烟草浸提液对大鼠体外培养成骨细胞的影响.docVIP

烟草浸提液对大鼠体外培养成骨细胞的影响 作者：王桂龙 侯玉东 石玲波 【摘要】 目的 探索烟草浸提液对体外培养成骨细胞的附着和生长的影响。方法 将周龄SD大鼠颅盖骨，剪成1.5 mm×1.5 mm大小组织块，先用0.25％胰蛋白酶消化20 min、0.1％Ⅱ型胶原酶消化30 min后，再对其进行培养。倒置显微镜观察细胞形态，并对其碱性磷酸酶(ALP)活性及矿化能力进行鉴定。实验分对照组和实验组，实验组按所加烟草浸提液浓度的不同分为0.8、1.6、3.2、6.4、12.5、25和50 g／L 7个浓度组，分别测定各实验组细胞的生长曲线。采用SPSS13.0软件包对数据进行双因素方差分析。结果 烟草浸体液对成骨细胞的附着和生长起抑制作用，并随其浓度的升高而增强。各处理组间存在显著差异（P＜0.01）；各组内的前3 d存在显著性差异（P＜0.01），但3.2～50 g／L组第4～7天间比较差异无统计学意义（P＞0.05）。结论 吸烟不利于口腔种植的临床愈合。 【关键词】 成骨细胞；原代培养；SD大鼠；烟草浸提液 【Abstract】 Objective To investigate the effects of smokeless tobacco extract(ST) on the attachment and growth of the rat’s osteoblasts in vitro.Methods The bone tissues from the calvaria in rats aged 1 week were cut into blocks of 1.5 mm × 1.5 mm.They were digested by 0.25％ trypsinase for 20 minutes and by 0.1％ collagenase II for 30 minutes and then the liquid supernatant was abandoned. The bone blocks were cultured to acquire isolated cells.The identification was conducted by cell morphology,histochemical staining of alkaline phosphatase (ALPase)and calcified nodules staining.The experiments were divided into control group and experiment groups.The experiment groups were divided into seven groups by the smokeless tobacco extract’s concentration: 0.8,1.6,3.2,6.4,12.5,25 and 50 g／L, then the cell growth curves were drawn. Statistical analysis was performed using SPSS13.0 software package for twoway ANOVA.Results ST inhibited attachment and growth of osteoblasts in all groups with a concentrationdependent manner. The difference among all of groups was significant（P＜0.01）; the difference between the same group was significant before 3 days（P＜0.01）, while the difference among 3.250 g／L groups was no significant after 4 days（P＞0.05）.Conclusion Smoking is harmful to the clinic healing of oral implant operation. 【Key words】 osteoblast,primary culture,rats,smokeless tobacco extract 目前，口腔种植技术因其独特的优势而获得了临床医师及患者的青睐，而骨结合则是其成功的基础。骨组织作为一种特殊的钙化组织，主要由成骨细胞完成并受体内多种因