| Objective:To investigate the effects of titanium spcimens with different surface character on cell proliferation and cytoskeleton spreading and mRNA expression of IL-6 and Cbfal in osteoblasts.Methods:The morphology and the elements analysis of the samples were inspected and detected by Scanning Electronic Microscopy and Energy Dispersive Spectrometer,the surface roughness and contact angle of the samples were measured by surface roughness measuring instrument and contact angle meter.MC3T3-El Sub-clone 14 cells were cultured on 3 different commercially pure titanium smooth pretreatment(PT),sandblasted and acid etched(SLA),anodic oxidation(AD) surfaces and cells were cultured on tissue culture polystyrene(TCPS) as control group. The proliferation was measured by MTT assay. The F-actin cytoskeleton of MC3T3 was evaluated by confocal laser scanning microscope after cultured with various concentrations of STE after 1.5 h and 6 h. The mRNA expression of IL-6 and Cbfal were quantified by quantitative real-time PCR.Results:Scratches were observed on PT samples'surfaces, multiple three dimensional structure were observed on SLA samples'surfaces,the AD samples exhibited porous structure on their surfaces.The elements of the 3 groups of spcimens were Ti;Ti,Al;Ti,O.The surface roughness of the 3 groups of spcimens were 0.18±0.01um,2.04±0.06um,0.18±0.02um(p<0.05,SLA vs. PT and AD),the contact angle of the 3 groups of spcimens were 54.47±3.33°,75.42±8.32°,38.91±4.00° (P<0.05).In regard to cell proliferation,the number of MC3T3 on the TCPS was the highest on every time point and the least on the SLA at the same time(P<0.05),AD group could promote cell proliferation relative to PT and SLA groups. After 1.5 h,F-actin network structure could barely seen on each group,more synapses could seen on SLA group,after 6 h,all the 3 groups cytoskeleton had started spread,the AD group showed a network structure.All the 3 groups of titanium spcimens could decrease mRNA expression of IL-6 and increase mRNA of Cbfal in MC3T3 cell lines(P<0.05) AD group could decrease mRNA expression of IL-6 and increase mRNA of Cbfal relative to PT and SLA groups and the same outcome was found in SLA group relative to PT group(P<0.05).Conclusions:Titanium spcimens with AD treated showed more obvious effects on promoting cell proliferation,decreasing mRNA espression of IL-6 and increasing mRNA of Cbfal in MC3T3 cell lines than other groups.Implats with AD treated might have some advantages in early osseointegration.Objective:To investigate the effects of different concentrations of smokeless tobacco extract on biological properties from osteoblast in order to identify possible pathological mechanisms that smoking affects osseointegration.Methods:MC3T3-E1 Sub-clone 14 cells were cultured in the presence of STE at 0 (control group),0.01,0.1,1,5,10 g/L. After 1 d,3 d,5 d,7 d,the proliferation was measured by MTT assay.The F-actin cytoskeleton of MC3T3 was evaluated by confocal laser scanning microscope after cultured with various concentrations of STE after 24 h.The mRNA expression of interleukin-6(IL-6) and core-binding factor al(Cbf?1) were quantified by quantitative real-time PCR(RT-qPCR) after cultured with various concentrations of STE after 48 h.Results:The MTT assay results showed that STE could inhibit MC3T3 proliferation at the concentrations of 0.01-10 g/L(P<0.05))5-10 g/L of STE could inhibite MC3T3 proliferation as the time prolonged(P<0.05). The cells'F-actin cytoskeleton in control group showed network structure and in the STE treatment groups F-actin cytoskeleton started changes and showed disruptive structures, as the concentration of STE increased, more significantly changes were observed. The STE increased the mRNA expression of IL-6 at the concentrations of 5 g/L and 10 g/L(P<0.05), decreased the mRNA expression of Cbfal at the concentrations of 0.1 g/L-10 g/L(P<0.05).Condusion:Tobacco may inhibit osteoblast proliferation, destroy F-actin cytoskeleton structure, increase the mRNA expression of IL-6 and decrease the mRNA expression of Cbfal,and inhibit cell differentiation and adhesion accordingly. We suggest that smoking is a disadvantage to osseointegration. |
PDF Full Text Request