Can leg ulcer fibroblasts phenotype be influenced by human amniotic membrane extract?
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Can leg ulcer fibroblasts phenotype be influenced by human amniotic membrane extract? H. Tauzin • S. Robin • P. Humbert • C. Viennet P. Saas • C. Courderot-Masuyer • P. Muret
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Received: 29 July 2013 / Accepted: 7 January 2014 / Published online: 11 February 2014 Ó Springer Science+Business Media Dordrecht 2014
Abstract Prevalence of leg ulcer in general population is important and new efficient treatments are now needed, especially for chronic leg ulcers. Human amniotic membrane (HAM) can be used as an alternative treatment for recalcitrant leg ulcers. The aim of this study is to investigate the effects of a HAM extract on normal fibroblasts (NF) and ulcer fibroblasts (UF). NF and UF were obtained from biopsies by explants technique. HAM extract was used at 10 lg of total proteins per ml. Single patient-matched NF and UF were compared, without or with HAM extract. Studied parameters were proliferation rate, retraction of free-floating lattices, alpha smooth muscle actin expression by flow cytometry, and synthesis of elastin, glycosaminoglycans (GAGs), pro-collagen I, MMP-1 and TIMP-1. Our results show that UF had a specific
phenotype compared to NF: low proliferation, high expression of alpha-SM actin and high synthesis of MMP-1, TIMP-1 and elastin. HAM extract significantly increased the synthesis of GAGs, pro-collagen I and MMP-1 in NF and decreased retraction of free lattices. HAM extract transiently increased UF proliferation, slowed down lattices retraction and decreased elastin synthesis. In conclusion, HAM extract has little effect on UF for the parameters studied and NF are more responsive than UF. However, clinical beneficial effect of HAM application on leg ulcers was previously observed and might rather be related to an action on keratinocytes and/or a modulation of the highly inflammatory environment of these chronic wounds. Keywords Leg ulcer Primary fibroblasts Human amniotic membrane Extract In vitro
H. Tauzin and S. Robin are co-authors of this study. H. Tauzin P. Humbert C. Viennet P. Saas P. Muret UMR 1098, SFR FED 4234, 25000 Besanc¸on, France H. Tauzin P. Humbert C. Viennet P. Saas P. Muret University of Franche-Comte, 25000 Besanc¸on, France H. Tauzin (&) S. Robin C. Courderot-Masuyer Bioexigence, 8 rue Alfred de Vigny, 25000 Besanc¸on, France e-mail: [email protected] P. Humbert Department of Dermatology, University Hospital of Besanc¸on, 25000 Besanc¸on, France
Introduction Clinical use of human amniotic membrane (HAM) on leg ulcers has been reported for the first time in 1910 (Davis 1910) and had a renewal in the 80s but randomized clinical studies are lacking. Nevertheless recent prospective clinical trials showed beneficial effects of cryopreserved HAM on recalcitrant leg ulcers: HAM transplantation is safe, promotes epithelial healing, suppresses fibrosis and significantly reduces pain (Mermet et al. 2007; Pesteil et al. 2007). The biological properties of HAM, among which anti-inflammatory,
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