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.
ABSTRACT
Gelatin is a natural biopolymer derived from collagen. Due to its many advantages, such as swelling capacity, biodegradability, biocompatibility, and commercial availability, gelatin is widely used in the field of pharmacy, medicine, and the food industry. Gelatin solutions easily form hydrogels during cooling, however, the materials are mechanically poor. To improve their properties, they are often chemically crosslinked. The cross-linking agents are divided into two groups: Zero-length and non-zero-length cross-linkers. In this study, gelatin was cross-linked by three different cross-linking agents: EDC-NHS, as a typically used cross-linker, and also squaric acid (SQ) and dialdehyde starch (DAS), as representatives of a second group of cross-linkers. For all prepared gelatin hydrogels, mechanical strength tests, thermal analysis, infrared spectroscopy, swelling ability, and SEM images were performed. The results indicate that the dialdehyde starch is a better cross-linking agent for gelatin than EDC-NHS. Meanwhile, the use of squaric acid does not give beneficial changes to the properties of the hydrogel.
