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Long‐term effect of platelet lysate on primary fibroblasts highlighted with a proteomic approach
Authors:Valentina Cipriani  Elia Ranzato  Valeria Balbo  Laura Mazzucco  Maria Cavaletto  Mauro Patrone
Affiliation:1. Department of Environment and Life Sciences, University of Piemonte Orientale, viale T. Michel 11, 15121 Alessandria, Italy;2. These two authors contributed equally to this work.;3. Department of Haematology and Blood Transfusion Medicine, Ospedale Santi Antonio e Biagio e Cesare Arrigo, Alessandria, Italy
Abstract:The use of platelets and platelet derivatives has acquired clinical relevance as a means of accelerating wound healing. Platelet beneficial effect is attributed to the release of growth factors and other bioactive substances able to regulate cellular activities. The purpose of this study was to evaluate the biological effects of platelet lysate (PL) on human primary skin fibroblasts. We studied cell viability, MAPK signalling and proteomic profile of fibroblasts exposed to a platelet lysate (PL) obtained from blood sample. Crystal violet and neutral red uptake assays showed the dose–response effects of PL on cell viability and metabolism at 3 and 6 days of exposure. Western blot demonstrated a more sustained activation of p38 than of ERK1/2. A proteomic approach was applied to identify soluble cellular components in primary fibroblasts that are differentially expressed in response to PL exposure. Protein identification was performed by mass spectrometry. The data demonstrate that human fibroblasts respond to PL exposure by modifying a number of proteins, related principally to stress response, metabolism and the cytoskeleton. Copyright © 2009 John Wiley & Sons, Ltd.
Keywords:human fibroblasts  cell signalling  platelet lysate  2D gel electrophoresis  proteome  MALDI–  TOF
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