Investigation Of Aminolevulnilic Acid Photodynamic Therapy On Proliferation And Apoptosis Of Fibroblast From Keloid

Objective:⑴To establish a reliable cultivation method for human keloid fibroblast（sKFB）andstudy the biological quale of the cells, provide a plat for in vitro reserch of keloid⑵Investigation ofALA-PDT on Proliferation of KFB,so that the mechanism whichALA-PDT affect KFB can be disclose to provide the theory base forcliniealprevention and treatment on keloid.⑶To investigate the effect ofALA-PDT on apoptosis of KFB and the caspase3、8、9protein levels irradiated by PDT in KBF.Analysis of the important apoptosispathways.Methods:⑴Primary culture of human keloid fibroblasts was performed with routine tissueblock adhering to the plate wall, study the biological quale of the cells,and KFB wereidentified by immunocytochemical staining.⑵KFB treated with ALA at different concentrations（0.5～8.0mmol/l）for5hourswere irradiated by red laser with energy density from1to64J/cm2，and detectedKFB survival after ALA-PDT by CCK8method.⑶KFB treated with ALA at4.0mmol/l concentration for5hours were irradiated byred laser with energy density at32J/cm2,then investigate the caspase3、8、9protein levels irradiated by PDT in KFB.Results:⑴KFB outgrow from tissue after5days, the cells have positive vimentin expressionby immunocytochemical staining and have mass tan granule in endochylema and bluecell nucleus.⑵Along with the ALA concentration and laser energy increase, the cells of theinhibition is also enhanced when the energy exceed8J/cm2. ⑶ALA-PDT can obviously increase the activation of caspase3、8、9protein levelsand increase the level of expression.Conclusions:⑴Tissue adherent method is a simple operation, low cost and can obtain highactivity, high purity of KFB. It can be used as a keloid in vitro experimentalresearch.⑵ALA-PDT can obviously inhibit the proliferation of KFB. Concentration and laserenergy are important factors affecting cell survival.⑶ALA-PDT can obviously induce apoptosis of KFB that is closely related with theactivity of caspase3、8、9protein. It indicate that Fas signaling pathway andmitochondrial signaling pathway might jointly participate in the process of apoptosis.