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.

Mesenchymal stem cell (MSC) and endothelial progenitor cell (EPC) growth and adhesion in six different bone graft substitutes

“…It is also known that MSC adhere to fibronectin [42] and that the fibronectin coating of b-TCP advances slightly the bone formation in vivo [43]. In the clinical reality, full plasma or autologous platelet-rich plasma [44] is an attractive alternative to pure fibronectin.…”

Integration of Blood Outgrowth Endothelial Cells in Dermal Fibroblast Sheets Promotes Full Thickness Wound Healing

“…According to this definition, two EPC types are distinguished: the early outgrowing and initially nonadherent fraction, which is clonally related to hematopoietic stem cells and does not incorporate into vessels, and the late outgrowing and immediately adherent fraction that lacks hematopoietic markers and does incorporate into vessels [11]. The latter cells are also referred to as ''blood outgrowth endothelial cells'' (BOEC) and have been derived from peripheral and/or cord blood [13][14][15][16]. They have been used for restoring blood flow in peripheral vascular disease models [17,18], for prevascularization of decellularized skin or bone tissue-engineered constructs [19,20] and as a vehicle for gene therapy [15,21].…”

Platelet-rich plasma to treat experimentally-induced skin wounds in animals: A systematic review and meta-analysis

“…Although the forest plot in Fig 3 showed two studies with not significant negative ES [ 51 , 53 ], two studies with null ES [ 30 , 46 ], seven studies with not significant positive ES [ 42 , 44 , 45 , 50 , 52 , 56 , 58 ] and four studies with significant positive ES [ 43 , 54 , 55 , 57 ]: the exclusion of each of the studies, in the sensitivity analysis, would have no relevant effect on the overall results of the meta-analysis supporting the efficacy of PRP for primary outcome. ( Fig 4 )…”