Diaphyseal Cortical Bone Research Articles

How to avoid genu recurvatum in leg-length discrepancy treated with tension-band plates. A volumetric magnetic resonance analysis

Aims and objectivesGenu recurvatum deformity after treatment of leg-length discrepancy (LLD) with tension-band plating is a recognized, but poorly described phenomenon in medical literature. The aim of this study was to evaluate clinical and radiological features of patients treated with tension-band plating for LLD assessing the development of a recurvatum deformity and its relationship to plate and screw disposition in a transversal plane, thus attempting to establish optimal plate positioning. Materials and methodsRetrospective study of children with LLD treated with tension-band plating. Primary endpoints were clinical and radiological knee recurvatum and anterior and posterior physeal areas measured drawing a line spanning from the lateral to the medial tension-band plates in the transverse plane using volumetric magnetic resonance imaging (vMRI). These findings were compared between patients with and without knee recurvatum. ResultsTwelve children (mean age 11.7 years) were included. Average follow-up was 2.6 years (1.5–5.0). Tension-band plating led to a significant reduction in LLD (mean, 15 mm). Six patients (50 %) developed clinical genu recurvatum (mean, 22°). According to vMRI, patients with genu recurvatum had a larger posterior to anterior physeal area ratio in both distal femur (1.6 versus 0.9, p < 0.05) and proximal tibial physes (2.2 versus 1.0, p < 0.05). ConclusionThe optimal position of the tension-band plates in distal femoral and proximal tibial physes should be in a point where a posterior to anterior physeal areas ratio is around 1.0, so as to achieve an even distribution of the physeal areas in the multidimensional physeal transverse plane. This point anatomically corresponds in the sagittal X-ray view to an imaginary line located just anterior to the posterior diaphyseal cortical bone on a true lateral radiograph for both femur and tibia.

Osteoblastgenic and Osteogenic Effects of KY-273 with CDK8/19 Inhibitory Activity in Bone Marrow Mesenchymal Stem Cells and Female Rats.

Osteoporosis is caused by imbalance between osteogenesis and bone resorption, thus, osteogenic drugs and resorption inhibitors are used for treatment of osteoporosis. The present study examined the effects of (R)-4-(1-hydroxyethyl)-3-{4-[2-(tetrahydropyran-4-yloxy)ethoxy]phenoxy}benzamide (KY-273), a diphenyl ether derivative, on CDK8/19 activity, osteoblast differentiation and femoral bone using micro-computed tomography in female rats. KY-273 potently inhibited CDK8/19 activity, promoted osteoblast differentiation with an increase in alkaline phosphatase (ALP) activity, and gene expression of type I collagen, ALP and BMP-4 in mesenchymal stem cells (ST2 cells). In female rat femur, ovariectomy decreased metaphyseal trabecular bone volume (Tb.BV), mineral content (Tb.BMC), yet had no effect on metaphyseal and diaphyseal cortical bone volume (Ct.BV), mineral content (Ct.BMC) and strength parameters (BSPs). In ovaries-intact and ovariectomized rats, oral administration of KY-273 (10 mg/kg/d) for 6 weeks increased metaphyseal and diaphyseal Ct.BV, Ct.BMC, and BSPs without affecting medullary volume (Med.V), but did not affect Tb.BV and Tb.BMC. In ovariectomized rats, alendronate (3 mg/kg/d) caused marked restoration of Tb.BV, Tb.BMC and structural parameters after ovariectomy, and increased metaphyseal but not diaphyseal Ct.BV, Ct.BMC, and BSPs. In ovaries-intact and ovariectomized rats, by the last week, KY-273 increased bone formation rate/bone surface at the periosteal but not the endocortical side. These findings indicate that KY-273 causes osteogenesis in cortical bone at the periosteal side without reducing Med.V. In conclusion, KY-273 has cortical-bone-selective osteogenic effects by osteoblastogenesis via CDK8/19 inhibition in ovaries-intact and ovariectomized rats, and is an orally active drug candidate for bone diseases such as osteoporosis in monotherapy and combination therapy.

Increased sizes and improved qualities of tibia bones by myostatin mutation in Japanese quail.

Production of large amounts of meat within a short growth period from modern broilers provides a huge economic benefit to the poultry industry. However, poor bone qualities of broilers caused by rapid growth are considered as one of the problems in the modern broilers industry. After discovery and investigation of myostatin (MSTN) as an anti-myogenic factor to increase muscle mass by targeted knockout in various animal models, additional positive effects of MSTN mutation on bone qualities have been reported in MSTN knockout mice. Although the same beneficial effects on muscle gain by MSTN mutation have been confirmed in MSTN mutant quail and chickens, bone qualities of the MSTN mutant birds have not been investigated, yet. In this study, tibia bones were collected from MSTN mutant and wild-type (WT) quail at 4months of age and analyzed by Micro-Computed Tomography scanning to compare size and strength of tibia bone and quality parameters in diaphysis and metaphysis regions. Length, width, cortical thickness, and bone breaking strength of tibia bones in the MSTN mutant group were significantly increased compared to those of the WT group, indicating positive effects of MSTN mutation on tibia bone sizes and strength. Furthermore, bone mineral contents and bone volume of whole diaphysis, diaphyseal cortical bone, whole metaphysis, and metaphyseal trabecular and cortical bones were significantly increased in the MSTN mutant group compared to the WT group, indicating increased mineralization in the overall tibia bone by MSTN mutation. Especially, higher bone mineral density (BMD) of whole diaphysis, higher total surface of whole metaphysis, and higher BMD, trabecular thickness, and total volume of metaphyseal trabecular bones in the MSTN mutant group compared to the WT group suggested improvements in bone qualities and structural soundness of both diaphysis and metaphysis regions with significant changes in trabecular bones by MSTN mutation. Taken together, MSTN can be considered as a potential target to not only increase meat yield, but also to improve bone qualities that can reduce the incidence of leg bone problems for the broiler industry.

Silk-Based Bioengineered Diaphyseal Cortical Bone Unit Enclosing an Implantable Bone Marrow toward Atrophic Nonunion Grafting.

Postnatal fracture healing of atrophic long bone diaphyseal nonunions remains a challenge for orthopedic surgeons. Paucity of autologous spongiosa has potentiated the use of tissue engineered bone grafts to improve success rates of bone marrow engraftment used in plate reosteosynthesis. Herein, the development and in vitro validation of a "sandwich-type" biofabricated diaphyseal cross-sectional unit, with an outer mechanically robust bioprinted cortical bone shell, encompassing an engineered bone marrow, are reported. Channelized silk fibroin blend sponges derived from Bombyx mori and Antheraea assama help in developing compartmentalized endosteum, exhibiting specialized osteoblasts (endosteal niche) and discontinuous endothelium (vascular niche). The cellular cross-talk between these two niches triggered via integrin-mediated cell adhesion, enables in preserving quiescence state of CD34+ /CD38- hematopoietic stem cells and their recycling in the engineered marrow. The outer cortical bone strut is developed through multimaterial microextrusion bioprinting strategy. Osteogenically primed mesenchymal stem cells-laden silk fibroin-nano-hydroxyapatite bioink is bioprinted alongside paramagnetic Fe-doped bioactive glass-polycaprolactone blend thermoplastic ink, reinforcing it for mechanical stability. Pulsed magnetic field actuation positively influences the osteogenic commitment and maturation of the bioprinted constructs via mechanotransductory route. Therefore, the assembled engineered marrow and bioprinted cortical shell hold promise as potential orthobiologic substitutes toward atrophic nonunion repairs.

Tibial cortical and trabecular variables together can pinpoint the timing of impact loading relative to menarche in premenopausal females.

Though relationships between limb bone structure and mechanical loading have provided fantastic opportunities for understanding the lives of prehistoric adults, the lives of children remain poorly understood. Our aim was to determine whether or not adult tibial skeletal variables retain information about childhood/adolescent loading, through assessing relationships between cortical and trabecular bone variables and the timing of impact loading relative to menarche in premenopausal adult females. Peripheral quantitative computed tomography was used to quantify geometric and densitometric variables from the proximal tibial diaphysis (66% location) and distal epiphysis (4% location) among 81 nulliparous young adult female controls and athletes aged 19-33 years grouped according to intensity of impact loading both pre- and post-menarche: (1) Low:Low (Controls); (2) High:Low; (3) High:High; (4) Moderate:Moderate; (5) Low:Moderate. ANCOVA was used to compare properties among the groups adjusted for age, stature, and body mass. Significant increases in diaphyseal total cross-sectional area and strength-strain index were documented among groups with any pre-menarcheal impact loading relative to groups with none, regardless of post-menarcheal loading history (p < .01). In contrast, significantly elevated distal trabecular volumetric bone mineral density was only documented among groups with recent post-menarcheal loading relative to groups with none, regardless of pre-menarcheal impact loading history (p < .01). The consideration of diaphyseal cortical bone geometric and epiphyseal trabecular bone densitometric variables together within the tibia can identify variation in pre-menarcheal and post-menarcheal impact loading histories among premenopausal adult females.

Compound deletion of thrombospondin-1 and -2 results in a skeletal phenotype not predicted by the single gene knockouts.

The trimeric thrombospondin homologs, TSP1 and TSP2, are both components of bone tissue and contribute in redundant and distinct ways to skeletal physiology. TSP1-null mice display increased femoral cross-sectional area and thickness due to periosteal expansion, as well as diminished matrix quality and impaired osteoclast function. TSP2-null mice display increased femoral cross-sectional thickness and reduced marrow area due to increased endosteal osteoblast activity, with very little periosteal expansion. Osteoblast lineage cells are reduced in TSP2-null mice, but not in TSP1-null. The functional effects of combined TSP1 and TSP2 deficiency remain to be elucidated. Here, we examined the spectrum of detergent soluble proteins in diaphyseal cortical bone of growing (6-week old) male and female mice deficient in both thrombospondins (double knockout (DKO)). Of 3429 detected proteins, 195 were differentially abundant in both male and female DKO bones. Physiologically relevant annotation terms identified by Ingenuity Pathway Analysis included "ECM degradation" and "Quantity of Monocytes." Manual inspection revealed that a number of proteins with shared expression among osteoclasts and osteocytes were reduced in DKO bones. To associate changes in protein content with phenotype, we examined 12-week old male and female DKO and WT mice. DKO mice were smaller than WT and in male DKO, femoral cross section area was reduced. Some of the male DKO femora also had a flattened, less circular cross-section. Male DKO bones were less stiff in bending and they displayed reduced ultimate load. Displacements at yield load and at max load were both elevated in male DKO. However, the ratios of post-yield to pre-yield displacements significantly diminished in DKO suggesting proportionally reduced post-yield behavior. Male DKO mice also exhibited reductions in trabecular bone mass, which were surprisingly associated with equivalent osteoblast numbers and accordingly increased osteoblast surface. Marrow-derived colony forming unit-fibroblastic was reduced in male and female DKO mice. Together our data suggest that when both TSP1 and TSP2 are absent, a unique, sex-specific bone phenotype not predicted by the single knockouts, is manifested.

Genetic variability affects the skeletal response to immobilization in founder strains of the diversity outbred mouse population

Mechanical unloading decreases bone volume and strength. In humans and mice, bone mineral density is highly heritable, and in mice the response to changes in loading varies with genetic background. Thus, genetic variability may affect the response of bone to unloading. As a first step to identify genes involved in bones' response to unloading, we evaluated the effects of unloading in eight inbred mouse strains: C57BL/6J, PWK/PhJ, WSB/EiJ, A/J, 129S1/SvImJ, NOD/ShiLtJ, NZO/HlLtJ, and CAST/EiJ. C57BL/6J and NOD/ShiLtJ mice had the greatest unloading-induced loss of diaphyseal cortical bone volume and strength. NZO/HlLtJ mice had the greatest metaphyseal trabecular bone loss, and C57BL/6J, WSB/EiJ, NOD/ShiLtJ, and CAST/EiJ mice had the greatest epiphyseal trabecular bone loss. Bone loss in the epiphyses displayed the highest heritability. With immobilization, mineral:matrix was reduced, and carbonate:phosphate and crystallinity were increased. A/J mice displayed the greatest unloading-induced loss of mineral:matrix. Changes in gene expression in response to unloading were greatest in NOD/ShiLtJ and CAST/EiJ mice. The most upregulated genes in response to unloading were associated with increased collagen synthesis and extracellular matrix formation. Our results demonstrate a strong differential response to unloading as a function of strain. Diversity outbred (DO) mice are a high-resolution mapping population derived from these eight inbred founder strains. These results suggest DO mice will be highly suited for examining the genetic basis of the skeletal response to unloading.

The cat as a small dog?—Comparison of trabecular and cortical bone microarchitecture of radius and ulna in cats and small dogs using microcomputed tomography

The forearms of dogs and cats do not only differ anatomically from each other, but there are also differences in prevalence of radius and ulna fractures between the two species. The prevalence of antebrachial fractures is 18.0% in dogs and 2.0–8.0% in cats. Many studies focus solely on the trabecular and cortical bone structure of dogs and the characteristics of the cat are often disregarded.The aim of this study was to evaluate the trabecular structure parameters [bone volume fraction per total volume (BV/TV), bone surface per total volume (BS/BV), trabecular number (Tb.N), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), connectivity density (Conn. D), degree of anisotropy (DA)] and the diaphyseal cortical bone density (Mean Density) of the antebrachium in cats and small dogs to visualise their differences.For this purpose, a total of 32 forearms of cats (n = 8) and small dogs (n = 8) were evaluated using microcomputed tomography and the findings were compared.The results of the study showed that cats had higher values for BV/TV, Tb.Th, Tb.Sp, DA and Mean Density and lower values for BS/BV, Tb.N and Conn.D at radius and ulna compared to dogs.According to the results of this study, the higher bone volume fraction (BV/TV), thicker trabeculae (Tb.Th), increased anisotropy (DA) and significantly higher diaphyseal cortical density (Mean Density) could contribute to the lower fracture risk of the antebrachium in cats compared to small dogs.

Cancellous Bone May Have a Greater Adaptive Strain Threshold Than Cortical Bone.

ABSTRACTStrain magnitude has a controlling influence on bone adaptive response. However, questions remain as to how and if cancellous and cortical bone tissues respond differently to varied strain magnitudes, particularly at a molecular level. The goal of this study was to characterize the time‐dependent gene expression, bone formation, and structural response of the cancellous and cortical bone of female C57Bl/6 mice to mechanical loading by applying varying load levels (low: −3.5 N; medium: −5.2 N; high: −7 N) to the skeleton using a mouse tibia loading model. The loading experiment showed that cortical bone mass at the tibial midshaft was significantly enhanced following all load levels examined and bone formation activities were particularly elevated at the medium and high loads applied. In contrast, for the proximal metaphyseal cancellous bone, only the high load led to significant increases in bone mass and bone formation indices. Similarly, expression of genes associated with inhibition of bone formation (e.g., Sost) was altered in the diaphyseal cortical bone at all load levels, but in the metaphyseal cortico‐cancellous bone only by the high load. Finite element analysis determined that the peak tensile or compressive strains that were osteogenic for the proximal cancellous bone under the high load were significantly greater than those that were osteogenic for the midshaft cortical tissues under the low load. These results suggest that the magnitude of the strain stimulus regulating structural, cellular, and molecular responses of bone to loading may be greater for the cancellous tissues than for the cortical tissues. © 2021 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

CLINICAL AND PATHOLOGIC FINDINGS OF AN OUTBREAK OF VITAMIN D3-RESPONSIVE METABOLIC BONE DISEASE IN HERON AND EGRET (FAMILY ARDEIDAE) CHICKS FED CAPELIN (MALLOTUS VILLOSUS).

An increase in cases of metabolic bone disease (MBD) in chicks of six species of heron and egret (family Ardeidae) was identified at a wildlife rehabilitation center in the spring and summer of 2018. The outbreak affected 34.3% of birds in care for four or more days during the first 3 mo of the study and was the most common reason for euthanasia during that time. Cases were characterized by lameness, increased flexibility of multiple long bones, angular deformities, and bone fractures. Gross postmortem examinations were conducted on 145 nestlings and fledglings that died or were euthanatized either because of MBD or for unrelated conditions. Histology was performed in four cases and three controls. Histologic findings were characterized by multiple lesions in the appendicular long bones, including variable elongation of the physis, retention of cartilage cores in the metaphyseal primary spongiosa, poorly mineralized osteoid seams within the primary spongiosa, thinning or lack of diaphyseal cortical bone compaction, and folding fractures typically propagating through the physis-metaphyseal interface. Folding fractures were often associated with focal metaphyseal fibroplasia. The parathyroid gland diameter of birds diagnosed postmortem with MBD in care was significantly larger than that of unaffected birds. The authors hypothesized that a dietary deficiency of vitamin D3 because of low levels in the bird's captive diet of capelin (Mallotus villosus) was the cause of the MBD. Starting in mid-July every chick's diet was supplemented with 714 IU oral vitamin D3/kg body weight per day, after which the number of birds developing MBD declined to a rate of 4.3%. This study characterizes the clinical, gross, radiographic, and histologic features of vitamin D3-responsive MBD in young herons and egrets and provides evidence to support the recommendation that captive birds on a diet of capelin be supplemented with vitamin D3, especially during growth.

Osteoblastic Wnt1 regulates periosteal bone formation in adult mice.

Compelling clinical data together with genetically modified mouse models have demonstrated that Wnt1 is a key Wnt ligand in bone metabolism, regulating both osteoblast activity and osteoclast differentiation. We have previously shown that deletion of Wnt1 in limb mesenchymal cells leads to severe ostepenic bone phenotype and spontaneous fractures very early after birth. However, the function of Wnt1 in mature skeleton remained unknown. To investigate the role of Wnt1 specifically in adult bone metabolism, we generated an osteoblast lineage-targeted inducible Wnt1 knockout mouse model using tetracycline-controlled Osterix-Cre mouse line (Osx-Cre). In this model, the Cre recombinase expression is suppressed by administering doxycycline (Dox) in drinking water. As expected, Wnt1Osx-/- mice without Dox developed spontaneous fractures early by 3weeks of age due to severe trabecular and cortical osteopenia. Administration of Dox to Wnt1Osx-Dox-/- and control mice until 4weeks of age suppressed Wnt1 deletion and completely prevented the fractures. Withdrawal of Dox led to deletion in Wnt1 allele but the fracture incidence progressively decreased in Wnt1Osx-Dox-/- mice at 8 or 12weeks of age (4 and 8weeks after Dox withdrawal). Interestingly, deletion of Wnt1 at 4weeks of age resulted only in a modest and transient trabecular osteopenia that was more pronounced in females and was normalized by 12weeks of age. However, diaphyseal cortical bone mass and cortical thickness in the femurs were significantly decreased in Wnt1Osx-Dox-/- mice of both genders. Mechanisticly, this was due to impaired periosteal bone formation. Based on our data, in addition to its essential role in early skeletal growth, Wnt1 is an important regulator of modeling-based bone formation and cortical thickness in adult mice.

Effect of dihydrotestosterone, 17-β-estrogen, genistein and equol on remodeling and morphology of bone in osteoporotic male rats during bone healing

IntroductionThe aim of this study was to investigate the effect of dihydrotestosterone (DHT), 17-β-estrogen (E2), genistein (GEN) and equol (EQ) on bone remodeling and bone morphology during healing of osteoporotic male rat tibiae. Materials and methods180 Sprague-Dawley male rats were divided in 5 groups of 36 animals. After orchidectomy (ORX) and development of osteoporosis, trepanation of the tibia was performed. Until the time of trepanation all groups received soya free food (SF), then food change occurred and treatment started. At day 95, 102 and 151, samples were taken and histomorphometry was performed to analyze changes in bone structure under treatment. At day 33 and 70 all animals received calcein respective alizarin for polychrome bone labeling. ResultsThe cortical bone was particularly affected. Treatment with DHT and E2 led to a significant long-term expansion of the thickness of the diaphyseal cortical bone, while the phytoestrogens EQ and GEN only had a positive short-term effect in this area. Only E2 preserved the trabecular bone for a limited time. In all groups, periosteal and endosteal bone areas showed the highest bone formation activity. The osteoporotic male injured bone shows a shift in mineral apposition rate (MAR) from periosteal to endosteal bone in the SF, DHT and E2 groups but not in the GEN and EQ phytohormones groups. An MAR decrease in trabecular bone formation was observed at day 70 in all groups except the E2 group. ConclusionWe conclude from our results that healing of cortical bone defects in a rat model of male osteoporosis are mainly influenced by the estrogen pathway. Nevertheless, effects via purely androgenic mechanisms can also be demonstrated. The role of a phytohormone therapy is only marginal and if only useful for a short-term supportive approach. The role of the periosteal to endosteal shift during male osteoporotic bone healing needs to be further examined.

Raman spectroscopy investigation shows that mineral maturity is greater in CD-1 than in C57BL/6 mice distal femurs after sexual maturity

ABSTRACT Purpose/Aim of the study mice are the most often used pre-clinical lab models for studying the pathologies of bone mineralization. However, recent evidence suggests that two of the most often used mice strains (C57BL/6J and CD-1) might show differences in the bone mineralization process. This study sought to investigate the main compositional properties of bone tissue between nonpathological C57BL/6J and CD-1 murine knee joints. Materials and Methods : to this end, medial and lateral condylar subchondral bones and the adjacent diaphyseal cortical bone of 13 murine femurs (n = 7 C57BL/6J and n = 6 CD-1 at eight weeks old, just after sexual maturation) were analyzed with ex vivo Raman spectroscopy. Results : regardless of the bone tissue analyzed, our results showed that CD-1 laboratory mice present a more mature mineral phase than C57BL/6J laboratory mice, but present no difference in maturity of the collagen phase. For both strains, the subchondral bone of the medial condylar and cortical bone from the diaphysis have similar compositional properties, and CD-1 presents less variation than C57BL/6J. Furthermore, we depict a novel parametric relationship between the crystallinity and carbonate-to-amide-I ratio that might help in deciphering the mineral maturation process that occurs during bone’s mineralization. Conclusions : Our results suggest that the timing of bone maturation might be different between non-pathological C57BL/6J and CD-1 murine knee femurs.

TSP1 and TSP2 deficiencies affect LOX protein distribution in the femoral diaphysis and pro-peptide removal in marrow-derived mesenchymal stem cells in vitro

ABSTRACTThrombospondin-1 and 2 have each been implicated in collagen fibrillogenesis. We addressed the possibility that deficits in lysyl oxidase (LOX) contribute to the extracellular matrix (ECM) phenotype of TSP-deficient bone. We examined detergent insoluble (mature cross-linked) and soluble (newly secreted) ECM fractions prepared from diaphyseal cortical bone. Detergent-insoluble hydroxyproline content, an indicator of cross-linked collagen content and LOX function, was reduced in female TSP-deficient bones. In male diaphyses, only TSP2 deficiency affected insoluble hydroxyproline content. Western blot suggested that removal of the LOX-pro-peptide (LOPP), an indication of LOX activation, was not affected by TSP status. Instead, the distribution of pro-LOX and mature LOX between immature and mature ECM was altered by TSP-status. LOX was also examined in primary marrow-derived mesenchymal stem cells (MSC) treated with ascorbate. Relative LOPP levels were elevated compared to WT in MSC conditioned medium from female TSP-deficient mice. When cells were serum starved to limit LOX pro-peptide removal, pro-LOX levels were elevated in TSP2-/- cells compared to wild-type. This phenotype was associated with a transient increase in BMP1 levels in TSP2-/- conditioned medium. TSP2 was detected in bone tissue and osteoblast cell culture. TSP1 was only detected in insoluble ECM prepared from WT diaphyseal bone samples. Our data suggest that the trimeric thrombospondins contribute to bone matrix quality by regulating the distribution of pro and mature LOX between newly secreted, immature ECM and mature, cross-linked ECM.

The effects of resistance training on bone mineral density and bone quality in type 2 diabetic rats.

Resistance training (RT) has been known to be effective in maintaining and improving bone strength, which is based on bone mineral density (BMD) and bone quality. However, it is not clear whether RT is effective in improving bone strength in patients with type‐2 diabetes mellitus (T2DM), who have a high risk of fracture. Therefore, we tested the effects of a 6‐week RT regimen using percutaneous electrical stimulation in T2DM model rats, male Otsuka Long‐Evans Tokushima Fatty (OLETF), and its control, Long‐Evans Tokushima Otsuka (LETO). After 6 weeks of RT, tibial BMD in RT legs was significantly higher than that in control (CON) legs in both groups. In diaphyseal cortical bone, bone area/tissue area, and cortical thickness was significantly increased in RT legs compared with CON legs in both groups. Cortical porosity was highly observed in OLETF compared with LETO, but RT improved cortical porosity in both groups. Interestingly, trabecular number, trabecular thickness and trabecular space as well as BMD and bone volume/tissue volume in proximal tibial metaphyseal trabecular bone were significantly improved in RT legs compared with CON legs in both groups. In contrast, connectivity density and structural model index were not affected by RT. These results indicate that the 6‐week RT regimen effectively increased BMD and improved bone quality in T2DM model rats as well as control rats. Therefore, RT may have the potential to improve bone strength and reduce fracture risk, even in patients with T2DM.

Anisotropy of elastic modulus and hydroxyapatite crystal strain in diaphyseal cortical bone

Anisotropy of elastic modulus and hydroxyapatite crystal strain in diaphyseal cortical bone

The androgen receptor is required for maintenance of bone mass in adult male mice

Previous studies evaluating the role of the androgen receptor (AR) for bone mass have used mouse models with global or tissue-specific lifelong inactivation of the AR. However, these mouse models have the AR inactivated already early in life and the relative roles of the AR during development, sexual maturation and in adult mice cannot be evaluated separately. The aim of the present study was to determine the specific roles of the AR in bone during sexual maturation and in adult mice. The AR was conditionally ablated at four (pre-pubertal) or ten (post-pubertal) weeks of age in male mice using tamoxifen-inducible Cre-mediated recombination. Both the pre-pubertal and the post-pubertal AR inactivation were efficient demonstrated by substantially lower AR mRNA levels in seminal vesicle, bone and white adipose tissue as well as markedly reduced weights of reproductive tissues when comparing inducible ARKO mice and control mice at 14 weeks of age. Total body BMD, as analyzed by DXA, as well as tibia diaphyseal cortical bone thickness and proximal metaphyseal trabecular bone volume fraction, as analyzed by μCT, were significantly reduced by both pre-pubertal and post-pubertal AR inactivation. These bone effects were associated with an increased bone turnover, indicating a high bone turnover osteoporosis. Pre-pubertal but not post-pubertal AR inactivation resulted in substantially increased fat mass. In conclusion, the AR is required for maintenance of both trabecular and cortical bone in adult male mice while AR expression during puberty is crucial for normal fat mass homeostasis in adult male mice.

Predicting Diaphyseal Cortical Bone Status Using Measures of Muscle Force Capacity

Muscle cross-sectional area (MCSA) is often used as a surrogate for the forces applied to bones during physical activity. Although MCSA is a strong predictor of cortical bone status, its use makes assumptions about the relationship between muscle size and force that are inaccurate. Furthermore, to measure MCSA and other muscle force surrogates typically requires expensive and/or radiative laboratory equipment. Thus, this study aimed to determine whether clinical laboratory- and field-based methodologies for measuring muscular force capacity accounted for similar variance in diaphyseal cortical bone status as a commonly used muscular force surrogate, MCSA, at the midtibia in young men and women. Healthy young adults (n = 142, 19.7 ± 0.7 yr old, 52.8% female) were assessed via peripheral quantitative computed tomography at the midtibia for cortical bone status and MCSA. Muscle force capacity was measured via Biodex dynamometer, Nottingham leg extensor power rig, and Vertec vertical jump. Regression analysis compared the independent variance predicted by each muscle force measure with that of MCSA, accounting for relevant confounders. MCSA, knee extension peak torque, and peak anaerobic power from vertical jump were independent predictors of select cortical structural outcomes (cortical thickness and area, periosteal and endosteal circumference, and estimated strength) accounting for up to 78.4% of the variance explained (all P < 0.05). However, cortical volumetric bone mineral density was unrelated to any measure or surrogate of muscle force capacity. MCSA is a strong independent predictor of cortical bone structure; however, both laboratory- and field-based measures of peak torque and/or peak anaerobic power are promising alternatives, explaining similar and sometimes greater variance than MCSA.

The effect of oral dabigatran etexilate on bone density, strength, and microstructure in healthy mice

Thrombin is a key component in the coagulation cascade where it converts factor V, VIII, XI, and fibrinogen. In addition to the abundant production of thrombin in the liver, osteoclasts synthesize and secrete thrombin as well. Osteoblasts express thrombin receptors, and it has been reported that thrombin stimulates the expression of RANKL relatively to OPG, resulting in greater osteoclast activation and bone degradation. Pradaxa (dabigatran etexilate, DE) is a new anticoagulant, which has recently been approved for clinical use. DE is a direct thrombin inhibitor with potential to modulate the RANKL/OPG ratio and thereby limit osteoclast activation and bone degradation. The purpose of the study was to investigate whether DE can increase bone density, bone strength, and bone microstructure in healthy male and female mice and to investigate whether the effect of DE is sex-dependent. Twenty-eight 14-week-old male C57BL/6 mice were stratified by weight into 4 groups: 1. Control 3weeks; 2. DE 3weeks; 3. Control 6weeks; 4. DE 6weeks. An identical study design was applied to twenty-four 14-week-old female C57BL/6 mice. Chow mixed with DE was offered ad libitum, resulting in a dose of 1.70mgDE/g body weight and 1.52mgDE/g body weight, to female and male mice, respectively. The animals were euthanized after 3 or 6weeks. Bone mineral density (aBMD) and bone mineral content (BMC) were evaluated with DEXA, 3D microstructural properties were determined with μCT, bone strength was determined with mechanical testing, and bone formation and resorption was evaluated with bone histomorphometry. In female mice, DE resulted in significant higher tibial aBMD values after 6weeks of intervention. Furthermore, DE significantly increased tibial diaphyseal cortical bone area and tissue area, which was accompanied by significantly increased strength of the tibial shaft. DE had no effect on femoral cortical bone or on femoral and vertebral trabecular 3D microstructure. Finally, bone histomorphometry showed that DE had no effect on MS/BS or Oc.S/BS. In male mice, no bone positive effects of DE were found in any of the parameters investigated. In conclusion, intervention with DE may result in a weak positive site specific effect at tibial cortical bone in female mice, and importantly, no major deleterious effects of DE on bone tissue were seen in either female or male mice despite the relatively high dose of DE used.

Longitudinal Effects of Single Hindlimb Radiation Therapy on Bone Strength and Morphology at Local and Contralateral Sites.

Radiation therapy (RTx) is associated with increased risk for late-onset fragility fractures in bone tissue underlying the radiation field. Bone tissue outside the RTx field is often selected as a "normal" comparator tissue in clinical assessment of fragility fracture risk, but the robustness of this comparison is limited by an incomplete understanding of the systemic effects of local radiotherapy. In this study, a mouse model of limited field irradiation was used to quantify longitudinal changes in local (irradiated) and systemic (non-irradiated) femurs with respect to bone density, morphology, and strength. BALB/cJ mice aged 12 weeks underwent unilateral hindlimb irradiation (4 × 5 Gy) or a sham procedure. Femurs were collected at endpoints of 4 days before treatment and at 0, 1, 2, 4, 8, 12, and 26 weeks post-treatment. Irradiated (RTx), Contralateral (non-RTx), and Sham (non-RTx) femurs were imaged by micro-computed tomography and mechanically tested in three-point bending. In both the RTx and Contralateral non-RTx groups, the longer-term (12- to 26-week) outcomes included trabecular resorption, loss of diaphyseal cortical bone, and decreased bending strength. Contralateral femurs generally followed an intermediate response compared with RTx femurs. Change also varied by anatomic compartment; post-RTx loss of trabecular bone was more profound in the metaphyseal than the epiphyseal compartment, and cortical bone thickness decreased at the mid-diaphysis but increased at the metaphysis. These data demonstrate that changes in bone quantity, density, and architecture occur both locally and systemically after limited field irradiation and vary by anatomic compartment. Furthermore, the severity and persistence of systemic bone damage after limited field irradiation suggest selection of control tissues for assessment of fracture risk or changes in bone density after radiotherapy may be challenging. © 2017 American Society for Bone and Mineral Research.