Calcified Tissue International

, Volume 103, Issue 2, pp 189–197 | Cite as

A Novel microCT Method for Bone and Marrow Adipose Tissue Alignment Identifies Key Differences Between Mandible and Tibia in Rats

  • Xavier CoutelEmail author
  • Cécile Olejnik
  • Pierre Marchandise
  • Jérôme Delattre
  • Hélène Béhal
  • Greet Kerckhofs
  • Guillaume Penel
Original Research


Bone homeostasis is influenced by the bone marrow adipose tissue (BMAT). BMAT distribution varies from one anatomical location in the skeleton to another. We developed an advanced microfocus computed tomography imaging and analysis protocol that allows accurate alignment of both the BMAT distribution and bone micro-architecture as well as calculation of the distance of the BMAT adipocytes from the bone surface. Using this protocol, we detected a different spatial BMAT distribution between the rat tibia and mandible: in the proximal metaphysis of the tibia a large amount of BMAT (~ 20% of the total BMAT) was located close to the bone surface (< 20 µm), whereas in the alveolar ridge ~ 30% of the total BMAT was located between 40 and 60 µm from the bone surface. In the alveolar ridge of rats, the trabecular bone volume was 48.3% higher compared to the proximal metaphysis of the tibia (p < 0.0001) and the percentage of adiposity determined to the relative marrow volume was lower (1.5%) compared to the proximal metaphysis of the tibia (9%, p = 0.0002). Interestingly, in the tibia a negative correlation was found between the percentage of adiposity in the total volume and the trabecular thickness (r =− 0.74, p = 0.037). The present study highlights that in comparison to tibial proximal metaphysis, the mandibular bone exhibits a massive trabecular network and a low BMAT content with almost no contact with the bone surface. These findings are of great interest because of the importance of the fat–bone interaction and its potential relevance to several resorptive bone diseases.


Rats Jaw bone Osmium staining Bone structure Micro-computed tomography imaging Adipocytes 



The authors thank the staff of the animal care facility (DHURE, Lille, France) for supplying the help on surgical procedures. This study was supported by the French Society of Rheumatology (SFR). GK acknowledges the Research Foundation—Flanders for her postdoctoral Grant (FWO/12R4315N) and her travel grant for a 6-months stay abroad in the PMOI lab.

Author Contributions

GP, CO, XC conceived and designed the experiments. XC, CO, PM, JD, GK performed the experiments (acquisition, analysis and/or interpretation of the data). XC drafted the manuscript. All authors critically revised the article and approved the final manuscript.


This study was supported by the French Society of Rheumatology.

Compliance with Ethical Standards

Conflict of interest

Xavier Coutel, Cécile Olejnik, Pierre Marchandise, Jérôme Delattre, Hélène Béhal, Greet Kerckhofs, and Guillaume Penel declare that they have no conflict of interest.

Human and Animal Rights and Informed Consent

All applicable international, national, and institutional guidelines for the care and use of animals were followed.


  1. 1.
    Rosen ED, Spiegelman BM (2014) What we talk about when we talk about fat. Cell 156:20–44. CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Lanske B, Rosen C (2017) Bone marrow adipose tissue: the first 40 years. J Bone Miner Res 32:1153–1156. CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Hardouin P, Marie PJ, Rosen CJ (2016) New insights into bone marrow adipocytes: report from the first European meeting on bone marrow adiposity (BMA 2015). Bone 93:212–215. CrossRefPubMedGoogle Scholar
  4. 4.
    van der Eerden B, van Wijnen A (2017) Meeting report of the 2016 bone marrow adiposity meeting. Adipocyte. PubMedPubMedCentralCrossRefGoogle Scholar
  5. 5.
    Scheller EL, Rosen CJ (2014) What’s the matter with MAT? Marrow adipose tissue, metabolism, and skeletal health. Ann N Y Acad Sci 1311:14–30. CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Craft CS, Scheller EL (2016) Evolution of the marrow adipose tissue microenvironment. Calcif Tissue Int 1–15.
  7. 7.
    Scheller EL, Doucette CR, Learman BS et al (2015) Region-specific variation in the properties of skeletal adipocytes reveals regulated and constitutive marrow adipose tissues. Nat Commun 6:7808. CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Akintoye SO, Lam T, Shi S et al (2006) Skeletal site-specific characterization of orofacial and iliac crest human bone marrow stromal cells in same individuals. Bone 38:758–768. CrossRefPubMedGoogle Scholar
  9. 9.
    Kozloff KM, Volakis LI, Marini JC, Caird MS (2010) Near-infrared fluorescent probe traces bisphosphonate delivery and retention in vivo. J Bone Miner Res 25:1748–1758. CrossRefPubMedGoogle Scholar
  10. 10.
    Yamada M, Matsuzaka T, Uetani M et al (1995) Normal age-related conversion of bone marrow in the mandible: MR imaging findings. Am J Roentgenol 165:1223–1228. CrossRefGoogle Scholar
  11. 11.
    Fazeli PK, Horowitz MC, MacDougald OA et al (2013) Marrow fat and bone—new perspectives. J Clin Endocrinol Metab 98:935–945. CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Hardouin P, Rharass T, Lucas S (2016) Bone marrow adipose tissue: to be or not to be a typical adipose tissue?. Front Endocrinol 7:.
  13. 13.
    Lips P, van Ginkel FC, Netelenbos JC (1985) Bone marrow and bone remodeling. Bone 6:343–344CrossRefPubMedGoogle Scholar
  14. 14.
    Schwartz AV (2015) Marrow fat and bone: review of clinical findings. Bone Res 6:40. CrossRefGoogle Scholar
  15. 15.
    Patsch JM, Li X, Baum T et al (2013) Bone marrow fat composition as a novel imaging biomarker in postmenopausal women with prevalent fragility fractures. J Bone Miner Res 28:1721–1728. CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Devlin MJ (2013) Bone marrow composition, diabetes, and fracture risk: more bad news for saturated fat. J Bone Miner Res 28:1718–1720. CrossRefPubMedGoogle Scholar
  17. 17.
    Paccou J, Hardouin P, Cotten A et al (2015) The role of bone marrow fat in skeletal health: usefulness and perspectives for clinicians. J Clin Endocrinol Metab 100:3613–3621. CrossRefPubMedGoogle Scholar
  18. 18.
    Pino AM, Miranda M, Figueroa C et al (2016) Qualitative aspects of bone marrow adiposity in osteoporosis. Front Endocrinol. CrossRefGoogle Scholar
  19. 19.
    Yeung DKW, Griffith JF, Antonio GE et al (2005) Osteoporosis is associated with increased marrow fat content and decreased marrow fat unsaturation: a proton MR spectroscopy study. J Magn Reson Imaging 22:279–285. CrossRefPubMedGoogle Scholar
  20. 20.
    Naveiras O, Nardi V, Wenzel PL et al (2009) Bone-marrow adipocytes as negative regulators of the haematopoietic microenvironment. Nature 460:259–263. CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Scheller EL, Cawthorn WP, Burr AA et al (2016) Marrow adipose tissue: trimming the fat. Trends Endocrinol Metab 27:392–403. CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Justesen J, Stenderup K, Ebbesen EN et al (2001) Adipocyte tissue volume in bone marrow is increased with aging and in patients with osteoporosis. Biogerontology 2:165–171CrossRefPubMedGoogle Scholar
  23. 23.
    During A, Penel G, Hardouin P (2015) Understanding the local actions of lipids in bone physiology. Prog Lipid Res 59:126–146. CrossRefPubMedGoogle Scholar
  24. 24.
    Mödder UI, Khosla S (2008) Skeletal stem/osteoprogenitor cells: current concepts, alternate hypotheses, and relationship to the bone remodeling compartment. J Cell Biochem 103:393–400. CrossRefPubMedGoogle Scholar
  25. 25.
    Clabaut A, Delplace S, Chauveau C et al (2010) Human osteoblasts derived from mesenchymal stem cells express adipogenic markers upon coculture with bone marrow adipocytes. Differentiation 80:40–45. CrossRefPubMedGoogle Scholar
  26. 26.
    Li M, Shen Y, Qi H, Wronski TJ (1996) Comparative study of skeletal response to estrogen depletion at red and yellow marrow sites in rats. Anat Rec 245:472–480.<472::AID-AR3>3.0.CO;2-UCrossRefPubMedGoogle Scholar
  27. 27.
    Du Z, Steck R, Doan N et al (2014) Estrogen deficiency associated bone loss in the maxilla: a methodology to quantify the changes in the maxillary intra-radicular alveolar bone in an ovariectomized rat osteoporosis model. Tissue Eng Part C Methods. PubMedCrossRefGoogle Scholar
  28. 28.
    Jiao K, Dai J, Wang M-Q et al (2010) Age- and sex-related changes of mandibular condylar cartilage and subchondral bone: a histomorphometric and micro-CT study in rats. Arch Oral Biol 55:155–163. CrossRefPubMedGoogle Scholar
  29. 29.
    Buie HR, Campbell GM, Klinck RJ et al (2007) Automatic segmentation of cortical and trabecular compartments based on a dual threshold technique for in vivo micro-CT bone analysis. Bone 41:505–515. CrossRefPubMedGoogle Scholar
  30. 30.
    Otsu N (1979) A threshold selection method from gray-level histograms. IEEE Trans Syst Man Cybern 9:62–66. CrossRefGoogle Scholar
  31. 31.
    Sengupta P (2013) The laboratory rat: relating its age with human’s. Int J Prev Med 4:624–630PubMedPubMedCentralGoogle Scholar
  32. 32.
    Sengupta S, Arshad M, Sharma S et al (2005) Attainment of peak bone mass and bone turnover rate in relation to estrous cycle, pregnancy and lactation in colony-bred Sprague–Dawley rats: suitability for studies on pathophysiology of bone and therapeutic measures for its management. J Steroid Biochem Mol Biol 94:421–429. CrossRefPubMedGoogle Scholar
  33. 33.
    Johnston BD, Ward WE (2015) The ovariectomized rat as a model for studying alveolar bone loss in postmenopausal women. BioMed Res Int 2015:1–12.
  34. 34.
    Kalu DN (1991) The ovariectomized rat model of postmenopausal bone loss. Bone Miner 15:175–191. CrossRefPubMedGoogle Scholar
  35. 35.
    Lelovas PP, Xanthos TT, Thoma SE et al (2008) The laboratory rat as an animal model for osteoporosis research. Comp Med 58:424–430PubMedPubMedCentralGoogle Scholar
  36. 36.
    Barngkgei I, Al Haffar I, Shaarani E et al (2016) Assessment of jawbone trabecular bone structure amongst osteoporotic women by cone-beam computed tomography: the OSTEOSYR project. J Investig Clin Dent 7:332–340. CrossRefPubMedGoogle Scholar
  37. 37.
    Mavropoulos A, Rizzoli R, Ammann P (2007) Different responsiveness of alveolar and tibial bone to bone loss stimuli. J Bone Miner Res 22:403–410. CrossRefPubMedGoogle Scholar
  38. 38.
    Elovic RP, Hipp JA, Hayes WC (1995) Ovariectomy decreases the bone area fraction of the rat mandible. Calcif Tissue Int 56:305–310CrossRefPubMedGoogle Scholar
  39. 39.
    Wronski TJ, Lowry PL, Walsh CC, Ignaszewski LA (1985) Skeletal alterations in ovariectomized rats. Calcif Tissue Int 37:324–328CrossRefPubMedGoogle Scholar
  40. 40.
    Devlin H, Whelton C (2015) Can mandibular bone resorption predict hip fracture in elderly women? A systematic review of diagnostic test accuracy. Gerodontology 32:163–168. CrossRefPubMedGoogle Scholar
  41. 41.
    Hassani-Nejad A, Ahlqwist M, Hakeberg M, Jonasson G (2013) Mandibular trabecular bone as fracture indicator in 80-year-old men and women. Eur J Oral Sci 121:525–531. CrossRefPubMedGoogle Scholar
  42. 42.
    Mavropoulos A, Ammann P, Bresin A, Kiliaridis S (2005) Masticatory demands induce region-specific changes in mandibular bone density in growing rats. Angle Orthod 75:625–630.[625:MDIRCI]2.0.CO;2PubMedGoogle Scholar
  43. 43.
    Mavropoulos A, Ödman A, Ammann P, Kiliaridis S (2010) Rehabilitation of masticatory function improves the alveolar bone architecture of the mandible in adult rats. Bone 47:687–692. CrossRefPubMedGoogle Scholar
  44. 44.
    Allen MR (2011) The effects of bisphosphonates on jaw bone remodeling, tissue properties, and extraction healing. Odontology 99:8–17. CrossRefPubMedGoogle Scholar
  45. 45.
    Choi DY, Sun KH, Won SY et al (2012) Trabecular bone ratio of the mandibular condyle according to the presence of teeth: a micro-CT study. Surg Radiol Anat 34:519–526. CrossRefPubMedGoogle Scholar
  46. 46.
    van Eijden TMGJ., van der Helm PN, van Ruijven LJ, Mulder L (2006) Structural and mechanical properties of mandibular condylar bone. J Dent Res 85:33–37. CrossRefPubMedGoogle Scholar
  47. 47.
    Bresin A, Kiliaridis S, Strid K-G (1999) Effect of masticatory function on the internal bone structure in the mandible of the growing rat. Eur J Oral Sci 107:35–44. CrossRefPubMedGoogle Scholar
  48. 48.
    Chappard D, Pascaretti-Grizon F, Gallois Y et al (2006) Medullar fat influences texture analysis of trabecular microarchitecture on X-ray radiographs. Eur J Radiol 58:404–410. CrossRefPubMedGoogle Scholar
  49. 49.
    Di Iorgi N, Rosol M, Mittelman SD, Gilsanz V (2008) Reciprocal relation between marrow adiposity and the amount of bone in the axial and appendicular skeleton of young adults. J Clin Endocrinol Metab 93:2281–2286. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Univ.Lille, Univ. Littoral Côte d’Opale, EA 4490 - PMOI – Physiopathologie des Maladies Osseuses InflammatoiresLilleFrance
  2. 2.Univ. Lille, CHU Lille, EA 2694 - Santé publique: épidémiologie et qualité des soins, Unité de Méthodologie et BiostatistiquesLilleFrance
  3. 3.Department of Development and Regeneration, Skeletal Biology and Engineering CenterKU LeuvenLeuvenBelgium
  4. 4.Prometheus, Division of Skeletal Tissue EngineeringKU LeuvenLeuvenBelgium

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