Advancing bone tissue engineering research using bone-derived cells laden in biomimetic gellan gum-based biomaterials

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.

Name thesis:degree_name

Tese de doutoramento em Engenharia de Tecidos, Medicina Regenerativa e Células Estaminais

Grantor

Universidade do Minho

Year dc:date.issued

2025

Author dc:creator

• Bastos, Ana Raquel Fernandes

Advisors dc:contributor.advisor

• Correlo, V. M.
• Reis, R. L.

• Bone cells
• Gellan Gum
• Hydrogels
• Hydroxyapatite
• Three-dimensional models
• Células ósseas
• Hidrogéis
• Hidroxiapatite
• Modelos tridimensionais

Statement dc:rights

• embargoedAccess (2 Years)

Licence dc:rights.uri

• http://creativecommons.org/licenses/by/4.0/

Language dc:language.iso

eng

Handle dc:identifier.uri

https://hdl.handle.net/1822/96268

OAI record oai:identifier

http://repositorium.sdum.uminho.pt/oai/request?verb=GetRecord&metadataPrefix=dim&identifier=oai%3Arepositorium.uminho.pt%3A1822%2F96268