Effect of Pepsin Digestion Time on The Properties of Temperature Sensitive Human Amniotic Membrane Derived Hydrogel

The human amniotic membrane (HAM) is one of the rare allograft tissues that are in use in clinical trials. Biocompatibility, antibacterial effect, low immunogenicity, and scar prevention are properties that have made HAM attractive for tissue engineering (TE) applications, for example, as a cell carrier, injectable hydrogel, and cell culture substrate. In this research, the effect of digestion time on the structure, gelation kinetics, rheological and biological properties of amniotic membrane-derived hydrogels was studied. The results determined that digestion with pepsin should be performed at least for 24 h.  Prolonging the digestion time to 72 h increased the shear modulus, fiber diameter, and gelation rate. Cytocompatibility assays with L929 fibroblast cells showed that the digestion time had no effect on the cell toxicity of the hydrogels. However, cell proliferation was improved due to preserved constitutive bioactive molecules. The results of this research can be used to develop amniotic membrane-derived hydrogels for TE applications.