Cross-linking of amniotic membranes

Human amniotic membrane was cross-linked with chemical and radiation methods to investigate the effect of cross-linking on its physicochemical and biodegradation properties. Radiation cross-linking was performed with gamma-ray and electron beam while chemical cross-linking was with glutaraldehyde (GA). Both gamma-ray and electron beam irradiation decreased the tensile strength and elongation at break of the amniotic membrane with an increase in the irradiation dose, whereas GA cross-linking had no effect on the tensile properties. This is probably due to the scission of collagen chains through irradiation. No significant change was observed on the water content of cross-linked amniotic membranes for any of the crosslinking methods and in marked contrast with cross-linking of a gelatin membrane. A permeation study revealed that protein permeation through the amniotic membrane was not influenced by the GA concentration at cross-linking. These findings are ascribed to the structure characteristic of the amniotic membrane. The membrane is composed of a fibrous mesh structure from an assemblage of collagen fibers. It is possible that cross-linking takes place in the interior of the fiber assembly without impairing the mesh structure, resulting in no change of the water content and protein permeability. In vitro degradation of cross-linked amniotic membranes revealed that radiation cross-linking appeared to be much less effective than GA cross-linking in retarding the degradation, probably because of low cross-linking densities. GA-cross-linked amniotic membranes were degraded more slowly as the GA concentration at cross-linking increased. When the GA-cross-linked amniotic membrane was subcutaneously implanted in the rat, the tissue response was mild, similar to that of the non-cross-linked native membrane.