SANTANNA, J.P. C.FARIA, R. R.ASSAD, I. P.PINHEIRO, C. C. G.AIELLO, V. D.Cyro AlbuquerqueRoberto BortolussiCESTARI, I. A.MAIZATO, M. J. S.HERNANDES, A. J.BUENO, D. F.FERNANDES, T. L.2023-01-012023-01-012022-02-05SANTANNA, J.P. C.; FARIA, R. R.; ASSAD, I. P.; PINHEIRO, C. C. G.; AIELLO, V. D.; ALBUQUERQUE-NETO, C.; BORTOLUSSI, R.; CESTARI, I. A.; MAIZATO, M. J. S.; HERNANDES, A. J.; BUENO, D. F.; FERNANDES, T. L. Tissue Engineering and Cell Therapy for Cartilage Repair: Preclinical Evaluation Methods. Tissue engineering. Part C, Methods, v. 28, n. 2, p. 73-82, feb. 2022.1937-3392https://repositorio.fei.edu.br/handle/FEI/4669A chondral injury is a limiting disease that can affect the quality of life and be an economic burden due to the cost of immediate treatment and loss in work productivity. If left untreated, such an injury may progress to osteoarthritis, a degenerative and debilitating joint disease characterized by pain and functional impairment. Mesenchymal stromal cells (MSCs), which have immune-modulatory properties and the ability to differentiate into chondroblasts and osteoblasts, are a predictable source for the treatment of cartilage injuries. This article presents tools to evaluate cartilage restoration by tissue engineering and cell therapy treatment in a translational and preclinical large animal model. In this controlled experimental study with 14 miniature pigs, a scaffold-free tissue engineering construct (TEC) derived from dental pulp and synovial MSCs for cartilage therapy was tested. Total thickness cartilage defects were performed in both posterior knees. The defect was left empty in one of the knees, and the other received the TEC. The tissue repair was morphologically assessed by magnetic resonance imaging (MRI) using the three-dimensional double echo steady-state (3D-DESS) sequence, and compositional assessment was carried out based on the T2 mapping technique. The osteochondral specimens were fixed for histopathology, decalcified, subjected to standard histological processing, sectioned, and stained with hematoxylin and eosin. The sections stained for immunohistochemical detection of collagen types were digested with pepsin and chondroitinase and incubated with antibodies against them. The mechanical evaluation involved analysis of Young's modulus of the cartilage samples based on the indentation and maximum compression test. In addition, a finite element model was used to simulate and characterize properties of the osteochondral block. At 6 months after surgery, there were no complications with the animals and the MRI, histological, immunohistochemical, and biomechanical evaluations proved to be effective and qualified to differentiate good quality chondral repair from inadequate repair tissue. The proposed methods were feasible and capable to properly evaluate the defect filled with TEC containing stromal cells after 6 months of follow-up in a large animal model for articular cartilage restoration. Impact Statement Articular chondral injuries are prevalent and represent an economic burden due to the cost of treatment. The engineering of cartilage tissue can promote the repair of chondral injuries and is dependent on selecting appropriate cells and biocompatible frameworks. In this article, methods for evaluation of a scaffold-free cell delivery system made from mesenchymal stromal cells were present in a translational study that allows further clinical safety and efficacy trials.Acesso RestritoArtigo10.1089/ten.TEC.2021.0208cartilage injuriescartilage restorationmagnetic resonance imagingmechanical evaluationmesenchymal stromal cellstissue engineering