AN ENZYMATIC METHOD TO OBTAIN A NEW SCAFFOLD FOR ENGINEERING CARTILAGE
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AN ENZYMATIC METHOD TO OBTAIN A NEW SCAFFOLD FOR ENGINEERING CARTILAGE Giraldo Gomez David M.1, Villegas Alvarez Fernando2, Garciadiego Cazares David3, Sotres Vega Avelina4, Piña Barba Maria C.1 1 Instituto de Investigaciones en Materiales, UNAM, Ciudad Universitaria, Circuito Exterior s/n, C.P. 04510, México D.F., México. 2 Departamento de Cirugía-Facultad de Medicina, UNAM, Ciudad Universitaria, Circuito Exterior s/n, C.P. 04510, México D.F., México. 3 Unidad de Ingeniería de Tejidos Terapia Celular y Medicina Regenerativa, Instituto Nacional de Rehabilitación, Calzada México Xochimilco Nº 289, C.P. 14389, México D.F., México. 4 Departamento de Investigación en Cirugía Experimental, Instituto Nacional de Enfermedades Respiratorias “Ismael Cossio Villegas”, Calzada de Tlalpan Nº 4502, C.P. 14080, México D.F., México
ABSTRACT The purpose of this study was to achieve a descellularized scaffold from cartilage tissue, which can be used as xenograft for cartilage tissue regeneration. This work presents the results obtained using one method to wash porcine trachea in order to remove cellular material from the extracellular matrix and to avoid the immune reaction using enzymatic detergent and partial enzymatic degradation with Deoxyribonuclease I (DNaseI), Ethylenediaminetetraacetic Acid (EDTA) and Trypsin. This treatment was qualitatively evaluated by Scanning Electron Microscopy (SEM), and H&E Stain (Histology), and quantitatively evaluated by DNA quantification. The thermal characterization of the descellularized scaffold was carried out using Termogravimetric Analysis (TGA) and Differential Scanning Calorimetry (DSC). The type of collagen obtained from the scaffold was determined through SDS-PAGE electrophoresis. When using Enzymatic Treatment (ET) to wash trachea tissue, it is possible to obtain an acellular xenograft; this procedure has the potential to avoid rejection reactions of the xenograft. INTRODUCTION The self-regeneration of cartilage is a very complicated process, this due to its low regenerative capacity and its low blood provision. However tissue engineering using biodegradable scaffolds and cell replacement therapy has become one of the promising approaches in the regeneration of cartilage [1-5]. The scaffolds provide a 3D structure that enables the cell growth while the differentiation function occurs. Numerous materials have been used to build these scaffolds, among which are those of natural origin as polylactic acid (PLA) polyglycolic acid (PGA), collagen type I, and descellularized cartilage matrix [2-10]. The descellularized matrices obtained from cartilage (ECM), which contain the naturals elements of cartilage with good biocompatibility for the growth of chondrocytes [11]. Theoretically, these scaffolds can be ideal for use in cartilage engineering, however, the used process to decellularization of the tissue is crucial, since all cellular material must be removed to avoid any type of immune response post-implantation, either way, the structural components from the ECM should be preserved to pr
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