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《Journal of Clinical Rehabilitative Tissue Engineering Research》 2009-28
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Effects of basic fibroblast growth factor transfection on canine gingival fibroblasts

Chen Xin-jian1,Yan Fu-hua1,Zhong Quan1,Zhao Xin1,Jiang Yi-ping2 1Stomatological Hospital Affiliated to Fujian Medical University,Fuzhou 350002,Fujian Province,China; 2Research Center of Cells and Developmental Engineering,Fujian Medical University, Fuzhou 350002, Fujian Province, China  
BACKGROUND:Studies have demonstrated that exogenous basic fibroblast growth factor (bFGF) has intensive effects to promote proliferation of gingival fibroblasts (GFs) cultured in vitro and the healing of gingival wounds. OBJECTIVE:To investigate the effects of bFGF gene transfection on the biological performance of Beagle canine GFs. DESIGN,TIME AND SETTING:An observation and comparison in vitro experiment regarding cells was accomplished in Centre of Cell Biology and Development of Fujian Medical University and Department of Comparative Medicine in Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA from April to September of 2008. MATERIALS:Four beagle dogs,male,12 months old,weighing 10-13 kg were used in this experiment. pIRES2-EGFP-bFGF plasmid containing full-length human bFGF gene cDNA was constructed and conserved by our institution. METHODS:Free gingiva of the 2nd,3rd and 4th premolars were excised from left upper jaw of Beagle dogs,rinsed with aseptic phosphate buffer four times,then cut into pieces and digested with 2.5 g/L pancreatin for 2 hours at 37 ℃. After the centrifugation and supernatant removal,DMEM containing 10% fetal bovine serum was added to incubate on 6-well plate with coverlips in 5% CO2 incubator at 37 ℃. Logarithmically growing cells were digested and passaged. GFs were transfected with pIRES2-EGFP-bFGF plasmid using liposome mediated method,while vacant plasmid transfection and un-transfection group served as controls. MAIN OUTCOME MEASURES:Proliferation and apoptosis feature of the GFs were evaluated by MTT and AOEB,respectively. The activity of alkaline phosphatase was assayed by chemical colorimetry. RESULTS:All of three groups cells entered log phrase on three days after transfection. MTT results showed that the proliferation of GFs transfected with bFGF was greater than cells transfected with vacant vector and untransfected cells (P 0.05). AO/EB dyeing showed the apoptosis rate of GFs transfected with bFGF was reduced compared with other two groups (P 0.05). After bGFG gene transfection,the ALP activity remained unchanged and there was no significant difference compared with untransfected cells. CONCLUSION:The transfection of bFGF gene to GFs can promote the proliferation of GFs and depress the apoptosis. No promotion is present with regard to the GFs differentiation.
【Fund】: 国家自然科学基金资助项目(34071892);; 福建省科技厅、福建医科大学科学研究发展基金资助项目(XZ04011)~~
【CateGory Index】: R780.2
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