Diabetic Covid-19 severity: Impaired glucose tolerance and pathologic bone loss.

Diabetes mellitus (DM), hypertension, and cardiovascular diseases (CVDs) are the leading chronic comorbidities that enhance the severity and mortality of COVID-19 cases. However, SARS-CoV-2 mediated deregulation of diabetes pathophysiology and comorbidity that links the skeletal bone loss remain unclear. We used both streptozocin-induced type 2 diabetes (T2DM) mouse and hACE2 transgenic mouse to enable SARS-CoV-2-receptor binding domain (RBD) mediated abnormal glucose metabolism and bone loss phenotype in mice. The data demonstrate that SARS-CoV-2-RBD treatment in pre-existing diabetes conditions in hACE2 (T2DM + RBD) mice results in the aggravated osteoblast inflammation and downregulation of Glucose transporter 4 (Glut4) expression via upregulation of miR-294-3p expression. The data also found increased fasting blood glucose and reduced insulin sensitivity in the T2DM + RBD condition compared to the T2DM condition. Femoral trabecular bone mass loss and bone mechanical quality were further reduced in T2DM + RBD mice. Mechanistically, silencing of miR-294 function improved Glut4 expression, glucose metabolism, and bone formation in T2DM + RBD + anti-miR-294 mice. These data uncover the previously undefined role of SARS-CoV-2-RBD treatment mediated complex pathological symptoms of diabetic COVID-19 mice with abnormal bone metabolism via a miRNA-294/Glut4 axis. Therefore, this work would provide a better understanding of the interplay between diabetes and SARS-CoV-2 infection.

Optimal radial force and size for palliation in gastroesophageal adenocarcinoma: a comparative analysis of current stent technology.

BACKGROUND: The optimal use of esophageal stents for malignant and benign esophageal strictures continues to be plagued with variability in pain tolerance, migration rates, and reflux-related symptoms. The aim of this study was to evaluate the differences in radial force exhibited by a variety of esophageal stents with respect to the patient's esophageal stricture. METHODS: Radial force testing was performed on eight stents manufactured by four different companies using a hydraulic press and a 5000 N force gage. Radial force was measured using three different tests: transverse compression, circumferential compression, and a three-point bending test. Esophageal stricture composition and diameters were measured to assess maximum diameter, length, and proximal esophageal diameter among 15 patients prior to stenting. RESULTS: There was a statistically significant difference in mean radial force for transverse compression tests at the middle (range 4.25-0.66 newtons/millimeter N/mm) and at the flange (range 3.32-0.48 N/mm). There were also statistical differences in mean radial force for circumferential test (ranged from 1.19 to 10.50 N/mm, p < 0.001) and the three-point bending test (range 0.08-0.28 N/mm, p < 0.001). In an evaluation of esophageal stricture diameters and lengths, the smallest median diameter of the stricture was 10 mm (range 5-16 mm) and the median proximal diameter normal esophagus was 25 mm (range 22-33 mm), which is currently outside of the range of stent diameters. CONCLUSIONS: Tested stents demonstrated significant differences in radial force, which provides further clarification of stent pain and intolerance in certain patients, with either benign or malignant disease. Similarly, current stent diameters do not successfully exclude the proximal esophagus, which can lead to obstructive-type symptoms. Awareness of radial force, esophageal stricture composition, and proximal esophageal diameter must be known and understood for optimal stent tolerance.

Mitochondrial epigenetics in bone remodeling during hyperhomocysteinemia.

Increased levels of homocysteine (Hcy), known as hyperhomocysteinemia (HHcy), is an independent risk factor of various diseases. Clinical studies report that people born with severe HHcy develop skeletal malformations with weaker bone. Studies also report that altered mitochondrial dynamics and altered epigenetics contribute to weaker bones and bone diseases. Although Hcy-induced mitochondrial dysfunction has been shown to affect bone metabolism, the role of mitochondrial epigenetics (mito-epigenetics) has not been studied in bones. The epigenetics in mitochondria is interesting as the mitochondrial genome size is small (16 kb) with fewer CpG, and without histones and introns. Recently, fascinating works on epigenetics along with the discovery of histone-like proteins in mitochondria are giving exciting areas for novel studies on mitochondria epigenetics. There are mutual cause and effect relationships between bone, mitochondria, Hcy, and epigenetics, but unfortunately, studies are lacking that describe the involvement of all these together in bone disease progression. This review describes the reciprocal relationships and mechanisms of Hcy-bone-mitochondria-epigenetics along with a short discussion of techniques which could be employed to assess Hcy-induced anomaly in bone, mediated through alterations in mito-epigenetics.

The role of homocysteine in bone remodeling.

Bone remodeling is a very complex process. Homocysteine (Hcy) is known to modulate this process via several known mechanisms such as increase in osteoclast activity, decrease in osteoblast activity and direct action of Hcy on bone matrix. Evidence from previous studies further support a detrimental effect on bone via decrease in bone blood flow and an increase in matrix metalloproteinases (MMPs) that degrade extracellular bone matrix. Hcy binds directly to extracellular matrix and reduces bone strength. There are several bone markers that can be used as parameters to determine how high levels of plasma Hcy (hyperhomocysteinemia, HHcy) affect bone such as: hydroxyproline, N-terminal collagen 1 telopeptides. Mitochondrion serves an important role in generating reactive oxygen species (ROS). Mitochondrial abnormalities have been identified during HHcy. The mechanism of Hcy-induced bone remodeling via the mitochondrial pathway is largely unknown. Therefore, we propose a mitochondrial mechanism by which Hcy can contribute to alter bone properties. This may occur both through generations of ROS that activate MMPs and could be extruded into matrix to degrade bone matrix. However, there are contrasting reports on whether Hcy affects bone density, with some reports in favour and others not. Earlier studies also found an alteration in bone biomechanical properties with deficiencies of vitamin B12, folate and HHcy conditions. Moreover, existing data opens speculation that folate and vitamin therapy act not only via Hcy-dependent pathways but also via Hcy-independent pathways. However, more studies are needed to clarify the mechanistic role of Hcy during bone diseases.

Exercise-Linked Skeletal Irisin Ameliorates Diabetes-Associated Osteoporosis by Inhibiting the Oxidative Damage-Dependent miR-150-FNDC5/Pyroptosis Axis.

Recent evidence suggests that physical exercise (EX) promotes skeletal development. However, the impact of EX on the progression of bone loss and deterioration of mechanical strength in mice with type 2 diabetic mellitus (T2DM) remains unexplored. In the current study, we investigated the effect of EX on bone mass and mechanical quality using a diabetic mouse model. The T2DM mouse model was established with a high-fat diet with two streptozotocin injections (50 mg/kg/body wt) in C57BL/6 female mice. The diabetic mice underwent treadmill exercises (5 days/week at 7-11 m/min for 60 min/day) for 8 weeks. The data showed that diabetes upregulated miR-150 expression through oxidative stress and suppressed FNDC5/Irisin by binding to its 3'-untranslated region. The decreased level of irisin further triggers the pyroptosis response in diabetic bone tissue. EX or N-acetyl cysteine or anti-miRNA-150 transfection in T2DM mice restored FNDC5/Irisin expression and bone formation. Furthermore, EX or recombinant irisin administration prevented T2DM-Induced hyperglycemia and improved glucose intolerance in diabetic mice. Furthermore, osteoblastic knockdown of Nlrp3 silencing (si-Nlrp3) or pyroptosis inhibitor (Ac-YVADCMK [AYC]) treatment restores bone mineralization in diabetic mice. Micro-computed tomography scans and mechanical testing revealed that trabecular bone microarchitecture and bone mechanical properties were improved after EX in diabetic mice. Irisin, either induced by skeleton or daily EX or directly administered, prevents bone loss by mitigating inflammasome-associated pyroptosis signaling in diabetic mice. This study demonstrates that EX-induced skeletal irisin ameliorates diabetes-associated glucose intolerance and bone loss and possibly provides a mechanism of its effects on metabolic osteoporosis.

Anterolateral and medial locking plate stiffness in distal tibial fracture model.

BACKGROUND: The purpose of this study was to compare the axial and torsional stiffness between anterolateral and medial distal tibial locking plates in a pilon fracture model. MATERIALS AND METHODS: The biomechanical stiffness of anterolateral or medial plated pilon fracture models was evaluated. Six Sawbones Composite Tibiae with a simulated pilon fracture representing varus or valgus comminution (OTA 43-A2.2) were plated with a Synthes 3.5-mm contoured LCP anterolateral or medial locking distal tibia plate. Load as a function of axial displacement and torque as a function of angular displacement were recorded. Each tibia was tested with a fracture wedge in place and removed with a medial and then anterolateral plate. RESULTS: Loading the tibial plateau medial to the central axis, no significant difference in mean stiffness between the anterolateral and medial plates was demonstrated with the fracture wedge in place. A significant difference was demonstrated with the wedge removed. Loading the plateau posterior to the central axis, no significant difference in mean stiffness between plates was demonstrated with the wedge in place or removed. With the wedge in place, there was a significant difference in mean torsional stiffness for clockwise rotation, but not counterclockwise rotation. With the wedge removed, no significant difference appeared in mean stiffness for clockwise and counterclockwise rotation. CONCLUSION: Distal tibia extra-articular fractures stabilized with anterolateral or medial locking plate constructs demonstrated no statistically significant difference in biomechanical stiffness in compression and torsion testing. CLINICAL RELEVANCE: We believe this study indicates the primary concern when treating a pilon fracture may be soft-tissue considerations. Further clinical studies are required before definitive changes can be recommended regarding pilon fracture fixation.

Exosomal lncRNA-H19 promotes osteogenesis and angiogenesis through mediating Angpt1/Tie2-NO signaling in CBS-heterozygous mice.

Rationale: Emerging evidence indicates that the growth of blood vessels and osteogenesis is tightly coordinated during bone development. However, the molecular regulators of intercellular communication in the bone microenvironment are not well studied. Therefore, we aim to investigate whether BMMSC-Exo promotes osteogenesis and angiogenesis via transporting lnc-H19 in the CBS- heterozygous mouse model. Methods: Using RT2 lncRNA PCR array screening, we identify a bone-specific, long noncoding RNA-H19 (lncRNA-H19/lnc-H19) in exosomes derived from bone marrow mesenchymal stem cells (BMMSC-Exo) during osteogenesis. Using bioinformatics analysis, we further discovered the seed sequence of miR-106a that could bind to lnc-H19. A luciferase reporter assay was performed to demonstrate the direct binding of miR-106a to the target gene angiopoietin 1 (Angpt1). We employed an immunocompromised Nude mouse model, to evaluate the effects of BMMSC-Exo on angiogenesis in vivo. Using a micro-CT scan, we monitored microstructural changes of bone in the experimental mice. Results: BMMSC-Exo possessed exosomal characteristics including exosome size, and typical markers including CD63, CD9, and TSD101. In vitro, BMMSC-Exo significantly promoted endothelial angiogenesis and osteogenesis. Mechanistic studies have shown that exosomal lnc-H19 acts as "sponges" to absorb miR-106 and regulate the expression of angiogenic factor, Angpt1 that activates lnc-H19/Tie2-NO signaling in mesenchymal and endothelial cells. Both of these effects on osteogenesis and angiogenesis are inhibited by antagonizing Tie2 signaling. Treatment of BMMSC-Exo also restored the bone formation and mechanical quality in vivo. Conclusion: These findings provide a novel insight into how the extracellular role of exosomal lnc-H19 affects osteogenesis and angiogenesis through competing endogenous RNA networks.

Probiotics Stimulate Bone Formation in Obese Mice via Histone Methylations.

Rationale: Manipulation of the gut microbiome can prevent pathologic bone loss. However, the effects of probiotics on mitochondrial epigenetic remodeling and skeletal homeostasis in the high-fat diet (HFD)-linked obesity remains to be explored. Here, we examined the impact of probiotics supplementation on mitochondrial biogenesis and bone homeostasis through the histone methylation mechanism in HFD fed obese mice. Methods: 16S rRNA gene sequencing was performed to study the microbiota composition in the gut and microbial dysbiosis in obese mouse model. High resolution (microPET/CT) imaging was performed to demonstrate the obese associated colonic inflammation. Obese-associated upregulation of target miRNA in osteoblast was investigated using a microRNA qPCR array. Osteoblastic mitochondrial mass was evaluated using confocal imaging. Overexpression of mitochondrial transcription factor (Tfam) was used to investigate the glycolysis and mitochondrial bioenergetic metabolism using Tfam-transgenic (Tg) mice fed on HFD. The bone formation and mechanical strength was evaluated by microCT analysis and three-point bending analysis. Results: High-resolution imaging (µ-CT) and mechanical testing revealed that probiotics induced a significant increase of trabecular bone volume and bone mechanical strength respectively in obese mice. Probiotics or Indole-3-propionic acid (IPA) treatment directly to obese mice, prevents gut inflammation, and improved osteoblast mineralization. Mechanistically, probiotics treatment increases mitochondrial transcription factor A (Tfam) expression in osteoblasts by promoting Kdm6b/Jmjd3 histone demethylase, which inhibits H3K27me3 epigenetic methylation at the Tfam promoter. Furthermore, Tfam-transgenic (Tg) mice, fed with HFD, did not experience obesity-linked reduction of glucose uptake, mitochondrial biogenesis and mineralization in osteoblasts. Conclusions: These results suggest that the probiotics mediated changes in the gut microbiome and its derived metabolite, IPA are potentially be a novel agent for regulating bone anabolism via the gut-bone axis.

A consideration of factors affecting palliative oral appliance effectiveness for obstructive sleep apnea: a scoping review.

STUDY OBJECTIVES: This scoping review allows physicians, researchers, and others interested in obstructive sleep apnea to consider effectiveness of oral appliances (OAs). The intent is to improve understanding of OA effectiveness by considering morphologic interaction in patients with obstructive sleep apnea. METHODS: Morphologic and biomechanical criteria for positional alterations of the mandible assessed success rates of OA appliances. Searches of databases (Medline, PubMed, The Cochrane Library, EBSCO) using terms: OA treatment effectiveness and positive and/or negative outcome predictors. Craniofacial predictors of OAs and obstructive sleep apnea biomechanical factors of anatomical traits associated with OA effectiveness were included. Databases searched radiographic cephalometric imaging for morphology/phenotypes and apnea-hypopnea index responses. Articles were excluded if title or abstract was not relevant or a case report. If the analysis did not report mean or standard deviation for apnea-hypoxia index, it was excluded. No language, age, or sex restrictions were applied. RESULTS: Analysis of 135 articles included in searched literature indicated alterations in musculature and pharyngeal airway structure through OA use. These alterations were individually unpredictable with wide variability 61.81% ± 12.29 (apnea-hypoxia index mean ± standard deviation). Morphologic variations as predictors were typically weak and idiosyncratic. Biomechanical factors and wide variations in the metrics of appliance application were unclear, identifying gaps in knowledge and practice of OAs. CONCLUSIONS: An integrated basis to identify morphologic and biomechanical elements of phenotypic expressions of sleep-disordered breathing in the design and application of OAs is needed. Current knowledge is heterogeneous and shows high variability. Identification of subgroups of patients with obstructive sleep apnea responding to OAs is needed.

The effect of modular tapered fluted stems on proximal stress shielding in the human femur.

The purpose of this study was to show a change in proximal femur surface strains following total hip arthroplasty and after the addition of BoneSource hydroxyapatite bone cement in the proximal region of an instrumented femur and to measure the surface strain on the proximal body. Seven third-generation composite femurs (Pacific Research Laboratories, Vashon, Wash) were instrumented with 12 uniaxial strain gages, 6 gages on the anterior face, and 6 gages on the posterior face of each femur. All femurs exhibited stress shielding since the strains in the proximal region were drastically reduced. There was a large decrease in strain in the mid-shaft region and small changes in strain in the distal region. The surface strains on the modular implant were relatively low.

In vivo micro-CT scanning of a rabbit distal femur: repeatability and reproducibility.

Before in vivo micro-CT scanning can be used to investigate femoral trabecular microarchitecture over time in rabbits, its repeatability and reproducibility must be demonstrated. To accomplish this, both distal femurs of two 6-month-old New Zealand white rabbits were scanned five times each in 1 day under different conditions (repeatability). Scanning was done at 28 microm isotropic voxel size to produce five image stacks of each femur. Three operators then followed a standard image processing protocol (reproducibility) to isolate two separate cubes from each anterior femoral condyle [total n = (8 cube sites)(5 scans)(3 operators) = 120]. Bone volume fraction (BV/TV) of the eight different cube sites (sample) ranged from 0.408 to 0.501 (mean: 0.453); trabecular thickness (Tb.Th) ranged from 158.1 to 185.5 microm (mean: 168.6 microm); and trabecular separation (Tb.Sp) ranged from 179.4 to 233.1 microm (mean: 204.7 microm). Using ANOVA and the variance component method, the total process variation was +/- 14.1% of the mean BV/TV of 0.453. The sample variation was +/- 13.9% (p < 0.001), the repeatability was +/- 2.1% (p < 0.001), and the reproducibility was +/- 0.1% (p > 0.05). Results were similar for Tb.Th and Tb.Sp. Though the contribution due to repeatability was statistically significant for each of the three indices, the natural sample differences were far greater than differences caused by repeated scanning under different conditions or by different operators processing the images. These findings suggest that in vivo micro-CT scanning of rabbit distal femurs was repeatable and reproducible and can be used with confidence to measure differences in trabecular bone microarchitecture at a single location in a longitudinal study design.

Impaction grafting for femoral component revision in a goat model using washed morselized cancellous allograft.

To determine whether washing morselized cancellous bone allograft in impaction grafting for revision hip arthroplasty would improve mechanical and biologic performance, left hip hemiarthroplasty with a collarless stem cemented into impacted morselized cancellous bone was performed in 22 goats. Washed allograft was used in the experimental group, and standard allograft was used in the control group. One of 11 experimental and 4 of 11 control implants were observed to be loose at 8 weeks. Washing allowed significantly more morselized cancellous bone to be placed in the experimental group compared to the control group (7.7+/-1.9 and 6.2+/-2.0 g, respectively, P<.05). Significantly less in vivo subsidence over the 8-week study period also was demonstrated in the experimental group compared to the control group (0.4+/-0.4 and 2.2+/-2.3 mm, respectively, P<.05). Angular motion during cyclic +/-1.5 Nm loading demonstrated significant differences between the 2 groups at time zero (2.67 degrees +/-1.02 degrees for the control group and 1.98 degrees +/-0.47 degrees for the experimental group, P<.05) and at 8 weeks (2.40 degrees +/-0.38 degrees for the control group and 1.74 degrees +/-0.55 degrees for the experimental group, P<.05). Histology showed little difference between the 2 groups, but there was a trend toward less inflammation in the experimental group.

Hydrogen sulfide epigenetically mitigates bone loss through OPG/RANKL regulation during hyperhomocysteinemia in mice.

Hydrogen sulfide (H2S) is a novel gasotransmitter produced endogenously in mammalian cells, which works by mediating diverse physiological functions. An imbalance in H2S metabolism is associated with defective bone homeostasis. However, it is unknown whether H2S plays any epigenetic role in bone loss induced by hyperhomocysteinemia (HHcy). We demonstrate that diet-induced HHcy, a mouse model of metabolite induced osteoporosis, alters homocysteine metabolism by decreasing plasma levels of H2S. Treatment with NaHS (H2S donor), normalizes the plasma level of H2S and further alleviates HHcy induced trabecular bone loss and mechanical strength. Mechanistic studies have shown that DNMT1 expression is higher in the HHcy condition. The data show that activated phospho-JNK binds to the DNMT1 promoter and causes epigenetic DNA hyper-methylation of the OPG gene. This leads to activation of RANKL expression and mediates osteoclastogenesis. However, administration of NaHS could prevent HHcy induced bone loss. Therefore, H2S could be used as a novel therapy for HHcy mediated bone loss.

Hydrogen Sulfide Promotes Bone Homeostasis by Balancing Inflammatory Cytokine Signaling in CBS-Deficient Mice through an Epigenetic Mechanism.

Previously, we have shown hyperhomocysteinemia (HHcy) to have a detrimental effect on bone remodeling, which is associated with osteoporosis. During transsulfuration, Hcy is metabolized into hydrogen sulfide (H2S), a gasotransmitter molecule known to regulate bone formation. Therefore, in the present study, we examined whether H2S ameliorates HHcy induced epigenetic and molecular alterations leading to osteoporotic bone loss. To test this mechanism, we employed cystathionine-beta-synthase heterozygote knockout mice, fed with a methionine rich diet (CBS+/- +Met), supplemented with H2S-donor NaHS for 8 weeks. Treatment with NaHS, normalizes plasma H2S, and completely prevents trabecular bone loss in CBS+/- mice. Our data showed that HHcy caused inhibition of HDAC3 activity and subsequent inflammation by imbalancing redox homeostasis. The mechanistic study revealed that inflammatory cytokines (IL-6, TNF-α) are transcriptionally activated by an acetylated lysine residue in histone (H3K27ac) of chromatin by binding to its promoter and subsequently regulating gene expression. A blockade of HDAC3 inhibition in CBS+/- mice by HDAC activator ITSA-1, led to the remodeling of histone landscapes in the genome and thereby attenuated histone acetylation-dependent inflammatory signaling. We also confirmed that RUNX2 was sulfhydrated by administration of NaHS. Collectively, restoration of H2S may provide a novel treatment for CBS-deficiency induced metabolic osteoporosis.

Biomechanics of the Injured Fibula Following Plate Fixation of a Concomitant Tibia Fracture To Fix or Not to Fix?

BACKGROUND: The mechanical role of supplemental fibula fixation in both bone lower leg fractures is not well defined. The benefit of fibula plate fixation in this context is controversial. The purpose of this study was to ascertain the mechanical contributions of the fibula under three conditions (intact, fractured, or plated fibula) following standard tibia locked plating. METHODS: A laboratory fracture model was created with 10 cadaveric legs (5 matched pairs) with no known history of lower extremity trauma or other musculoskeletal conditions. A both bone lower leg fracture was simulated by performing distal osteotomies, 3 to 5 cm above the tibia plafond, leaving a bony defect to simulate an unstable fracture (AOTrauma OTA classification 43-A3). Coronal and sagittal gauge-pins were placed above and below the fracture sites to measure relative displacement across three planes of motion. Axial and torsional loads were applied to the leg under the following conditions: tibia intact and fibula intact (control 1), tibia fracture and fibula intact, tibia fracture and fibula fracture (control 2), and the three conditions of primary interest: tibia plated and fibula intact, tibia plated and fibula fracture (osteotomy), and tibia plated and fibula plated. The load applied for level 1 was 75 N of axial compression and 0.3 Nm of torque, and the load for level 2 was 175 N of axial compression and 1.3 Nm of torque. RESULTS: There were significant differences in motion across the fracture site of the injured leg when the tibia was not plated compared with an intact and plated tibia, p < 0.05. However, when the tibia was plated, there were no significant differences in fracture motion when the fibula was left either intact, osteotomized, or underwent supplemental plate fixation, p > 0.05. This was true regardless of the loads applied. CONCLUSION: The mechanical stability of supplemental fibula fixation in a both bone lower leg fracture model was not significantly improved from standalone distal tibia fixation in this laboratory model. The clinical effects of these findings are yet to be demonstrated.

Assessment of Distal Radioulnar Joint Stability After Reconstruction With the Brachioradialis Wrap.

BACKGROUND: The brachioradialis (BR) wrap technique is an option to restore the stability of the distal radioulnar joint (DRUJ). The technique capitalizes on the BR's advantageous insertion point on the radial styloid and the ability of the BR to be harvested with minimal to no deficit. The tendon can then be wrapped around the radius and ulna, tunneling under the pronator quadratus and extensor compartments and secured back into its insertion to provide stability. In this cadaveric study, we used micro-computed tomography (CT) to assess the stability restored by this procedure. METHODS: Axial CT scans were taken of cadaveric specimens (n = 10) in 3 different positions (neutral, 60° pronation, and 60° supination) to establish the baseline measurements of each DRUJ. Surgical disruption of the dorsal and volar ligaments of each DRUJ then simulated a destabilizing injury and the specimens were scanned again. The specimens then underwent the BR wrap procedure and were scanned once more. Degree of ulnar subluxation with respect to the Sigmoid notch was determined using the modified radioulnar line method. RESULTS: The mean percentages of subluxation in the neutral position for the normal, injured, and reconstructed DRUJ were 22.4±4.9%, 56.2±12.9%, and 29.0±6.5%, respectively. In 60° pronation, these values were 15.4±4.7%, 53.5±15.0%, and 36.5±11.8%, respectively. In 60° supination, these values were 18.6±2.5%, 69.7±20.5%, and 31.9±8.7%, respectively. CONCLUSIONS: Values differed significantly between normal and injured conditions in all positions. No significant difference was noted between normal and reconstructed conditions, suggesting reconstruction improves DRUJ biomechanics and more closely approximates normal stability.

Inhibition of CaMKK2 Enhances Fracture Healing by Stimulating Indian Hedgehog Signaling and Accelerating Endochondral Ossification.

Approximately 10% of all bone fractures do not heal, resulting in patient morbidity and healthcare costs. However, no pharmacological treatments are currently available to promote efficient bone healing. Inhibition of Ca2+ /calmodulin (CaM)-dependent protein kinase kinase 2 (CaMKK2) reverses age-associated loss of trabecular and cortical bone volume and strength in mice. In the current study, we investigated the role of CaMKK2 in bone fracture healing and show that its pharmacological inhibition using STO-609 accelerates early cellular and molecular events associated with endochondral ossification, resulting in a more rapid and efficient healing of the fracture. Within 7 days postfracture, treatment with STO-609 resulted in enhanced Indian hedgehog signaling, paired-related homeobox (PRX1)-positive mesenchymal stem cell (MSC) recruitment, and chondrocyte differentiation and hypertrophy, along with elevated expression of osterix, vascular endothelial growth factor, and type 1 collagen at the fracture callus. Early deposition of primary bone by osteoblasts resulted in STO-609-treated mice possessing significantly higher callus bone volume by 14 days following fracture. Subsequent rapid maturation of the bone matrix bestowed fractured bones in STO-609-treated animals with significantly higher torsional strength and stiffness by 28 days postinjury, indicating accelerated healing of the fracture. Previous studies indicate that fixed and closed femoral fractures in the mice take 35 days to fully heal without treatment. Therefore, our data suggest that STO-609 potentiates a 20% acceleration of the bone healing process. Moreover, inhibiting CaMKK2 also imparted higher mechanical strength and stiffness at the contralateral cortical bone within 4 weeks of treatment. Taken together, the data presented here underscore the therapeutic potential of targeting CaMKK2 to promote efficacious and rapid healing of bone fractures and as a mechanism to strengthen normal bones. © 2018 American Society for Bone and Mineral Research.

Scapular thickness--implications for fracture fixation.

The purpose of this study was to measure and map scapula osseous thickness to identify the optimal areas for internal fixation. Eighteen (9 pairs) scapulae from 2 female and 7 male cadavers were used. After harvest and removal of all soft tissues, standardized measurement lines were made based on anatomic landmarks. For consistency among scapulae, measurements were taken at standard percentage intervals along each line approximating the distance between two consecutive reconstruction plate screw holes. Two-mm-diameter drill holes were made at each point, and a standard depth gauge was used to measure thickness. The glenoid fossa (25 mm) displayed the greatest mean osseous thickness, followed by the lateral scapular border (9.7 mm), the scapula spine (8.3 mm), and the central portion of the body of the scapula (3.0 mm). To optimize screw purchase and internal fixation strength, the lateral border, the lateral aspect of the base of the scapula spine, and the scapula spine itself should be used for anatomic sites of internal fixation of scapula fractures.

Micro-computed tomography assisted distal femur metaphyseal blunt punch compression for determining trabecular bone strength in mice.

Shorter generation time and the power of genetic manipulation make mice an ideal model system to study bone biology as well as bone diseases. However their small size presents a challenge to perform strength measurements, particularly of the weight-bearing cancellous bone in the murine long bones. We recently developed an improved method to measure the axial compressive strength of the cancellous bone in the distal femur metaphysis in mice. Transverse micro-computed tomography image slices that are 7µm thick were used to locate the position where the epiphysis-metaphysis transition occurs. This enabled the removal of the distal femur epiphysis at the exact transition point exposing the full extent of metaphyseal trabecular bone, allowing more accurate and consistent measurement of its strength. When applied to a murine model system consisting of five month old male wild-type (WT) and Ca(2+)/calmodulin dependent protein kinase kinase 2 (CaMKK2) knockout (KO) Camkk2(-/-) mice that possess recorded differences in trabecular bone volume, data collected using this method showed good correlation between bone volume fraction and strength of trabecular bone. In combination with micro-computed tomography and histology, this method will provide a comprehensive and consistent assessment of the microarchitecture and tissue strength of the cancellous bone in murine mouse models.

The effect of wire plane tilt and olive wires on proximal tibia fracture fragment stability and fracture site motion.

To analyze the effect of the tilt angle relationship between the crossed wire plane and the bone axis on the stiffness of fine wire external fixation, load-deformation behavior was compared across different tilt angles (0 degree, 10 degrees, and 20 degrees) of the plane containing crossed smooth or olive wires under identical conditions of central axial compression, medial compression-bending, posterior compression-bending, posteromedial compression-bending, and torsion. Stiffness values were calculated from the load-deformation and torque-angle curves. A tilt angle of 20 degrees with olive wires provided significantly greater stiffness compared to smooth wires at any angle in any loading condition (p < 0.05). A tilt angle of 20 degrees with olive wires was also significantly more stiff than a tilt angle of 0 degree with olive wires in any loading condition. In torsion, olive wires with 10 degrees and 20 degrees tilt were not significantly different, while in posterior bending olive wires with 10 degrees tilt were significantly stiffer than olive wires with 0 degree or 20 degrees tilt. With smooth wires, tilting the wire plane caused a decrease in stiffness in posterior bending, posteromedial bending, and torsion. Overall, the use of olive wires in conjunction with tilting the wire plane enhances the fixation stiffness for proximal tibia fractures while allowing more options for wire configurations that avoid neurovascular and musculotendinous structures, and wounds.