Chinese Journal of Integrative Medicine

, Volume 18, Issue 4, pp 276–282 | Cite as

In vitro and in vivo effects of puerarin on promotion of osteoblast bone formation

  • Ming-yu Zhang (张明宇)
  • Hui Qiang (强 辉)
  • Hua-qing Yang (杨华清)
  • Xiao-qian Dang (党晓谦)
  • Kun-zheng Wang (王坤正)
Original Article



To assess the effect of puerarin, a natural flavonoid found in Chinese Pueraria Lobata (Wild.) Ohwi, on promotion of new bone formation.


Osteoblasts isolated from calvarial of newborn rats were cultured in vitro in the presence of puerarin at various concentrations. The viability of osteoblasts and alkaline phosphotase activity and mineral node formation were determined. In addition, osteoblasts seeded in the β-tricaclium phosphate scalfolds as bone substitute were implanted in rat dorsal muscles. Half -of the recipient rats received intramuscular injection of puerarin at 10 mg/(kg·d) for 7 days. Osteogenesis was analyzed by examining the histology after 4 weeks of implantation.


The viability of osteoblasts treated with puerarin at either 40 or 80 μmol/L was significantly higher than that of the control (P<0.05 and P<0.01, respectively). Alkaline phosphatase and mineral modules were significantly increased in osteoblasts cultured with puerarin at 40 or 80 mol/L when compared with that of the untreated cells. The puerarin-treated rats had a higher rate of bone formation in the osteoblast implants than the control rats (6.35% vs. 1.32%, respectively, P<0.05).


Puerarin was able to affect osteoblast proliferation and differentiation, and promote the new bone formation in osteoblast implants.


puerarin osteoblast β-tricalcium phosphate new bone formation 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Hirokazu K, Takaaki T, Masaaki C, Takahiro K. Repair of segmental bone defects in rabbit tibiae using a complex of b-tricalcium phosphate, type I collagen, and fibroblast growth factor-2. Biomaterials 2006;27:5118–5126.CrossRefGoogle Scholar
  2. 2.
    Kneser U, Schaefer DJ, Polykandriotis E, Horch RE. Tissue engineering of bone: the reconstructive surgeon’s point of view. J Cell Mol Med 2006;10:7–19.PubMedCrossRefGoogle Scholar
  3. 3.
    Smith LA, Liu XH, Ma PX. Nano-fibrous scaffolds for tissue engineering. Soft Matter 2008;4:2144–2149.PubMedCrossRefGoogle Scholar
  4. 4.
    Liang G, Yang Y, Oh S, Ong JL, Zheng C, Ran J, et al. Ectopic osteoinduction and early degradation of recombinant human bone morphogenetic protein-2-loaded porous beta-tricalcium phosphate in mice. Biomaterials 2005;26:4265–4271.PubMedCrossRefGoogle Scholar
  5. 5.
    Langer R, Vacanti JP. Tissue engineering. Science 1993;260:920–926.PubMedCrossRefGoogle Scholar
  6. 6.
    Eppley BL, Pietrzak WS, Blanton MW. Allograft and alloplastic bone substitutes: a review of science and technology for the craniomaxillofacial surgeon. J Craniofac Surg 2005;16:981–989.PubMedCrossRefGoogle Scholar
  7. 7.
    Kneser U, Schaefer DJ, Polykandriotis E, Horch RE. Tissue engineering of bone: the reconstructive surgeon’s point of view. J Cell Mol Med 2006;10:7–19.PubMedCrossRefGoogle Scholar
  8. 8.
    Anderson JB, Garner SC. Phytoestrogens and bone. Baillieres Clin Endocrinol Metab 1998;12:543–557.PubMedCrossRefGoogle Scholar
  9. 9.
    Wu HQ, Guo HN, Wang HQ, Chang MZ, Zhang GL, Zhao YX. Protective effect and mechanism of puerarin on learning-memory disorder after global cerebral ischemiareperfusion injury in rats. Chin J Integr Med 2009;15:54–59.PubMedCrossRefGoogle Scholar
  10. 10.
    Wang XX, Wu J, Chiba H, Umegaket K, Yamada K, Ishimi Y. Puerariae radix prevents bone loss in ovariectomized mice. J Bone Miner Metab 2003;21:268–275.PubMedCrossRefGoogle Scholar
  11. 11.
    Blarir, HC, Jordan SE, Peterson TG, Stephenal B. Variable effects of ty-rosine kinase inhibitor on avian osteoclastic activity and reduction of bone loss in ovariectiomized rats. J Cell Biochem 1996;61:629–637.CrossRefGoogle Scholar
  12. 12.
    Heidi D, Natasj VV, Erna D, Maeyer Ronald V, Etienne S, Leo DR, et al. Isolation, proliferation and differentiation of osteoblastic cells to study cell/biomaterial interactions: comparison of different isolation techniques and source. Biomaterials 2004;25:757–768.CrossRefGoogle Scholar
  13. 13.
    Yuan J, Cui L, Zhang WJ, Liu W, Cao Y. Repair of canine mandibular bone defects with bone marrow stromal cells and porous beta-tricalcium phosphate. Biomaterials 2007;28:1005–1013.PubMedCrossRefGoogle Scholar
  14. 14.
    Zhou Q, Fu T. Effect of puerarin on the healing osteoporotic fracture in ovariectomized rats. Chin J Clin Rehabilit 2006;10:45–47.Google Scholar
  15. 15.
    Habibovic P, Yuan H, van der Valk CM, Meijer G, van Blitterswijk CA, de Groot K. 3D microenvironment as essential element for osteoinduction by biomaterials. Biomaterials 2005;26:3565–3575.PubMedCrossRefGoogle Scholar
  16. 16.
    Dang ZC, Audinot V, Papapoulos SE, Boutin JA, Löwik CWGM. Peroxisome proliferator-activated receptor g (PPARg) as a molecular target for the soy phytoestrogen genistein. J Biol Chem 2003;278:962–967.PubMedCrossRefGoogle Scholar
  17. 17.
    Dang ZC, Löwik CW. The balance between concurrent activation of ERs and PPARs determines daidzeininduced osteogenesis and adipogenesis. J Bone Miner Res 2004;19:853–861.PubMedCrossRefGoogle Scholar
  18. 18.
    Viereck V, Gründker C, Blaschke S, Siggelkow H, Emons G, Hofbauer LC. Phytoestrogen genistein stimulates the production of osteoprotegerin by human trabecular osteoblasts. J Cell Biochem 2002;84:725–735.PubMedCrossRefGoogle Scholar
  19. 19.
    Notoya M, Tsukamoto Y, Nishimura H, Woo JT, Nagai K, Lee IS, et al. Quercetin, a flavonoid, inhibits the proliferation, differentiation, and mineralization of osteoblasts in vitro. Eur J Pharmacol 2004;485:89–96.PubMedCrossRefGoogle Scholar
  20. 20.
    Garcia T, Roman S, Jackson A, Thellhaber J, Connolly T, Spinella-Jaegle S, et al. Behavior of osteoblast, adipocyte, and myoblast markers in genome-wide expression analysis of mouse calvaria primary osteoblasts in vitro. Bone 2002;31:205–211.PubMedCrossRefGoogle Scholar
  21. 21.
    Whyte MP. Hypophosphatasia and the role of alkaline phosphatase in skeletal mineralization. Endocr Rev 1994;15:439–461.PubMedGoogle Scholar
  22. 22.
    Fedarko NS, Bianco P, Vetter U, Gehron Robey P. Human bone cell enzyme expression and cellular heterogeneity: correlation of alkaline phosphatase enzyme activity with cell cycle. J Cell Physiol 1990;144:115–121.PubMedCrossRefGoogle Scholar
  23. 23.
    Cowles EA, DeRome ME, Pastizzo G, Brailey LL, Gronowicz GA. Mineralization and expression of matrix proteins during in vivo bone development. Calcif Tissue Int 1998;62:74–82.PubMedCrossRefGoogle Scholar
  24. 24.
    Maenoa S, Nikia Y, Matsumotoa H, Morioka H, Yatabe T, Funayama A, et al. The effect of calcium ion concentration on osteoblast viability, proliferation and differentiation in monolayer and 3D culture. Biomaterials 2005;26:4847–4855.CrossRefGoogle Scholar
  25. 25.
    Xu SL, Li DC, Xie YZ, Lu JX, Dai KR. The growth of stem cells within β-TCP scaffolds in a fluid-dynamic environment. Mater Sci Engin 2008;C28:164–170.Google Scholar

Copyright information

© Chinese Association of the Integration of Traditional and Western Medicine and Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Ming-yu Zhang (张明宇)
    • 1
  • Hui Qiang (强 辉)
    • 2
  • Hua-qing Yang (杨华清)
    • 3
  • Xiao-qian Dang (党晓谦)
    • 4
  • Kun-zheng Wang (王坤正)
    • 4
  1. 1.Department of Sport Injurythe Affiliated Red Cross Hospital of Medical College of Xi’an Jiaotong UniversityXi’anChina
  2. 2.Department of OrthopedicsShaanxi Provincial People’s HospitalXi’anChina
  3. 3.Department of Orthopedicsthe Second Hospital of Tsinghua UniversityBeijingChina
  4. 4.Department of Orthopedicsthe Second Affiliated Hospital of Medical College of Xi’an Jiaotong UniversityXi’anChina

Personalised recommendations