title: Anti-fibrotic effects of fresh and cryopreserved human amniotic membrane in a rat liver fibrosis model.
abstract: 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.
authors.affiliation: Centro di Ricerca E. Menni, Fondazione Poliambulanza Istituto Ospedaliero, Via Bissolati 57, 25124, Brescia, Italy.
authors.forename: Emanuele
authors.initials: E
authors.lastname: Ricci
authors.forename: Graziella
authors.initials: G
authors.lastname: Vanosi
authors.forename: Andrea
authors.initials: A
authors.lastname: Lindenmair
authors.forename: Simone
authors.initials: S
authors.lastname: Hennerbichler
authors.forename: Anja
authors.initials: A
authors.lastname: Peterbauer-Scherb
authors.forename: Susanne
authors.initials: S
authors.lastname: Wolbank
authors.forename: Anna
authors.initials: A
authors.lastname: Cargnoni
authors.forename: Patrizia Bonassi
authors.initials: PB
authors.lastname: Signoroni
authors.forename: Marino
authors.initials: M
authors.lastname: Campagnol
authors.forename: Christian
authors.initials: C
authors.lastname: Gabriel
authors.forename: Heinz
authors.initials: H
authors.lastname: Redl
authors.forename: Ornella
authors.initials: O
authors.lastname: Parolini
mesh_terms: D000199:Actins / Q000378:metabolism; D000650:Amnion / Q000637:transplantation*; D000818:Animals; D001652:Bile Ducts / Q000473:pathology; D003094:Collagen / Q000378:metabolism; D015925:Cryopreservation; D003893:Desmin / Q000378:metabolism; D004195:Disease Models, Animal; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D053539:Keratin-19 / Q000378:metabolism; D008026:Ligation; D008099:Liver / Q000473:pathology; D008103:Liver Cirrhosis / Q000473:pathology / Q000628:therapy*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013053:Spectrophotometry
chemical_list: D000199:Actins; D003094:Collagen; D003893:Desmin; D005353:Fibronectins; D053539:Keratin-19; C498862:Acta2 protein, rat