RESUMEN
BACKGROUND: The use of extracellular matrix (ECM) derived from decellularized tissue is increasingly frequent in regenerative medicine and tissue engineering. However, it is recognized that currently used decellularization procedures have negative effects on ECM integrity. The objective of this study was to investigate the impact of a decellularization protocol with a new detergent on the ECM integrity of porcine pericardium (PP) compared with other traditional detergents. MATERIALS AND METHODS: Fresh PP were decellularized by sodium deoxycholate in combination with Triton X-100 (SDT), sodium dodecyl sulfate (SDS), and octyl-glucopyranoside (OGP), respectively. Histologic analysis and scanning electron microscopy were performed to confirm the removal of cells and to examine the structure of ECM. DNA content was examined by the method of DNA extraction. Mechanical properties and biochemical compositions of ECM were also studied. RESULTS: Histologic analysis and DNA determination demonstrated that SDS and OGP completely removed the cells, and the major ECM structure was preserved well for PP treated with 1% (wt/vol) OGP but disrupted for PP treated with SDS; whereas treatment with SDT was insufficient to remove cells from PP. Uniaxial tensile tests showed that PP decellularized by OGP had similar mechanical properties to native PP, whereas the mechanical properties of PP decellularized by SDS and SDT decreased. The biochemical compositions of PP decellularized by OGP were also well conserved, except that glycosaminoglycans markedly decreased. Moreover, the results obtained in the MTT study further indicated that the cytotoxicity of PP decellularized by OGP was significantly lower than that decellularized by SDS and SDT. CONCLUSION: It is suggested that the environmentally friendly and nontoxic OGP can be used as a decellularizing agent. The OGP method could achieve both complete removal of cells from native PP and preservation of the matrix structure; thus, it might be a suitable approach to preparation of tissue engineering heart valve scaffold.
Asunto(s)
Detergentes/farmacología , Glucósidos/farmacología , Pericardio/efectos de los fármacos , Andamios del Tejido , Animales , Línea Celular , Microscopía Electrónica de Rastreo , Porcinos , Resistencia a la Tracción , Pruebas de ToxicidadRESUMEN
Biologic scaffold materials composed of extracellular matrix (ECM) are typically obtained in processes that involve decellularization of tissues or organs. Decellularized tissues and organs have been successfully used in a variety of tissue engineering/regenerative medicine applications. Preservation of the complex composition and three-dimensional ultrastructure of the ECM is highly desirable but it is recognized that all methods of decellularization result in disruption of the structure and potential loss of composition. The efficiency of cell removal from a tissue is dependent on the origin of the tissue and the physical, chemical, and enzymatic methods that are used. Each of these treatments affects the biochemical composition, tissue ultrastructure, and mechanical behavior of the remaining ECM scaffold, and all of the treatment methods affect the host response to the material as well. Tissue decellularization with preservation of ECM integrity and bioactivity can be optimized by making correct decisions regarding the agents and techniques utilized during processing. In this paper, the most commonly used decellularization methods are described, and consideration given to the effects of these methods upon the biologic scaffold material and recently described antigen removal strategy are presented.