Wound Healing Society

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Bioactivity Of A Cryopreserved Morselized Amnion Suspension
Morakot Likhitpanichkul, Christopher Cowan, Sita Damaraju, Ankur Gandhi, Sunil Saini.
Integra LifeSciences, Corp, Plainsboro, NJ, USA.

BACKGROUND - Human amniotic allografts have been used for treatment of acute and chronic wounds as the amniotic tissue inherently contains growth factors and cytokines that can modulate wound healing processes. Cryopreserved morselized amnion suspension (MAS) is a flowable form of amniotic tissue used for treating wounds in hard to reach locations. Cryopreservation is used to reduce loss of bioactivity from the morselization process. In this study, we aimed to assess the bioactivity of cryopreserved MAS in vitro. METHODS - Cell proliferation assessment, utilizing BrdU colorimetric assay, and cell migration assessment, using a transwell plate setup, were performed on human mesenchymal stem cells (hMSCs) treated with MAS. An inflammation assay was performed with lipopolysaccharide (LPS) activated peripheral blood mononuclear cells (PBMCs) to evaluate inhibition of pro-inflammatory cytokine production after addition of MAS. RESULTS - Results showed hMSC proliferation was significantly increased 60-180% in MAS treated group compared to control after 72 hours (n=4 per group, 3 lots of MAS, p<0.001). The number of hMSCs migrated through the transwell was found to significantly increase by 8-10 folds with MAS treatment after 24 hours (n=4 per group, 3 lots of MAS, p<0.001). The expression of pro-inflammatory cytokines produced by LPS-stimulated PBMC was found to significantly reduce by more than 90% for TNF-alpha, IL-1beta, and IL-6, and by 75% for IL-8 (n=4 per group; 5 lots of MAS, p<0.001). CONCLUSIONS - These results confirm that factors contained in MAS are bioactive and could be beneficial in providing the appropriate environment for wound treatment.

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