Leucocyte and Platelet‐rich Fibrin: a carrier of autologous multipotent cells for regenerative medicine

Abstract

The wound healing is a complex process wherein inflammation, proliferation and regeneration evolve according to a spatio‐temporal pattern from the activation of coagulation cascade to the formation of a plug clot including fibrin matrix, blood‐borne cells and cytokines/growth factors. Creating environments conducive to tissue repair, the haemoderivatives are commonly proposed for the treatment of hard‐to‐heal wounds. Here, we explored in vitro the intrinsic regenerative potentialities of a leucocyte‐ and platelet‐rich fibrin product, known as CPL‐MB, defining the stemness grade of cells sprouting from the haemoderivative. Using highly concentrated serum‐based medium to simulate wound conditions, we isolated fibroblast‐like cells (CPL‐CMCs) adhering to plastic and showing stable in vitro propagation, heterogeneous stem cell expression pattern, endothelial adhesive properties and immunomodulatory profile. Due to their blood derivation and expression of CXCR4, CPL‐CMCs have been suggested to be immature cells circulating in peripheral blood at quiescent state until activation by both coagulation event and inflammatory stimuli such as stromal‐derived factor 1/SDF1. Expressing integrins (CD49f, CD103), vascular adhesion molecules (CD106, CD166), endoglin (CD105) and remodelling matrix enzymes (MMP2, MMP9, MMP13), they showed a transendothelial migratory potential besides multipotency. Taken together, our data suggested that a standardized, reliable and economically feasible blood product such as CPL‐MB functions as an artificial stem cell niche that, under permissive conditions, originate ex vivo immature cells that could be useful for autologous stem cell‐based therapies.

Regenerative Potential of Platelet-Rich Fibrin Releasate Combined with Adipose Tissue–Derived Stem Cells in a Rat Sciatic Nerve Injury Model

“…PRFr can be used in a variety of clinical applications, based on the premise that higher GF content should promote better healing. Interestingly, beside the enrichment in platelets and leukocytes, the entrapment of stem-like cells with high regenerative potential within the fibrin network has recently been acknowledged 35 , providing an even more solid basis for the use of PRF in regenerative medicine 17 . Literature records concerning the nerve regeneration with stem cell and platelet-rich fibrin therapies are currently limited.…”

Biological and Mechanical Properties of Platelet-Rich Fibrin Membranes after Thermal Manipulation and Preparation in a Single-Syringe Closed System

“…Serum squeezed out from the PRF clot, called hyperacute serum, has higher cell proliferative effect on bone marrow mesenchymal stem cells (MSCs), osteoblasts and osteoarthritic chondrocyte cells than PRP [ 16 , 17 ]. After removing the hyperacute serum fraction, the remaining PRF membrane is a three-dimensional, biocompatible, biodegradable scaffold, which can slowly and sustainably release bioactive molecules, which facilitate cell adhesion and proliferation [ 18 , 19 ]. PRF membrane can provide an appropriate cell-niche for most cell types, because the outer layer allows contact of cells and ECM molecules through the surface molecules of the membrane.…”

From Platelet-Rich Plasma to Advanced Platelet-Rich Fibrin: Biological Achievements and Clinical Advances in Modern Surgery

“…There are not enough in vivo studies to support that, when using the platelet concentrates, we are also using M2 (alternatively activated macrophages [anti-inflammatory molecule producer])-polarized macrophages instead of M1 (classically activated macrophages [pro-inflammatory molecules producer]). 38,39 Tissue repairing requires a strategic collaboration of all leukocytes, epithelial and endothelial cells, fibroblasts, and platelets. In Choukron's A-PRF, the number of leukocytes includes more neutrophils that also can contribute to a proor anti-inflammatory state of the macrophages modulating tissue regeneration.…”

“…4 Recent in vitro studies show that L-PRF ability to potentiate angiogenesis and vasculogenesis at the injury site is mediated not only by the liberation of bioactive molecules but also by the presence of hematopoietic stem cells and endothelial progenitor cells. 39 The second-generation concentrates L-PRF and A-PRF have a similar level of platelets and cytokines. However, A-PRF releases a significantly higher quantity of growth factors TGF-β, PDGF, VEGF, and chemotactic molecules CCL-5 and eotaxin, when compared with traditional PRF.…”