PDF(Pubmed)
Abstract：
One of the most important medical interventions for individuals with heart valvular disease is heart valve replacement, which is not without substantial challenges, particularly for pediatric patients. Due to their biological properties and biocompatibility, natural tissue-originated scaffolds derived from human or animal sources are one type of scaffold that is widely used in tissue engineering. However, they are known for their high potential for immunogenicity. Being free of cells and genetic material, decellularized xenografts, consequently, have low immunogenicity and, thus, are expected to be tolerated by the recipient\'s immune system. The scaffold ultrastructure and ECM composition can be affected by cell removal agents. Therefore, applying an appropriate method that preserves intact the structure of the ECM plays a critical role in the final result. So far, there has not been an effective decellularization technique that preserves the integrity of the heart valve\'s ultrastructure while securing the least amount of genetic material left. This study demonstrates a new protocol with untraceable cells and residual DNA, thereby maximally reducing any chance of immunogenicity. The mechanical and biochemical properties of the ECM resemble those of native heart valves. Results from this study strongly indicate that different critical factors, such as ionic detergent omission, the substitution of Triton X-100 with Tergitol, and using a lower concentration of trypsin and a higher concentration of DNase and RNase, play a significant role in maintaining intact the ultrastructure and function of the ECM.
摘要：
心脏瓣膜疾病患者最重要的医疗干预措施之一是心脏瓣膜置换术，这并非没有实质性的挑战，特别是儿科患者。由于其生物学特性和生物相容性,来源于人或动物的天然组织来源的支架是一种广泛用于组织工程的支架。然而,它们以其高的免疫原性潜力而闻名。没有细胞和遗传物质，去细胞化异种移植物,因此,具有低免疫原性，因此，预计受者的免疫系统会耐受。细胞去除剂可影响支架超微结构和ECM组成。因此,应用适当的方法来保持ECM结构的完整性在最终结果中起着至关重要的作用。到目前为止,目前还没有一种有效的去细胞化技术，既能保护心脏瓣膜超微结构的完整性，又能确保留下的遗传物质最少。这项研究证明了一种新的协议，具有无法追踪的细胞和残留的DNA，从而最大限度地减少任何免疫原性的机会。ECM的机械和生化特性类似于天然心脏瓣膜。这项研究的结果强烈表明，不同的关键因素，如离子洗涤剂遗漏，用Tergitol取代TritonX-100，使用较低浓度的胰蛋白酶和较高浓度的DNase和RNase，在维持ECM的超微结构和功能完整中起重要作用。
