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In Vitro Bioactivity Of Tri-layer Placental Allograft Membrane
Sita M. Damaraju, Paul Bonvallet, Heli Modi, Morakot Likhitpanichkul, Ankur Gandhi, Sunil Saini.
Integra Lifesciences, Corp, Plainsboro, NJ, USA.

Application of placental tissue allografts has the potential to be an efficacious treatment for chronic wounds due to its inherent reservoir of growth factors, cytokines and extracellular matrix molecules. Processing of these placental tissues is essential for long term storage and “off the shelf “availability but without compromising the functionality of inherent bioactive factors. In this study, the bioactivity of a novel, dehydrated tri-layer placental allograft membrane (TPAM), consisting of amnion, chorion and amnion, was evaluated in vitro. TPAM was homogenized and extracted in media overnight at 4oC. Bioactivity of TPAM extracts was evaluated in a wound healing scratch assay with adult fibroblasts to assess cell migration. Chemotactic migration assay was performed with human mesenchymal stem cells (hMSCs) in a 24 well Trans-well plate setup. Inflammation assay was performed with lipopolysaccharide activated peripheral blood mononuclear cells (PBMCs) to determine whether TPAM could reduce the expression of pro-inflammatory cytokines. Adult fibroblasts were seeded on TPAM to evaluate cell attachment and proliferation over 7 days in culture. TPAM promoted enhanced adult fibroblasts and hMSCs migration, which was significantly higher than controls (n=4 per group; p<0.05). Addition of TPAM extracts to PBMCs significantly reduced the expression of pro-inflammatory mediators by more than 90% for TNF-alpha, IL-1beta, and IL-6, and by 40% for IL-8 (n=4 per group; p<0.05). In addition, TPAM promoted cell attachment and proliferation with adult fibroblasts in culture over a 7-day period. These results confirm that TPAM contains factors which are bioactive and could aid in providing the appropriate environment to close chronic wounds.

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