Effects of chemical and physical methods on decellularization of murine skeletal muscles

Autor(es)

Miranda, Carla M. F. C. ^[1] ; Leonel, Luciano C. P. C. ^[1] ; Canada, Rafael R. ^[2] ; Maria, Durvanei A. ^[3] ; Miglino, Maria Angelica ^[1] ; del Sol, Mariano ^[4] ; Lobo, Sonja E. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Fac Med Vet & Zootecnia, Programa Posgrad Anat Anim Domest & Silvestres, Dept Cirurgia, Disciplina Anat, Av Prof Dr Orlando Marques de Paiva 87, BR-05508270 Sao Paulo, SP - Brazil

^[2] Univ Sao Judas Tadeu, Grad Ciencias Biol, Av Pereira Barreto 1479, BR-09751000 Sao Bernardo Do Campo, SP - Brazil

^[3] Inst Butantan, Lab Biol Mol, Av Vital Brasil 1500, BR-05503900 Sao Paulo, SP - Brazil

^[4] Univ La Frontera, Dept Ciencias Basicas, Francisco Salazar 1145, Temuco 4811230 - Chile

• Tipo de documento: Artigo Científico
• Fonte: Anais da Academia Brasileira de Ciências; v. 93, n. 2 2021.
• Citações Web of Science: 0

Resumo

Abstract Volumetric 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. (AU)

• Processo FAPESP: 14/50844-3 - Matriz extracelular na saúde e matriz placentária na regeneração de tecidos
• Beneficiário: Maria Angelica Miglino
• Modalidade de apoio: Auxílio à Pesquisa - Temático