School of Pharmaceutical Sciences, Jilin University, 1163 Xinmin Street, Changchun, Jilin Province, 130021, People's Republic of China.
China-Japan Union Hospital, Jilin University, Changchun, People's Republic of China.
Biol Trace Elem Res. 2020 Jan;193(1):226-233. doi: 10.1007/s12011-019-01684-9. Epub 2019 Mar 14.
Fluoride accumulates and is toxic to bones. Clinical bone lesions occur in a phased manner, being less severe early in the natural course of skeletal fluorosis. Previous research rarely focused on osteocyte, osteoclast, and osteoblast at the same time, although these three types of cells are involved in the process of fluorosis. In this study, commitment of bone cells was performed according to their respective characteristics. Osteocyte-like cells were verified by protein expression of sclerostin (SOST) in IDG-SW3 cell culture with mineral medium. Positive tartrate-resistant acid phosphatase (TRACP) staining, characteristic of osteoclasts, is observed in RAW264.7 cells after administration of RANKL. We successfully purified a high percentage (94%) of bone mesenchymal stem cells (BMSCs) co-expressing CD34 and CD44. Parallel studies were performed to observe cell viability and apoptosis rates in osteocyte, osteoclast, and osteoblast like cells by using MTT and Annexin V FITC assays. Our results demonstrated that osteocytes have a strong tolerance to high fluoride concentrations, while osteoclasts are more sensitive to changes of fluoride dose. The range of anabolic action of fluoride concentration on osteoblast was narrow. Notably, fluoride exposure aggravated apoptosis of osteocyte and osteoclast induced by administration of PTH and TGF-β, respectively. In short, three types of bone cells display disparate responses to fluoride exposure and to PTH- and TGF-β-induced apoptosis.
氟化物会在体内积累并对骨骼产生毒性。临床骨骼病变呈阶段性发生,在氟骨症自然病程的早期,病变程度较轻。既往研究很少同时关注成骨细胞、破骨细胞和成骨细胞,尽管这三种细胞都参与了氟中毒过程。在这项研究中,根据各自的特点对骨细胞进行了定向诱导。在 IDG-SW3 细胞矿化培养基中培养的成骨细胞样细胞通过骨钙素(SOST)的蛋白表达得到验证。在给予 RANKL 后,RAW264.7 细胞中观察到抗酒石酸酸性磷酸酶(TRACP)染色阳性,这是破骨细胞的特征。我们成功地纯化了 94%高表达 CD34 和 CD44 的骨髓间充质干细胞(BMSCs)。通过 MTT 和 Annexin V FITC 检测,我们平行研究了成骨细胞、破骨细胞和成骨细胞样细胞的细胞活力和细胞凋亡率。我们的结果表明,成骨细胞对高氟浓度具有较强的耐受性,而破骨细胞对氟剂量变化更为敏感。氟浓度对成骨细胞的合成作用范围较窄。值得注意的是,氟暴露加重了 PTH 和 TGF-β分别诱导的成骨细胞和破骨细胞凋亡。总之,三种类型的骨细胞对氟暴露和 PTH 和 TGF-β诱导的凋亡的反应不同。