Oral Radiology

, Volume 27, Issue 1, pp 8–16 | Cite as

Inhibitory effect of maritime pine bark extract (Pycnogenol®) on deterioration of bone structure in the distal femoral epiphysis of ovariectomized mice

  • Tomoko Takano
  • Yusuke KozaiEmail author
  • Ryota Kawamata
  • Hiromi Wakao
  • Takashi Sakurai
  • Isamu Kashima
Original Article



To evaluate the inhibitory effects of maritime pine bark extract (Pycnogenol®) on the deterioration of bone mineral density (BMD) and trabecular structure due to osteoporosis in ovariectomized (OVX) mice.

Materials and methods

Five-week-old OVX ICR mice were divided into three groups: (1) OVX mice given Pycnogenol (Pycnogenol), (2) sham-operated mice (sham), and OVX mice not given Pycogenol (OVX control). All mice received standard feed; drinking water was provided ad libitum, with tap water for the sham and OVX control groups, and water containing Pycnogenol (120 mg/L) for the Pycnogenol group. Mice were housed for 3 months under these conditions, and then the femurs were resected and blood samples collected. The BMD of the distal femoral epiphysis was analyzed by peripheral quantitative computed tomography. Micro-computed tomography was also performed to evaluate the three-dimensional structure. Deterioration of BMD and trabecular structure was compared between the groups.


The Pycnogenol group showed a reduced loss of BMD compared to the OVX control group, which led to a significantly higher trabecular BMD in the former group. Additionally, surface area, number, content and complexity of the trabeculae, intertrabecular distance, and trabecular connectivity were all preserved in the Pycnogenol group. Pycnogenol thus significantly prevented trabecular architectural deterioration.


Our findings suggest that Pycnogenol may be useful in preventing BMD loss and trabecular architectural deterioration in osteoporosis.


Pycnogenol Osteoporosis Bone quality Bone mineral density Matrix metalloproteinases 


  1. 1.
    Drehsen G. From ancient pine bark uses to Pycnogenol. In: Packer L, Miramatzu M, Yoshikawa T, editors. Antioxidant food supplements in human health. New York: Academic Press; 1999. p. 311–22.CrossRefGoogle Scholar
  2. 2.
    Packer L, Rimbach G, Virgili F. Antioxidant activity and biologic properties of a procyanidin-rich extract from pine (Pinus maritime) bark, Pycnogenol. Free Radic Biol Med. 1999;27:704–24.PubMedCrossRefGoogle Scholar
  3. 3.
    Rohdewald P. A review of the French maritime pine bark extract (Pycnogenol®), a herbal medication with a diverse clinical pharmacology. Int Clin Pharmacol Ther. 2002;40:158–68.Google Scholar
  4. 4.
    Hosseini S, Pishnamazi S, Sadrzadeh SMH, Farid R, Watson RR. Pycnogenol® in the management of asthma. J Med Food. 2001;4:201–9.PubMedCrossRefGoogle Scholar
  5. 5.
    Lau BHS, Riesen SK, Truong KP, Lau EW, Rohdewald P, Barreta RA. Pycnogenol® as an adjunct in the management of childhood asthma. J Asthma. 2004;41:825–32.PubMedCrossRefGoogle Scholar
  6. 6.
    Sime S, Reeve VE. Protection from inflammation, immunosuppression and carcinogenesis induced by UV radiation in mice by topical Pycnogenol. Photochem Photobiol. 2004;79:193–8.PubMedCrossRefGoogle Scholar
  7. 7.
    Cisar P, Jany R, Waczulikova I, Sumegova K, Muchova J, Vojtassak J, et al. Effect of pine bark extract (Pycnogenol®) on symptoms of knee osteoarthritis. Phytother Res. 2008;22:1087–92.PubMedCrossRefGoogle Scholar
  8. 8.
    Grimm T, Schäfer A, Högger P. Antioxidant activity and inhibition of matrix metalloproteinases by metabolites of maritime pine bark extract (Pycnogenol®). Free Radic Biol Med. 2004;36:811–22.PubMedCrossRefGoogle Scholar
  9. 9.
    Grimm T, Chovanová Z, Muchová J, Sumegova K, Liptakova A, Durackova A, et al. Inhibition of NF-kB activation and MMP-9 secretion by plasma of human volunteers after ingestion of maritime pine bark extract (Pycnogenol®). J Inflamm. 2006;3:1–6.CrossRefGoogle Scholar
  10. 10.
    Gack S, Vallon R, Schmidt J, Grigoriadis A, Tuckermann J, Schenkel J, et al. Expression of interstitial collagenase during skeletal development of the mouse is restricted to osteoblast-like cells and hypertrophic chondrocytes. Cell Growth Differ. 1995;6:759–67.PubMedGoogle Scholar
  11. 11.
    Johansson N, Saarialho-Kere U, Airoka K, Herva R, Nissinen L, Westermarck J, et al. Collagenase-3 (MMP-13) is expressed by hypertrophic chondrocytes, periosteal cells, and osteoblasts during human fetal bone development. Dev Dyn. 1997;208:387–97.PubMedCrossRefGoogle Scholar
  12. 12.
    Chin JR, Werb Z. Matrix metalloproteinases regulate morphogenesis, migration and remodeling of epithelium, tongue skeletal muscle and cartilage in the mandibular arch. Development. 1997;124:1519–30.PubMedGoogle Scholar
  13. 13.
    Tezuka K, Nemoto K, Tezuka Y, Sato T, Ikeda Y, Kobori M, et al. Identification of matrix metalloproteinase 9 in rabbit osteoclasts. J Biol Chem. 1994;269:1506–9.Google Scholar
  14. 14.
    Katsunuma N. Molecular mechanisms of bone collagen degradation in bone resorption. J Bone Miner Metab. 1997;15:1–8.CrossRefGoogle Scholar
  15. 15.
    Tezuka K, Tezuka Y, Maejima A, Sato T, Nemoto K, Kamioka Y, et al. Molecular cloning of a possible cysteine proteinase predominantly expressed in osteoclasts. J Biol Chem. 1994;269:1106–9.PubMedGoogle Scholar
  16. 16.
    Ferreti JL. Peripheral quantitative computed tomography for evaluating structural and mechanical properties of small bone. In: An YH, Draughn RA, editors. Mechanical testing of bone and the bone-implant interface. Boca Raton: CRC Press; 2000. p. 390–2.Google Scholar
  17. 17.
    RATOC System Engineering Co. Ltd. TRI/3D-BON. Basic operation manual. Tokyo, Japan: RATOC System Engineering Co Ltd. 2002.Google Scholar
  18. 18.
    Parfitt AM, Matthews CHE, Villanueva AR, Kleerekoper M, Frame B, Rao DS. Relationships between surface, volume, and thickness of iliac trabecular bone in aging and in osteoporosis. J Clin Invest. 1983;72:1396–409.PubMedCrossRefGoogle Scholar
  19. 19.
    Odgaad A. Three-dimensional methods fore quantification of cancellous bone architecture. Bone. 1997;20:315–28.CrossRefGoogle Scholar
  20. 20.
    Feldkamp LA, Goldstein SA, Parfitt AM, Jesion G, Kleerekoper M. The direct examination of three-dimensional bone architecture in vitro by computed tomography. J Bone Miner Res. 1989;4:3–11.PubMedCrossRefGoogle Scholar
  21. 21.
    Ikuta A, Kumasaka S, Kashima I. Quantitative analysis using the star volume method applied to skeleton patterns extracted with a morphological filter. J Bone Miner Metab. 2000;18:271–7.PubMedCrossRefGoogle Scholar
  22. 22.
    Kumasaka S, Kiyohara S, Takahashi T, Asai H, Kashima I. Morphologically extracted trabecular skeleton superimposed upon digital radiograph structure. J Bone Miner Metab. 2000;18:208–11.PubMedCrossRefGoogle Scholar
  23. 23.
    Vesterby A, Gunndersen HJG, Melsen F. Star volume of marrow space and trabeculae of the first lumber vertebra: sampling efficiency and biological variation. Bone. 1989;10:7–13.PubMedCrossRefGoogle Scholar
  24. 24.
    Nakamura K, Matsubara M, Asai H, Koyama A, Fujikawa T, Kashima I. Mathematical morphology for extraction of bone trabecural pattern: preliminary investigation of quantitative analysis using the star volume. J Jpn Soc Bone Morphom. 1999;9:45–51.Google Scholar
  25. 25.
    Garrahan NJ, Mellish RW, Compston JE. A new method for the two-dimensional analysis of bone structure in human iliac crest biopsies. J Microsc. 1986;142:341–9.PubMedGoogle Scholar
  26. 26.
    Croucher PI, Garrahan NJ, Compston JE. Assessment of cancellous bone structure: comparison of strut analysis, trabecular bone pattern factor, and marrow space star volume. J Bone Miner Res. 1996;11:955–61.PubMedCrossRefGoogle Scholar
  27. 27.
    Sinaki M, Itoi E, Wahner HW, Wollan P, Gelzcer R, Mullan BP, et al. Stronger back muscles reduce the incidence of vertebral fractures; a prospective 10 year follow-up of postmenopausal women. Bone. 2002;30:836–41.PubMedCrossRefGoogle Scholar
  28. 28.
    Steiner E, Jergas M, Genant H. Radiology of osteoporosis. In: Marcus R, Feldman D, Kelsey J, editors. osteoporosis. San Diego: Academic Press; 1996. p. 1019–54.Google Scholar
  29. 29.
    Arnold JS. Trabecular patterns and shapes in aging and osteoporosis. Metab Bone Dis Rel Res. 1980;2S:297–308.Google Scholar
  30. 30.
    Mosekilde L. Age-related changes in vertebral trabecular bone architecture assessed by a new method. Bone. 1988;9:247–50.PubMedCrossRefGoogle Scholar
  31. 31.
    Thomsen JS, Ebbesen EN, Mosekilde LI. Age-related differences between thinning of horizontal and vertical trabeculae in human lumbar bone as assessed by a new computerized method. Bone. 2002;31:136–42.PubMedCrossRefGoogle Scholar
  32. 32.
    Singh YM, Nagrath AR, Maini PS. Changes in trabecular pattern of the upper end of the femur as an index of osteoporosis. J Bone Joint Surg Am. 1970;52:457–67.PubMedGoogle Scholar
  33. 33.
    Ciarelli TE, Fyhrie DP, Schaffler MB, Goldstein SA. Variations in three-dimensional cancellous bone architecture of the proximal femur in female hip fractures and in controls. J Bone Miner Res. 2000;15:32–40.PubMedCrossRefGoogle Scholar
  34. 34.
    Homminga J, McCreadie BR, Ciarelli TE, Weinans H, Goldstein SA, Huiskes R. Cancellous bone mechanical properties from normals and patients with hip fractures differ on the structure level, not on the bone hard tissue level. Bone. 2002;30:759–64.PubMedCrossRefGoogle Scholar
  35. 35.
    Ito M, Ikeda K, Nishiguchi M, Shindo H, Uetani M, Hosoi T, et al. Multi-detector-row CT imaging of vertebral microstructure for evaluation of facture risk. J Bone Miner Res. 2005;20:1828–36.PubMedCrossRefGoogle Scholar
  36. 36.
    Kinney JH, Ladd AJ. The relationship between three-dimensional connectivity and the elastic properties of trabecular bone. J Bone Miner Res. 1998;13:839–45.PubMedCrossRefGoogle Scholar
  37. 37.
    Dempster DW. Exploiting and bypassing the bone remodeling cycle to optimize the treatment of osteoporosis. J Bone Miner Res. 1997;12:1152–4.PubMedCrossRefGoogle Scholar
  38. 38.
    Tsugawa N, Shiraki M, Suhara Y, Kamao M, Tanaka K, Okano T. Vitamin K status of healthy Japanese women; age-related vitamin K requirement for γ-carboxylation of osteocalcin. Am Clin Nutr. 2006;83:380–6.Google Scholar
  39. 39.
    Ichikawa T, Horie IK, Ikeda K, Blumberg B, Inoue S. Steroid and xenobiotic receptor SXR mediates vitamin K2-activated transcription of extracellular matrix-related genes and collagen accumulation in osteoblastic cells. J Biol Chem. 2006;281:16927–34.PubMedCrossRefGoogle Scholar

Copyright information

© Japanese Society for Oral and Maxillofacial Radiology and Springer 2011

Authors and Affiliations

  • Tomoko Takano
    • 1
  • Yusuke Kozai
    • 2
    Email author
  • Ryota Kawamata
    • 2
  • Hiromi Wakao
    • 2
  • Takashi Sakurai
    • 2
  • Isamu Kashima
    • 2
  1. 1.Division of Dentistry for Special Patients, Department of Clinical Care MedicineKanagawa Dental CollegeYokosukaJapan
  2. 2.Division of Radiology, Department of Maxillofacial Diagnostic ScienceKanagawa Dental CollegeYokosukaJapan

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