The Assay Of Bfgf In Vitreous After Cryocoagulation

Objective To investigate the relationship of cryocoagulation and basic fibroblast growth factor(bFGF) and observe the effect and possible mechanism of bFGF in proliferative vitreoretinopathy(PVR) by measuring the assay of bFGF in vitreous of patients with primary rhegmatogenous retinal detachment (the Control Group) and patients with recurred rhegmatogenous retinal detachment who had been undergone cryocoagulation(the Experimental Group).We expect to decrease the rate of PVR and recurred rhegmatogenous retinal detachment,and finally improve the retinal resetting rate.Methods Analyze records of 62 patients with rhegmatogenous retinal detachment who hospitalized in our department during the period between March,2009 and July,2010. 28 cases in the experimental group who underwent recurred rhegmatogenous retinal detachment applied scleral buckling associated with Cryocoagulation. 34 cases in the control group with rhegmatogenous retinal detachment applied vitrectomy for the first time. All the cases were taken primary vitreous liquid before vitrectomy.And got supernate after centrifugate. Then a two-site enzyme-linked immunosorbent assay was applied to quantitate the level of bFGF in 34 cases'(34 eyes) vitreous of control group and 28 cases (28 eyes) of experimental group.There are purified bFGF standard substances in the kit.The monoclonal antibody which is specific to bFGF situates at the subface of the ELISA pick-up plate. Add the sample or standard substance into the micropore which peridium the monoclonal antibody.Then bFGF combines with monoclonal antibody and bFGF antibody which are tagged by Horseradish Peroxidase.Then produces antibody-antigen enzyme labelled antibody compounds.They are chromogenic after thorough washing with 3,3',5,5'-Tetramethyl benzidine.Under the catalysis of HRP enzyme,TMB translate into blue color and finally yellow color with the effect of acid.Apply ELISA microwell plate spectrodensitometry instrument to measure the Optical Density Value of the sample under light-wave of 450nm.The deeper the color is,the larger OD value is and higher content of bFGF.Use bFGF standard sample making a standard curve whose density is from 0~40ng/L and homologous with its OD value.Compare each OD value of the sample with the standard curve or the linear regression equation and calculate the content of bFGF in the sample.Results 1.The bFGF level in vitreous: the Experimental Group: PVR B:(7.99±1.41)ng/L,PVR C:(12.18±3.01)ng/L,PVR D:(19.45±4.10)ng/L;the Control Group: PVR A:(3.57±1.28)ng/L ,PVR B:(5.51±1.07)ng/L,PVR C:(8.32±1.45)ng/L,PVR D:(13.55±3.08)ng/L; 2.The distinction between the same PVR(above PVR B) level in Experimental Group and Control Group had statistical significance(P﹤0.05).And bFGF contents of experimental group are all higher than the control group; 3.The distinction between PVRB and PVR C, PVR C and PVR D in Experimental group has statistical significance(P=0.003<0.01, P=0.014<0.05). 4.The corrected vision when discharged from hospital:24 cases(85.7%)improved in experimental group,3 cases(10.7%)improved 2~3 lines,21 cases (75%)lower than 2 lines and the best corrected vision is 0.4;In the control group, 29 cases(85.3%)improved in experimental group,5 cases(14.7%)improved 2~3 lines,24 cases(70.6%)lower than 2 lines and the best corrected vision is 0.8. 5.The anatomical reduction rate:25 cases(89.3%)in experimerntal group and 32 cases(94.1%)in control group acquire the reattachment of retina;The final rate of the reattachment of retina:27 cases(96.4%)in experimental group and 33 cases(97.1%)in the control group;The distinction of the anatomical reduction rate between the two groups has no statistical significance(P>0.05).Conclusions Cryocoagulation could increase the bFGF content in vitreous;bFGF has close relationship with the severity degree of PVR.