The Orientation of Nanoscale Apatite Platelets in Relation to Osteoblastic–Osteocyte Lacunae on Trabecular Bone Surface
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The orientation of nanoscale mineral platelets was quantitatively evaluated in relation to the shape of lacunae associated with partially embedded osteocytes (osteoblastic–osteocytes) on the surface of deproteinised trabecular bone of adult sheep. By scanning electron microscopy and image analysis, the mean orientation of mineral platelets at the osteoblastic–osteocyte lacuna (Ot.Lc) floor was found to be 19° ± 14° in the tibia and 20° ± 14° in the femur. Further, the mineral platelets showed a high degree of directional coherency: 37 ± 7 % in the tibia and 38 ± 9 % in the femur. The majority of Ot.Lc in the tibia (69.37 %) and the femur (74.77 %) exhibited a mean orientation of mineral platelets between 0° and 25°, with the largest fraction within a 15°–20° range, 17.12 and 19.8 % in the tibia and femur, respectively. Energy dispersive X-ray spectroscopy and Raman spectroscopy were used to characterise the features observed on the anorganic bone surface. The Ca/P (atomic %) ratio was 1.69 ± 0.1 within the Ot.Lc and 1.68 ± 0.1 externally. Raman spectra of NaOCl-treated bone showed peaks associated with carbonated apatite: ν1, ν2 and ν4 PO4 3−, and ν1 CO3 2−, while the collagen amide bands were greatly reduced in intensity compared to untreated bone. The apatite-to-collagen ratio increased considerably after deproteinisation; however, the mineral crystallinity and the carbonate-to-phosphate ratios were unaffected. The ~19°–20° orientation of mineral platelets in at the Ot.Lc floor may be attributable to a gradual rotation of osteoblasts in successive layers relative to the underlying surface, giving rise to the twisted plywood-like pattern of lamellar bone.
KeywordsBone Biomineralisation Osteoblastic–osteocyte Lacuna Trabecular bone Electron microscopy Raman spectroscopy
This study was supported by the Swedish Research Council (Grant K2015-52X-09495-28-4), the BIOMATCELL VINN Excellence Center of Biomaterials and Cell Therapy, the Region Västra Götaland, an ALF/LUA Grant, the IngaBritt and Arne Lundberg Foundation, the Dr. Felix Neubergh Foundation, Promobilia, the Hjalmar Svensson Foundation, and the Materials Science Area of Advance at Chalmers and the Department of Biomaterials, University of Gothenburg.
Compliance with Ethical Standards
Conflict of Interest
Furqan A. Shah, Ezio Zanghellini, Aleksandar Matic, Peter Thomsen, and Anders Palmquist declare that they have no conflict of interest.
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
Human and Animal Rights and Informed Consent
All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. This article does not contain any studies with human participants performed by any of the authors.
- 12.Mann S, Webb J, Williams RJP (1989) Biomineralization: chemical and biochemical perspectives. VCH, WeinheimGoogle Scholar
- 17.Pazzaglia UE, Congiu T, Marchese M, Dell’Orbo C (2010) The shape modulation of osteoblast-osteocyte transformation and its correlation with the fibrillar organization in secondary osteons: a SEM study employing the graded osmic maceration technique. Cell Tissue Res 340:533–540CrossRefPubMedGoogle Scholar
- 21.Boyde A (1972) Scanning electron microscope studies of bone. In: Bourne GH (ed) The biochemistry and physiology of bone. Academic Press Inc, New YorkGoogle Scholar
- 22.Rezakhaniha R, Agianniotis A, Schrauwen JTC, Griffa A, Sage D, Bouten CVC, van de Vosse FN, Unser M, Stergiopulos N (2012) Experimental investigation of collagen waviness and orientation in the arterial adventitia using confocal laser scanning microscopy. Biomech Model Mechanobiol 11:461–473CrossRefPubMedGoogle Scholar
- 25.Dooley KA (2011) Raman spectroscopic studies of bone biomechanical function and development in animal models. University of MichiganGoogle Scholar
- 30.Pritchard JJ (1972) The Osteoblast. In: Bourne GH (ed) The biochemistry and physiology of bone. Academic Press Inc, New YorkGoogle Scholar
- 41.Mahamid J, Aichmayer B, Shimoni E, Ziblat R, Li C, Siegel S, Paris O, Fratzl P, Weiner S, Addadi L (2010) Mapping amorphous calcium phosphate transformation into crystalline mineral from the cell to the bone in zebrafish fin rays. Proc Natl Acad Sci USA 107:6316–6321PubMedCentralCrossRefPubMedGoogle Scholar