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Teleost fish possess calcium carbonate otoliths located in separate chambers (utriculus, sacculus, and lagena) of their membranous labyrinth. This study analyzed the surface topography of the sagittal otolith of the Pacific sardine (Sardinops sagax) and the daily and annual increments in these otoliths. The otolith surface, characterized by laser scanning confocal microscopy for the first time, consisted of a system of prominent ridges and valleys (grooves), but it is unclear whether these structures are functional or represent time-resolving markings reflecting growth periodicity. Within the first-year volume, daily increments, each consisting of an incremental (more mineralized) and a discontinuous (less mineralized) zone, were resolved by optical microscopy and backscattered electron (BSE) imaging in the scanning electron microscope (SEM). Daily growth increments could, however, not be resolved in volumes formed after the first year, presumably because otolith growth markedly slows down and spacing of incremental markings narrows in older fish. Throughout otolith growth, the crystalline network continues across the discontinuous zones. Fluorochrome labeling provides additional information on growth after the first year. Compared with optical and BSE imaging, synchrotron microComputed Tomography of intact otoliths (with 0.69 µm volume elements) was less able to resolve daily increments; X-ray phase contrast reconstructions provided more detail than reconstructions with absorption contrast. Future research directions are proposed. [ABSTRACT FROM AUTHOR]

Shark vertebrae and their centra (vertebral bodies) are high-performance structures able to survive millions of cycles of high amplitude strain despite lacking a repair mechanism for accumulating damage. Shark centra consist of mineralized cartilage, a biocomposite of bioapatite (bAp), and collagen, and the nanocrystalline bAp's contribution to functionality remains largely uninvestigated. Using the multiple detector energy-dispersive diffraction (EDD) system at 6-BM-B, the Advanced Photon Source, and 3D tomographic sampling, the 3D functionality of entire centra were probed. Immersion in ethanol vs phosphate-buffered saline produces only small changes in bAp d -spacing within a great hammerhead centrum. EDD mapping under in situ loading was performed an entire blue shark centrum, and 3D maps of bAp strain showed the two structural zones of the centrum, the corpus calcareum and intermedialia, contained opposite-signed strains approaching 0.5%, and application of ~8% nominal strain did not alter these strain magnitudes and their spatial distribution. [ABSTRACT FROM AUTHOR]

We analyzed pedicle bone from roe bucks that had died around antler casting or shortly before or during the rutting period. Pedicles obtained around antler casting were highly porous and showed signs of intense osteoclastic activity that had caused the formation of an abscission line. Following the detachment of the antler plus a portion of pedicle bone, osteoclastic activity in the pedicles continued for some time, and new bone was deposited onto the separation plane of the pedicle stump, leading to partial pedicle restoration. Pedicles obtained around the rutting period were compact structures. The newly formed, often very large secondary osteons, which had filled the resorption cavities, exhibited a lower mineral density than the persisting older bone. The middle zones of the lamellar infilling frequently showed hypomineralized lamellae and enlarged osteocyte lacunae. This indicates a deficiency in mineral elements during the formation of these zones that occurred along with peak antler mineralization. We suggest that growing antlers and compacting pedicles compete for mineral elements, with the rapidly growing antlers being the more effective sinks. The competition between the two simultaneously mineralizing structures is probably more severe in Capreolus capreolus than in other cervids. This is because roe bucks regrow their antlers during late autumn and winter, a period of limited food and associated mineral supply. The pedicle is a heavily remodeled bone structure with distinct seasonal variation in porosity. Pedicle remodeling differs in several aspects from the normal bone remodeling process in the mammalian skeleton. [ABSTRACT FROM AUTHOR]

Laboratory microcomputed tomography (microCT) and synchrotron microCT imaged intact human second metacarpal bones (mc2) from two UK archeological sites: Ancaster (3rd to 4th century CE) and Wharram Percy (11th to 14th century CE). Two female mc2 were studied from each of three age at death cohorts (young, 18–29 years; middle, 30–49 years; old ≥50 years) along with a modern control mc2. The present investigation is complementary with an X‐ray scattering study of the same mc2 where the authors found no age‐at‐death‐related changes in carbonated apatite lattice parameters and found collagen D‐period peaks in the small angle regime in a minority of the mc2. This led the present authors to ask whether microCT could assign mc2 to the age cohort estimated by dental wear and whether material between bioerosion porosity and apparently free of diagenetic changes correlated with presence of strong D period peaks. Lab‐microCT derived values of bone volume fraction BV/TV (bone volume BV divided by total volume TV) for distal and proximal metaphyses provided age estimates that agree with those of dental wear. Cortical microstructure corroborated the BV/TV determination. Synchrotron microCT revealed significant diagenesis in all of the Wharram Percy and two of the Ancaster mc2; the resulting microstructural changes were attributed to microbial attack. The Ancaster mc2 whose microstructure matched that of the modern mc2 had D‐period peaks with intensities matching the modern bone. MicroCT with different voxel sizes was shown, therefore, to be very useful in age determination and in the assessment of 3D diagenetic changes. [ABSTRACT FROM AUTHOR]

High‐energy, wide‐angle x‐ray scattering (WAXS, x‐ray diffraction) and small‐angle x‐ray scattering (SAXS) were used to study intact human second metacarpal bones (mc2) from two UK archeological sites. A novel method correcting for irregular mass distribution was applied in these transmission geometry experiments done at beamline 1‐ID of the Advanced Photon Source. The authors asked whether there were age‐at‐death‐related changes in carbonated apatite (cAp) lattice parameters and whether SAXS could detect collagen D‐period peaks in the archeological mc2. For each of the two sites, Ancaster and Wharram Percy in England, six female mc2s were studied; for each site, two were from each of three age‐at‐death cohorts (young, 18–29 years; middle, 30–49 years; old ≥50 years) along with a modern control mc2. The Rietveld method was applied to the WAXS patterns to provide precise lattice parameter values. The cAp lattice parameters did not correlate with age‐at‐death estimated from dental wear. From WAXS and the 00.2 diffraction peak widths, four archeological mc2s possessed coherently scattering domain lengths (crystallite c‐axis sizes) that matched that of the modern mc2; SAXS revealed the same four archeological mc2 had D‐period peak intensities equivalent to that of the modern mc2. The other eight archeological mc2s had significantly larger crystallite sizes (than the modern mc2) and weak or absent D‐period peaks, differences attributed to diagenetic changes. Based on these data, the authors suggest that WAXS 00.2 peak width and SAXS D‐period peak intensity can be used with intact bones to select those likely to retain largely unaltered tissue nanostructure, which might be required for other analyses. Taken as a whole, the results suggest detecting age‐related deterioration in nanostructural features may be difficult in bone showing significant bioerosion. [ABSTRACT FROM AUTHOR]

Background: After spinal surgery and other orthopaedic procedures, most patients receive opioids for pain, leading to potential complications such as pseudarthrosis and opioid abuse associated with long-term use. As an alternative, the endocannabinoid system has been shown to have antinociceptive activity, while contributing to bone homeostasis via the CB1 and CB2 cannabinoid receptors. This study evaluates the impact of the cannabinoid receptor agonist WIN55,212-2 (WIN55) on osteogenic differentiation in vitro as well as bone regeneration and spinal fusion in a preclinical rat model.Methods: Primary rat bone marrow stromal cells were cultured in standard or osteogenic media and exposed to vehicle alone or WIN55. Runx2 and Alkaline phosphatase (Alpl) were quantified via qPCR (quantitative real-time polymerase chain reaction), followed by assessment of ALP activity and matrix mineralization. For in vivo evaluation, 45 female Sprague Dawley rats (n = 15 per group) underwent L4-L5 posterolateral spinal fusion with bilateral placement of collagen scaffolds preloaded with low-dose rhBMP-2 (recombinant human bone morphogenetic protein-2; 0.5 μg/implant). Postoperatively, rats received the vehicle alone or 0.5 or 2.5 mg/kg WIN55 via daily intraperitoneal injections for 5 days. Bone regeneration and spinal fusion were assessed using radiography, manual palpation-based fusion scoring, microcomputed tomography imaging, and histology.Results: mRNA expression levels of Runx2 and Alp were similar among cells treated with vehicle alone and WIN55. Likewise, exposure to WIN55 did not inhibit ALP activity or bone matrix mineralization. In this animal model, no significant differences were found among groups with regard to mean fusion score, fusion rate, or new bone volume.Conclusions: WIN55 showed no adverse impact on osteogenic differentiation, bone regeneration, and spinal fusion. This supports that cannabinoid receptor agonists should be further investigated as a potential alternative approach for postoperative analgesia following spinal fusion and other orthopaedic procedures requiring bone-healing.Clinical Relevance: The identification of alternative treatments for postoperative pain following orthopaedic surgical procedures is crucial in combating the ongoing opioid abuse crisis. The endocannabinoid system may represent a viable alternative target for addressing orthopaedic postoperative pain. [ABSTRACT FROM AUTHOR]