Produção de hidrogel para reconstrução de cartilagens com potencial aplicação em saúde humana e animal

Abstract

The increase in life expectancy and, consequently, the aging of the world population are accompanied by an increase in the number of joint diseases, such as osteoarthritis. Osteoarthritis is a degenerative disease of extracellular matrix components, with a palliative treatment strategy and without cure until now. In this scenario, tissue engineering becomes a potential area for the development of new osteoarthritis treatments, since its main purpose is to recover tissue in a degenerative process through the use of synthetic or biological biomaterials. For this reason, the present study aims to establish a decellularization protocol for later production of a biomaterial (hydrogel) derived from an extracellular matrix of cartilaginous tissue. Also, to evaluate the chondrogenic induction capacity of this matrix in conditioned medium. For this purpose, the hyaline cartilages of bovine tracheas were subjected to two different physical-chemical treatments to remove cellular content. Quantification of DNA content, biochemical and morphological analyzes were performed in order to assess the efficiency and quality of the developed protocol. For the evaluation of the induction of chondrogenic differentiation, morphologyical analysis and proliferation rate were performed. The first protocol was able to significantly remove the DNA content (approximately 62.5%) without altering the amount of glycosaminoglycans in the matrix. However, type II collagen showed alteration in its fibrillar suprastructure and significant reduction when exposed to the decalcifying agent for longer (approximately 65.2% and 55.9%). The second decellularization protocol was able to significantly remove the DNA content of chondrocytes (approximately 91.96%) without changing the amount of glycosaminoglycans and type II collagen in the cartilaginous matrix. However, type II collagen showed changes in its fibrillar suprastructure. The decellularized extracellular matrix was able to induce the differentiation of stem cells from human exfoliated deciduous teeth in culture observed by the morphological change (from spindle to spherical shape) and its proliferation increased when in the presence of lesser matrix. Therefore, the protocols developed for decellularization of bovine tracheal cartilage were able to reduce the DNA content without losing essential components of the extracellular matrix and with potential application in tissue engineering in the treatment of osteoarthritis. In addition, the matrix showed good biocompatibility with the stem cells under study, stimulating the differentiation of these stem cells to cells similar to chondrocytes, without commercial chemical inducers, needing further study to characterize this differentiation and its long-term proliferation.