Effect of concentrated growth factors on function and Wnt3a expression of human periodontal ligament cells in vitro

Platelets. 2017 May;28(3):281-286. doi: 10.1080/09537104.2016.1213381. Epub 2016 Sep 6.

Abstract

Concentrated growth factors (CGFs), the new generation of platelet concentrate products, appear to exhibit superior potential for tissue regeneration. However, there are only a few studies supporting this. This study was designed to investigate the effect of CGFs on proliferation and alkaline phosphatase (ALP) activity of human periodontal ligament cells (hPDLCs) in vitro. Furthermore, as bone homeostasis is fundamentally controlled by Wnt-mediated signals, we also investigated Wnt3a expression of hPDLCs after treatment of CGFs. hPDLCs and CGFs were obtained from the same volunteer. CGFs or combination of recombined human TGF-β1 (rhTGF-β1) and PDGF-AB (rhPDGF-AB) were added to hPDLCs in different concentrations. The rate of proliferation was analyzed by an MTT assay. ALP activity was assessed using p-NPP assay. Quantitative RT-PCR was used to evaluate the gene expression of Wnt3a. In a range of concentrations, CGFs significantly promoted the proliferation of hPDLCs in a dose-dependent manner. ALP activity was also enhanced by CGFs in a dose-dependent and time-dependent manner. The stimulatory effect of CGFs was much greater than rhTGF-β1 and rhPDGF-AB combination. Quantitative RT-PCR results showed that Wnt3a mRNA expression was increased at 24 h in hPDLCs treated by CGFs. CGFs can enhance hPDLCs proliferation and ALP activity and may have great potential in clinical and biotechnological applications.

Keywords: Alkaline phosphatase activity; Wnt3a; cell proliferation; concentrated growth factors; periodontal ligament cells.

MeSH terms

  • Alkaline Phosphatase / genetics
  • Alkaline Phosphatase / metabolism
  • Blood Platelets / chemistry*
  • Cell Differentiation / drug effects
  • Cell Proliferation / drug effects
  • Complex Mixtures / chemistry
  • Dose-Response Relationship, Drug
  • Gene Expression Regulation
  • Humans
  • Intercellular Signaling Peptides and Proteins / isolation & purification
  • Intercellular Signaling Peptides and Proteins / pharmacology*
  • Periodontal Ligament / cytology
  • Periodontal Ligament / drug effects*
  • Periodontal Ligament / metabolism
  • Platelet-Derived Growth Factor / pharmacology*
  • Primary Cell Culture
  • Recombinant Proteins / pharmacology
  • Transforming Growth Factor beta1 / pharmacology*
  • Wnt3A Protein / agonists
  • Wnt3A Protein / genetics*
  • Wnt3A Protein / metabolism

Substances

  • Complex Mixtures
  • Intercellular Signaling Peptides and Proteins
  • Platelet-Derived Growth Factor
  • Recombinant Proteins
  • Transforming Growth Factor beta1
  • WNT3A protein, human
  • Wnt3A Protein
  • platelet-derived growth factor AB
  • Alkaline Phosphatase