Molecular and Cellular Biochemistry

, Volume 340, Issue 1–2, pp 73–80 | Cite as

Comparative effects of a novel plant-based calcium supplement with two common calcium salts on proliferation and mineralization in human osteoblast cells

  • Ram Sudheer Adluri
  • Lijun Zhan
  • Manashi Bagchi
  • Nilanjana Maulik
  • Gautam Maulik


Calcium is an essential mineral to support bone health and serves as a major therapeutic intervention to prevent and delay the incidence of osteoporosis. Many individuals do not obtain the optimum amount of calcium from diets and depend on bioavailable calcium supplements. The present study was conducted to examine the effect of a novel plant-based calcium supplement, derived from marine algae, and contains high levels of calcium, magnesium, and other bone supporting minerals [commercially known as AlgaeCal (AC)], on proliferation, mineralization, and oxidative stress in cultured human osteoblast cells, and compared with inorganic calcium carbonate and calcium citrate salts. Cultured human fetal osteoblast cells (hFOB 1.19) were treated with AC (0.5 mg/ml, fixed by MTT assay), calcium carbonate, or calcium citrate. These cells were harvested after 4 days of treatment for ALP activity, PCNA expression, and DNA synthesis, and 2 days for Ca2+ deposition in the presence and absence of vitamin D3 (5 nM). The ability of AC to reduce H2O2 (0.3 mM)-induced oxidative stress was assessed after 24 h of treatment. ALP activity was significantly increased with AC treatment when compared to control, calcium carbonate, or calcium citrate (4.0-, 2.0-, and 2.5-fold, respectively). PCNA expression (immunocytochemical analysis), DNA synthesis (4.0-, 3.0-, and 4.0-fold, respectively), and Ca2+ deposition (2.0-, 1.0-, and 4.0-fold, respectively) were significantly increased in AC-treated cells when compared with control, calcium carbonate, or calcium citrate treatment. These markers were further enhanced following additional supplementation of vitamin D3 in the AC-treated group cells. AC treatment significantly reduced the H2O2-induced oxidative stress when compared to calcium carbonate or calcium citrate (1.5- and 1.4-fold, respectively). These findings suggest that AC may serve as a superior calcium supplement as compared to other calcium salts tested in the present study. Hence, AC may be developed as a novel anti-osteoporotic supplement in the near future.


Calcium supplement Osteoporosis Mineralization Proliferation Oxidative stress DNA synthesis 



This study was supported by M/s. AlgaeCal, Vancouver, Canada. We thank T. Mahesh for his help in finalizing all the figures.


  1. 1.
    Scott JC (1997) Epidemiology of osteoporosis. J Clin Rheumatol 3:9–13CrossRefPubMedGoogle Scholar
  2. 2.
    Lanham-New SA (2008) Importance of calcium, vitamin D and vitamin K for osteoporosis prevention and treatment. Proc Nutr Soc 67:163–176CrossRefPubMedGoogle Scholar
  3. 3.
    Bonewald ML, Johnson LF (2008) Osteocytes, mechanosensing and Wnt signaling. Bone 42:606–615CrossRefPubMedGoogle Scholar
  4. 4.
    Hamdy RC (2009) Fracture risk assessment in postmenopausal women. Rev Endocr Metab DisordGoogle Scholar
  5. 5.
    Forrest CM, Mackay GM, Oxford L, Stoy N, Stone LG, Darlington TW (2006) Kynurenine pathway metabolism in patients with osteoporosis after 2 years of drug treatment. Clin Exp Pharmacol Physiol 33:1078–1087CrossRefPubMedGoogle Scholar
  6. 6.
    Yang S, Madyastha P, Bingel S, Ries L, Key W (2001) A new superoxide-generating oxidase in murine osteoclasts. J Biol Chem 276:5452–5458CrossRefPubMedGoogle Scholar
  7. 7.
    Muthusami S, Ramachandran I, Muthusamy B, Vasudevan G, Prabhu V, Subramaniam V, Jagadeesan S, Narasimhan A (2005) Ovariectomy induces oxidative stress and impairs bone antioxidant system in adult rats. Clin Chim Acta 360:81–86CrossRefPubMedGoogle Scholar
  8. 8.
    Subramaniam M, Jalal SM, Rickard DJ, Harris SA, Bolander TC, Spelsberg ME (2002) Further characterization of human fetal osteoblastic hFOB 1.19 and hFOB/ER alpha cells: bone formation in vivo and karyotype analysis using multicolor fluorescent in situ hybridization. J Cell Biochem 87:9–15CrossRefPubMedGoogle Scholar
  9. 9.
    Galli C, Passeri G, Cacchioli A, Gualini G, Ravanetti F, Elezi GM, Macaluso E (2009) Effect of laser-induced dentin modifications on periodontal fibroblasts and osteoblasts: a new in vitro model. J Periodontol 80:1648–1654CrossRefPubMedGoogle Scholar
  10. 10.
    de Souza Malaspina TS, Zambuzzi WF, dos Santos CX, Campanelli AP, Laurindo FR, Sogayar JM, Granjeiro MC (2009) A possible mechanism of low molecular weight protein tyrosine phosphatase (LMW-PTP) activity modulation by glutathione action during human osteoblast differentiation. Arch Oral Biol 54:642–650CrossRefPubMedGoogle Scholar
  11. 11.
    Jhaveri A, Walsh SJ, Wang Y, McCarthy G, Gronowicz M (2008) Therapeutic touch affects DNA synthesis and mineralization of human osteoblasts in culture. J Orthop Res 26:1541–1546CrossRefPubMedGoogle Scholar
  12. 12.
    Bi Y, Nielsen KL, Kilts TM, Yoon AMAK, Wimer HF, Greenfield EM, Heegaard MF, Young AM (2006) Biglycan deficiency increases osteoclast differentiation and activity due to defective osteoblasts. Bone 38:778–786CrossRefPubMedGoogle Scholar
  13. 13.
    Choi EM, Kim YS, Lee GH (2009) Protective effects of dehydrocostus lactone against hydrogen peroxide-induced dysfunction and oxidative stress in osteoblastic MC3T3-E1 cells. Toxicol In Vitro 23:862–867CrossRefPubMedGoogle Scholar
  14. 14.
    Coelho MH, Fernandes MJ (2000) Human bone cell cultures in biocompatibility testing. Part II: effect of ascorbic acid, beta-glycerophosphate and dexamethasone on osteoblastic differentiation. Biomaterials 21:1095–1102CrossRefPubMedGoogle Scholar
  15. 15.
    Stoimenov T, Helleday I (2009) PCNA on the crossroad of cancer. Biochem Soc Trans 37:605–613CrossRefPubMedGoogle Scholar
  16. 16.
    Heaney RP (2009) Dairy and bone health. J Am Coll Nutr 28(Suppl 1):82S–90SPubMedGoogle Scholar
  17. 17.
    Frassetto LA, Morris RC Jr, Sellmeyer A, Sebastian DE (2008) Adverse effects of sodium chloride on bone in the aging human population resulting from habitual consumption of typical American diets. J Nutr 138:419S–422SPubMedGoogle Scholar
  18. 18.
    Ilich JE, Kerstetter JZ (2000) Nutrition in bone health revisited: a story beyond calcium. J Am Coll Nutr 19:715–737PubMedGoogle Scholar
  19. 19.
    Ryder KM, Shorr RI, Bush AJ, Kritchevsky SB, Harris T, Stone K, Cauley FA, Tylavsky J (2005) Magnesium intake from food and supplements is associated with bone mineral density in healthy older white subjects. J Am Geriatr Soc 53:1875–1880CrossRefPubMedGoogle Scholar
  20. 20.
    Jugdaohsingh R, Anderson SH, Tucker KL, Elliott H, Kiel DP, Thompson JJ, Powell RP (2002) Dietary silicon intake and absorption. Am J Clin Nutr 75:887–893PubMedGoogle Scholar
  21. 21.
    Bae YJ, Kim JY, Choi MK, Chung MH, Kim YS (2008) Short-term administration of water-soluble silicon improves mineral density of the femur and tibia in ovariectomized rats. Biol Trace Elem Res 124:157–163CrossRefPubMedGoogle Scholar
  22. 22.
    Rico H, Gallego-Lago JL, Hernandez ER, Villa LF, Sanchez-Atrio A, Seco JJ, Gervas C (2000) Effect of silicon supplement on osteopenia induced by ovariectomy in rats. Calcif Tissue Int 66:53–55CrossRefPubMedGoogle Scholar
  23. 23.
    Nielsen FH, Hunt CD, Mullen JR, Hunt LM (1987) Effect of dietary boron on mineral, estrogen, and testosterone metabolism in postmenopausal women. FASEB J 1:394–397PubMedGoogle Scholar
  24. 24.
    Beattie HS, Peace JH (1993) The influence of a low-boron diet and boron supplementation on bone, major mineral and sex steroid metabolism in postmenopausal women. Br J Nutr 69:871–884CrossRefPubMedGoogle Scholar
  25. 25.
    Hunt CD (1994) The biochemical effects of physiologic amounts of dietary boron in animal nutrition models. Environ Health Perspect 102(Suppl 7):35–43CrossRefPubMedGoogle Scholar
  26. 26.
    New SA, Robins SP, Campbell MK, Martin JC, Garton MJ, Bolton-Smith C, Grubb DA, Lee DM, Reid SJ (2000) Dietary influences on bone mass and bone metabolism: further evidence of a positive link between fruit and vegetable consumption and bone health? Am J Clin Nutr 71:142–151PubMedGoogle Scholar
  27. 27.
    Macdonald HM, New SA, Golden MH, Campbell DM, Reid MK (2004) Nutritional associations with bone loss during the menopausal transition: evidence of a beneficial effect of calcium, alcohol, and fruit and vegetable nutrients and of a detrimental effect of fatty acids. Am J Clin Nutr 79:155–165PubMedGoogle Scholar
  28. 28.
    New SA (2003) Intake of fruit and vegetables: implications for bone health. Proc Nutr Soc 62:889–899PubMedGoogle Scholar
  29. 29.
    Prynne CJ, Mishra GD, O’Connell MA, Muniz G, Laskey MA, Yan L, Prentice F, Ginty A (2006) Fruit and vegetable intakes and bone mineral status: a cross sectional study in 5 age and sex cohorts. Am J Clin Nutr 83:1420–1428PubMedGoogle Scholar
  30. 30.
    Tucker KL, Hannan MT, Chen H, Cupples LA, Wilson DP, Kiel PW (1999) Potassium, magnesium, and fruit and vegetable intakes are associated with greater bone mineral density in elderly men and women. Am J Clin Nutr 69:727–736PubMedGoogle Scholar
  31. 31.
    Tucker KL, Chen H, Hannan MT, Cupples LA, Wilson PW, Felson DP, Kiel D (2002) Bone mineral density and dietary patterns in older adults: the Framingham Osteoporosis Study. Am J Clin Nutr 76:245–252PubMedGoogle Scholar
  32. 32.
    McGartland CP, Robson PJ, Murray LJ, Cran GW, Savage MJ, Watkins DC, Rooney CA, Boreham MM (2004) Fruit and vegetable consumption and bone mineral density: the Northern Ireland Young Hearts Project. Am J Clin Nutr 80:1019–1023PubMedGoogle Scholar
  33. 33.
    Moore CE, Murphy MF, Holick MM (2005) Vitamin D intakes by children and adults in the United States differ among ethnic groups. J Nutr 135:2478–2485PubMedGoogle Scholar
  34. 34.
    Liu ZG, Zhang R, Li C, Ma X, Liu L, Wang QB, Mei JP (2009) The osteoprotective effect of Radix Dipsaci extract in ovariectomized rats. J Ethnopharmacol 123:74–81CrossRefPubMedGoogle Scholar
  35. 35.
    Nielsen FH (1991) Nutritional requirements for boron, silicon, vanadium, nickel, and arsenic: current knowledge and speculation. FASEB J 12:2661–2667Google Scholar
  36. 36.
    Vaaro P, Saari E, Paaso A, Koivistoinen P (2008) Strontium in Finnish foods. Int J Vitam Nutr Res 52:342–350Google Scholar

Copyright information

© Springer Science+Business Media, LLC. 2010

Authors and Affiliations

  • Ram Sudheer Adluri
    • 1
  • Lijun Zhan
    • 1
  • Manashi Bagchi
    • 2
  • Nilanjana Maulik
    • 1
  • Gautam Maulik
    • 3
  1. 1.Molecular Cardiology and Angiogenesis Laboratory, Department of SurgeryUniversity of Connecticut Health CenterFarmingtonUSA
  2. 2.NutriTodayBostonUSA
  3. 3.Department of Cancer Biology, Dana Farber Cancer InstituteHarvard Medical SchoolBostonUSA

Personalised recommendations