Properties of viable lyopreserved amnion are equivalent to viable cryopreserved amnion with the convenience of ambient storage

• Published in: PloS one 2018-10, Vol.13 (10), p.e0204060-e0204060
• Main Authors: Dhall, Sandeep, Sathyamoorthy, Malathi, Kuang, Jin-Qiang, Hoffman, Tyler, Moorman, Matthew, Lerch, Anne, Jacob, Vimal, Sinclair, Steven Michael, Danilkovitch, Alla
• Format: Article
• Language: English
• Subjects: Amnion; Amniotic membrane; Angiogenesis; Animal models; Antiinfectives and antibacterials; Biology and Life Sciences; Care and treatment; Clinical outcomes; Clinical trials; Cryopreservation; Diabetes; Diabetes mellitus; Equivalence; Extracellular matrix; Granulation; Growth factors; Hypoxia; Inflammation; Low temperature; Medical research; Medicine and Health Sciences; Mesenchyme; Methods; Microorganisms; Properties; Properties (attributes); Research and Analysis Methods; Secretion; Shelf life; Stem cell transplantation; Stem cells; Storage; Structure; Structure-function relationships; Temperature requirements; Tumor necrosis factor-α; Vascular endothelial growth factor; Wounds
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0204060

Human amniotic membrane (AM) has a long history of clinical use for wound treatment. AM serves as a wound protective barrier maintaining proper moisture. AM is anti-inflammatory, anti-microbial and antifibrotic, and supports angiogenesis, granulation tissue formation and wound re-epithelialization. These properties of AM are attributed to its native extracellular matrix, growth factors, and endogenous cells including mesenchymal stem cells. Advances in tissue preservation have helped to overcome the short shelf life of fresh AM and led to the development of AM products for clinical use. Viable cryopreserved amnion (VCAM), which retains all native components of fresh AM, has shown positive outcomes in clinical trials for wound management. However, cryopreservation requires ultra-low temperature storage and shipment that limits widespread use of VCAM. We have developed a lyopreservation technique to allow for ambient storage of living tissues. Here, we compared the structural, molecular, and functional properties of a viable lyopreserved human amniotic membrane (VLAM) with properties of VCAM using in vitro and in vivo wound models. We found that the structure, growth factors, and cell viability of VLAM is similar to that of VCAM and fresh AM. Both, VCAM and VLAM inhibited TNF-α secretion and upregulated VEGF expression in vitro under conditions designed to mimic inflammation and hypoxia in a wound microenvironment, and resulted in wound closure in a diabetic mouse chronic wound model. Taken together, these data demonstrate that VLAM structural and functional properties are equivalent to VCAM but without the constraints of ultra-low temperature storage.