title: A plasma-based biomatrix mixed with endothelial progenitor cells and keratinocytes promotes matrix formation, angiogenesis, and reepithelialization in full-thickness wounds.
abstract: In search of an autologous vascularized skin substitute, we treated full-thickness wounds (FTWs) with autologous platelet-rich plasma gel (APG) in which we embedded endothelial progenitor cells (EPCs) and basal cell keratinocytes (KCs). We cultivated autologous KCs in low-serum conditions and expanded autologous EPCs from venous blood. FTWs (n = 55) were created on the backs of four pigs, covered with wound chambers, and randomly assigned to the following treatments: (1) APG, (2) APG + KCs, (3) APG + EPCs, (4) APG + KCs + EPCs, and (5) saline. All wounds were biopsied to measure neovascularization (lectin Bandeiraea Simplicifolia-1 (BS-1), alpha smooth muscle actin [alphaSMA], and membrane type 1 matrix metalloproteinase (MT1-MMP)), matrix deposition (fibronectin, collagen type I/III, and alphavbeta3), and reepithelialization. Wound fluids were analyzed for protein expression. All APG-treated wounds showed more vascular structures (p < 0.001), and the addition of EPCs further improved neovascularization, as confirmed by higher lectin, alphaSMA, and MT1-MMP. APG groups had higher collagen I/III (p < 0.05), alphavbeta3, and fibronectin content (p < 0.001), and they exhibited higher concentrations of platelet-derived growth factor subunit bb, basic fibroblast growth factor, hepatocyte growth factor, insulin growth factor-1, transforming growth factor-beta1 and -beta3, matrix metalloproteinase-1 and -z9, and tissue-inhibiting matrix metalloproteinase-1 and -2. Applying APG + KCs resulted in the highest reepithelialization rates (p < 0.001). No differences were found for wound contraction by planimetry. In this porcine FTW model, APG acts as a supportive biomatrix that, along with the embedded cells, improves extracellular matrix organization, promotes angiogenesis, and accelerates reepithelialization.
authors.affiliation: Department of Plastic and Reconstructive Surgery, KU Leuven University Hospital, Katholieke Universiteit Leuven, Leuven, Belgium.
authors.forename: Pieter
authors.initials: P
authors.lastname: Vermeulen
authors.forename: Stijn
authors.initials: S
authors.lastname: Dickens
authors.forename: Karlien
authors.initials: K
authors.lastname: Degezelle
authors.forename: Stefaan
authors.initials: S
authors.lastname: Van den Berge
authors.forename: Benoit
authors.initials: B
authors.lastname: Hendrickx
authors.forename: Jan Jeroen
authors.initials: JJ
authors.lastname: Vranckx
mesh_terms: D000818:Animals; D042783:Endothelial Cells / Q000166:cytology*; D004848:Epithelium / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D007150:Immunohistochemistry; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology*; D018919:Neovascularization, Physiologic; D053657:Platelet-Rich Plasma / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology*; D034421:Sus scrofa; D014947:Wounds and Injuries / Q000473:pathology*
chemical_list: D036341:Intercellular Signaling Peptides and Proteins