Injectable thermosensitive hydrogel combined with human amniotic fluid stem cells differentiated into chondrocytes for articular cartilage regeneration

• Grant number: 21/14738-8
• Support Opportunities: Regular Research Grants
• Start date: May 01, 2022
• End date: August 31, 2024
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Ibsen Bellini Coimbra
• Grantee: Ibsen Bellini Coimbra
• Host Institution: Faculdade de Ciências Médicas (FCM). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated researchers: Angela Maria Moraes ; Carolina Coli Zuliani
• Associated scholarship(s): 22/09275-1 - Cultivation and chondrogenic differentiation of stem cells in 3D model, BP.TT

Abstract

Articular cartilage, damaged by trauma, sports activities or chronic diseases such as osteoarthritis (OA), has a low capacity for regeneration. So far, there is no effective drug or therapy for the definitive treatment, those often evolving to surgical procedures such as arthroplasty. Currently, the aim is to develop a therapy combining the use of biomaterials obtained by tissue engineering combined with regenerative medicine techniques with the use of stem cells for the regeneration of articular cartilage. Therefore, in this study, we intend to differentiate into chondrocytes stem cells (SC) present in human amniotic fluid (AF), due to their multipotentiality and plasticity in three-dimensional culture system based on non-adhesive micromolded hydrogel molds by 3D printing. This system allows deposits of uniform cellular spheroids and adaptations that will then be combined with a thermo-responsive hydrogel composed of methylcellulose and carboxymethyl chitosan. Both biomaterials were developed at the Faculty of Chemical Engineering at UNICAMP. SC from AF, obtained from routine amniocentesis, will be isolated and characterized by flow cytometry. The production of spheroids will be standardized and then evaluated through the production of extracellular matrix using the cartilage-specific histological stains (Masson's Trichrome, Picrossirius Red, Alcian Blue and Hematoxylin & Eosin), the production of collagen II and aggrecan through immunohistochemistry and gene expression by qRT-PCR. The thermosensitive spheroid-hydrogel set will be studied for its cytotoxicity, cell viability by means of confocal microscopy, and morphology by scanning electron microscopy (SEM). Subsequently, it is expected that the results from the study are favorable and that they open up new fronts both for studies in experimental models of chondral lesions in animals and for the implantation of this therapy through intra-articular infiltrations. In the future, this treatment could avoid open surgeries, allowing the patient to recover quickly with the formation of hyaline tissue and, in addition, to be financially viable for the incorporation of this technology by the Unified Health System (SUS) since it is a totally national technology. (AU)