Leukocyte-Poor Platelet-Rich Plasma-Derived Growth Factors Enhance Human Fibroblast Proliferation In Vitro

Clin Orthop Surg. 2018 Jun;10(2):240-247. doi: 10.4055/cios.2018.10.2.240. Epub 2018 May 18.

Abstract

Background: Leukocyte-poor platelet-rich plasma (LP-PRP) from peripheral blood is currently used as a concentrated source of growth factors to stimulate repair at sites of soft tissue injury. Fibroblasts are primary mediators of wound healing. Thus, we aimed to assess the positive effect of LP-PRP on human fibroblast proliferation in vitro.

Methods: LP-PRP was prepared from 49 donors. The fibroblasts were seeded, and at 24 hours after seeding, 1 × 107/10 µL LP-PRP was added once to each well. The cells were harvested 10 times during study period at our planned points, and we examined cell proliferation using the water-soluble tetrazolium salt-1 assay. We collected the supernatants and measured the amount of growth factors such as platelet-derived growth factor (PDGF)-AB/BB, insulin-like growth factor-1 (IGF-1), transforming growth factor-β1 (TGF-β1), and vascular endothelial growth factor (VEGF), which are known to be involved in wound healing processes, by multiplex assay.

Results: Human fibroblasts treated with LP-PRP showed a significant increase in proliferation when compared to untreated controls (p < 0.001 at days 4, 6, and 8). Multiplex cytokine assays revealed various secretion patterns. PDGF-AB/BB appeared at early time points and peaked before fibroblast proliferation. IGF-1 and TGF-β1 secretion gradually increased and peaked on days 4 and 6 post-treatment. The early VEGF concentration was lower than the concentration of other growth factors but increased along with cell proliferation.

Conclusions: Platelets in LP-PRP release growth factors such as PDGF, IGF-1, TGF-β1 and VEGF, and these growth factors have a promoting effect for human fibroblast proliferation, one of the important mediators of wound healing. These results suggest that growth factors derived from LP-PRP enhance the proliferation of human fibroblast.

Keywords: Fibroblast; Platelet-derived growth factor; Platelet-rich plasma; Transforming growth factor-β1.

MeSH terms

  • Cell Culture Techniques
  • Cell Proliferation / drug effects*
  • Cells, Cultured
  • Fibroblasts / cytology
  • Fibroblasts / drug effects*
  • Humans
  • Platelet-Derived Growth Factor / analysis
  • Platelet-Derived Growth Factor / pharmacology*
  • Platelet-Rich Plasma / chemistry*
  • Platelet-Rich Plasma / cytology
  • Transforming Growth Factor beta / analysis
  • Transforming Growth Factor beta / pharmacology*

Substances

  • Platelet-Derived Growth Factor
  • Transforming Growth Factor beta