Leukocyte-and Platelet-Rich Fibrin for enhanced tissue repair: an in vitro study characterizing cellular composition, growth factor kinetics and transcriptomic insights

Mol Biol Rep. 2024 Sep 4;51(1):954. doi: 10.1007/s11033-024-09890-y.

Abstract

Background: Leukocyte- and platelet-rich fibrin (L-PRF) is an autologous platelet concentrate, prepared by centrifugation of blood and consisting of a dense fibrin network with incorporated leukocytes and platelets. This study aims to perform an in-depth analysis of the cells, growth factors, and transcriptome of L-PRF.

Methods and results: Fresh, 1 week and 2 weeks cultured human L-PRF membranes and liquid L-PRF glue were characterized on cellular and transcriptional level using flow cytometry (n = 4), single-cell RNA sequencing (n = 5) and RT-qPCR. Growth factor kinetics were investigated using ELISA (EGF, VEGF, PDGF-AB, TGF-β1, bFGF). L-PRF contained a large number of viable cells (fresh 97.14 ± 1.09%, 1 week cultured 93.57 ± 1.68%), mainly granulocytes in fresh samples (53.9 ± 19.86%) and T cells in cultured samples (84.7 ± 6.1%), confirmed with scRNA-seq. Monocytes differentiate to macrophages during 1 week incubation. Specifically arterial L-PRF membranes were found to release significant amounts of VEGF, EGF, PDGF-AB and TGF-β1.

Conclusion: We characterized L-PRF using in vitro experiments, to obtain an insight in the composition of the material including a possible mechanistic role for tissue healing. This was the first study characterizing L-PRF at a combined cellular, proteomic, and transcriptional level.

Keywords: Autologous; Characterization; Leukocyte and platelet-rich fibrin; Platelet concentrate.

MeSH terms

  • Blood Platelets / metabolism
  • Cells, Cultured
  • Humans
  • Intercellular Signaling Peptides and Proteins* / genetics
  • Intercellular Signaling Peptides and Proteins* / metabolism
  • Kinetics
  • Leukocytes* / metabolism
  • Platelet-Rich Fibrin* / metabolism
  • Transcriptome* / genetics
  • Wound Healing / genetics

Substances

  • Intercellular Signaling Peptides and Proteins