Jiaoming Dong1, Yu Li, Xiumei Mo. 1. College of Chemistry, Chemical Engineering and Biotechnology, Dong hua University, Shanghai, China.
Abstract
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.
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.