Ricci Emanuele, Vanosi Graziella, Lindenmair Andrea, Hennerbichler Simone, Peterbauer-Scherb Anja, Wolbank Susanne, Cargnoni Anna, Signoroni Patrizia Bonassi, Campagnol Marino, Gabriel Christian, Redl Heinz, Parolini Ornella
Centro di Ricerca E. Menni, Fondazione Poliambulanza Istituto Ospedaliero, Via Bissolati 57, 25124, Brescia, Italy.
Cell Tissue Bank. 2013 Sep;14(3):475-88. doi: 10.1007/s10561-012-9337-x. Epub 2012 Aug 29.
The human amniotic membrane (hAM), thanks to its favorable properties, including anti-inflammatory, anti-fibrotic and pro-regenerative effects, is a well-known surgical material for many clinical applications, when used both freshly after isolation and after preservation. We have shown previously that hAM patching is a potential approach to counteract liver fibrosis. Indeed, when fresh hAM was used to cover the liver surface of rats with liver fibrosis induced by the bile duct ligation (BDL) procedure, the progression and severity of fibrosis were significantly reduced. Since cryopreservation enables safety and long-term storage of hAM but may influence its functional properties, here we compared the anti-fibrotic effects of fresh and cryopreserved hAM in rats with BDL-induced liver fibrosis. After BDL, the rat liver was covered with a piece of fresh or cryopreserved hAM, or left untreated. Six weeks later, the degree of liver fibrosis was assessed histologically using the Knodell and the METAVIR scoring systems. Digital image analysis was used to quantify the percentage of the areas of each liver section displaying ductular reaction, extracellular matrix (ECM) deposition, activated myofibroblasts and hepatic stellate cells (HSCs). Liver collagen content was also determined by spectrophotometric technique. The degree of liver fibrosis, ductular reaction, ECM deposition, and the number of activated myofibroblasts and HSCs were all significantly reduced in hAM-treated rats compared to control animals. Fresh and cryopreserved hAM produced the same anti-fibrotic effects. These findings indicate that cryopreservation maintains the anti-fibrotic properties of hAM when used as a patch to reduce the severity of liver fibrosis.
人羊膜（hAM）因其具有抗炎、抗纤维化和促再生等良好特性，无论是在分离后立即使用还是保存后使用，都是一种在许多临床应用中广为人知的外科材料。我们之前已经表明，hAM贴片是对抗肝纤维化的一种潜在方法。事实上，当用新鲜的hAM覆盖胆管结扎（BDL）手术诱导肝纤维化的大鼠肝脏表面时，纤维化的进展和严重程度显著降低。由于冷冻保存能够实现hAM的安全长期储存，但可能会影响其功能特性，因此我们在此比较了新鲜和冷冻保存的hAM对BDL诱导的大鼠肝纤维化的抗纤维化作用。BDL术后，用一片新鲜或冷冻保存的hAM覆盖大鼠肝脏，或不进行处理。六周后，使用Knodell和METAVIR评分系统通过组织学评估肝纤维化程度。使用数字图像分析来量化每个肝切片中显示小胆管反应、细胞外基质（ECM）沉积、活化肌成纤维细胞和肝星状细胞（HSCs）的区域百分比。肝胶原含量也通过分光光度技术测定。与对照动物相比，hAM治疗的大鼠肝纤维化程度、小胆管反应、ECM沉积以及活化肌成纤维细胞和HSCs的数量均显著降低。新鲜和冷冻保存的hAM产生相同的抗纤维化作用。这些发现表明，当用作贴片以减轻肝纤维化严重程度时，冷冻保存可维持hAM的抗纤维化特性。
