Anti-fibrotic effects of fresh and cryopreserved human amniotic membrane in a rat liver fibrosis model
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Anti-fibrotic effects of fresh and cryopreserved human amniotic membrane in a rat liver fibrosis model Emanuele Ricci • Graziella Vanosi • Andrea Lindenmair • Simone Hennerbichler • Anja Peterbauer-Scherb • Susanne Wolbank • Anna Cargnoni • Patrizia Bonassi Signoroni Marino Campagnol • Christian Gabriel • Heinz Redl • Ornella Parolini
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Received: 9 May 2012 / Accepted: 10 August 2012 / Published online: 29 August 2012 Ó Springer Science+Business Media B.V. 2012
Abstract The human amniotic membrane (hAM), thanks to its favorable properties, including antiinflammatory, 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
E. Ricci A. Cargnoni P. B. Signoroni O. Parolini (&) Centro di Ricerca E. Menni, Fondazione Poliambulanza Istituto Ospedaliero, Via Bissolati 57, 25124 Brescia, Italy e-mail: [email protected]; [email protected] G. Vanosi Dipartimento di Scienze Cliniche Veterinarie-Sezione di Radiologia Clinica e Sperimentale, Facolta` di Medicina Veterinaria, Universita` degli Studi di Milano, 20133 Milan, Italy A. Lindenmair S. Wolbank H. Redl Ludwig Boltzmann Institute for Experimental and Clinical Traumatology/AUVA Research Centre, Austrian Cluster for Tissue Regeneration, 1200 Vienna, Austria A. Lindenmair S. Hennerbichler A. Peterbauer-Scherb C. Gabriel Red Cross Blood Transfusion Service of Upper Austria, Austrian Cluster for Tissue Regeneration, 4017 Linz, Austria M. Campagnol Facolta` di Medicina Veterinaria, Central Animal Facility, Universita` degli Studi di Milano, 20133 Milan, Italy
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 antifibr
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