Sodium selenite promotes osteoblast differentiation via the WNT/β-catenin signaling pathway

Document Type : Original Article


1 Institute for Skeletal Aging & Orthopedic Surgery, Hallym University-Chuncheon Sacred Heart Hospital, Chuncheon-si, 24252, Gangwon-do, Republic of Korea

2 Institute for Skeletal Aging and Orthopedic Surgery, Hallym University-Chuncheon Sacred Heart Hospital, Chuncheon, Korea;Department of Bio-Informatics, School of Computer and Information Sciences, Galgotias University,

3 Institute for Skeletal Aging and Orthopedic Surgery, Hallym University-Chuncheon Sacred Heart Hospital, Chuncheon-si, Gangwon-do, Republic of Korea


Objective: Osteoporosis is regarded as a silent disorder affecting bone slowly, leading to an increased risk of fractures. Lately, selenium has been found to be associated with the acquisition and maintenance of bone health by affecting the bone remodeling process. However, the mechanism of action of selenium on bone is poorly understood. Here, the protective effects of sodium selenite on the differentiation process of osteoblasts as well as under oxidative stress-induced conditions were evaluated.
Materials and Methods: MC3T3-E1 cells, were treated with a various doses (0, 0.1, 0.2, 0.4, 0.8, 1.6, 3.2, 6.4 ug/ml) of sodium selenite. Cell viability and cytotoxicity were observed by MTT and LDH assay. The osteogenic activity and expression level of osteogenic markers were confirmed through ALP activity, real-time RT-PCR and sirius red staining. Role of sodium selenite and involvement of WNT signaling was assessed by Axin-2 reporter assay and western blotting.
Results: It was observed that the sodium selenite could promote the ALP activity and collagen synthesis in pre-osteoblasts. Moreover, sodium selenite increased the mRNA expression levels of osteogenic transcriptional factors, such as runt-related transcription factor 2 (Runx2) and osterix (OSX). In addition, the terminal differentiation markers, such as osteocalcin (OCN), and collagen 1α (Col1α), were also increased. Treatment of sodium selenite recused the H2O2-induced inhibition of osteoblastic differentiation of pre-osteoblasts cells. Furthermore, sodium selenite restored the H2O2 repressed β-catenin stability and axin-2 reporter activity in MC3T3-E1 cells implicating involvement of WNT signaling pathway.
Conclusion: It may be concluded that selenite can stimulate bone formation and rescue the oxidative repression of osteogenesis by activating WNT signaling pathway and may act as a potential therapeutic intervention for osteoporosis.


  1. Rachner TD, Khosla S, Hofbauer LC. Osteoporosis: now and the future. Lancet. 2011; 377(9773): 1276-1287.
  2. Anthamatten A, Parish A. Clinical update on osteoporosis. J Midwifery Womens Health. 2019; 64(3): 265-275.
  3. Reid IR. Short-term and long-term effects of osteoporosis therapies. Nat Rev Endocrinol. 2015; 11(7): 418-428.
  4. Pacifici R. Cytokines, estrogen, and postmenopausal osteoporosis-- the second decade. Endocrinology. 1998; 139(6): 2659-2661.
  5. Zhou X, Yuan W, Xiong x, zhang z, liu j, zheng y, et al. Ho-1 in bone biology: potential therapeutic strategies for osteoporosis. Front Cell Dev Biol. 2021; 9: 791585.
  6. Luo G, Li F, Li X, Wang ZG, Zhang B. TNFalpha and RANKL promote osteoclastogenesis by upregulating RANK via the NFkappaB pathway. Mol Med Rep. 2018; 17(5): 6605-6611.
  7. Manolagas SC. From estrogen-centric to aging and oxidative stress: a revised perspective of the pathogenesis of osteoporosis. Endocr Rev. 2010; 31(3): 266-300.
  8. Sharma AR, Nam JS. Kaempferol stimulates WNT/beta-catenin signaling pathway to induce differentiation of osteoblasts. J Nutr Biochem. 2019; 74: 108228.
  9. Jagga S, Sharma AR, Kim EJ, Nam JS. Isoflavone-enriched whole soy milk powder stimulates osteoblast differentiation. J Food Sci Technol. 2021; 58(2): 595-603.
  10. Ross JA, Kasum CM. Dietary flavonoids: bioavailability, metabolic effects, and safety. Annu Rev Nutr. 2002; 22: 19-34.
  11. Kocijan R, Klaushofer K, Misof BM. Osteoporosis therapeutics 2020. Handb Exp Pharmacol. 2020; 262: 397-422.
  12. Baron R, Kneissel M. WNT signaling in bone homeostasis and disease: from human mutations to treatments. Nat Med. 2013; 19(2): 179-192.
  13. Karner CM, Long F. Wnt signaling and cellular metabolism in osteoblasts. Cell Mol Life Sci. 2017; 74(9): 1649-1657.
  14. Cross CE, Halliwell B, Borish ET, Pryor WA, Ames BN, Saul RL, et al. Oxygen radicals and human disease. Ann Intern Med. 1987; 107(4): 526-545.
  15. Forman HJ, Torres M. Redox signaling in macrophages. Mol Aspects Med. 2001; 22(4-5): 189-216.
  16. Nordberg J, Arner ES. Reactive oxygen species, antioxidants, and the mammalian thioredoxin system. Free Radic Biol Med. 2001; 31(11): 1287-1312.
  17. Liu H, Bian W, Liu S, Huang K. Selenium protects bone marrow stromal cells against hydrogen peroxide-induced inhibition of osteoblastic differentiation by suppressing oxidative stress and ERK signaling pathway. Biol Trace Elem Res. 2012; 150(1-3): 441 450.
  18. Yang Y, Su Y, Wang D, Chen Y, Wu T, Li G, et al. Tanshinol attenuates the deleterious effects of oxidative stress on osteoblastic differentiation via Wnt/FoxO3a signaling. Oxid Med Cell Longev. 2013; 2013: 351895.
  19. Liang D, Xiang L, Yang M, Zhang X, Guo B, Chen Y, et al. ZnT7 can protect MC3T3-E1 cells from oxidative stress-induced apoptosis via PI3K/Akt and MAPK/ERK signaling pathways. Cell Signal. 2013; 25(5): 1126-1135.
  20. Lean JM, Jagger CJ, Kirstein B, Fuller K, Chambers TJ. Hydrogen peroxide is essential for estrogen-deficiency bone loss and osteoclast formation. Endocrinology. 2005; 146(2): 728-735.
  21. Wang Y, Xie D, Li J, Long H, Wu J, Wu Z, et al. Association between dietary selenium intake and the prevalence of osteoporosis: a cross-sectional study. BMC Musculoskelet Disord. 2019; 20(1): 585.
  22. Moreno-Reyes R, Egrise D, Neve J, Pasteels JL, Schoutens A. Selenium deficiency-induced growth retardation is associated with an impaired bone metabolism and osteopenia. J Bone Miner Res. 2001; 16(8): 1556-1563.
  23. Sun JY, Hou YJ, Fu XY, Fu XT, Ma JK, Yang MF, et al. Seleniumcontaining protein from selenium-enriched spirulina platensis attenuates cisplatin-induced apoptosis in MC3T3-E1 mouse preosteoblast by inhibiting mitochondrial dysfunction and ROS-mediated oxidative damage. Front Physiol. 2018; 9: 1907.
  24. Lee SS, Sharma AR, Choi BS, Jung JS, Chang JD, Park S, et al. The effect of TNFalpha secreted from macrophages activated by titanium particles on osteogenic activity regulated by WNT/BMP signaling in osteoprogenitor cells. Biomaterials. 2012; 33(17): 4251-4263.
  25. Nam JS, Sharma AR, Jagga S, Lee DH, Sharma G, Nguyen LT, et al. Suppression of osteogenic activity by regulation of WNT and BMP signaling during titanium particle induced osteolysis. J Biomed Mater Res A. 2017; 105(3): 912-926.
  26. Allan CB, Lacourciere GM, Stadtman TC. Responsiveness of selenoproteins to dietary selenium. Annu Rev Nutr. 1999; 19: 1-16.
  27. Maleki N, Safavi A, Doroodmand MM. Determination of selenium in water and soil by hydride generation atomic absorption spectrometry using solid reagents. Talanta. 2005; 66(4): 858-862.
  28. Zhang J, Munger RG, West NA, Cutler DR, Wengreen HJ, Corcoran CD. Antioxidant intake and risk of osteoporotic hip fracture in Utah: an effect modified by smoking status. Am J Epidemiol. 2006; 163(1): 9-17.
  29. Sun L, Yu F, Xu Z, Zeng X, Ferreri M, Han B. Alteration of osteocalcin mRNA expression in ovine osteoblasts in dependence of sodium fluoride and sodium selenite medium supplementation. Acta Biol Hung. 2010; 61(1): 52-63.
  30. Finkel T, Holbrook NJ. Oxidants, oxidative stress and the biology of ageing. Nature. 2000; 408(6809): 239-247.
  31. Bai XC, Lu D, Bai J, Zheng H, Ke ZY, Li XM, et al. Oxidative stress inhibits osteoblastic differentiation of bone cells by ERK and NFkappaB. Biochem Biophys Res Commun. 2004; 314(1): 197-207.
  32. Xu ZS, Wang XY, Xiao DM, Hu LF, Lu M, Wu ZY, et al. Hydrogen sulfide protects MC3T3-E1 osteoblastic cells against H2O2- induced oxidative damage-implications for the treatment of osteoporosis. Free Radic Biol Med. 2011; 50(10): 1314-1323.
  33. Mody N, Parhami F, Sarafian TA, Demer LL. Oxidative stress modulates osteoblastic differentiation of vascular and bone cells. Free Radic Biol Med. 2001; 31(4): 509-519.
  34. Kim WK, Meliton V, Bourquard N, Hahn TJ, Parhami F. Hedgehog signaling and osteogenic differentiation in multipotent bone marrow stromal cells are inhibited by oxidative stress. J Cell Biochem. 2010; 111(5): 1199-1209.
  35. Sendur OF, Turan Y, Tastaban E, Serter M. Antioxidant status in patients with osteoporosis: a controlled study. Joint Bone Spine. 2009; 76(5): 514-518.
  36. Muthusami S, Ramachandran I, Muthusamy B, Vasudevan G, Prabhu V, Subramaniam V, et al. Ovariectomy induces oxidative stress and impairs bone antioxidant system in adult rats. Clin Chim Acta. 2005; 360(1-2): 81-86.
  37. Ebert R, Ulmer M, Zeck S, Meissner-Weigl J, Schneider D, Stopper H, et al. Selenium supplementation restores the antioxidative capacity and prevents cell damage in bone marrow stromal cells in vitro. Stem Cells. 2006; 24(5): 1226-1235.
  38. McCubrey JA, Lahair MM, Franklin RA. Reactive oxygen speciesinduced activation of the MAP kinase signaling pathways. Antioxid Redox Signal. 2006; 8(9-10): 1775-1789.
  39. Guo C, Yang RJ, Jang K, Zhou XL, Liu YZ. Protective effects of pretreatment with quercetin against lipopolysaccharide-induced apoptosis and the inhibition of osteoblast differentiation via the MAPK and Wnt/beta-Catenin pathways in MC3T3-E1 cells. Cell Physiol Biochem. 2017; 43(4): 1547-1561.