Solomon A, Wajngarten M, Alviano F, Anteby I, Elchalal U, Pe'er J, Levi-Schaffer F
Department of Ophthalmology, Hadassah University Hospital, The Hebrew University--Hadassah Faculty of Medicine, Jerusalem, Israel.
Clin Exp Allergy. 2005 Jul;35(7):941-8. doi: 10.1111/j.1365-2222.2005.02285.x.
The amniotic membrane (AM), which is the innermost layer of the placenta, was shown to possess anti-inflammatory and anti-fibrotic properties in various in vitro and clinical studies.
To evaluate the anti-fibrotic and anti-inflammatory effects of the AM matrix (AMM) on human conjunctival and lung fibroblasts in an in vitro system that tests fibrotic and inflammatory responses at the effector stages of allergic inflammation.
Human conjunctival or lung fibroblasts were seeded on plastic or on the stromal aspect of the AM, which was mounted on plastic inserts. Sonicates of human peripheral blood eosinophils activated with lipopolysaccharide (LPS), or human mast cell (HMC-1) leukaemia cell sonicates, were added to sub-confluent fibroblast monolayers. Proliferation of the sub-confluent fibroblasts was assessed using the [3H]-thymidine incorporation assay. The production of transforming growth factor (TGF)-beta1, granulocyte-macrophage colony-stimulating factor (GM-CSF) and IL-8 in conjunctival or lung fibroblasts was measured in conditioned media from these cultures by ELISA.
After 4 days in culture, the [3H]-thymidine incorporation assay indicated a reduced proliferation of activated conjunctival and lung fibroblasts when cultured directly on the AMM. The production of both TGF-beta1 and IL-8 was significantly suppressed in activated conjunctival fibroblasts cultured on the AMM compared with those cultured on plastic, while the production of both TGF-beta1 and GM-CSF was decreased in human lung fibroblast cultured on the AMM.
The AMM is capable of suppressing fibrotic responses in an in vitro system of effector stages of ocular allergic inflammation. These data may provide a basis for exploring matrix components in the AM for the treatment of allergic eye disease.
羊膜是胎盘的最内层，在各种体外和临床研究中显示具有抗炎和抗纤维化特性。
在一个测试过敏性炎症效应阶段纤维化和炎症反应的体外系统中，评估羊膜基质（AMM）对人结膜和肺成纤维细胞的抗纤维化和抗炎作用。
将人结膜或肺成纤维细胞接种在塑料上或安装在塑料插入物上的羊膜基质面上。将用脂多糖（LPS）激活的人外周血嗜酸性粒细胞超声裂解物或人肥大细胞（HMC-1）白血病细胞超声裂解物添加到亚汇合的成纤维细胞单层中。使用[3H]-胸腺嘧啶核苷掺入法评估亚汇合成纤维细胞的增殖。通过ELISA在这些培养物的条件培养基中测量结膜或肺成纤维细胞中转化生长因子（TGF）-β1、粒细胞-巨噬细胞集落刺激因子（GM-CSF）和IL-8的产生。
培养4天后，[3H]-胸腺嘧啶核苷掺入法表明，直接在AMM上培养时，活化的结膜和肺成纤维细胞的增殖减少。与在塑料上培养的相比，在AMM上培养的活化结膜成纤维细胞中TGF-β1和IL-8的产生均显著受到抑制，而在AMM上培养的人肺成纤维细胞中TGF-β1和GM-CSF的产生均减少。
AMM能够在眼部过敏性炎症效应阶段的体外系统中抑制纤维化反应。这些数据可能为探索羊膜中的基质成分用于治疗过敏性眼病提供依据。
