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. 2007 Apr;40(2):241-52.
doi: 10.1111/j.1365-2184.2007.00430.x.

The effect of different platelet-rich plasma concentrations on proliferation and differentiation of human periodontal ligament cells in vitro

J Han  1 , H X Meng, J M Tang, S L Li, Y Tang, Z B Chen
Affiliations

The effect of different platelet-rich plasma concentrations on proliferation and differentiation of human periodontal ligament cells in vitro

J Han et al. Cell Prolif. 2007 Apr.

Abstract

Objectives: The use of platelets and platelet products has become increasingly popular clinically as a means of accelerating endosseous wound healing. It is likely that growth factors released by activated platelets at the site of injury play a role in periodontal regeneration by regulating cellular activity. The purpose of this study was to evaluate the biological effects of platelet-rich plasma (PRP) on human periodontal ligament cells (hPDLCs) in vitro.

Materials and methods: Primary cultures of hPDLCs were obtained from healthy premolars. PRP was isolated by two-step centrifugation. Two main growth factors present in the thrombin-activated PRP (platelet-derived growth factor [PDGF-AB] and transforming growth factor-beta1 [TGF-beta1]) were evaluated using ELISA assay. Activated PRP or the combination of recombined human TGF-beta1 (rhTGF-beta1) and PDGF-AB (rhPDGF-AB) were added to hPDLCs in different concentrations to assess cell proliferation and osteogenic differentiation.

Results: PRP contained high levels of TGF-beta1 and PDGF-AB. Cell attachment, proliferation and ALP activity were enhanced by addition of PRP or rhTGF-beta1 and rhPDGF-AB combination to the cell cultures, while the stimulatory potency of PRP was much greater than the latter. These stimulatory effects presented in a dose-dependant manner, it seemed that PRP with 50~100 ng/ml TGF-beta1 was an ideal concentration.

Conclusions: PRP can enhance hPDLC adhesion, proliferation and induce the differentiation of hPDLC into mineralized tissue formation cell; thereby contribute to the main processes of periodontal tissue regeneration. For economical and biological reasons, PRP has more clinical beneficial than analogous growth factors.

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Figures

Figure 1
Figure 1
Effect of different PRP concentrations on hPDLC proliferation. The result represents the means ± SD of three replicates. Control group, serum‐free DMEM medium; PRP, serum‐free DMEM medium with the supernatant of activated platelet‐rich plasma; growth factor, serum‐free DMEM medium containing the combination of rhTGF‐β1 and rhPDGF‐AB. *P < 0.05 vs. control group; **P < 0.05 PRP group vs. growth factor group; ***P < 0.05 vs. other concentration subgroups among PRP group.
Figure 2
Figure 2
Effect of different PRP concentrations on ALP activity of hPDLC. The result represents the means ± SD of three replicates. Control group, serum‐free DMEM medium; PRP, serum‐free DMEM medium with the supernatant of activated platelet‐rich plasma; growth factor, serum‐free DMEM medium containing the combination of rhTGF‐β1 and rhPDGF‐AB. *P < 0.05 vs. control group; **P < 0.05 PRP group vs. growth factor group; ***P < 0.05 vs. other concentration subgroups among PRP group.
Figure 3
Figure 3
Effect of PRP on hPDLC attachment. The result represents the means ± SD of three replicates. Control group, serum‐free DMEM medium; PRP, 12.5% (v/v) platelet‐rich plasma in serum‐free DMEM medium. The end concentration of TGF‐β1 and PDGF‐AB was 50 ng/ml and 17 ng/ml, respectively. *P < 0.05 PRP group vs. control group.
Figure 4
Figure 4
Time‐dependent effect of PRP on hPDLC proliferation. The result represents the means ± SD of three replicates. Control group, serum‐free DMEM medium; PRP, 12.5% (v/v) platelet‐rich plasma in serum‐free DMEM medium, the end concentration of TGF‐β1 and PDGF‐AB was 50 ng/ml and 17 ng/ml, respectively; growth factor, serum‐free DMEM medium containing the combination of 50 ng/ml rhTGF‐β1 and 17 ng/ml rhPDGF‐AB.
Figure 5
Figure 5
Time‐dependent effect of PRP on ALP activity of hPDLC. The result represents the means ± SD of three replicates. Control group, serum‐free DMEM medium; PRP, 12.5% (v/v) platelet‐rich plasma in serum‐free DMEM medium, the end concentration of TGF‐β1 and PDGF‐AB was 50 ng/ml and 17 ng/ml, respectively; growth factor, serum‐free DMEM medium containing the combination of 50 ng/ml rhTGF‐β1 and 17 ng/ml rhPDGF‐AB. *P < 0.05 vs. control group; **P < 0.05 PRP group vs. GF group.
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