Produção Nacional
Revisado por pares
Effects of chemical and physical methods on decellularization of murine skeletal muscles
2021; Brazilian Academy of Sciences; Volume: 93; Issue: 2 Linguagem: Inglês
10.1590/0001-3765202120190942
ISSN1678-2690
AutoresC.M.F.C. Miranda, Luciano Leonel, Rafael R. Cañada, Durvanei Augusto Maria, María Angélica Miglino, Mariano del Sol, Sonja Ellen Lobo,
Tópico(s)Additive Manufacturing and 3D Printing Technologies
ResumoVolumetric muscle loss causes functional weakness and is often treated with muscle grafts or implant of biomaterials. Extracellular matrices, obtained through tissue decellularization, have been widely used as biological biomaterials in tissue engineering. Optimal decellularization method varies among tissues and have significant impact on the quality of the matrix. This study aimed at comparing the efficacy of four protocols, that varied according to the temperature of tissue storage and the sequence of chemical reagents, to decellularize murine skeletal muscles. Tibialis anterior muscles were harvested from rats and were frozen at -20°C or stored at room temperature, followed by decellularization in solutions containing EDTA + Tris, SDS and Triton X-100, applied in different sequences. Samples were analyzed for macroscopic aspects, cell removal, decrease of DNA content, preservation of proteins and three-dimensional structure of the matrices. Processing protocols that started with incubation in SDS solution optimized removal of cells and DNA content and preserved the matrix ultrastructure and composition, compared to those that were initiated with EDTA + Tris. Freezing the samples before decellularization favored cell removal, regardless of the sequence of chemical reagents. Thus, to freeze skeletal muscles and to start decellularization with 1% SDS solution showed the best results.
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