Inhibition of human scar fibroblast cells growth by margarita liquid / 中国组织工程研究

ABSTRACT

BACKGROUND:Study shows that margarita liquid has effect on promoting histiocyte regeneration and removing scars.OBJECTIVE:To observe the effects of margarita liquid on the proliferation of fibroblasts in human skin scar tissues.DESIGN,TIME AND SETTING:A grouping contrast observational experiment was performed in the Experiment Centre of Guangxi Medical University from September to December in 2008.MATERIALSScar tissue samples were obtained from patients in the Department of Plastic Surgery,Guangxi Medical University.Margarita liquid was the digest of margarita purchased from Beihai Gofar Marine Biological Industry Co.,Ltd and rich in multi-amino acids,polypeptides,vitamin,mineral matters and natural enzymes.METHODS:Human skin scar cell line was established by removing non-fibroblasts through repeated primary culture and serial subcultivation of scar fibroblasts with reference to Veelken method. The 3-5 generation human skin scar fibroblasts on exponential phase of growth were made into single cell suspension by trypsin digestion which was then inoculated on plastic 96-well cell culture plate,with the density of 0.5×10~4/well as well as 100 μL cell suspension and 100 μL DMEM medium in each well. After culture for 24 hours,primary medium was discarded. The grouping of the experiment200 μL mediums with 125.00,62.50,31.30,15.60,7.80,3.90,1.95,0.98 mg/L margarita liquid were used respectively in margarita liquid group;200 μL pure medium was added into each well in control group.MAIN OUTCOME MEASURES:MTT and TUNEL assay were used to examine the proliferation and the apoptosis of fibroblastsrespectively.RESULTS:The 50% inhibiting concentration (IC_(50)) of margarita liquid on fibroblasts was 15 mg/L. Margarita liquid at any other concentration but 0.98 mg/L was effective in inhibiting fibroblast growth in a dose-dependent way,i.e. the higher margarita liquidconcentration,the higher inhibition ratio on fibroblast growth. Fibroblasts cultured with 15 mg/L (IC_(50)50) margarita liquid had got reduced volume,lessened cytoplasm,decreased density,increased apoptosis rate and buffy colour. Fibroblasts in control group were large,rich in cytoplasm and compact. Apoptotic index was higher in the margarita liquid group than in the blank control group CONCLUSION:Margarita liquid could inhibit the proliferation of skin scar fibroblasts cultured in vitro and induce the apoptosis of them.