printable bioactive ink to meet the increasing need of on-demand bone tissue models. 3D bone tissue analogues were printed and supported SaOs-2 cell viability and osteogenesis in vitro, which highlights the osteoconductive and osteogenic properties of the composite inks.","A saúde óssea enfrenta desafios críticos, tais como a cicatrização lenta de fraturas, a osteoporose e infeções ósseas. A osteoporose, que causa mais de 8,9 milhões de fraturas anualmente, sobrecarrega o sistema de saúde global e é atualmente tratada com bisfosfonatos, enxertos ósseos e fatores de crescimento. No entanto, esses tratamentos falham frequentemente devido à necessidade de intervenções repetidas, eficácia limitada e aos fatores complexos subjacentes à doença. Esta tese aborda o desafio relacionado com o desenvolvimento de tratamentos ineficazes para distúrbios ósseos como a osteoporose, focando-se no desenvolvimento em laboratório de modelos tridimensionais (3D) de tecido ósseo. Estes modelos, altamente realistas, podem posteriormente ser usados para melhorar a compreensão das condições ósseas e avaliar a eficácia de potenciais novas terapias. A utilização de hidrogéis esponjosos de Goma Gelana (GG) em engenharia de tecidos tem um elevado potencial devido às suas semelhanças com a matriz extracelular, nomeadamente o seu alto teor de água, microestrutura, desempenho mecânico e adesão celular. Nesta tese, agentes bioativos e osteocondutivos foram introduzidos nos hidrogéis esponjosos poliméricos, tais com a lactoferrina (Lf) e hidroxiapatite (HAp). Em particular, a adição de HAp estimulou a diferenciação e mineralização dos osteoblastos em osteócitos, sendo selecionada para os estudos seguintes. De seguida, os hidrogéis esponjosos de GG/HAp foram moldados em forma de anel (externo) e um disco (interno) e cultivados com células estaminais derivadas de medula óssea humana (HBM-MSCs) (anel externo) e com HBM-MSCs e células endoteliais (disco interno) para construir um modelo 3D que represente anatomicamente o osso cortical e trabecular. Após atingirem a osteogénese e a maturidade das estruturas semelhantes a capilares, respetivamente, foram acoplados de forma a formarem um modelo integrado 3D de osso vascularizado, dando origem a uma plataforma adequada para estudar a fisiologia óssea e avaliação de novas terapias. A inflamação aguda foi induzida neste modelo através da suplementação com Fator de Necrose Tumoral alfa (TNF-α) para simular a indução inicial de doenças ósseas inflamatórias, como a osteoporose, e avaliar o seu impacto na formação vascular, osteogénese e inflamação. Os resultados mostraram que o TNF-α não afetou a viabilidade celular nem alterou significativamente a expressão dos marcadores angiogénicos e osteogénicos. No entanto, os níveis de citocinas pró-inflamatórias aumentaram proporcionalmente com a quantidade de TNF-α e diminuíram com o tempo de cultura, sugerindo a resolução gradual da fase aguda da inflamação. Por fim, a composição/teor de polímero GG/HAp foi ajustado para obter uma “ink” bioativa de forma a dar resposta à crescente necessidade de desenvolver modelos de tecido ósseos altamente reprodutíveis. Análogos de tecido ósseo 3D foram impressos e suportaram a viabilidade e osteogénese das células SaOs-2 in vitro, destacando as propriedades osteocondutivas e osteogénicas das “inks” compósitas."]},{"key":"dc:title","label":"Title","values":["Advancing bone tissue engineering research using bone-derived cells laden in biomimetic gellan gum-based biomaterials"]}]}],"canonical_facts":{"dc:contributor.advisor":["Correlo, V. M.","Reis, R. L."],"dc:creator":["Bastos, Ana Raquel Fernandes"],"dc:date.accessioned":["2025-07-02T10:04:44Z"],"dc:date.issued":["2025-04-15"],"dc:description.abstract":["Bone health is critically challenged by issues such as slow fracture healing, osteoporosis, and bone infections. Osteoporosis, causing over 8.9 million fractures annually, burdens global healthcare system and is currently managed with bisphosphonates, bone grafts, and growth factors. However, these treatments frequently fall short due to the need for repeated interventions, limited effectiveness, and the disease’s complex underlying factors. This thesis tackles the challenge of developing ineffective treatments for bone disorders like osteoporosis by focusing on the development of three-dimensional (3D) bone-engineered tissue models. These highly realistic models can be further used to enhance our understanding of bone conditions and evaluate the efficacy of potential new therapies. Gellan Gum (GG) spongy-like hydrogels are recognized in tissue engineering for finely represent the extracellular matrix owed to their high-water content, microstructure, mechanical performance, and cell adhesiveness. In this thesis, bioactive and osteoconductive cues were introduced in spongy-like hydrogels polymeric by adding lactoferrin (Lf) and hydroxyapatite (HAp). Particularly, the addition of HAp stimulated osteoblasts to osteocyte differentiation and mineralization, being selected for further studies. Subsequently, GG/HAp spongy-like hydrogels were shaped into an outer ring and an inner disc and cultured with human bone marrow-derived stem cells (HBM-MSCs) (outer ring) and with both HBM-MSCs and endothelial cells (inner disc) to build a 3D tissue model that anatomically represents the cortical and spongy bone. Once the sole constructs achieved osteogenesis and capillary-like structures maturity, they were assembled and formed an integrated 3D vascular-bone-like tissue model, offering a suitable platform for studying bone physiology and evaluating therapies. Acute inflammation was induced in this model through TNF-α supplementation to mimic the initial trigger of inflammatory bone diseases like osteoporosis and further evaluate its impact on vascular assembly, osteogenesis, and inflammation. Findings showed that TNF-α did not impact cell viability or significantly change the expression of angiogenic and osteogenic markers. However, it increased pro-inflammatory cytokine levels in a dose-dependent manner and inversely proportional to the culture time, suggesting its resolution along the time. Lastly, the GG/HAp polymer(s) content was tailored to obtain a printable bioactive ink to meet the increasing need of on-demand bone tissue models. 3D bone tissue analogues were printed and supported SaOs-2 cell viability and osteogenesis in vitro, which highlights the osteoconductive and osteogenic properties of the composite inks.","A saúde óssea enfrenta desafios críticos, tais como a cicatrização lenta de fraturas, a osteoporose e infeções ósseas. A osteoporose, que causa mais de 8,9 milhões de fraturas anualmente, sobrecarrega o sistema de saúde global e é atualmente tratada com bisfosfonatos, enxertos ósseos e fatores de crescimento. No entanto, esses tratamentos falham frequentemente devido à necessidade de intervenções repetidas, eficácia limitada e aos fatores complexos subjacentes à doença. Esta tese aborda o desafio relacionado com o desenvolvimento de tratamentos ineficazes para distúrbios ósseos como a osteoporose, focando-se no desenvolvimento em laboratório de modelos tridimensionais (3D) de tecido ósseo. Estes modelos, altamente realistas, podem posteriormente ser usados para melhorar a compreensão das condições ósseas e avaliar a eficácia de potenciais novas terapias. A utilização de hidrogéis esponjosos de Goma Gelana (GG) em engenharia de tecidos tem um elevado potencial devido às suas semelhanças com a matriz extracelular, nomeadamente o seu alto teor de água, microestrutura, desempenho mecânico e adesão celular. Nesta tese, agentes bioativos e osteocondutivos foram introduzidos nos hidrogéis esponjosos poliméricos, tais com a lactoferrina (Lf) e hidroxiapatite (HAp). Em particular, a adição de HAp estimulou a diferenciação e mineralização dos osteoblastos em osteócitos, sendo selecionada para os estudos seguintes. De seguida, os hidrogéis esponjosos de GG/HAp foram moldados em forma de anel (externo) e um disco (interno) e cultivados com células estaminais derivadas de medula óssea humana (HBM-MSCs) (anel externo) e com HBM-MSCs e células endoteliais (disco interno) para construir um modelo 3D que represente anatomicamente o osso cortical e trabecular. Após atingirem a osteogénese e a maturidade das estruturas semelhantes a capilares, respetivamente, foram acoplados de forma a formarem um modelo integrado 3D de osso vascularizado, dando origem a uma plataforma adequada para estudar a fisiologia óssea e avaliação de novas terapias. A inflamação aguda foi induzida neste modelo através da suplementação com Fator de Necrose Tumoral alfa (TNF-α) para simular a indução inicial de doenças ósseas inflamatórias, como a osteoporose, e avaliar o seu impacto na formação vascular, osteogénese e inflamação. Os resultados mostraram que o TNF-α não afetou a viabilidade celular nem alterou significativamente a expressão dos marcadores angiogénicos e osteogénicos. No entanto, os níveis de citocinas pró-inflamatórias aumentaram proporcionalmente com a quantidade de TNF-α e diminuíram com o tempo de cultura, sugerindo a resolução gradual da fase aguda da inflamação. Por fim, a composição/teor de polímero GG/HAp foi ajustado para obter uma “ink” bioativa de forma a dar resposta à crescente necessidade de desenvolver modelos de tecido ósseos altamente reprodutíveis. Análogos de tecido ósseo 3D foram impressos e suportaram a viabilidade e osteogénese das células SaOs-2 in vitro, destacando as propriedades osteocondutivas e osteogénicas das “inks” compósitas."],"dc:identifier.uri":["https://hdl.handle.net/1822/96268"],"dc:language.iso":["eng"],"dc:relation":["info:eu-repo/grantAgreement/FCT/POR_NORTE/PD%2FBD%2F143043%2F2018/PT","COVID/BD/153027/2022"],"dc:rights":["embargoedAccess (2 Years)"],"dc:rights.uri":["http://creativecommons.org/licenses/by/4.0/"],"dc:subject":["Bone cells","Gellan Gum","Hydrogels","Hydroxyapatite","Three-dimensional models","Células ósseas","Hidrogéis","Hidroxiapatite","Modelos tridimensionais"],"dc:title":["Advancing bone tissue engineering research using bone-derived cells laden in biomimetic gellan gum-based biomaterials"],"dc:type":["doctoralThesis"],"thesis:degree_name":["Tese de doutoramento em Engenharia de Tecidos, Medicina Regenerativa e Células Estaminais"],"thesis:institution_name":["Universidade do Minho"]},"updated_at":"2026-08-21T16:46:39Z"}