Culture of mesenchymal stem cells derived from equine synovial membrane in alginate hydrogel microcapsules

Abstract

Background: Mesenchymal stem cells derived from the synovial membrane (MSCSM) have a greater potential for joint regeneration, besides the capacity for chondrogenic differentiation, since they are a source closer to the chondrocytes. This study aimed to cultivate and evaluate viability and differentiation of MSCSM encapsulated in a three-dimensional alginate hydrogel (HA) scaffold. Samples of the synovial membrane of the metatarsophalangeal joint of 4 horses were collected by astroscopic surgery. These were subjected to enzymatic digestion, isolated mesenchymal cells, cultured in monolayers and encapsulated at various concentrations, 104; 204; 504; 105; 205 cells in 1.5% sodium alginate solution. The gelatinization process was carried out and cultured for 4 weeks. Viability and cell proliferation were performed by dissolving the microcapsules and counting with trypan blue. The ratio of live cells and total live cells at intervals 0, 7, 14, 21 and 28 days was analyzed. Results: For the evaluation of differentiation, histological sections stained with hematoxylin and eosin and toluidine blue were performed. There was no statistical difference in the proportion of live cells between groups over the 28 days. The group of 105 cells obtained a higher total number of living cells at the end of the experiment. Through the histological analysis it was possible to observe at 7 days a low amount of spherical cells with chondrocyte characteristics. On day 21, chondrogenic differentiation became evident, with pericellular and territorial matrix production. Conclusions: This study demonstrated the efficiency of HA as a scaffold for MSCSM and the chondrogenic differentiation, promising for use in the treatment of joint injuries in horses.