초록접수 현황
| 17F-232 | 구연 발표 |
Effect of Different washing Methods on the Decellularization Process Based on Sodium Dodecyl Sulfate for Porcine Pulmonary Artery Wall
Hyungtae Kim, MD, Si Chan Sung, MD, Kwang Ho Choi, MD, Young Suk Kim
Department of Thoracic and Cardiovascular Surgery, Pusan National University Yangsan Hospital, Pusan National University College of Medicine, Gyeongsangnam-do, Republic of Korea
Purpose : To determine whether residual cytotoxic SDS is responsible for the low levels of cell repopulation, we have used two washing methods to remove residual SDS and performed extensive biochemical, mechanical, and structural analyses to determine the effect of SDS-based decellularization for porcine pulmonary artery (PA) wall.
Methods : Fresh porcine PA walls were decellurized using SDS and sodium deoxycholic acid (SDC). Decellularized tissues were rinsed with three different washing techniques (phosphate buffered saline (PBS), pH 9 buffer solution, and 75% ethanol (EtOH)). Histologic, biochemical, and mechanical analyses were performed. Implantations into subcutaneous tissue of rats and patch implantation into carotid artery of dogs were performed as in vivo studies.
Results : The decellularization protocol based on SDS and SDC was able to remove cells effectively. The major extracellular matrix (ECM) structure (collagen, elastic fiber, and glycosaminoglycan) were properly preserved under 75% EtOH washing technique. Significantly reduced residual SDS content was identified in the EtOH-washed tissue compared with other methods (0.19±0.03 ug/mg in EtOH-washed tissue, 4.60±0.83 ug/mg in PBS-wahed tissue, and 2.50±1.05 ug/mg in pH 9-washed tissue). No significant difference in mechanical strength and cell viability test was observed as comparing with other methods. In the rat model study, no acute rejection and massive calcification were recognized. In vivo study, we were able to identify an ingrowing intimal tissue in EtOH-washed group.
Conclusion : EtOH wahing technique on the decellularization process based on SDS for the porcine PA wall can reduce residual SDS content and preserve ECM structure, and also could enhance cell repopulation after re-implantation.
Methods : Fresh porcine PA walls were decellurized using SDS and sodium deoxycholic acid (SDC). Decellularized tissues were rinsed with three different washing techniques (phosphate buffered saline (PBS), pH 9 buffer solution, and 75% ethanol (EtOH)). Histologic, biochemical, and mechanical analyses were performed. Implantations into subcutaneous tissue of rats and patch implantation into carotid artery of dogs were performed as in vivo studies.
Results : The decellularization protocol based on SDS and SDC was able to remove cells effectively. The major extracellular matrix (ECM) structure (collagen, elastic fiber, and glycosaminoglycan) were properly preserved under 75% EtOH washing technique. Significantly reduced residual SDS content was identified in the EtOH-washed tissue compared with other methods (0.19±0.03 ug/mg in EtOH-washed tissue, 4.60±0.83 ug/mg in PBS-wahed tissue, and 2.50±1.05 ug/mg in pH 9-washed tissue). No significant difference in mechanical strength and cell viability test was observed as comparing with other methods. In the rat model study, no acute rejection and massive calcification were recognized. In vivo study, we were able to identify an ingrowing intimal tissue in EtOH-washed group.
Conclusion : EtOH wahing technique on the decellularization process based on SDS for the porcine PA wall can reduce residual SDS content and preserve ECM structure, and also could enhance cell repopulation after re-implantation.
책임저자: Si Chan Sung
Department of Thoracic and Cardiovascular Surgery, Pusan National University Yangsan Hospital, Pusan National University College of Medicine, Gyeongsangnam-do, Republic of Korea
발표자: Hyungtae Kim, E-mail : 2719k@naver.com