Development of Biological Scaffolds for Pediatric Patients having Growth Potential

已经研究了基于脱细胞基质的工程化移植物，以使目前的生物假体具有更高的耐久性和生长潜力，且免疫原性更低。Determination通常用于去除细胞，而脱细胞依赖于它们从其表面渗透到组织中，并且在大的或硬的tissues.METHODS猪组织的心脏瓣膜、主动脉、心肌、骨骼肌和气管在无菌条件下分离。立即通过980 Mpa（10，000 atm）冷等静压（CIP）处理，然后在4 ℃下清洗（PowerGraft处理）。组织学观察、PERV检测、DNA检测、磷脂检测和生物力学研究结果表明：CIP处理10 min，洗涤7 d，气管软骨等组织完全无细胞。压榨应用于整个组织，并且在该过程中不使用洗涤剂。在处理的组织中未检测到PERV。据报道，超过600 MPa的CIP可以灭活大多数病毒。DNA含量低于1 μ g/ml，磷脂含量低于0.5mg/g。胶原和弹性蛋白纤维在脱细胞组织中保持良好，生物力学性能无明显变化。结论PowerGraft处理可在短时间内完全去除猪细胞和PERV。这可以为组织再生提供更安全的脱细胞组织。

OBJECTIVESTissue engineered grafts based on acellular matrices have been studied to give more durability with growth potential and less immunogenicity to the current bioprostheses. Detergents are commonly used for removal of the cells, whereas the decellularization depends on their permeation in the tissue from its surface and may not be achieved completely in large or hard tissues.METHODSPorcine tissues of the heart valves, aorta, cardiac muscle, skeletal muscle, and trachea were isolated under sterile condition. They were treated immediately by cold isostatic pressing (CIP) of 980 Mpa (10,000 atm) followed by washing at 4℃ (PowerGraft treatment). They were then subjected to histological study, porcine endogeneoua retrovirus (PERV) assay, DNA assay, phospholipids assay, and biomechanical study.RESULTSThe tissues including tracheal cartilage were completely cell free when they were treated by the CIP for 10 min and washing for 7 days. The pressing is applied to the whole of the tissue and no detergents are used in the process. There was no PERV detected from the tissue treated. It has been reported that viruses are mostly inactivated by the CIP more than 600 MPa. The amount of DNA and phospholipids were lower than 1 μg/ml and 0.5 mg/g, respectively. The collagen and elastin fibers were well maintained in the acellular tissue and there were no significant changes in biomechanical properties.CONCLUSIONSPorcine cells and PERV were removed completely in a short period by the PowerGraft treatment. This may have more secure acellular tissues for the tissue regeneration.