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Retention of Endogenous Viable Cells Enhances the Anti-Inflammatory Activity of Cryopreserved Amnion.

Duan-Arnold Y, Gyurdieva A, Johnson A, Uveges TE, Jacobstein DA, Danilkovitch A - Adv Wound Care (New Rochelle) (2015)

Bottom Line: Approach: The tissue composition of int-hAM and dev-hAM was compared with fresh hAM through histology and cell viability analysis.Compared with dev-hAM, int-hAM showed significantly greater downregulation of TNF-α and IL-1α, upregulation of PGE2 and IL-10, and stronger inhibition of collagenase.For the first time, we show that viable endogenous cells significantly augment the anti-inflammatory activity of cryopreserved hAM.

View Article: PubMed Central - PubMed

Affiliation: Osiris Therapeutics, Inc., Columbia, Maryland.

ABSTRACT

Objective: Human amniotic membrane (hAM) has been used to treat wounds for more than 100 years. However, widespread use of fresh hAM has been limited due to its short shelf life and safety concerns. To overcome these concerns, different preservation methods have been introduced. The majority of these methods result in devitalized hAM (dev-hAM). Recently, we developed a cryopreservation method that retains all hAM components intact (int-hAM), including viable endogenous cells. To understand the advantages of retaining viable cells in preserved hAM, we compared the anti-inflammatory properties of int-hAM and dev-hAM. Approach: The tissue composition of int-hAM and dev-hAM was compared with fresh hAM through histology and cell viability analysis. We also evaluated the ability of int-hAM and dev-hAM to regulate tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α), and IL-10 release when co-cultured with immune cells; to produce prostaglandin E2 (PGE2) on TNF-α stimulation; and to inhibit proteases. Results: Int-hAM maintained the structural and cellular integrity of fresh hAM. Int-hAM had >80% cell viability post-thaw and remained viable for at least a week in culture. Viable cells were not detected in dev-hAM. Compared with dev-hAM, int-hAM showed significantly greater downregulation of TNF-α and IL-1α, upregulation of PGE2 and IL-10, and stronger inhibition of collagenase. Innovation and Conclusion: A new cryopreservation method has been developed to retain all native components of hAM. For the first time, we show that viable endogenous cells significantly augment the anti-inflammatory activity of cryopreserved hAM.

No MeSH data available.


Related in: MedlinePlus

Effects of human amniotic membrane (hAM) on human peripheral blood mononuclear cell (hPBMC)-induced anti-inflammatory factor interleukin-10 (IL-10) release. IL-10 levels in conditioned medium were measured after 24 h co-cultures of lipopolysaccharide (LPS)-stimulated hPBMCs with viable intact cryopreserved hAM (int-hAM) and devitalized cryopreserved hAM (dev-hAM). Negative (Neg) and positive (Pos) controls were unstimulated and LPS-activated hPBMCs, respectively. Data shown represent the mean±SD of one experiment performed in triplicate. Student's t-test was used for statistical analysis. ***p<0.001.
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Related In: Results  -  Collection


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f4: Effects of human amniotic membrane (hAM) on human peripheral blood mononuclear cell (hPBMC)-induced anti-inflammatory factor interleukin-10 (IL-10) release. IL-10 levels in conditioned medium were measured after 24 h co-cultures of lipopolysaccharide (LPS)-stimulated hPBMCs with viable intact cryopreserved hAM (int-hAM) and devitalized cryopreserved hAM (dev-hAM). Negative (Neg) and positive (Pos) controls were unstimulated and LPS-activated hPBMCs, respectively. Data shown represent the mean±SD of one experiment performed in triplicate. Student's t-test was used for statistical analysis. ***p<0.001.

Mentions: Activation of hPBMCs by LPS was used to detect the production of anti-inflammatory cytokine IL-10. When LPS-activated hPBMCs were incubated with TNF-α pretreated tissues, we observed an increase in IL-10 release (255±81 pg/mL) and a decrease in IL-10 release (86±20 pg/mL) relative to the positive control (174±27 pg/mL) for int-hAM and dev-hAM, respectively (Fig. 4).


Retention of Endogenous Viable Cells Enhances the Anti-Inflammatory Activity of Cryopreserved Amnion.

Duan-Arnold Y, Gyurdieva A, Johnson A, Uveges TE, Jacobstein DA, Danilkovitch A - Adv Wound Care (New Rochelle) (2015)

Effects of human amniotic membrane (hAM) on human peripheral blood mononuclear cell (hPBMC)-induced anti-inflammatory factor interleukin-10 (IL-10) release. IL-10 levels in conditioned medium were measured after 24 h co-cultures of lipopolysaccharide (LPS)-stimulated hPBMCs with viable intact cryopreserved hAM (int-hAM) and devitalized cryopreserved hAM (dev-hAM). Negative (Neg) and positive (Pos) controls were unstimulated and LPS-activated hPBMCs, respectively. Data shown represent the mean±SD of one experiment performed in triplicate. Student's t-test was used for statistical analysis. ***p<0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4529089&req=5

f4: Effects of human amniotic membrane (hAM) on human peripheral blood mononuclear cell (hPBMC)-induced anti-inflammatory factor interleukin-10 (IL-10) release. IL-10 levels in conditioned medium were measured after 24 h co-cultures of lipopolysaccharide (LPS)-stimulated hPBMCs with viable intact cryopreserved hAM (int-hAM) and devitalized cryopreserved hAM (dev-hAM). Negative (Neg) and positive (Pos) controls were unstimulated and LPS-activated hPBMCs, respectively. Data shown represent the mean±SD of one experiment performed in triplicate. Student's t-test was used for statistical analysis. ***p<0.001.
Mentions: Activation of hPBMCs by LPS was used to detect the production of anti-inflammatory cytokine IL-10. When LPS-activated hPBMCs were incubated with TNF-α pretreated tissues, we observed an increase in IL-10 release (255±81 pg/mL) and a decrease in IL-10 release (86±20 pg/mL) relative to the positive control (174±27 pg/mL) for int-hAM and dev-hAM, respectively (Fig. 4).

Bottom Line: Approach: The tissue composition of int-hAM and dev-hAM was compared with fresh hAM through histology and cell viability analysis.Compared with dev-hAM, int-hAM showed significantly greater downregulation of TNF-α and IL-1α, upregulation of PGE2 and IL-10, and stronger inhibition of collagenase.For the first time, we show that viable endogenous cells significantly augment the anti-inflammatory activity of cryopreserved hAM.

View Article: PubMed Central - PubMed

Affiliation: Osiris Therapeutics, Inc., Columbia, Maryland.

ABSTRACT

Objective: Human amniotic membrane (hAM) has been used to treat wounds for more than 100 years. However, widespread use of fresh hAM has been limited due to its short shelf life and safety concerns. To overcome these concerns, different preservation methods have been introduced. The majority of these methods result in devitalized hAM (dev-hAM). Recently, we developed a cryopreservation method that retains all hAM components intact (int-hAM), including viable endogenous cells. To understand the advantages of retaining viable cells in preserved hAM, we compared the anti-inflammatory properties of int-hAM and dev-hAM. Approach: The tissue composition of int-hAM and dev-hAM was compared with fresh hAM through histology and cell viability analysis. We also evaluated the ability of int-hAM and dev-hAM to regulate tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α), and IL-10 release when co-cultured with immune cells; to produce prostaglandin E2 (PGE2) on TNF-α stimulation; and to inhibit proteases. Results: Int-hAM maintained the structural and cellular integrity of fresh hAM. Int-hAM had >80% cell viability post-thaw and remained viable for at least a week in culture. Viable cells were not detected in dev-hAM. Compared with dev-hAM, int-hAM showed significantly greater downregulation of TNF-α and IL-1α, upregulation of PGE2 and IL-10, and stronger inhibition of collagenase. Innovation and Conclusion: A new cryopreservation method has been developed to retain all native components of hAM. For the first time, we show that viable endogenous cells significantly augment the anti-inflammatory activity of cryopreserved hAM.

No MeSH data available.


Related in: MedlinePlus