The Study Of Celecoxib On The Expression Of MMP-2,MMP-9on Corneal Neovascularization After Thermal Burn In Rabbit

Objective：1. To observe the effect of celecoxib on the growth of cornealneovascularization after burn and to explore the role of celecoxib in theinhibition of corneal neovascularization.2. To compare effects of amniotic membrane transplantation andcelecoxib to corneal neovascularization after thermal burn,provide atheoretical basis for the clinical u se of celecoxib treatment of cornealneovascularization.3. To evaluate topical use of celecoxib on the expression of MMP-2andMMP-9in corneal neovascularization secondary to thermal burn,explore thepossible mechanism of corneal neovascularization inhib ition.Methods：1.We randomly divided28healthy New Zealand white rabbits to fourgroups (A,B,C,D):group A,control group of neovascularization (n=8);groupB,celecoxib group (8mg,n=8);group C,celecoxib group (12mg,n=8);Dgroup,control group (n=4).Neovascul arization models were developed by rabbitcorneal burn in three groups (A,B,and C).Histological morphology,growthcondition and area of corneal neovascularization among these groups werecompared at day4,day7and day14under slit lamp microscope.Softwar epackage SPSS19.0were used for statistical analysis.Numerical data wasdescribed as X±S.The areas of blood vessels at each time point were comparedby analysis of variance.Statistical significance was set at α=0.05.2. We randomly divided24healthy New Zealand white rabbits to threegroups:group A,control group of neovascularization;group B,celecoxib group(n=8);group C,amniotic membrane transplantation group (n=8).Neovascularization models were developed by rabbit corneal burn inthree groups (A,B,and C).Histological morphology,growth condition and areaof corneal neovascularization among these groups were compared at day15and day30under slit lamp microscope.Software package SPSS19.0were usedfor statistical analysis.Numerical data was described as X±S.The areas ofblood vessels at each time point were compared by analysis of variance.Statistical significance was set at α=0.05.3. We randomly divided44healthy New Zealand white rabbits to threegroups:group A,control group of neovascularization (n=20);group B,celecoxibgroup (n=20);group C,control group (n=4).Neovascularization models weredeveloped by rabbit corneal burn in groups A and B.Rabbits were sacrificed atday1,day4,day7,day14and day28.The cornea was extracted and divided totwo equal parts,one part for fixation in neutral formalin and another forhomogenization to detect MMP-2,MMP-9in the supernatant.Software packageSPSS19.0were used for statistical analysis.Numerical data was describedas X±S.Concentrations of MMP-2and MMP-9were compared by students’test.The areas of blood vessels at each time point were compared by analysisof variance,and Pearson correlation by coefficient analysis.Statisticalsignificance was set at α=0.05.Results：1.On4,7,14days after the establishment of models,the areas of newblood vessel were (X±S):（11.32±1.11） mm2、(38.49±4.64)mm2、(43.30±4.39)mm2in neovascular control group;(9.69±1.30)mm2、(31.15±4.85)mm2、(37.19±5.27)mm2in celecoxib group of8mg;(8.33±1.23)mm2、(30.31±4.93)mm2、(36.69±3.54)mm2in celecoxib groupof12mg.No neovascularization was found in normal group.There wasstatistically significant difference among the three groups.Neovascularizationarea in celecoxib group of8mg or12mg was significantly lower than controlgroup (P<0.05).There was no difference between the two celecoxib group (P>0.05) but statistically significant difference among three groups at di fferenttime points (P<0.05).2.On15and30days after model establishment,the areas of new blood vessel were (X±S):（43.22±4.35） mm2、(37.53±4.57)mm2in neovascularcontrol group;36.80±3.60） mm2、(28.61±3.93)mm2in celecoxib group;（37.07±5.2）mm2、(30.43±5.17)mm2in amniotic membrane transplantationgroup.There was statistically significant difference among the threegroups.Neovascularization area in celecoxib group and amniotic membranetransplantation group was significantly lower than control group(P<0.05).There was no difference between these two groups (P>0.05) butstatistically significant difference among three groups at different time points(P<0.05).3.At different time point,MMP-2concentrations in the neovascularcontrol group were （19.687±7.583） μg/L、（40.728±4.258） μg/L、（101.314±12.695）μg/L、（83.724±9.848）μg/L、（34.986±7.318）μg/L andMMP-9concentrations were（15.137±3.806） μg/L、（32.583±7.513） μg/L、（76.798±8.495）μg/L、（53.063±8.248）μg/L、（34.596±7.469）μg/L;MMP-2concentrations in the celecoxib group were （13.610±5.527） μg/L、（34.505±10.201）μg/L、（78.546±15.042）μg/L、（59.460±4.826）μg/L、（29.291±5.668）μg/L and MMP-9concentrations were（13.016±2.752）μg/L、（22.759±5.005）μg/L、（56.932±7.823） μg/L、（36.266±5.024） μg/L、（23.876±5.917）μg/L.There was statistically significant difference between MMP-2concentrations in the two groups (F=42.093， P=0.000),at different time point(F=181.405， P=0.000) and significant interaction effect between time andlocation (F=4.598， P=0.002);MMP-9concentrations in the two groups werestatistically different (F=66.889， P=0.000) as well as at different time points(F=154.135， P=0.000) and significant interaction effect between time andlocation (F=4.484， P=0.003).Positively correlation in between was found(r=0.902， P=0.000).Conclusions:1.Topical use of celecoxib has the role of neovascularization inhibitionafter corneal thermal burn.2.There was no significant difference between topical use of celecoxiband amniotic membrane transplantation for neovascularization inhibition af tercorneal thermal burn. 3.The mechanism of angiogenesis inhibition after topical use ofcelecoxib for corneal thermal burn may be associated with the activation andexpression of MMP-2and MMP-9.