Effects Of Cigarette Smoke Condensate And Nicotine On Growth Status And Cytokine Expression Of Human Gingival Fibroblast On Titanium Plate

Objective:With the development of dental implant technology and biomaterials,dental implant has become the first choice with dentition defect and dentition defect.The success of implant needs not only good osseointegration,but also the establishment of a stable functional implant soft tissue in the early stage.Smoking and periodontal pocket formation,attachment loss,bone loss and tooth loss are closely related,this study of human gingival fibroblasts were cultured on the titanium plate,by observing the effect of smoke condensation extraction(CSC)and nicotine to fiber cells of titanium material surface on gum and to explore the mechanism of implant failure theory of tobacco root interface.Methods:HGFs were cultured using tissue-explant technique.Cellular morphology and cell growth were observed and photographed under inverted microscope.The third passage human HGFs were taken to characterize the cell lineage by immunocytochemical staining and measure cells growth curve.The 1 mm thickness,diameter 10 mm titanium plate was polished by metallographic sandpaper level by level.Titanium plate surface morphology was observed by Stereo microscope.surface roughness of titanium plate was observed by surface roughness measuring instrument and define roughness with Ra value.The number of cell was tested by MTT of titanium plate surface RT-PCR to evaluate the mRNA level of fibronectin.Results: 1.SEM showed nicotine would contribute to cell death in 16ug/ml while CSC in 250ug/ml and improve cell viability.2.CSC and nicotine would stimulate cell spread in 50ug/ml and 3.21ug/ml respectively.Besides that,CSC restrained cell spread in 150ug/ml,but nicotine did not show similar result with same concentration.Both could speed ?-actin production and CSC performed better.3.CSC would hold-up cell spread and survival.In addition,CSC might interfere myofibroblasts differentiation.CSC could significantly change gingival mesenchyme viability,spread and fibroblast differentiation.When removed stimulator,cell showed no difference to normal cell.4.After being cultured for 2h,cell adhesion was evaluated by MTT and colorimetric analysis was performed at 492 nm.CSC and nicotine would restrain HGF adhesion in titanium,and with increasing of CSC and nicotine concentration,lower and lower cell would adhere to titanium disk.Conclusions:MTT test evaluated nicotine and CSC impact to HGFs adhesion and nicotine would reduce HGFs adhesion.ELISA and RT-PCR were performed to access the nicotine and CSC effect to Fn synthesis by HGFs.The result confirmed that nicotine and CSC held-up Fn synthesis by HGFs in concentration-dependent manners.With increasing of 2 stimulators' concentration,Fn synthesis decreased correspondently.Therefore,it is illustrated that nicotine concentration would affect HGFs growth,aggravate periodontitis and jeopardize periodontium attachment.