赖灵妍, 贾莹, 丁雪, 胡笳, 陈波. 氟染毒SD大鼠不同阶段颌骨内部环境和抗剪切力力学特性的变化[J]. 环境与职业医学, 2023, 40(1): 95-100. DOI: 10.11836/JEOM22253
引用本文: 赖灵妍, 贾莹, 丁雪, 胡笳, 陈波. 氟染毒SD大鼠不同阶段颌骨内部环境和抗剪切力力学特性的变化[J]. 环境与职业医学, 2023, 40(1): 95-100. DOI: 10.11836/JEOM22253
LAI Lingyan, JIA Ying, DING Xue, HU Jia, CHEN Bo. Time-course changes of intraosseous environment and shear resistance mechanical properties of jawbone in SD rats with fluorosis[J]. Journal of Environmental and Occupational Medicine, 2023, 40(1): 95-100. DOI: 10.11836/JEOM22253
Citation: LAI Lingyan, JIA Ying, DING Xue, HU Jia, CHEN Bo. Time-course changes of intraosseous environment and shear resistance mechanical properties of jawbone in SD rats with fluorosis[J]. Journal of Environmental and Occupational Medicine, 2023, 40(1): 95-100. DOI: 10.11836/JEOM22253

氟染毒SD大鼠不同阶段颌骨内部环境和抗剪切力力学特性的变化

Time-course changes of intraosseous environment and shear resistance mechanical properties of jawbone in SD rats with fluorosis

  • 摘要: 背景

    目前关于氟骨症的研究大部分集中在四肢骨、躯干骨,有关颌骨损害的研究甚少。颌骨是口腔疾病发生和治疗不可避免的结构基础。

    目的

    通过慢性氟染毒观察大鼠颌骨内环境与最大抗剪切力载荷(LSFmax)变化,初步探讨氟染毒对颌骨力学性能的影响。

    方法

    筛选实验:选取48只SD雄性大鼠随机分成对照组及50、150、250 mg·L−1染氟组,每组12只。染氟组通过喂饲不同浓度的氟化钠水溶液进行造模,对照组饮用贵州地区自来水。再根据饲氟周期(观察时间点0、2、4及6个月)分为4个亚组,每组3只。利用电子万能力学试验机测定颌骨LSFmax,天狼星红染色及免疫组化半定量分析I型胶原(Col1)、骨形成相关因子Runt转录因子2(Runx2)。正式实验:选取12只SD雄性大鼠随机均分为染氟组及对照组,染氟组大鼠以150 mg·L−1氟化钠水溶液饲氟5个月,对照组饮用贵州地区自来水。利用电子万能力学测试仪测定颌骨LSFmax,利用抗酒石酸酸性磷酸酶(TRAP)染色对破骨细胞计数,利用天狼星红染色及免疫组织化学法半定量检测Col1、Runx2、骨形态发生蛋白2(BMP-2)、碱性磷酸酶(ALP)、组织蛋白酶K(Cath k)表达,利用微计算机断层扫描技术(Micro CT)观测骨小梁微结构。

    结果

    筛选实验:对照组及50 mg·L−1染氟组LSFmax在第2个月达峰值,50 mg·L−1染氟组LSFmax在第4个月达谷值;150 mg·L−1染氟组在第4个月LSFmax高于第6个月(P<0.05);250 mg·L−1染氟组在各时间点LSFmax波动均无统计学意义。同一时间点,各组间LSFmax比较未表现具有统计学意义的差异。50 mg·L−1、150 mg·L−1及250 mg·L−1染氟组Col1从第2个月开始均高于0时间点(P<0.05)。Runx2随浓度、时间变化未表现具有统计学意义的差异。正式实验:以150 mg·L−1氟浓度饲氟5个月后,大鼠LSFmax大于对照组(P<0.05);Col1、Runx2、BMP2、ALP、Cath K表达均高于对照组(P<0.05);破骨细胞计数和骨小梁微结构指标变化未表现具有统计学意义的差异。

    结论

    慢性氟染毒能够增强颌骨抗剪切力学性能。

     

    Abstract: Background

    At present, a large number of reports focus on the bones of limbs and trunk, while there are few studies on the effect of fluorosis on jawbone which is the inevitable structural basis for the development and treatment of oral diseases.

    Objective

    To preliminarily investigate the effect of fluoride exposure on the mechanical properties of jawbone by observing the changes in the intraosseous environment and the maximum load against shearing force (LSFmax) of the jawbone in rats with chronic fluoride treatment.

    Methods

    Screening experiment: 48 SD male rats were randomly divided into a control group and three fluoride exposure groups (50, 150, and 250 mg·L−1 fluoride concentration), 12 rats in each group. The fluoride exposure groups were molded by feeding different concentrations of sodium fluoride solution, and the control group drank tap water from Guizhou area. Each group was further divided into 4 subgroups with 3 animals each according to observation time points after 0, 2, 4, and 6 months. The LSFmax of the jawbone was measured with an electronic universal ergometer, the expression of type I collagen (Col1) was shown by Sirius red staining, and the expression of runt-related transcription factor 2 (Runx2) was determined semi-quantitatively by immunohistochemistry at selected time points. Formal experiment: 12 male SD rats were randomly divided into a fluoride exposure group and a control group. The fluoride exposure group were fed with 150 mg·L−1 sodium fluoride solution, and the control group drank tap water from Guizhou. After feeding with fluoride for 5 months, the ergometer was used to measure the LSFmax of the jawbone. Osteoclasts were counted after tartrate resistant acid phosphatase (TRAP) staining. Col1, Runx2, bone morphogenetic protein 2 (BMP-2), alkaline phosphatase (ALP), and cathepsin K (Cath K) were detected semi-quantitatively by immunohistochemistry expression and Sirius red staining. Micro computed tomography (Micro CT) was used to observe the trabecular bone microstructure.

    Results

    Screening experiment: The LSFmax of the control group and the 50 mg·L−1 fluoride exposure group reached the peak value at the 2nd month, and the LSFmax of the 50 mg·L−1 fluoride exposure group reached the valley value at the 4th month. The LSFmax of the 150 mg·L−1 fluoride exposure group at the 4th month was higher than that at the 6th month (P<0.05). There was no significant difference in the LSFmax at each time point in the 250 mg·L−1 fluoride exposure group. At the same time point, there was no statistically significant difference in LSFmax among the groups. The Col1 levels of the 50 mg·L−1, 150 mg·L−1, and 250 mg·L−1 fluoride exposure groups were higher than the time point 0 from the 2nd month (P<0.05). The Runx2 showed no statistically significant difference by concentration or time. Formal experiment: After feeding with 150 mg·L−1 fluoride for 5 months, the LSFmax of the fluoride exposure group was greater than that of the control group (P<0.05). The expressions of Col1, Runx2, BMP2, ALP, and Cath K in the fluorosis exposure group were higher than those in the control group (P<0.05). There were no statistically significant differences in osteoclast count or indicators of bone trabecular microstructure.

    Conclusion

    Chronic fluoride exposure may increase the shear strength of jaw bone.

     

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