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Nicotine Increases Osteoblast Activity of Induced Bone Marrow Stromal Cells in a Dose-Dependent Manner: An in vitro Cell Culture Experiment

机译:尼古丁以剂量依赖性方式增加诱导的骨髓基质细胞的成骨细胞活性:体外细胞培养实验

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摘要

Previous studies by our group showed that nicotine delivered via a transdermal nicotine patch significantly enhanced posterior spinal fusion rates in rabbits. Nicotine transdermal patches provide a steady serum level; there may be a dose-dependent effect of nicotine on posterior spinal fusion. In an in vitro cell culture model of rabbit bone marrow–derived osteoblast-like cells, cells were exposed to different concentrations of nicotine (0, 20, 40, 80 ng/mL and 10, 100, 250 μg/mL). Wells were stained with an alkaline phosphatase (ALP) staining kit to determine ALP enzyme activity. Cells were stained with Von Kossa for mineralization. A two-way analysis of variance (ANOVA) using dose and time as variables showed significant differences among groups; post hoc analysis showed that the 100-μg/mL dose of nicotine significantly enhanced ALP activity over controls. A one-way ANOVA using dose as the variable showed that the 100- and 250-μg/mL doses had significantly greater mineralization than controls. Dose-response analysis revealed a statistically significant effect of nicotine dose on ALP activity and Von Kossa activity. The effects of nicotine on spinal fusion may be dose-dependent and due to stimulation of osteoblastic activity. Nicotine may not be responsible for the inhibited bone healing observed in smokers.
机译:我们小组先前的研究表明,通过透皮尼古丁贴剂递送的尼古丁可显着提高兔子的后路脊柱融合率。尼古丁透皮贴剂可提供稳定的血清水平;尼古丁对后路脊柱融合可能存在剂量依赖性作用。在兔骨髓来源的成骨细胞样细胞的体外细胞培养模型中,细胞暴露于不同浓度的尼古丁(0、20、40、80 ng / mL和10、100、250μg/ mL)。用碱性磷酸酶(ALP)染色试剂盒对孔进行染色以确定ALP酶的活性。细胞用Von Kossa染色以矿化。使用剂量和时间作为变量的方差(ANOVA)双向分析显示组之间存在显着差异。事后分析表明,与对照组相比,100μg/ mL剂量的尼古丁可显着增强ALP活性。使用剂量作为变量的单向方差分析显示100和250μg/ mL剂量的矿化度明显高于对照。剂量反应分析显示,尼古丁剂量对ALP活性和Von Kossa活性具有统计学意义。尼古丁对脊柱融合的影响可能是剂量依赖性的,并且是由于刺激了成骨细胞活性。尼古丁可能与吸烟者观察到的骨骼愈合受抑制无关。

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