Department of Veterinary Science, College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University, Chuncheon 200-701, Republic of Korea.
College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk 28644, Republic of Korea.
J Biosci Bioeng. 2019 Nov;128(5):613-621. doi: 10.1016/j.jbiosc.2019.04.017. Epub 2019 May 22.
Decellularized esophageal matrices are ideal scaffolds for esophageal tissue engineering. Unfortunately, in order to improve transplantation possibilities, they require modification to reduce their degradation rate and immunogenicity. To date, no modifying agent has been approved to overcome these limitations. The objective of this study was to evaluate the ability of silver nanoparticles (AgNPs) to improve the structural stability and biocompatibility of decellularized rat esophagi. AgNPs have the advantage over currently used agents in that they bind with collagen fibers in a highly ordered manner, via non-covalent binding mechanisms forming multiple binding sites, while other agents provide only two-point connections between collagen molecules. Rat esophagi were decellularized, loaded with 5 μg/mL of AgNPs (100 nm), and then treated with an immobilization-complex buffer composed of ethyl carbodiimide hydrochloride and N-hydroxysuccinimide (EDC/NHS). Then, they were evaluated in terms of ultra-structural morphology, water uptake, in vitro resistance to enzymatic and thermal degradation, indentation strength, in vitro anti-calcification, cytocompatibility with rat bone marrow derived stromal cells (rat-BMSCs), angiogenic properties, and in vivo biocompatibility, and compared to scaffolds modified using glutaraldehyde and EDC/NHS complex buffer alone. AgNP-modified scaffolds showed an improved ultrastructure, good water uptake, and considerable resistance against in vitro degradation and indentation, and a high resistance against in vitro calcification. Moreover, they were cytocompatible for allogeneic rat-BMSCs. Additionally, AgNPs did not alter the angiogenic properties of the modified scaffolds and decreased host immune responses after their subcutaneous implantation. The structural properties and biocompatibility of decellularized esophageal matrices could be improved by conjugation with AgNPs.
脱细胞食管基质是食管组织工程的理想支架。然而，为了提高移植的可能性，需要对其进行修饰以降低降解率和免疫原性。迄今为止，还没有批准任何修饰剂来克服这些限制。本研究旨在评估纳米银颗粒（AgNPs）改善脱细胞大鼠食管结构稳定性和生物相容性的能力。与目前使用的试剂相比，AgNPs 具有优势，因为它们通过非共价结合机制以高度有序的方式与胶原纤维结合，形成多个结合位点，而其他试剂仅在胶原分子之间提供两点连接。将大鼠食管脱细胞，负载 5μg/mL 的 AgNPs（100nm），然后用包含盐酸碳二亚胺和 N-羟基琥珀酰亚胺（EDC/NHS）的固定复合物缓冲液处理。然后，根据超微结构形态、吸水率、体外抵抗酶解和热降解的能力、压痕强度、体外抗钙化能力、与大鼠骨髓基质细胞（大鼠-BMSCs）的细胞相容性、血管生成特性和体内生物相容性进行评估，并与单独使用戊二醛和 EDC/NHS 复合缓冲液修饰的支架进行比较。AgNP 修饰的支架显示出改善的超微结构、良好的吸水率以及对体外降解和压痕的显著抵抗力，并且对体外钙化具有很高的抵抗力。此外，它们对同种异体大鼠-BMSCs 具有细胞相容性。此外，AgNPs 不会改变修饰支架的血管生成特性，并在皮下植入后降低宿主免疫反应。通过与 AgNPs 缀合，可以改善脱细胞食管基质的结构特性和生物相容性。
