J4 ›› 2011, Vol. 37 ›› Issue (1): 84-88.

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Effect of platelet-rich plasma on proliferation of human dermal fibroblasts in vitro

WANG Yue1| MA Ying-zhi2,3|ZHU Ze3|SU Xue-jing3|ZHOU Yan-min1   

  1. 1.Department of Implant Center|Stomatology Hospital|Jilin University|Changchun 130021|China;2.Clinic of Navigate Air Force University|Changchun 130022|China;3.Key Laboratory of Pathophysiology|Ministry of Education|Norman Bethune College of Medicine|Jilin University|Changchun 130021|China
  • Received:2010-10-12 Online:2011-01-28 Published:2011-01-28

Abstract:

Objective To evaluate the effects of platelet-rich plasma (PRP) on proliferation of human dermal fibroblasts (hDFbs) in vitro and investigate the mechanism of PRP promoting wound healing. Methods hDFbs and PRP  were separated from a healthy adult donor. PRP was prepared by a double centrifugation technique. In the proliferation experiment ainverted phase contrast microscope was used to observe the growth of cells after treated with different concentrations PRP,such as 0,12.5%,25.0%,50.0% and 100.0%. Immunocytochemistry was used to evaluate the expressions of platelet-derived growth factor (PDGF) treated with 50.0% PRP in different volumes.Fluorescent coloration was used to observe the  proliferation of the cells which adhered to the titanium material. Cell cycle of hDFbs was detected by flow cytometry. The cell proliferation was also evaluated by CCK-8 assay. Results The microscope investigation showed that the number of cells in 100%PRP group was higher than those in other groups. The significant increase in PDGF expression was observed when treated with  30 μL PRP,but the IOD in 10 μL PRP group was higher than that in 20 μL PRP group(836.27±21.15  vs  794.35±30.26,P<0.05). The fluorescent coloration showed that PRP  improved the cell quantity on the material compared with  control group. The flow cytometry  showed that PRP  stimulated the cells to enter the S phage to copy DNA. In the 2nd day after treated with PRP the percent of the cells  in S phage in  PRP group was  higher than that in control group (34.41%  vs  22.00%,P<0.05);In the 8th day the  percent of  the cells  in  G0G1 phage  was  higher than that in control group (95.07%  vs  89.70%,P<0.05 ).The result of  CCK-8 showed  that the  absorbency number in 100.0% PRP group was significantly higher than that in 12.5%group (1.13±0.05  vs  0.75±0.04,P<0.05) in the 4th day. Conclusion High concentration PRP has a positive influence on hDFbs proliferation, but it isn’t dose-dependent. An appropriate concentration PRP can promote the proliferation of hDFbs.

Key words: human dermal fibroblasts;platelet-rich plasma;proliferation;culture techniques;cell cycle

CLC Number: 

  • R446.6