Experimental Study On Comparison Of Two Kinds Of Platelet Rich Fibrin On The Proliferation Speed And Quality Of Human Gingival Fibroblasts

Objective:By comparing two kinds of platelet rich fibrin,advanced platelet rich fibrin and chrounkron‘s platelet rich fibrin,cultured in vitro with the human gingival fibroblast,to discover its influence on the cell's growth speed and quality.Analysis the advantages and disadvantages of the two biological materials which from autologous blood.To provide a theoretical basis for the clinical choice of material to repair oral soft tissue defects.Methods:1.The obtation and authentication of the humans gingival fibroblast :collect 8 gingival samples from the vonlunteer who need to extract the third molar.Then cultured the samples in MEM to get the primary generation cells.The vimentin and cytokeratin antibodies were applied to identify the third generation fibroblasts with Immunohistochemistry.2.Preparation the A-PRF and PRF membrance:draw blood to the specific vitro from the vonlunteers,then centrifuged in 1500 rpm,14min and 2700 rpm,12min in centrifuge to get the PRF gel.Do the standard way to get the membrance,and longitudinal cut them into four pieces,weight them to the same.3.Separate all into 3 groups,while setting the cell concentration to 3*104/ml.A-PRF Group :flat the A-PRF membrance on the bottom of culture plate,add 1000 ul cell suspension on it.PRF Group :flat the PRF membrance on the bottom of culture plate,add 1000 ul cell suspension on it.Blank group :add 1000 ul cell suspension.4.Detect the cell proliferation on 1,3,5,7day with CCK-8.5.Detect the COL1A1 on 1,3,5,7day with RT-PCR.6.Detect the structure of the membrance on 1,3,5,7day.7.Statistical analysis the result.Results: 1.Cell culture:8 samples were obtained and cultured well.On the 7day of tissue culture,some cells immigrated from the tissue block.They were in clear boundary.On the 15 day it can be observed lots cells dissociated into a radial shape.2.Immunocytochemical assay was used to observe the expression of vimentin and cytokeration protein.3.CCK-8 assay showed no Statistical significance on the first day;on the third day,A-PRF group and PRF group had significant difference(P < 0.05)APRF group,PRF group and blank control group had significant difference(P < 0.05),on the fifth day,A-PRF group and PRF group had significant difference(P<0.05),PRF group and blank control group had significant difference(P < 0.05),A-PRF group and blank control group had specific significant difference(P < 0.01),on the seventh day,A-PRF group and blank control group had significant difference(P < 0.05),PRF group and blank control group had specific significant difference(P <0.01),A-PRF group,and PRF group had no significant difference(P >0.05).4.PCR to test the COL1A1: showed no Statistical significance on the first day(P > 0.05);on the third day,A-PRF group,and PRF group had significant difference(P < 0.05),A-PRF group,PRF group and blank control group had significant difference(P < 0.05),on the fifth day,A-PRF group,PRF group and blank control group had significant difference(P < 0.05),PRF group and blank control group had no significant difference(P>0.05),on the seventh day,A-PRF group,PRF group and blank control group had significant difference(P < 0.05),PRF group and blank control group had no significant difference(P > 0.05),PRF group and blank control group had specific significant difference(P < 0.01),A-PRF group,and PRF group had no significant difference(P>0.05).5.SEM observation :the fibroblast were in better condition on the surface of A-PRF than PRF.The structure of A-PRF membrance was more steady,and showed less changes,with a strong clot.The cells connected with each other and growing into lamellar.