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Favorable Modulation Of Inflammation And Immunity Is Achieved With Both Single And Tri-layer Amniotic Membrane Allografts
Victoria Stefanelli, Paul Bonvallet, Sita Damaraju, Qiaoling Lin, Heli Modi, Sunil Saini, Ankur Gandhi.
Integra LifeSciences, Plainsboro, NJ, USA.

BACKGROUND: The benefits of amniotic-based products across various regenerative applications are owed to a multitude of factors including cytokines, growth factors, and extracellular matrix components that synergistically function to accelerate the healing process. Innately immune-privileged, these materials are also associated with anti-inflammatory, anti-microbial, and immune-modulatory functions essential for both effective wound healing and minimization of scarring. The purpose of the present study was to elucidate the full range of anti-inflammatory and immunomodulatory capabilities of both a single-layer dehydrated amniotic membrane allograft (DAMA) and a tri-layer placental allograft membrane (TPAM) such that their contributions towards improved wound healing outcomes could be better understood.
METHODS: The anti-inflammatory functionalities of DAMA and TPAM were evaluated via cytokine analysis following PBMC activation with lipopolysaccharide in the presence of DAMA or TPAM extracts. An M1/M2 macrophage polarization assay was implemented to explore immune-modulation specifically through the assessment of CCL22, CCL18, PDGF, IL-13, TNF-alpha, IFN-gamma, IL-6, and RANTES. Macrophage polarization was conducted using both direct physical contact of the cells with DAMA/TPAM as well as soluble DAMA/TPAM extracts alone. Polarization outcomes were compared to non-stimulated macrophages as well as those in contact with non-immune-privileged collagen constructs. Finally, the effect of DAMA/TPAM extracts on T cell stimulation was explored using standard assays.
RESULTS: Cytokine analysis indicates that amniotic extracts possess a significant ability to reduce pro-inflammatory molecule secretion including TNF-alpha, IL-1beta, IL-10, MIP-1a, and IL-6 by over 90% in PBMC compared to controls (n=4 per group; p<0.05). Macrophage polarization analysis suggests that exposure to both DAMA and TPAM (n= 3 per group) modulates an M2-preferred macrophage response as a consequence of both soluble factors alone as well as physical contact with constructs. T cell responses were also favorably altered through exposure to amniotic extracts.
CONCLUSIONS: The ability of DAMA and TPAM to constructively influence both inflammatory and immunomodulatory processes therefore likely constitutes a substantial part of their overall effectiveness in wound remediation.


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