The analysis of gelatin methacryloyl hydrogel constructs loaded with chondrocytes in the regeneration of osteochondral tissue in the knee joint.

• Grant number: 24/04270-7
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Start date: September 30, 2024
• End date: March 29, 2025
• Field of knowledge: Engineering - Biomedical Engineering - Bioengineering
• Principal Investigator: Renata Neves Granito
• Grantee: Amanda Sardeli Alqualo Assaf
• Supervisor: Veerle Bloemen
• Host Institution: Instituto de Saúde e Sociedade (ISS). Universidade Federal de São Paulo (UNIFESP). Campus Baixada Santista. Santos , SP, Brazil
• Institution abroad: University of Leuven, Leuven (KU Leuven), Belgium
• Associated to the scholarship: 22/13535-9 - Additive manufacturing applied in the development a new bioglass scaffold of the SiO2 - Na2O - CaO - P2O5 system, developed from marine sponge biosilica: biological evaluation in vitro and in vivo, BP.MS

Abstract

Hyaline cartilage is crucial for the proper functioning of the knee joint, as it provides the necessary support for movement, stability and lubrication in key compartments of the joint. Its avascular and aneural characteristics make it susceptible to mechanical damage caused by injury, trauma, inflammation and degenerative conditions such as osteoarthritis. As a result, joint diseases have a substantial impact on increasing hospitalization costs and decreasing quality of life for millions of individuals around the world. In this context, tissue engineering has sought effective strategies for regenerating osteochondral tissues by combining cells with biodegradable biomaterials that mimic the native tissue microenvironment. An original study by Prof. Bloemen and her team at KU Leuven previously evaluated three-dimensional constructs engineered with chondroblasts derived from human iPSCs in gelatin methacrylate (GelMA) hydrogels, demonstrating that the constructs support cell survival and proliferation, as well as the in vitro formation of cartilage-like tissue. In addition, the team proved that ectopic implantation of these constructs promotes the formation of hyaline cartilage-like tissue. In this study, we intend to advance these investigations by evaluating tissue regeneration after orthotopic implantation of cell-loaded hydrogels in the rat knee. Therefore, after a regenerative period of 16 weeks, the main focus will be on understanding the roles of the biomaterial, the loaded cells and their interaction with the host cells at the implantation site; in other words, on identifying the contribution of the construct and the host to the formation of neotissue. The aim would be to identify which part of the neo-tissue is human (contribution of the implanted cells) or murine (contribution of the host), as well as which part of the hydrogel is still present after explantation (contribution of the biomaterial). In addition, as an interface with the study carried out in Brazil, a new 3D bioprinting ink will be developed using marine bioglass, sodium alginate and GelMA, since this biomaterial showed good results in terms of cell viability and proliferation. Analyses of gene expression by qPCR, histology (H&E, Alcian Blue, Safranin O, Toluidine Blue, Masson's Trichrome), immunohistochemistry (type I collagen, type II collagen, osteocalcin) and microCT analysis will be carried out for the GelMA loaded with chondrocytes and viability and gene expression analyses by qPCR for the bioprinted scaffold to assess osteoconductivity. This approach increases the translatability of research results to clinical applications, making it an important tool in biomedical research. Consequently, it has the significant potential to advance the future treatment of osteochondral defects, with the ultimate goal of improving the quality of life of individuals facing compromised joint conditions.