Abstract:
Human amniotic membranes (hAMs) obtained during cesarean sections have proven to be clinically useful as an interesting biomaterial in a wide range of tissue engineering applications such as ocular surface reconstruction, burn treatments, chronic wounds, or bedsore ulcers. It presents antimicrobial properties, promotes epithelization, reduces inflammation and angiogenesis, contains growth factors, and constitutes the reservoir of stem cells. However, variability in hAM stiffness and its fast degradation offers an explanation for the poor clinical applications and reproducibility. In addition, the preparatory method of hAM for clinical use can affect its mechanical properties, and these differences can influence its application. As a directly applied biomaterial, the hAM should be available in a ready-to-use manner in clinical settings. In the present study, we performed an analysis to improve the mechanical properties of hAM by the addition of various reagents used as protein cross-linkers: EDC/NHS, PEG-dialdehyde, PEG-NHS, dialdehyde starch, and squaric acid. The effect of hAM modification using different cross-linking agents was determined via infrared spectroscopy, thermal analyses, mechanical properties analyses, enzymatic degradation, and cytotoxicity tests. The use of PEG-dialdehyde, PEG-NHS, dialdehyde starch, and squaric acid increases the mechanical strength and elongation at the breaking point of hAM, while the addition of EDC/NHS results in material stiffening and shrinkage. Also, the thermal stability and degradation resistance were evaluated, demonstrating higher values after cross-linking. Overall, these results suggest that modification of human amniotic membrane by various reagents used as protein cross-linkers may make it easier to use hAM in clinical applications, and the presented study is a step forward in the standardization of the hAM preparation method.
摘要:
在剖宫产期间获得的人羊膜(hAMs)已被证明在广泛的组织工程应用中作为一种有趣的生物材料在临床上是有用的,例如眼表重建。烧伤治疗,慢性伤口,或褥疮溃疡。它具有抗菌性能,促进上皮形成,减少炎症和血管生成,含有生长因子,并构成干细胞的储库。然而,hAM硬度的变异性及其快速降解为临床应用和可重复性差提供了解释。此外,临床使用的hAM的制备方法会影响其机械性能,这些差异会影响其应用。作为一种直接应用的生物材料,在临床环境中,hAM应该可以随时使用.在本研究中,我们进行了一项分析,通过添加用作蛋白质交联剂的各种试剂来改善hAM的机械性能:EDC/NHS,PEG-二醛,PEG-NHS,二醛淀粉,还有方酸.通过红外光谱测定了使用不同交联剂的hAM改性效果,热分析,力学性能分析,酶促降解,和细胞毒性测试。PEG-二醛的使用,PEG-NHS,二醛淀粉,和方酸增加机械强度和伸长率在断裂点的ham,而EDC/NHS的添加导致材料硬化和收缩。此外,评估了热稳定性和耐降解性,交联后显示更高的值。总的来说,这些结果表明,通过用作蛋白质交联剂的各种试剂修饰人羊膜可能使hAM在临床应用中更容易使用,提出的研究是hAM制备方法标准化的一步。