Protocol optimization for abtaining lyophilized human amniotic membrane extract

Abstract

The amniotic membrane (AM) is the inner amniotic fluid-filled membrane which forms a cavity around the embryo. AM has long been used in the treatment of venous ulcers and burns. Freshly extracted AM has been shown to possess the highest effect, thus making the extraction method costly and severely limiting its possible usage. To counter this problem, several extraction methods have been developed aimed at preserving the growth factors present in AM and maintaining the AM in a sterile condition. The use of lyophilized AM extract could help preserve its active properties which is associated with the presence of a variety of cytokines necessary for wound-healing. However, there are no standard methods for obtaining amniotic-membrane extracts to date. We chose two available methods in which the resulting extract had the highest regenerative properties. Comparative analyses of these two methods were not carried out. Hence, in order to determine which method gave AM lyophilisates with the highest cytokine composition, we determined the cytokine content of the obtained lyophilisates with reference to that of the native membrane.