Specific Bone Surface Research Articles

Preventive effect of teriparatide on medication-related osteonecrosis of the jaw in rats

This study aimed to investigate the preventive effect of teriparatide (TPD) administration on medication-related osteonecrosis of the jaw (MRONJ) before tooth extraction due to periodontal lesions in bilaterally ovariectomized female rats treated with zoledronic acid. Thirty skeletally mature Sprague–Dawley rats were randomly divided into three groups: control (CONT, n = 10), zoledronic acid (ZA, n = 10), and zoledronic acid and teriparatide (ZA-TPD, n = 10). The rats were sacrificed 8 weeks after tooth extraction. Micro-computed tomography analysis of the tibia showed that bone mineral density was highest in the CONT, followed by that in the ZA and ZA-TPD groups (CONT/ZA, p = 0.009; CONT/ZA-TPD, p < 0.001; ZA/ZA-TPD, p < 0.001). In the trabecular bone analysis of the extraction site, significant differences in specific bone surface (CONT/ZA, p = 0.010; CONT/ZA-TPD, p = 0.007; ZA/ZA-TPD, p = 0.002) and trabecular thickness (CONT/ZA-TPD, p = 0.002; ZA/ZA-TPD, p = 0.002) were observed. Histological analyses of the extraction sites revealed characteristic MRONJ lesions in the ZA group. Osteonecrosis, inflammatory cells, and sequestrum were less frequently observed in the ZA-TPD group than in the ZA group. In conclusion, TPD administration before tooth extraction helped reduce the occurrence of MRONJ in rats treated with zoledronic acid, confirming its preventative effects.

Icariside II-Loaded Calcium Phosphate Cement Scaffolds Combined with Bone Marrow Mesenchymal Stem Cells for Osteogenesis

Purpose: Previous studies have demonstrated the osteogenic effects of icariside II (ICSII), which is a metabolic product of the prenylated active flavonol icariin (ICA) from the roots of Epimedium. However, the in vivo osteogenic effects of ICSII remain unclear. Thus, in this study, we evaluated the osteogenic effects of ICSII in vivo. Methods: Complexes of calcium phosphate cement (CPC) and bone marrow mesenchymal stem cells (BMSCs) with or without ICSII were subcutaneously implanted into nude mice (ectopic osteogenesis test) and into tooth sockets in beagles after maxillary canine tooth extraction (in situ osteogenesis test). The samples were harvested at different time points, and the in vivo osteogenic effect of the ICSII on the BMSCs was evaluated by histology, microcomputed tomography (micro-CT) and the bone mineralization apposition rate (MAR). Results: The proliferation and viability of BMSCs in the ICSII-loaded CPC scaffold were significantly increased (P &lt; 0.01). The new bone area and MAR in the CPC+BMSC+ICSII group were greater than those in the CPC+BMSC group (P &lt; 0.05), but there was no significant difference in markers evaluated by immunohistochemistry and integrated optical density (IOD) analysis (P &gt; 0.05), with the exception of Runx-2 expression in the CPC+BMSC+ICSII group. After 2 months, the bone mineral content (BMC) and specific bone surface (bone surface divided by bone volume, BS/BV) were significantly increased (P &lt; 0.05) in the CPC+BMSC+ICSII group compared with the CPC+BMSC group. The bone mineral density (BMD), bone volume divided by total volume (BV/TV), and trabecular number (Tb.N) were increased in the CPC+BMSC+ICSII group, but the differences were not significant (P &gt; 0.05). Conclusions: Our results suggest that ICSII can likely induce bone formation by BMSCs and be used as a promising factor for building scaffold composites in bone tissue engineering.

The association between paraspinal muscle parameters and vertebral pedicle microstructure in patients undergoing lumbar fusion surgery.

Lumbar fusion surgery has become a standard procedure in spine surgery and commonly includes the posterior placement of pedicle screws. Bone quality is a crucial factor that affects pedicle screw purchase. However, the relationship between paraspinal muscles and the bone quality of the pedicle is unknown. The aim of the study was to determine the relationship between paraspinal muscles and the ex vivo bony microstructure of the lumbar pedicle. Prospectively, collected data of patients undergoing posterior lumbar fusion for degenerative spinal conditions was analyzed. Pre-operative lumbar magnetic resonance imaging (MRI) scans were evaluated for a quantitative assessment of the cross-sectional area (CSA), functional cross-sectional area (fCSA), and the proportion of intramuscular fat (FI) for the psoas muscle and the posterior paraspinal muscles (PPM) at L4. Intra-operative bone biopsies of the lumbar pedicle were obtained and analyzed with microcomputed tomography (µCT) scans. The following cortical (Cort) and trabecular (Trab) bone parameters were assessed: bone volume fraction (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), connectivity density (CD), bone-specific surface (BS/BV), apparent density (AD), and tissue mineral density (TMD). A total of 26 patients with a mean age of 59.1years and a mean BMI of 29.8kg/m2 were analyzed. fCSAPPM showed significant positive correlations with BV/TVTrab (ρ = 0.610; p < 0.001), CDTrab (ρ = 0.679; p < 0.001), Tb.NTrab (ρ = 0.522; p = 0.006), Tb.ThTrab (ρ = 0.415; p = 0.035), and ADTrab (ρ = 0.514; p = 0.007). Cortical bone parameters also demonstrated a significant positive correlation with fCSAPPM (BV/TVCort: ρ = 0.584; p = 0.002; ADCort: ρ = 0.519; p = 0.007). FIPsoas was negatively correlated with TMDCort (ρ = - 0.622; p < 0.001). This study highlights the close interactions between the bone microstructure of the lumbar pedicle and the paraspinal muscle morphology. These findings give us further insights into the interaction between the lumbar pedicle microstructure and paraspinal muscles.

P11. The correlation between paraspinal muscle parameters and vertebral pedicle microstructure in patients undergoing lumbar fusion

P11. The correlation between paraspinal muscle parameters and vertebral pedicle microstructure in patients undergoing lumbar fusion

Mechanistic PK-PD model of alendronate treatment of postmenopausal osteoporosis predicts bone site-specific response.

Alendronate is the most widely used drug for postmenopausal osteoporosis (PMO). It inhibits bone resorption, affecting osteoclasts. Pharmacokinetics (PK) and pharmacodynamics (PD) of alendronate have been widely studied, but few mathematical models exist to simulate its effect. In this work, we have developed a PK model for alendronate, valid for short- and long-term treatments, and a mechanistic PK-PD model for the treatment of PMO to predict bone density gain (BDG) at the hip and lumbar spine. According to our results, at least three compartments are required in the PK model to predict the effect of alendronate in both the short and long terms. Clinical data of a 2-year treatment of alendronate, reproduced by our PK-PD model, demonstrate that bone response is site specific (hip: 7% BDG, lumbar spine: 4% BDG). We identified that this BDG is mainly due to an increase in tissue mineralization and a decrease in porosity. The difference in BDG between sites is linked to the different loading and dependence of the released alendronate on the bone-specific surface and porosity. Osteoclast population diminishes quickly within the first month of alendronate treatment. Osteoblast population lags behind but also falls due to coupling of resorption and formation. Two dosing regimens were studied (70 mg weekly and 10 mg daily), and both showed very similar BDG evolution, indicating that alendronate accumulates quickly in bone and saturates. The proposed PK-PD model could provide a valuable tool to analyze the effect of alendronate and to design patient-specific treatments, including drug combinations.

Microarchitecture and morphology of bone tissue over a wide range of BV/TV assessed by micro-computed tomography and three different threshold backgrounds.

The microarchitecture of bone both results from and in turn affects the remodelling process. Bone-specific surface, for instance, is one of these important microarchitectural parameters because remodelling is also considered to be a surface-mediated phenomenon (Berli etal.[1]). An understanding of these structural parameters across the widest possible range of porosity is essential to illuminating how bone reacts to disease, in different skeletal sites and in either its cancellous or cortical forms. 112 samples from an elephant femur were examined by micro-computed tomography (μCT), 31 of which contained both mineralised and demineralised tissue. A critical factor in all scans is setting the correct threshold (with background the surrounding medium) and hence 3 different backgrounds were used: air, water and collagen. The effect of the 3 background thresholds on the physical characteristics of bone (BS/TV, BS/BV, TbSp, TbTh, Dmat, vs BV/TV) was then determined. The results showed that using a threshold set by the collagen background had a profound effect on the histomorphometry bone parameters when assessed by μCT. However, the differences between air and water were not significant, suggesting that comparable data can be produced in a laboratory environment when scanning porous bone samples under either wet or dry conditions- counter to common belief. Determining which is more suitable, air or water, in laboratory and in clinical μCT imaging is important to improve the quality and relevance of biomechanics research. The data with collagen as the threshold were illuminating as they showed that remodelling rates and the relative organic to mineral presence varied with BV/TV, concurring with some other recent studies [2,3,4].

The interplay between BMU activity linked to mechanical stress, specific surface and inhibitory theory dictate bone mass distribution: Predictions from a 3D computational model

Bone mechanical and biological properties are closely linked to its internal tissue composition and mass distribution, which are in turn governed by the purposeful action of the basic multicellular units (BMUs). The orchestrated action of osteoclasts and osteoblasts, the resorbing and forming tissue cells respectively, in BMUs is responsible for tissue maintenance, repair and adaptation to changing load demands through the phenomenon known as remodelling. In this work, a computational mechano-biological model of bone remodelling based on the inhibitory theory and a new scheme of bone resorption introduced previously in a 2D model, is extended to a 3D model of the real external geometry of a femur under normal walking loads. Starting from a uniform apparent density (ratio of tissue local mass to total local volume) distribution, the BMU action can be shown to lead naturally to an internal density distribution similar to that of a real bone, provided that the initial density value is high enough to avoid unrealistic final mass deposition in zones of high energy density and excessive damage. Physiological internal density values are reached throughout the whole 3D geometry, and at the same time a ‘boomerang’-like relationship between apparent and material density (ratio of tissue mass to tissue volume) emerges naturally under the proposed remodelling scheme. It is also shown here that bone-specific surface is a key parameter that determines the intensity of BMU action linked to the mechanical and biological requirements. Finally, by engaging in simulations of bone in disuse, we were able to confirm the appropriate selection of the model parameters. As an example, our results show good agreement with experimental measurements of bone mass on astronauts a fact that strengthens our belief in the insightful nature of our novel 3D computational model.

Volumetric analysis of the periodontal microstructure under antiresorptive therapy: an experimental study in rabbits.

The objective of this study wasto volumetrically assess changes in the periodontal microstructure under antiresorptive therapy. Microtomographic scans from a total of 9 Dutch Belted rabbits having been randomly allocated to either the intravenous administration of amino-bisphosphonate (zoledronic acid) (Za) (n = 5) or a negative control group (nZa) (n = 4) were obtained at 10months following a repeated drug administration. A quantification of the periodontal space thickness (P.Th) of both maxillary and mandibular most posterior premolars, as well as of the 2nd molars was performed. Bone micromorphometry was assessed by means of bone volume per total volume (BV/TV), the bone mineral density (BMD), trabecular thickness (Tb.Th), trabecular number (Tb.N), bone surface (BS), and the specific bone surface (BS/BV). Za was associated with significantly higher P.Th (P = 0.010), which was most pronounced in the upper jaw. Bone micromorphometry revealed no significant differences among the two groups, i.e., Za and nZa, for all the investigated parameters. Volumetric analysis revealed that antiresorptive therapy was associated with periodontal space widening, whereas major effects on the bone micro-morphology could not be observed. A deep understanding of specific periodontal and alveolar bone alterations in patients under antiresorptive therapy might help to prevent the onset of medication-related osteonecrosis of the jaw.

The Role of Surface Chemistry in the Osseointegration of PEEK Implants.

Poly(etheretherketone) (PEEK) implants suffer from poor osseointegration because of chronic inflammation. In this study, we hypothesized that adding NH2 and COOH groups to the surface of PEEK could modulate macrophage responses by altering protein adsorption and improve its osseointegration. NH2 and COOH-functionalized PEEK surfaces induced pro- and anti-inflammatory macrophage responses, respectively, and differences in protein adsorption patterns on these surfaces were related to the varied inflammatory responses. The macrophage responses to NH2 surfaces significantly reduced the osteogenic differentiation of mesenchymal stem cells (MSCs). MSCs cultured on NH2 surfaces differentiated less than those on COOH surfaces even though NH2 surfaces promoted the most mineralization in simulated body fluid solutions. After 14 days in rat tibia unicortical defects, the bone around NH2 surfaces had thinner trabeculae and higher specific bone surface than the bone around unmodified implants; surprisingly, the NH2 implants significantly increased bone-binding over the unmodified implants, while COOH implants only showed a trend for increasing bone-binding. Taken together, these results suggest that both mineral-binding and immune responses play a role in osseointegration, and PEEK implant integration may be improved with mixtures of these two functional groups to harness the ability to reduce inflammation and bind bone strongly.

Evaluation of the effect of vitamin D3 on mandibular condyles in an ovariectomized mouse model: a micro-CT study

BackgroundThis study aimed to investigate the effect of ovariectomy and vitamin D3 on bone microstructure; this effect was examined in three regions of interest at one femoral and two mandibular sampling sites bone in an ovariectomized mouse model.MethodsThirty-six week-old female mice were randomly divided into three groups: 10 subjects were given oral cholecalciferol (vitamin D3) daily for 6 weeks after undergoing bilateral ovariectomy (D3 group), while 10 ovariectomized subjects (OVX) and 10 subjects who underwent a sham operation (SHAM) received peanut oil daily during the investigation. After extermination, the left hemimandible and femur were removed and scanned by micro-CT. The bone micromorphology parameters were analyzed and the BMD was calculated.ResultsThe bone volume fraction (BV/TV) was significantly lower in the trabecular bone of the mandibular condyle in the OVX group than in the SHAM and D3 groups. Also there was a significant difference between the SHAM and D3 groups. The specific bone surface (BS/BV) was significantly higher in the OVX and D3 groups than in the SHAM group. Trabecular thickness (Tb.Th) was significantly higher in the SHAM group, and the trabecular bone pattern factor (Tb.Pf) was significantly higher in the OVX group than in the other two groups. Bone mineral density (BMD) of the femur and the mandible was significantly lower in the OVX group than in the SHAM and D3 groups.ConclusionsOur results show that ovariectomy causes a significantly weaker bone microstructure in the mandibular condyle, where the protective effect of vitamin D3 resulted in a partial resorption.

Soluble epoxide hydrolase inhibitor can protect the femoral head against tobacco smoke exposure-induced osteonecrosis in spontaneously hypertensive rats

Exposure to tobacco smoke (TS) has been considered a risk factor for osteonecrosis of the femoral head (ONFH). Soluble epoxide hydrolase inhibitors (sEHIs) have been found to reduce inflammation and oxidative stress in a variety of pathologies. This study was designed to assess the effect of sEHI on the development of ONFH phenotypes induced by TS exposure in spontaneously hypertensive (SH) rats. SH and normotensive Wistar Kyoto (WKY) rats were exposed to filtered air (FA) or TS (80 mg/m3 particulate concentration) 6 h/day, 3 days/week for 8 weeks. During this period, sEHI was delivered through drinking water at a concentration of 6 mg/L. Histology, immunohistochemistry, and micro-CT morphometry were performed for phenotypic evaluation. As results, TS exposure induced significant increases in adipocyte area, bone specific surface (BS/BV), and trabecular separation (Tb.SP), as well as significant decreases in bone mineral density (BMD), percent trabecular area (Tb.Ar), HIF-1a expression, bone volume fraction (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) in both SH and WKY rats. However, the protective effects of sEHI were mainly observed in TS-exposed SH rats, specifically in the density of osteocytes, BMD, Tb.Ar, HIF-1a expression, BV/TV, BS/BV, Tb.N, and Tb.SP. Our study confirms that TS exposure can induce ONFH especially in SH rats, and suggests that sEHI therapy may protect against TS exposure-induced osteonecrotic changes in the femoral head.

The Novel Role of Checkpoint Inhibitor PD-1H/VISTA in Osteoclast Activation and Multiple Myeloma Bone Disease

IntroductionMultiple myeloma (MM)-induced bone disease remains one of its most devastating complications, caused by increased bone resorption by overactivated osteoclasts coupled with impaired bone formation. MM cells produce osteoclast-activating factors that induce osteoclast activation and extensive bone resorption. Our previous work demonstrated that matrix metalloproteinase 13 (MMP-13) is a critical osteoclastogenic factor that is highly secreted by MM cells (Fu J etc. JCI. 2016). We also identified that the checkpoint inhibitor, programmed death-1 homolog (PD-1H/VISTA), serves as the MMP-13 receptor in osteoclasts and mediates MMP-13-dependent osteoclastogenic function which is largely blocked in Pd-1h -/-osteoclasts (Fu J etc. ASH 2019, 2020). While the inhibitory role of PD-1H/VISTA in T-cells has recently been described (ElTanbouly MA etc. Science 2020), its cellular binding proteins remain unclear, and its role in osteoclast activation andMM bone disease have not been addressed.Methods and ResultsTo identify its interacting proteins, PD-1H-His 6 recombinant protein was expressed in mouse bone marrow mononuclear cells, the associated proteins pulled down by Ni-NTA agarose beads from cell lysates and identified by mass spectrometry. Functional annotation charting of the 75 proteins enriched in PD-1H pull-down samples (with signal ratio of PD-1H-His 6 pull-down vs control >2) indicated that almost 30% of the interacting targets were either cytoskeletal or cytoskeleton-associated proteins.Given that the F-actin cytoskeleton undergoes dynamic reorganization during osteoclast differentiation and plays critical roles in bone resorption, we further addressed the role of PD-1H in F-actin cytoskeleton regulation. Initially, osteoclasts form F-actin-rich adhesive structures, termed podosomes. At later stages, podosomes collectively rearrange into clusters and rings and finally into sealing belts to mediate osteoclast spreading, migration and bone resorption (Teitelbaum SL. Ann N Y Acad Sci. 2011). By confocal immunofluorescence microscopy, we found that PD-1H co-localized with F-actin podosome clusters, rings and sealing belts during osteoclast differentiation (Figure 1A). The functional role of PD-1H in F-actin cytoskeleton reorganization was addressed using Pd-1h -/- osteoclast wherein Pd-1h knockout lead to the disruption of podosome clusters at early stages relative to WT controls, while at later stages, Pd-1h -/-osteoclasts exhibited significantly fewer F-actin rings and belts (Figure 1B). Further, binding of MMP-13 to PD-1H increased the number of osteoclasts forming F-actin rings and belts, as well as the size of F-actin belts, which was blocked in Pd-1h -/- osteoclasts.To determine the role of PD-1H in the development of myeloma-induced lytic bone lesions, 5TGM1 myeloma cells were bilaterally intratibially injected into Pd-1h wtRag2 -/- or Pd-1h -/-Rag2 -/- mice (n=10) to induce lytic bone lesions. Three weeks following intratibial 5TGM1 injection, tibiae were harvested for micro-computed tomography. Subsequent quantitative histomorphology analyses of the trabecular and cortical bones confirmed that the knockout of Pd-1h reduced MM-induced bone destruction with significantly less decrease in trabecular bone volume (Tb. [BV/TV]), trabecular bone number (Tb. N.), trabecular bone thickness (Tb. Th.), as well as less increase in trabecular bone spacing (Tb. Sp.) and bone specific surface (Tb. [BS/BV]) compared to Pd-1h wtRag2 -/- mice. Similar effects were observed in cortical bone with less decrease in cortical bone thickness (CT. Th.), cortical bone area fraction (CT. [BA/TA]), and cortical tissue mineral density (CT. TMD) in 5TGM1 bearing Pd-1h -/-Rag2 -/- mice vs Pd-1h wtRag2 -/- mice (Table 1).ConclusionsTaken together, our study, for the first time, reveals the novel role of checkpoint inhibitor, PD-1H/VISTA, in osteoclasts and myeloma bone disease. PD-1H associates with cytoskeleton proteins and regulates the F-actin cytoskeleton reorganization which is critical for osteoclast bone resorption activity. Further, PD-1H mediates MMP-13-induced osteoclast fusion, F-actin belts formation, and osteoclast activation. Pd-1h -/-in recipient mice significantly impairs MM-induced bone loss, demonstrating that PD-1H/VISTA plays a critical role in MM bone disease. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Microcomputed tomographic analysis of bone microarchitecture after sinus augmentation with hyaluronic matrix: a case-control study.

The aim of this study was to analyze trabecular microarchitecture of augmented sinuses with hyaluronic matrix and xenograft by microcomputed tomography, and to investigate whether hyaluronic matrix has an effect on the newly formed bone quality. Thirteen patients undergoing maxillary sinus augmentation were included in this split-mouth study. Right and left sinus sites were randomly assigned to test and control group. In test group, the sinus was grafted with hyaluronic matrix and xenograft; in control group, only with xenograft. Four months after augmentation, bone samples were harvested during implant placement and analyzed for the following trabecular microarchitecture parameters using microcomputed tomography: bone volume (BV), total volume (TV), bone volume fraction (BV/TV), bone surface (BS), specific bone surface (BS/BV), bone surface density (BS/TV), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), trabecular pattern factor (Tb.Pf), and fractal dimension (FD). There was statistically significant difference only for BS/TV parameter between two groups. BS/TV was higher in hyaluronic matrix group compared with control group. Addition of hyaluronic matrix to xenograft may enhance bone quality in terms of bone surface density. However, more research investigating the microstructural variation of augmented sinuses is needed with a greater sample.

Mesenchymal stem cells and platelet-rich plasma-impregnated polycaprolactone-\u03b2 tricalcium phosphate bio-scaffold enhanced bone regeneration around dental implants

BackgroundFinding a material that supports bone regeneration is the concern for many investigators. We supposed that a composite scaffold of poly(ε) caprolactone and β-tricalcium phosphate (PCL-TCP) would entail desirable characteristics of biocompatibility, bioresorbability, rigidity, and osteoconductivity for a proper guided bone regeneration. Furthermore, the incorporation of mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP) would boost the bone regeneration. We conducted this study to evaluate the bone regeneration capacity of PCL-TCP scaffold that is loaded with MSCs and PRP.Materials and methodsFive miniature pigs received 6 implants in 6 created-mandibular bony defects in the right and left lower premolar areas. The bony defects were managed according to the following three groups: the PCL-TCP scaffold loaded with MSCs and PRP (MSCs+PRP+PCL-TCP) group (n = 10), PCL-TCP scaffold loaded with PRP (PRP+PCL-TCP) group (n = 10), and PCL-TCP scaffold group (n = 10). After 12 weeks, the bone regeneration was assessed using fluorochrome bone labeling, μCT bone morphogenic analysis, and histomorphometric analysis.ResultsAll of the three groups supported the bone regeneration around the dental implants. However, the PCL-TCP scaffold loaded with MSCs and PRP (MSCs+PRP+PCL-TCP) group showed non-significant higher bone surface, bone specific surface, and bone surface density than the other two groups as revealed by the μCT bone morphogenic analysis. Histologically, the same group revealed higher bone-implant contact ratio (BIC) (p = 0.017) and new bone height formation (NBH, mm) (p = 0.0097) with statistically significant difference compared to the PCL-TCP scaffold group.ConclusionsPCL-TCP scaffold is compatible for bone regeneration in bone defects surrounding dental implants. Moreover, the incorporation of MSCs and PRP optimized the bone regeneration process with respect to the rate of scaffold replacement, the height of the regenerated bone, and implant stability.

A temporal study on musculoskeletal morphology and metabolism in hibernating Daurian ground squirrels (Spermophilus dauricus).

Hibernators provide a natural model to study the mechanisms underlying the prevention of disuse-induced musculoskeletal deterioration. Currently, however, these mechanisms remain poorly understood. Here, we investigated changes in morphology and metabolic indices in the hindlimb skeletal muscle and bone of Daurian ground squirrels (Spermophilus dauricus) during different periods of hibernation, and further explored the possible mechanisms involved in the musculoskeletal maintenance of hibernators after prolonged inactivity. Results showed that, compared with levels in the summer active group (SA), almost all morphological indices of skeletal muscle and bone, including muscle mass, muscle fiber cross-sectional area, bone mass, bone length, and bone mechanical properties, were unchanged in the different periods of hibernation. Only a few microstructural parameters of bone showed deterioration in the post-hibernation group (POST), including increased specific bone surface (+71%), decreased trabecular thickness (-43%), and decreased average cortical thickness (-51%) in the tibia, and increased trabecular separation (+60%) in the femur. Furthermore, most examined metabolic indices involved in muscle protein turnover and bone remodeling were unchanged, except for several indices in the inter-bout arousal group (IBA), i.e., increase in the phosphorylation of eukaryotic initiation factor 4E binding protein 1 (4E-BP1) (IBA vs. SA, +80%) in the vastus medialis muscle, increase in chymotrypsin-like activity (IBA vs. SA, +62%) in the tibialis anterior muscle, increase in osteoblast number (IBA vs. SA, +110%; IBA vs. torpor (TOR), +68%) and osteoclast number (IBA vs. TOR, +105%) per bone surface in the tibia, and increase in osteoclast surface per bone surface (IBA vs. TOR, +128%) in the femur. The above evidence demonstrates that the musculoskeletal morphology of squirrels was largely preserved, and musculoskeletal metabolism was generally maintained after prolonged hibernation inactivity. These findings suggest that the well-maintained musculoskeletal metabolism may be a vital mechanism underlying the preservation of the musculoskeletal system during hibernation. The coincident up-regulation of several metabolic indicators during IBA indicates that musculoskeletal metabolism may be relatively active during this period; however, its role in musculoskeletal maintenance during hibernation needs further clarification.

Trabecular Bone Assessment Using Magnetic-Resonance Imaging: A Pilot Study.

The aim of this study was to assess trabecular bone morphology via magnetic-resonance imaging (MRI) using microcomputed tomography (µCT) as the control group. Porcine bone samples were scanned with T1-weighted turbo spin echo sequence imaging, using TR 25 ms, TE 3.5 ms, FOV 100 × 100 × 90, voxel size 0.22 × 0.22 × 0.50 mm, and scan time of 11:18. µCT was used as the control group with 80 kV, 125 mA, and a voxel size of 16 µm. The trabecular bone was segmented on the basis of a reference threshold value and morphological parameters. Bone volume (BV), Bone-volume fraction (BvTv), Bone specific surface (BsBv), trabecular thickness (TbTh), and trabecular separation (TbSp) were evaluated. Paired t-test and Pearson correlation test were performed at p = 0.05. MRI overestimated BV, BvTv, TbTh, and TbSp values. BsBv was the only parameter that was underestimated by MRI. High statistical correlation (r = 0.826; p < 0.05) was found for BV measurements. Within the limitations of this study, MRI overestimated trabecular bone parameters, but with a statistically significant fixed linear offset.

Quantification of 3D microstructural parameters of trabecular bone is affected by the analysis software.

Over the last decades, the use of high-resolution imaging systems to assess bone microstructural parameters has grown immensely. Yet, no standard defining the quantification of these parameters exists. It has been reported that different voxel size and/or segmentation techniques lead to different results. However, the effect of the evaluation software has not been investigated so far. Therefore, the aim of this study was to compare the bone microstructural parameters obtained with two commonly used commercial software packages, namely IPL (Scanco, Switzerland) and CTan (Bruker, Belgium). We hypothesized that even when starting from the same segmented scans, different software packages will report different results. Nineteen trapezia and nineteen distal radii were scanned at two resolutions (20 μm voxel size with microCT and HR-pQCT 60 μm). The scans were segmented using the scanners' default protocol. The segmented images were analyzed twice, once with IPL and once with CTan, to quantify bone volume fraction (BV/TV), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), trabecular number (Tb.N) and specific bone surface (BS/BV). Only small differences between IPL and CTan were found for BV/TV. For Tb.Th, Tb.Sp and BS/BV high correlations (R2≥0.99) were observed between the two software packages, but important relative offsets were observed. For microCT scans, the offsets were relative constant, e.g., around 15% for Tb.Th. However, for the HR-pQCT scans the mean relative offsets ranged over the different bone samples (e.g., for Tb.Th from 14.5% to 19.8%). For Tb.N, poor correlations (0.43≤R2≤0.81) for all tested cases were observed. We conclude that trabecular bone microstructural parameters obtained with IPL and CTan cannot be directly compared except for BV/TV. For Tb.Th, Tb.Sp and BS/BV, correction factors can be determined, but these depend on both the image voxel size and specific anatomic location. The two software packages did not produce consistent data on Tb.N. The development of a universal standard seems desirable.

When Lithics Hit Bones: Evaluating the Potential of a Multifaceted Experimental Protocol to Illuminate Middle Palaeolithic Weapon Technology

Recent zooarchaeological and isotope analyses have largely settled the debate surrounding Neanderthal hunting capacities, repeatedly demonstrating their successful acquisition of large ungulates. Nevertheless, the functional identification of individual tools as hunting weapons remains a methodological challenge. In-depth studies have focussed mainly on small subsets of lithic artefacts from selected assemblages assessing features of breakage patterns, retouch, shape and use wear. Studies focussing on associated hunting lesions are rarer and often focus on reconstructing very specific bone surface marks encountered in the archaeological record. This study aims to add to our understanding of the formation and characteristics of projectile impact marks (PIMs) on bone through a series of highly monitored, replicative experiments, using thrusting and throwing spears with replica Levallois points into two wild pig carcasses. In total, 152 shots were made, and for each a series of attributes was recorded, including velocity and location of impact. Subsequent quantitative analyses focussed on understanding the various factors underlying the formation of different types of projectile impact marks. These experiments demonstrate that PIM formation results from the properties of both the impacting projectile and bone element. PIMs can signal impacts caused by different delivery methods but only on some parts of the skeleton. These results are contextualised in relation to the occurrence and recognition of Palaeolithic PIMs and patterns of Neanderthal behaviour. These experiments are only a first step in improving the recognition of these signatures in the archaeological record and providing better insights into understanding of the mechanisms of Neanderthal hunting.

Protective Effects of Methoxsalen Supplementation on Chronic Alcohol-Induced Osteopenia and Steatosis in Rats.

Osteopenia or osteoporosis occurs frequently in alcoholics and patients with alcoholic fatty liver disease. Methoxsalen (MTS), 8-methoxypsoralen, improved osteoporosis in ovariectomized and diabetic mouse models; however, its effects on alcohol-induced osteopenia and steatosis have not been reported. This study examined the effects of MTS on alcohol-induced bone loss and steatosis. Rats in the alcohol groups were fed a Liber-DeCarli liquid diet containing 36% of its calories as alcohol. MTS was at 0.005% in their diet, while alendronate (positive control; 500 μg/kg BW/day) was administered orally for eight weeks. The pair-fed group received the same volume of isocaloric liquid diet containing dextrin-maltose instead of alcohol as the alcohol control group consumed the previous day. In the alcohol-fed rats, the MTS and alendronate increased the bone volume density, bone surface density and trabecular number, while the bone specific surface, trabecular separation and structure model index were decreased in the tibia. MTS down-regulated tibial tartrate-resistant acid phosphatase 5 (TRAP) expression compared to the alcohol control group. MTS or alendronate prevented chronic alcohol-induced hepatic lipid accumulation and the triglyceride level in the alcohol-fed rats by decreasing the lipogenic enzyme activities and increasing the fatty acid oxidation enzyme activities. MTS reduced significantly the serum levels of alcohol, TRAP and tumor necrosis factor-α compared to the alcohol control group. Overall, these results suggest that MTS is likely to be an alternative agent for alcoholic osteopenia and hepatosteatosis.

BMI-related subchondral bone microstructural changes in knee osteoarthritis: overloading or metabolic disturbance?

Purpose: Obesity is one of the major risk factors for knee osteoarthritis (KOA), and it may contribute to KOA possibly through two main mechanisms: increased mechanical loading, as well as elevated low-grade systemic inflammation. There has been a growing interest in the relationship between body weight index (BMI), and subchondral trabecular bone (SCTB) microarchitectural changes in KOA, e.g. at the medial compartment. This study aims to fill the research gap, describing the correlation of SCTB microarchitectural changes in lateral compartment with BMI, ultimately to probe the predominant underlying mechanism. Methods: Samples of tibial plateaus from 54 (14 male, 40 female) patients with late stage KOA who were undergoing total knee arthroplasty (TKA) were collected. The samples were divided into three groups, according to the patients’ BMI (lean, BMI < 25; overweight, 25 < BMI < 30; obese, BMI > 30). The region of interest (ROI) was drawn manually for every sample (Figure 1). Three-dimension microarchitectural parameters in both medial and lateral compartments were obtained with the help of micro-CT analysis. Results: In lateral compartment (Figure 2), positive correlations were found between BMI and bone volume fraction (BV/TV) (r = 0.392, p=0.003) and trabecular thickness (Tb.Th) (r = 0.449, p=0.01). Negative correlation was found between BMI and specific bone surface (BS/BV) (r = -0.379, p=0.005) and trabecular pattern factor (Tb.Pf) (r= -0.353, p=0.009). In medial compartment, unlike previous researches, our study did not describe a significant correlation between microarchitectural parameters and BMI. Conclusions: This is the first study showing that BMI is related to microarchitectural changes in lateral compartments of late stage KOA patients. It can be interpreted from the results that bone density and quality increase with BMI. Mechanical loading will increase bone density while low-grade inflammation will erode bone, decreasing its quality and quantity. Our data imply that mechanical loading is a major mechanism to account for the deterioration of SCTB in lateral compartment in the late stage of KOA.The conflicting findings with previous researches in the medial compartment might be due to long waiting list in southern China. Patients usually receive TKA after medial compartment is fully worn out. We assume that in our patients population the KOA is in a critical end-stage, after the collapse of the medial compartment, so the lateral compartment’s mechanical loading is increased, which leads to more severe SCTB sclerosis.View Large Image Figure ViewerDownload Hi-res image Download (PPT)