Novel Insights Into the Architecture of Macro and Microstructures in Cattle Ossa Cordis.

Ossa cordis, bones located within the heart trigones, are often classified as heterotopic or ectopic bones. Despite their high prevalence in cattle and some other bovids, little is known about their structure or development. Scanning electron microscopy, X-ray microtomography, gross dissections, and measurements showed the anatomical locations, prevalence, shapes, and measurements of the cardiac bones in both Egyptian Baladi cattle and Holstein-Friesians. All cattle (n = 12) had an Ossa cordis dextrum (average = 50.70 × 20.91 × 5.40 mm). Additionally, 80% Egyptian Baladi and 57% Holstein-Friesian had a smaller Ossa cordis sinistrum (average = 24.94 × 12.75 × 4.12 mm). Egyptian Baladi Ossa cordis were smaller than observed in Holstein-Friesians. Energy-dispersive X-ray analysis showed the elemental constitution (carbon, oxygen, calcium, nitrogen, phosphorus, sodium, and magnesium) of Ossa cordis and Cartilago cordis. These imaging techniques, plus four histological stains (hematoxylin and eosin, Crossman's trichrome, Alcian blue with Van Gieson, and Sirius Red) and microscopy, demonstrated osteoblasts, osteocytes, osteoclasts, astrocytes, blood vessels, bone marrow, lamellar and woven bone, cortical bone, trabeculations with pores and canaliculi, and fibrous components including collagen in the Ossa cordis dextrum and sinistrum. Hyaline cartilage and fibrocartilage (chondrocytes and cartilage matrix) were found within and surrounding the Ossa cordis. These findings were additionally compared against other cattle breeds and species.

Structural hierarchy confers error tolerance in biological materials.

Structural hierarchy, in which materials possess distinct features on multiple length scales, is ubiquitous in nature. Diverse biological materials, such as bone, cellulose, and muscle, have as many as 10 hierarchical levels. Structural hierarchy confers many mechanical advantages, including improved toughness and economy of material. However, it also presents a problem: Each hierarchical level adds a new source of assembly errors and substantially increases the information required for proper assembly. This seems to conflict with the prevalence of naturally occurring hierarchical structures, suggesting that a common mechanical source of hierarchical robustness may exist. However, our ability to identify such a unifying phenomenon is limited by the lack of a general mechanical framework for structures exhibiting organization on disparate length scales. Here, we use simulations to substantiate a generalized model for the tensile stiffness of hierarchical filamentous networks with a nested, dilute triangular lattice structure. Following seminal work by Maxwell and others on criteria for stiff frames, we extend the concept of connectivity in network mechanics and find a similar dependence of material stiffness upon each hierarchical level. Using this model, we find that stiffness becomes less sensitive to errors in assembly with additional levels of hierarchy; although surprising, we show that this result is analytically predictable from first principles and thus potentially model independent. More broadly, this work helps account for the success of hierarchical, filamentous materials in biology and materials design and offers a heuristic for ensuring that desired material properties are achieved within the required tolerance.

Comparison of bone microstructures via high-resolution peripheral quantitative computed tomography in patients with different stages of chronic kidney disease before and after starting hemodialysis.

Chronic kidney disease (CKD) negatively affects bone strength; however, the osteoporotic conditions in patients with CKD are not fully understood. Moreover, the changes in bone microstructure between pre-dialysis and dialysis are unknown. High-resolution peripheral quantitative computed tomography (HR-pQCT) reveals the three-dimensional microstructures of the bone. We aimed to evaluate bone microstructures in patients with different stages of CKD. This study included 119 healthy men and 40 men admitted to Nagasaki University Hospital for inpatient education or the initiation of hemodialysis. The distal radius and tibia were scanned with HR-pQCT. Patient clinical characteristics and bone microstructures were evaluated within 3 months of initiation of hemodialysis (in patients with CKD stage 5 D), patients with CKD stage 4-5, and healthy volunteers. Cortical bone parameters were lower in the CKD group than in healthy controls. Tibial cortical and trabecular bone parameters (cortical thickness, cortical area, trabecular volumetric bone mineral density, trabecular-bone volume fraction, and trabecular thickness) differed between patients with CKD stage 5 D and those with CKD stage 4-5 (p < 0.01). These differences were also observed between patients with CKD stage 5 and those with CKD stage 5 D (p < 0.017), but not between patients with CKD stage 4 and those with CKD stage 5, suggesting that the bone microstructure rapidly changed at the start of hemodialysis. Patients with CKD stage 5 D exhibited tibial microstructural impairment compared with those with CKD stage 4-5. HR-pQCT is useful for elucidation of the pathology of bone microstructures in patients with renal failure.

Multicolor quantitative confocal imaging cytometry.

Multicolor 3D quantitative imaging of large tissue volumes is necessary to understand tissue development and organization as well as interactions between distinct cell types in situ. However, tissue imaging remains technically challenging, particularly imaging of bone and marrow. Here, we describe a pipeline to reproducibly generate high-dimensional quantitative data from bone and bone marrow that may be extended to any tissue. We generate thick bone sections from adult mouse femurs with preserved tissue microarchitecture and demonstrate eight-color imaging using confocal microscopy without linear unmixing. We introduce XiT, an open-access software for fast and easy data curation, exploration and quantification of large imaging data sets with single-cell resolution. We describe how XiT can be used to correct for potential artifacts in quantitative 3D imaging, and we use the pipeline to measure the spatial relationship between hematopoietic cells, bone matrix and marrow Schwann cells.

Netrin-1 regulates ERK1/2 signaling pathway and autophagy activation in wear particle-induced osteoclastogenesis.

BACKGROUND: Artificial joint replacement surgery is often accompanied by osteolysis induced aseptic loosening around the prosthesis. Wear particles from joint replacement are thought to be one of the main factors leading to local inflammation and osteolysis at the prosthesis site. The aim of this study was to investigate the molecular mechanism of osteoclast formation and dissolution induced by wear particles and the potential roles of Netrin-1, the ERK1/2 pathway and autophagy activation in this process. METHODS: The messenger RNA levels in cells and tissues were detected with real-time quantitative PCR. The western blotting was used to detect the expression of proteins. A CCK-8 kit was used to detect the viability of RAW 264.7 cells. Moreover, an air pouch model of bone resorption was established. Immunohistochemistry was used to detect the expression of TRAP and Netrin-1 in rat bone tissue. Cell culture supernatants were collected in the rat air pouch model of bone resorption, and the levels of RANKL and OPG were detected with enzyme-linked immunosorbent assay. The protein levels of TRAP and Netrin-1 in bone tissue were examined by immunohistochemistry. RESULTS: Titanium wear particles induced osteoclast formation and autophagy activation. Moreover, blocking autophagy suppressed the osteoclastogenesis after exposure to wear particles in vitro. The activation of the ERK1/2 pathway and the overexpression of Netrin-1 were both found to play important roles in osteoclastogenesis mediated by autophagy. Moreover, 3-MA effectively decreased the secretion of proinflammatory cytokines mediated by wear particles. CONCLUSION: Blockade of autophagy inhibits the osteoclastogenesis and inflammation induced by wear particles, thus potentially providing novel treatment strategies for abnormal osteoclastogenesis and aseptic prosthesis loosening induced by wear particles.

Laser fabrication of structural bone: surface morphology and biomineralization assessment.

The current work explores the surface morphology of the laser-ablated bone using Yb-fiber coupled Nd:YAG laser (λ = 1064 nm) in continuous wave mode. As the laser-ablated region contains physiochemically modified carbonized and nonstructural region, it becomes unknown material for the body. Thus, biomineralization on such a laser-ablated region was assessed by in vitro immersion test in noncellular simulated body fluid. The presence of hydroxyapatite was detected in the precipitated mineral product using scanning electron microscopy equipped with energy dispersive spectroscopy, and X-ray diffraction analysis. The effect of varying laser parameters on distribution of surface morphology features was identified and its corresponding effect on biomineralization was studied.

Efficacy for Whitlockite for Augmenting Spinal Fusion.

Whitlockite (WH) is the second most abundant inorganic component of human bone, accounting for approximately 25% of bone tissue. This study investigated the role of WH in bone remodeling and formation in a mouse spinal fusion model. Specifically, morphology and composition analysis, tests of porosity and surface area, thermogravimetric analysis, an ion-release test, and a cell viability test were conducted to analyze the properties of bone substitutes. The MagOss group received WH, Group A received 100% beta-tricalcium phosphate (ß-TCP), Group B received 100% hydroxyapatite (HAp), Group C received 30% HAp/70% ß-TCP, and Group D received 60% HAp/40% ß-TCP (n = 10 each). All mice were sacrificed 6 weeks after implantation, and micro-CT, hematoxylin and eosin (HE) staining, and Masson trichome (MT) staining and immunohistochemistry were performed. The MagOss group showed more homogeneous and smaller grains, and nanopores (<500 nm) were found in only the MagOss group. On micro-CT, the MagOss group showed larger fusion mass and better graft incorporation into the decorticate mouse spine than other groups. In the in vivo experiment with HE staining, the MagOss group showed the highest new bone area (mean: decortication group, 9.50%; A, 15.08%; B, 15.70%; C, 14.76%; D, 14.70%; MagOss, 22.69%; p < 0.0001). In MT staining, the MagOss group demonstrated the highest new bone area (mean: decortication group, 15.62%; A, 21.41%; B, 22.86%; C, 23.07%; D, 22.47%; MagOss, 26.29%; p < 0.0001). In an immunohistochemical analysis for osteocalcin, osteopontin, and CD31, the MagOss group showed a higher positive area than other groups. WH showed comparable bone conductivity to HAp and ß-TCP and increased new bone formation. WH is likely to be used as an improved bone substitute with better bone conductivity than HAp and ß-TCP.

Genetically Modified Ferritin Nanoparticles with Bone-Targeting Peptides for Bone Imaging.

Bone homeostasis plays a major role in supporting and protecting various organs as well as a body structure by maintaining the balance of activities of the osteoblasts and osteoclasts. Unbalanced differentiation and functions of these cells result in various skeletal diseases, such as osteoporosis, osteopetrosis, and Paget's disease. Although various synthetic nanomaterials have been developed for bone imaging and therapy through the chemical conjugation, they are associated with serious drawbacks, including heterogeneity and random orientation, in turn resulting in low efficiency. Here, we report the synthesis of bone-targeting ferritin nanoparticles for bone imaging. Ferritin, which is a globular protein composed of 24 subunits, was employed as a carrier molecule. Bone-targeting peptides that have been reported to specifically bind to osteoblast and hydroxyapatite were genetically fused to the N-terminus of the heavy subunit of human ferritin in such a way that the peptides faced outwards. Ferritin nanoparticles with fused bone-targeting peptides were also conjugated with fluorescent dyes to assess their binding ability using osteoblast imaging and a hydroxyapatite binding assay; the results showed their specific binding with osteoblasts and hydroxyapatite. Using in vivo analysis, a specific fluorescent signal from the lower limb was observed, demonstrating a highly selective affinity of the modified nanoparticles for the bone tissue. These promising results indicate a specific binding ability of the nanoscale targeting system to the bone tissue, which might potentially be used for bone disease therapy in future clinical applications.

Enhanced BMP-2-Mediated Bone Repair Using an Anisotropic Silk Fibroin Scaffold Coated with Bone-like Apatite.

The repair of large bone defects remains challenging and often requires graft material due to limited availability of autologous bone. In clinical settings, collagen sponges loaded with excessive amounts of bone morphogenetic protein 2 (rhBMP-2) are occasionally used for the treatment of bone non-unions, increasing the risk of adverse events. Therefore, strategies to reduce rhBMP-2 dosage are desirable. Silk scaffolds show great promise due to their favorable biocompatibility and their utility for various biofabrication methods. For this study, we generated silk scaffolds with axially aligned pores, which were subsequently treated with 10× simulated body fluid (SBF) to generate an apatitic calcium phosphate coating. Using a rat femoral critical sized defect model (CSD) we evaluated if the resulting scaffold allows the reduction of BMP-2 dosage to promote efficient bone repair by providing appropriate guidance cues. Highly porous, anisotropic silk scaffolds were produced, demonstrating good cytocompatibility in vitro and treatment with 10× SBF resulted in efficient surface coating. In vivo, the coated silk scaffolds loaded with a low dose of rhBMP-2 demonstrated significantly improved bone regeneration when compared to the unmineralized scaffold. Overall, our findings show that this simple and cost-efficient technique yields scaffolds that enhance rhBMP-2 mediated bone healing.

Visualization of woven bone structure through analysis of biopsy specimens using synchrotron radiation and conventional X-ray microcomputed tomography.

This study explores the application of synchrotron radiation and conventional microcomputed tomography (SR-µCT and C-µCT, respectively) in evaluating bone-biopsy specimens. Bone-biopsy specimens were obtained using a trephine bur during bone-graft removal for implant placement six months after performing a maxillary sinus bone-graft procedure. Image data of specimens were obtained using SR-µCT and C-µCT. SR-µCT was performed using the 6C biomedical imaging beamline at the Pohang Accelerator Laboratory with a monochromatic X-ray beam of 23 keV, and C-µCT was performed using a table-top CT scanner (Skyscan 1272). Reconstruction images obtained using the two methods were qualitatively compared with 2D images evaluated under 3D visualization. The SR-µCT images, especially of the new-bone-graft-woven-bone formation, were less noisy and sharper than the C-µCT images. To evaluate the new-bone-graft-woven-bone formation, only the SR-µCT images showed areas of new bone (NB) formation with bone substitute (BS; Bio-Oss) and woven bone (WB) contact, and correctly visualized true 3D structures of bone formation. Hence, µCT techniques are non-destructive and can provide detailed images of bone biopsy. In particular, SR-µCT can be used to obtain improved image quality with contrast of NB, BS and WB, demonstrating a level of detail comparable with bone formation. SR-µCT could be an unbiased 3D alternative for imaging WB formation and for high-throughput analysis.

Abnormal lacuno-canalicular network and negative correlation between serum osteocalcin and Cobb angle indicate abnormal osteocyte function in adolescent idiopathic scoliosis.

Adolescent idiopathic scoliosis (AIS) is a prevalent spinal deformity occurring during peripubertal growth period that affects 1-4% of adolescents globally without clear etiopathogenetic mechanism. Low bone mineral density is an independent and significant prognostic factor for curve progression. Currently, the cause underlying low bone mass in AIS remains elusive. Osteocytes play an important role in bone metabolism and mineral homeostasis, but its role in AIS has not been studied. In the present study, iliac bone tissues were harvested from 21 patients with AIS (mean age of 14.3 ± 2.20 yr old) with a mean Cobb angle of 55.6 ± 10.61° and 13 non-AIS controls (mean age of 16.5 ± 4.79 yr old) intraoperatively. Acid-etched scanning electron microscopy (SEM) images of AIS demonstrated abnormal osteocytes that were more rounded and cobblestone-like in shape and were aligned in irregular clusters with shorter and disorganized canaliculi. Further quantitative analysis with FITC-Imaris technique showed a significant reduction in the canalicular number and length as well as an increase in lacunar volume and area in AIS. SEM with energy-dispersive X-ray spectroscopy analysis demonstrated a lower calcium-to-phosphorus ratio at the perilacunar/canalicular region. Moreover, microindentaion results revealed lower values of Vickers hardness and elastic modulus in AIS when compared with controls. In addition, in the parallel study of 99 AIS (27 with severe Cobb angle of 65.8 ± 14.1° and 72 with mild Cobb angle of 26.6 ± 9.1°) with different curve severity, the serum osteocalcin level was found to be significantly and negatively associated with the Cobb angle. In summary, the findings in this series of studies demonstrated the potential link of abnormal osteocyte lacuno-canalicular network structure and function to the observed abnormal bone mineralization in AIS, which may shed light on etiopathogenesis of AIS.-Chen, H., Zhang, J., Wang, Y., Cheuk, K.-Y., Hung, A. L. H., Lam, T.-P., Qiu, Y., Feng, J. Q., Lee, W. Y. W., Cheng, J. C. Y. Abnormal lacuno-canalicular network and negative correlation between serum osteocalcin and Cobb angle indicate abnormal osteocyte function in adolescent idiopathic scoliosis.

Characterization of the Robinow syndrome skeletal phenotype, bone micro-architecture, and genotype-phenotype correlations with the osteosclerotic form.

Robinow syndrome (RS) is a genetically heterogeneous skeletal dysplasia with recent reports suggesting an osteosclerotic form of the disease. We endeavored to investigate the full spectrum of skeletal anomalies in a genetically diverse cohort of RS patients with a focus on the bone micro-architecture. Seven individuals with molecularly confirmed RS, including four with DVL1 variants and single individuals with variants in WNT5A, ROR2, and GPC4 underwent a musculoskeletal focused physical examination, dual-energy X-ray absorptiometry (DEXA) scan, and high-resolution peripheral quantitative computed tomography (HR-pQCT). Skeletal examination revealed variability in limb shortening anomalies consistent with recent reports. DEXA scan measures revealed increased total body bone mineral density (BMD) (3/7), cranial BMD (5/7), and non-cranial BMD (1/7). Cranial osteosclerosis was only observed in DVL1-RS (4/4) and GPC4-RS (1/1) subjects and in one case was complicated by choanal atresia, bilateral conductive hearing loss, and cranial nerve III, VI, and VII palsy. HR-pQCT revealed a unique pattern of low cortical BMD, increased trabecular BMD, decreased number of trabeculations, and increased thickness of the trabeculations for the DVL1-RS subjects. The spectrum of skeletal anomalies including the micro-architecture of the bones observed in RS has considerable variability with some osteosclerosis genotype-phenotype correlations more frequent with DVL1 variants.

Contribution to understand the biomineralization of bones.

INTRODUCTION: The goal is to propose a material scientific hypothesis for the atomic arrangement of calcium phosphates during the mineralization of bones. MATERIALS AND METHODS: It was reached by the analysis of bones of healthy and osteoporotic rats using analytical transmission electron microscopic methods. RESULTS: Electron diffraction patterns show hydroxyapatite (HAP) as dominant phase within the mineralized areas. In the electron energy loss spectrum, a double peak of the phosphorous L-edge seems to be a characteristic feature of the phosphorous binding in biological HAP. The hypothesis bases on periodic features on the collagen surface which agree with distances between oxygen atoms in the (200) plane of octacalcium phosphate (OCP). Bridge pillars for the HAP network consist of OCP coupled with a half unit cell on collagen by oxygen-hydrogen bridges. Possibly, the metastable OCP bridges are only a transient step, while the mineralization is starting. OCP and HAP couple by similar distances of calcium atoms in an interface close to the (100) planes of the OCP and the HAP network. To reach the perfect overlap of the equidistant Ca atoms, the HAP network has to be rotated by 22.5° around the a-axis, 11.5° around the c-axis of HAP, and 10.1° around an axis perpendicular to a and c. CONCLUSIONS: A supercell based on this idea is able to explain the dominance of HAP in the electron diffraction patterns, the arrangement of the (002) lattice planes perpendicular to the collagen fiber axis, and sections of high-resolution TEM images.

Evolution of surface morphology of Er:YAG laser-machined human bone.

The extensive research on the laser machining of the bone has been, so far, restricted to drilling and cutting that is one- and two-dimensional machining, respectively. In addition, the surface morphology of the laser machined region has rarely been explored in detail. In view of this, the current work employed three-dimensional laser machining of human bone and reports the distinct surface morphology produced within a laser machined region of human bone. Three-dimensional laser machining was carried out using multiple partially overlapped pulses and laser tracks with a separation of 0.3 mm between the centers of consecutive laser tracks to remove a bulk volume of the bone. In this study, a diode-pumped pulse Er:YAG laser (λ = 2940 nm) was employed with continuously sprayed chilled water at the irradiation site. The resulting surface morphology evolved within the laser-machined region of the bone was evaluated using scanning electron microscopy, energy dispersive spectroscopy, and X-ray micro-computed tomography. The distinct surface morphology involved cellular/channeled scaffold structure characterized by interconnected pores surrounded by solid ridges, produced within a laser machined region of human structural bone. Underlying physical phenomena responsible for evolution of such morphology have been proposed and explained with the help of a thermokinetic model.

Investigation of the mechanism of gamma irradiation effect on bovine bone.

Radiation sterilization is an effective method of bone sterilization prior to bone graft transplantation. Gamma irradiation affects the biological and mechanical properties of bone; depending on the dose of radiation. The effect of gamma irradiation on bone mechanical properties is an unwanted phenomenon. However the mechanism of the effect of irradiation on bone mechanical properties is not properly understood. In this research paper the mechanism of the effect of gamma irradiation on bovine bone is investigated using scanning electron microscopy, energy-dispersive X-rays spectroscopy and Fourier transform infrared spectroscopy techniques. Gamma irradiation affects the mineral and fiber composition of bovine bone. The mineral content of bone especially calcium, magnesium and phosphorus decrease with increasing dose of gamma radiation. At Nano-level gamma irradiation alter amide I, amide II and amide III collagen contents. High dose gamma irradiation induces collagen cross-linking reaction in bone and degrades bone properties.

The Association of Vitamin D in Youth and Early Adulthood with Bone Mineral Density and Microarchitecture in Early Adulthood.

This study aimed to describe the association of vitamin D status at different stages of growth with bone measures in adolescence and early adulthood. There were 415 participants followed from age 8 to 16, and 201 further followed to age 25. Areal bone mineral density (BMD) at the spine, hip and total body was measured by dual-energy X-ray absorptiometry at ages 16 and 25, and tibial and radial trabecular and cortical bone microarchitecture by high resolution peripheral quantitative computerised tomography at age 25. Serum 25-hydroxyvitamin D (25OHD) concentrations were measured at ages 8, 16 and 25. Multivariable linear regression was used to analyse the association of 25OHD concentrations at three timepoints with bone measures at ages 16 and 25. The proportion of participants with vitamin D deficiency (< 50 nmol/L) was 11%, 43% and 41% at three timepoints, respectively. Serum 25OHD concentrations at age 8 were not significantly associated with any bone measures at age 16 or 25. Serum 25OHD concentrations at age 16 had a significant association with higher BMD at nearly all sites at ages 16 and 25 as well as lower radial porosity and more compact trabecular microarchitecture (higher density, increased number and reduced separation) at both the radius and tibia at age 25. Serum 25OHD concentrations at age 25 were only associated with hip BMD. Higher vitamin D concentrations in adolescence, to a lesser extent at age 25, have beneficial associations with BMD and bone microarchitecture in early adulthood. Optimising vitamin D status particularly during adolescence should be a priority.

Prospective Follow-Up of Cortical Interruptions, Bone Density, and Micro-structure Detected on HR-pQCT: A Study in Patients with Rheumatoid Arthritis and Healthy Subjects.

OBJECTIVES: The purpose of the study was to prospectively investigate change (repair or progression) in the number, surface area and volume of cortical interruptions, bone density (vBMD) and micro-structural parameters assessed by high-resolution peripheral quantitative computed tomography (HR-pQCT) in finger joints of patients with rheumatoid arthritis (RA) treated with synthetic disease modifying anti-rheumatic drugs (sDMARDs) and/or biologic DMARDs (bDMARDs) over a 1-year follow-up period, and in comparison with healthy subjects (HS). METHODS: Thirty-two patients with RA (221 joints, 53% on bDMARDs) and 32 HS (117 joints) were assessed at baseline and after 1 year using semi-automatic analysis of HR-pQCT images. Mean changes (group level) and the proportion of joints (joint level) with changes beyond the least significant change were calculated. RESULTS: At baseline, 530 interruptions were identified in patients, and 136 in HS. The mean of the interruption parameters did not significantly change in either group Mean vBMD decreased more in patients than in HS (- 4.4 versus - 1.1 mgHA/cm3, respectively). In patients versus HS, proportionally more joints showed repair in interruption volume (6.6% versus 1.7%, respectively) and loss of vBMD (26.7% versus 12.9%, respectively). In patients on sDMARDs versus patients on bDMARDs, proportionally more joints showed progression in the number of interruptions and loss of vBMD (6.1% versus 1.8% and 31.3% versus 17.2%, respectively). CONCLUSIONS: HR-pQCT is able to quantify bone repair and progression. Cortical interruption-, vBMD-, and micro-structure were impaired in RA, of which vBMD and micro-structure further deteriorated, particularly in patients on sDMARDs.

Bone Microstructure in Response to Vitamin D3 Supplementation: A Randomized Placebo-Controlled Trial.

Vitamin D supplementation is often used in the prevention and treatment of osteoporosis, but the role of vitamin D has lately been questioned. We aimed to investigate the effect of 3 months of daily vitamin D3 supplementation (70 µg [2800 IU] vs. placebo) initiated in winter months on bone health. This study is a double-blinded placebo-controlled randomized trial. Bone health was assessed by bone turnover markers, DXA, HRpQCT, and QCT scans. The participants were 81 healthy postmenopausal women with low 25(OH)D (< 50 nmol/l) and high PTH levels (> 6.9 pmol/l) at screening. Vitamin D3 supplementation significantly increased levels of 25(OH)D and 1,25(OH)2D by 59 nmol/l and 19 pmol/l, respectively, whereas PTH was reduced by 0.7 pmol/l (all p < 0.0001). Compared with placebo, vitamin D3 did not affect bone turnover markers, aBMD by DXA or trabecular bone score. Vitamin D3 increased trabecular vBMD (QCT scans) in the trochanter region (0.4 vs. - 0.7 g/cm3) and the femoral neck (2.1 vs. - 1.8 g/cm3) pall < 0.05. HRpQCT scans of the distal tibia showed reduced trabecular number (- 0.03 vs. 0.05 mm-1) and increased trabecular thickness (0.001 vs. - 0.005 mm), as well as an improved estimated bone strength as assessed by failure load (0.1 vs. - 0.1 kN), and stiffness (2.3 vs. - 3.1 kN/mm pall ≤ 0.01). Changes in 25(OH)D correlated significantly with changes in trabecular thickness, stiffness, and failure load. Three months of vitamin D3 supplementation improved bone strength and trabecular thickness in tibia, vBMD in the trochanter and femoral neck, but did not affect aBMD.

The Shisa3 knockout mouse exhibits normal bone phenotype.

Wnt signaling is important for both skeletal development and bone disease, with Wnt inhibitory factors playing critical roles in bone metabolism. SHISA3 blocks the maturation and transportation of Frizzled receptors to the cell surface, thereby inhibiting the Wnt/ß-catenin signaling pathway in lung cancer. However, the function of Shisa3 in bone biology remains uninvestigated. This study found that Shisa3 was strongly expressed in the calvarial bones of mice, especially in osteoblasts. In addition, adenovirus-mediated gene transfer of murine Shisa3 significantly inhibited Wnt3a-induced nuclear translocation of ß-catenin and mRNA expression of the Wnt target gene Axin2. In bone phenotype assessments of Shisa3 knockout (Shisa3 KO) mice, micro-computed tomography, mRNA expressions of osteoblast markers, and skeletal preparations all displayed no significant differences compared with Shisa3 wild-type mice. mRNA expression analysis of canonical Wnt signaling target genes (Axin2, Lef1, Dkk1, and Tnfrsf11b) in calvarial bones at P0.5 also revealed no significant findings. In Axin2Cre/ERT2 knock-in mice, the number of Axin2-expressing cells in the calvariae of Shisa3 KO and control mice were comparable. Thus, there appears to be a redundancy in the function of Shisa3 in bone development, likely with other Shisa family members.

Towards a Connectomic Description of the Osteocyte Lacunocanalicular Network in Bone.

PURPOSE OF REVIEW: Osteocytes are the most abundant bone cells. They are completely encased in mineralized tissue, sitting inside lacunae that are connected by a multitude of canaliculi. In recent years, the osteocyte network has been shown to fulfill endocrine functions and to communicate with a number of other organs. This review addresses emerging knowledge on the connectome of the lacunocanalicular network in different types of bone tissue. RECENT FINDINGS: Recent advances in three-dimensional imaging technology started to reveal parameters that are well known from general theory to characterize the function of networks, such as network density, degree of nodes, or shortest path length through the network. The connectome of the lacunocanalicular network differs in some aspects between lamellar and woven bone and seems to change with age. More research is needed to relate network structure to function, such as intercellular transport or communication and its role in mechanosensation, as well as to understand the effect of diseases.