Distal Femoral Metaphysis Research Articles

A Novel Approach to Lower-limb Axial Alignment Analysis: A CT Study.

Purpose:To analyze the torsion of the lower extremities in a healthy cohort and to determine the contribution of different segments of the femur and tibia to the torsion of both bones.Methods:In this cross-sectional study, 32 patients with nonjoint or bone-related symptoms were analyzed by CT angiography. Lower-limb torsion, femoral torsion, proximal femoral torsion, femoral shaft torsion, distal femoral torsion, tibial torsion, proximal tibial torsion, and distal tibial torsion were measured.Results:The median total limb torsion was 25° external torsion, with the median femoral torsion being −9° and the median tibial torsion 30°. Both femoral metaphyses had internal torsion, with the internal torsion of the proximal metaphysis being approximately three times greater than that of the distal femoral metaphysis. The shaft was found to compensate with an external torsion of approximately two-thirds of the internal torsion of both femoral metaphyses. The proximal metaphysis of the tibia accounted for approximately one-third of the external torsion, with the segment from the distal to the tibial tubercle accounting for the remaining two-thirds of the tibial torsion.Conclusions:The diaphysis and distal metaphysis are the major contributors to external torsion of the tibia, whereas the proximal metaphysis is the major contributor to the internal torsion of the femur.

Chronic administration of Cl-amidine, a pan-peptidylarginine deiminase inhibitor, does not reverse bone loss in two different murine models of osteoporosis.

Recent in vitro studies have shown a role for the peptidyl-arginine deiminases (PADs) in bone resorption. However, it is unknown whether these enzymes are involved in bone loss in vivo. Thus, we evaluated the antiresorptive effect of a pan-PAD inhibitor in two murine models of osteoporosis: (a) primary osteoporosis induced by ovariectomy (OVX); and (b) secondary osteoporosis associated to Type-1 diabetes induced by streptozotocin (STZ, 50 mg/kg, i.p., five daily administrations). Five weeks after OVX and 15 weeks after injections of STZ, mice received daily administrations of Cl-amidine (3 or 10 mg/kg, i.p.) or vehicle for 30 consecutive days. At the end of the treatment, femur and vertebra were harvested for microCT analysis. Blood samples were collected for determination of antibodies against cyclic citrullinated peptides (anti-CCP) by enzyme-linked immunosorbent assay. Serum levels of anti-CCP antibodies from diabetic mice were not significantly different compared to control mice. However, a significant loss of both trabecular bone at the femoral neck and cortical bone at the femoral diaphysis was found in diabetic mice, and Cl-amidine did not reverse the diabetes-induced bone loss. Mice with OVX had significantly lower serum levels of anti-CCP compared to mice with sham surgery. OVX resulted in significant loss of both trabecular bone at the L5 vertebra and distal femoral metaphysis. Cl-amidine did not block the OVX-induced bone loss. Our results suggest that chronic treatment with Cl-amidine at the doses and period of time administered is not long enough to inhibit bone loss in two different murine models of osteoporosis.

Thermoneutral housing attenuates premature cancellous bone loss in male C57BL/6J mice.

Mice are a commonly used model to investigate aging-related bone loss but, in contrast to humans, mice exhibit cancellous bone loss prior to skeletal maturity. The mechanisms mediating premature bone loss are not well established. However, our previous work in female mice suggests housing temperature is a critical factor. Premature cancellous bone loss was prevented in female C57BL/6J mice by housing the animals at thermoneutral temperature (where basal rate of energy production is at equilibrium with heat loss). In the present study, we determined if the protective effects of thermoneutral housing extend to males. Male C57BL/6J mice were housed at standard room temperature (22°C) or thermoneutral (32°C) conditions from 5 (rapidly growing) to 16 (slowly growing) weeks of age. Mice housed at room temperature exhibited reductions in cancellous bone volume fraction in distal femur metaphysis and fifth lumbar vertebra; these effects were abolished at thermoneutral conditions. Mice housed at thermoneutral temperature had higher levels of bone formation in distal femur (based on histomorphometry) and globally (serum osteocalcin), and lower global levels of bone resorption (serum C-terminal telopeptide of type I collagen) compared to mice housed at room temperature. Thermoneutral housing had no impact on bone marrow adiposity but resulted in higher abdominal white adipose tissue and serum leptin. The overall magnitude of room temperature housing-induced cancellous bone loss did not differ between male (current study) and female (published data) mice. These findings highlight housing temperature as a critical experimental variable in studies using mice of either sex to investigate aging-related changes in bone metabolism.

Structural and metabolic features of the femur of rats after the implantation of glass crystalline material

In vivo research is an important link in the development of osteoplastic materials.Aim: to study the structural features and markers of bone metabolism after an implantation into the distal metaphysis of the rat femur of glass crystalline calcium phosphate material (GCM). Methods: hole defects in the distal metaphysis of the femur were performed in 28 white laboratory male rats (age 4 months, weight 200–260 g). GCM cylindrical samples with a diameter of 1 mm and a height of 3 mm were placed in defects. A comparison group for biochemical studies was 5 intact rats. Animals were sacrificed 7, 14, 28, and 90 days after surgery. The histological studies of the bone structure around the implant and assessment of osseointegration were performed. In the blood serum the activity of alkaline phosphatase, the content of chondroitin sulfates, urea, the activity of alanine aminotransferase were determined.Results: 7 days after surgery the relative content of bone tissue around implants was (28.59 ± 1.33) %, after 14 days it increased by 2.71 times (p < 0.001). After 30 days bone tissue was located around the entire perimeter of the implant. Bioresorption of the material continued throughout the observation period. A significant increase in the activity of the alkaline phosphatase in blood serum compared with the rate of intact rats was shown from the 7th to the 30th day of observation. The maximum increase of chondroitin sulfates was recorded on the 7th day. The content of urea in the blood serum and the activity of alanine aminotransferase did not statistically significantly differ in terms of the experiment or from indices of intact animals.Conclusions: the studied material is biocompatible, has osteoinductive and osteoconductive qualities. After implanting it into the bone, bone repair is not impaired, and lamellar bone tissue is formed for the final study period (90 days). A characteristic feature of the material is its gradual resorption until the 90th day of observation.

Biomechanical and Bone Material Properties of Schnurri-3 Null Mice.

ABSTRACT Schnurri‐3 (Shn3) is an essential regulator of postnatal skeletal remodeling. Shn3‐deficient mice (Shn3–/–) have high bone mass; however, their bone mechanical and material properties have not been investigated to date. We performed three‐point bending of femora, compression tests of L3 vertebrae. We also measured intrinsic material properties, including bone mineralization density distribution (BMDD) and osteocyte lacunae section (OLS) characteristics by quantitative backscatter electron imaging, as well as collagen cross‐linking by Fourier transform infrared microspectroscopy of femora from Shn3–/– and WT mice at different ages (6 weeks, 4 months, and 18 months). Moreover, computer modeling was performed for the interpretation of the BMDD outcomes. Femora and L3 vertebrae from Shn3–/– aged 6 weeks revealed increased ultimate force (2.2‐ and 3.2‐fold, p < .01, respectively). Mineralized bone volume at the distal femoral metaphysis was about twofold (at 6 weeks) to eightfold (at 4 and 18 months of age) in Shn3–/– (p < .001). Compared with WT, the average degree of trabecular bone mineralization was similar at 6 weeks, but increased at 4 and 18 months of age (+12.6% and +7.7%, p < .01, respectively) in Shn3–/–. The analysis of OLS characteristics revealed a higher OLS area for Shn3–/– versus WT at all ages (+16%, +23%, +21%, respectively, p < .01). The collagen cross‐link ratio was similar between groups. We conclude that femora and vertebrae from Shn3–/– had higher ultimate force in mechanical testing. Computer modeling demonstrated that in cases of highly increased bone volume, the average degree of bone matrix mineralization can be higher than in WT bone, which was actually measured in the older Shn3–/– groups. The area of 2D osteocyte lacunae sections was also increased in Shn3‐deficiency, which could only partly be explained by larger remnant areas of primary cortical bone. © 2019 The Authors. JBMR Plus published by Wiley Periodicals, Inc. on behalf of American Society for Bone and Mineral Research.

Effects of the combination of low-level laser therapy and anionic polymer membranes on bone repair.

In view of the limitations of bone reconstruction surgeries using autologous grafts as a gold standard, tissue engineering is emerging as an alternative, which permits the fabrication and improvement of scaffolds to stimulate osteogenesis and angiogenesis, processes that are essential for bone repair. Polymers are used to mimic the extracellular bone matrix and support cell growth. In addition, bone neoformation can be induced by external factors such as laser irradiation, which stimulates bone metabolism. The objective of this study was to evaluate the regeneration of bone defects using collagen and elastin membranes derived from intestinal serosa and bovine auricular cartilage combined with low-level laser application. Thirty-six Wistar rats were operated to create a 3-mm defect in the distal metaphysis of the left femur and divided into six groups: G1 (control, no treatment); G2 (laser); G3 (elastin graft), G4 (elastin+laser); G5 (collagen graft); G6 (collagen+laser). The animals were sacrificed 6weeks after surgery and the femurs were removed for analysis of bone repair. Macroscopic and radiological results showed the absence of an infectious process in the surgical area. This was confirmed by histological analysis, which revealed no inflammatory infiltrate. Histomorphometry showed that the formation of new bone started from the margins of the bone defect and its volume was greater in elastin+laser and collagen+laser. We conclude that newly formed bone in the graft area was higher in the groups that received the biomaterials and laser. The collagen and elastin matrices showed biocompatibility.

Establishment of a clinically relevant large animal model to assess the healing of metaphyseal bone.

Despite the high incidence of metaphyseal bone fractures in patients, the mechanisms underlying the healing processes are poorly understood due to the lack of suitable experimental animal models. Hence, the present study was conducted to establish and characterise a clinically relevant large-animal model for metaphyseal bone healing. Six female adult Merino sheep underwent full wedge-shaped osteotomy at the distal left femur metaphysis. The osteotomy was stabilised internally with a customised anatomical locking titanium plate that allowed immediate post-operative full-weight bearing. Bone healing was evaluated at 12 weeks post-fracture relative to the untouched right femur. Histological and quantitative micro-computed tomography results revealed an increased mineralised bone mass with a rich bone microarchitecture. New trabeculae healed by direct intramembranous ossification, without callus and cartilaginous tissue formation. Stiffness at the cortical and trabecular regions was comparable in both groups. Functional morphological analysis of the osteocyte lacunae revealed regularly arranged spherically shaped lacunae along with the canalicular network. Bone surface biochemical analysis using time-of-flight secondary-ion mass spectrometry showed high and homogeneously distributed levels of calcium and collagenous components. Ultrastructure imaging of the new trabeculae revealed a characteristic parallel arrangement of the collagen fibrils, evenly mineralised by the dense mineral substance. The specialised bone cells were also characterised by their unique structural features. Bone remodelling in the fractured femur was evident in the higher expression levels of prominent bone formation and resorption genes. In conclusion, the novel metaphyseal fracture model is beneficial for studying healing and treatment options for the enhancement of metaphyseal bone defects.

Correlation between diffusion tensor imaging parameters of the distal femoral physis and adjacent metaphysis, and subsequent adolescent growth.

Diffusion-tensor imaging (DTI) depicts the movement of water through columns of cartilage and newly formed bone and provides information about velocity of growth and growth potential. To determine the correlation between DTI tractography parameters of the distal femoral physis and metaphysis and the height change after DTI in pubertal and post-pubertal children. We retrospectively analyzed DTI images of the knee in 47 children with a mean age of 14.1years in a 2-year period. In sagittal echoplanar DTI studies, regions of interest were placed in the femoral physis. Tractography was performed using a fractional anisotropy threshold of 0.15 and a maximum turning angle of 40°. The sample was divided to assess short-term and long-term growth after DTI. Short-term growth (n=25) was the height change between height at MRI and 1year later. Long-term growth (n=36) was the height gain between height at MRI and at the growth plateau. For the short-term group, subjects with larger tract volume (R2=0.40) and longer track lengths (R2=0.38) had larger height gains (P<0.01). For the long-term group, subjects with larger tract volume (R2=0.43) and longer track lengths (R2=0.32) had a larger height gain at the growth plateau (P<0.01). Intra- and inter-observer variability were good-excellent. Follow-up data of growth 1year after DTI evaluation and at skeletal maturity confirms that DTI parameters are associated with the amount of post-imaging growth.

Adipose-derived stem cell sheets accelerate bone healing in rat femoral defects.

In the present study, we investigated whether both adipose-derived stem cell (ADSC) and osteogenic-induced ADSC sheets could promote bone healing in a rat distal femoral metaphysis bone defect model. A through-hole defect of 1 mm diameter was drilled into each distal femur of 12 week old rats. Forty-five rats were randomly assigned to three groups: (1) control group; (2) ADSC sheet group; or (3) osteogenic-induced ADSC sheet group. We evaluated each group by analysis of computerized tomography scans every week after the surgery, histological analysis, and DiI labeling (a method of membrane staining for post implant cell tracing). Radiological and histological evaluations showed that a part of the hole persisted in the control group at four weeks after surgery, whereas the hole was restored almost completely by new bone formation in both sheet groups. The mean value of bone density (in Houndsfield units) for the bone defect area was significantly higher in both sheet groups than that in the control group (p = 0.05) at four weeks postoperative. A large number of osteocalcin positive osteoblasts were observed at the area of bone defect, especially in the osteogenic-induced ADCS sheet group. DiI labeling in the newly formed bone showed that each sheet had differentiated into bone tissue at four weeks after surgery. The ADSC and the osteogenic-induced ADSC sheets promoted significantly quicker bone healing in the bone defect. Moreover, the osteogenic-induced ADSC sheet may be more advantageous for bone healing than the ADSC sheet because of the higher number of osteocalcin positive osteoblasts via the transplantation.

N-(3-methoxybenzyl)-(9Z,12Z,15Z)-octadecatrienamide promotes bone formation via the canonical Wnt/β-catenin signaling pathway.

Osteoporosis is characterized by low bone mineral density and microarchitectural deterioration of bone tissue. N-(3-methoxybenzyl)-(9Z,12Z,15Z)-octadecatrienamide (MBOC) is one of the macamides isolated from Maca (Lepidium meyenii Walp.), a cruciferous plant from the Andes of Peru. In this study, C3H/10T1/2 mesenchymal stem cells were treated with MBOC in osteogenic induction medium. An ovariectomized (OVX) mouse model was used to investigate the effect of 1-month MBOC treatment on the prevention of postmenopausal osteoporosis. Remarkably, trabecular thickness, trabecular number, and bone volume/tissue volume of the distal femoral metaphysis were significantly increased in OVX+MBOC mice compared with OVX mice, as revealed by microcomputed tomography analysis. Trabecular separation was decreased in OVX+MBOC mice compared with OVX mice. Consistently, MBOC increased the levels of osteocalcin and runt-related transcription factor 2 in OVX mice, as well as the expression of runt-related transcription factor 2, osterix, and alkaline phosphatase in C3H/10T1/2 cells. Mechanistically, MBOC activates the canonical Wnt/β-catenin signaling pathway via inhibiting phosphorylation of GSK-3β at Tyr216 and maintaining β-catenin expression. Collectively, the current study demonstrates the robustness of MBOC in the induction of mesenchymal stem cells osteogenic differentiation and consequent bone formation, suggesting that MBOC may be a potentially effective drug to treat postmenopausal osteoporosis.

Modulation of bone turnover by Cissus quadrangularis after ovariectomy in rats.

In women, age-related bone loss is associated with increased risk of bone fracture. Existing therapies are associated with severe side effects; thus, there is a need to find alternative medicines with less or optimal side effects. Cissus quadrangularis (CQ), an Ayurvedic medicine used to enhance fracture healing, was tested for its bone protective properties and studied to discern the mechanism by which it is beneficial to bone. Female Sprague Dawley rats were either sham operated or ovariectomized and were fed CQ for 3months. Several biochemical markers, cytokines and hormones were assayed. Femur, tibia and lumbar vertebrae were subjected to pQCT and µCT densitometry. MC3T3 cells were cultured, treated with CQ and used to analyze miRNA content and subjected to qPCR for gene expression analysis related to bone metabolism. CQO rats showed protected bone mass and microarchitecture of trabecular bone in the distal femoral metaphysis and the proximal tibial metaphysis. The lumbar vertebrae, however, showed no significant changes. Serum protein expression levels of P1NP increased and Trap5b and CTX levels decreased with in vivo CQ treatment. Some influence on the anti- and pro-inflammatory markers was also observed. Significantly high level of estradiol in the CQO rats was observed. In vitro expression of a few genes related to bone metabolism showed that osteocalcin increased significantly. The other genes-collagen I expression, SPP1, BMP2, DCAT1-decreased significantly. Certain miRNA that regulate bone turnover using the BMP pathway and Wnt signaling pathways were upregulated by CQ. qPCR after acute treatment with CQ showed significantly increased levels of osteocalcin and decreased levels of Wnt/β catenin antagonist DCAT1. Overall, CQ protected the microarchitecture of the long bones from ovariectomy-induced bone loss. This may be because of decreased inflammation and modulation through the BMP and Wnt signaling pathways. We conclude that CQ is a potential therapeutic agent to treat postmenopausal osteoporosis with no side effects.

Short-term antidepressant treatment has long-lasting effects, and reverses stress-induced decreases in bone features in rats

Antidepressants are among the most-prescribed class of drugs in the world and though weight gain is a common outcome of antidepressant treatment, that effect is not well understood. We employed an animal model comprised of 2 weeks of chronic restraint stress with antidepressant treatment, followed by diet-induced obesity. We showed that short-term antidepressant treatment had long-lasting effects, not only leading to weight gain, but also enhancing trabecular and cortical bone features in rats; therefore, weight gain in this model was different from that of the classic diet-induced obesity. Late in the post-restraint recovery period, antidepressant-treated animals were significantly heavier and had better bone features than saline-treated controls, when assessed in the distal femoral metaphysis. The propensity to gain weight might have influenced the rate of catch-up growth and bone allometry, as heavier animals treated with fluoxetine also had enhanced bone features when compared to non-stressed animals. Therefore, short-term antidepressant treatment ameliorated the long-term effects of stress on body growth and bone. Growth and bone structural features were associated with leptin levels, and the interaction between leptin levels and antidepressant was significant for bone mineral content, suggesting that short-term antidepressants in the context of long-term diet-induced obesity modified the role of leptin in bone formation. To our knowledge this is the first study reporting that short-term antidepressant treatment has long-lasting effects in restoring the effects of chronic stress in body weight and bone formation. Our findings may be relevant to the understanding and treatment of osteoporosis, a condition of increasing prevalence due to the aging population.

Comparison of the removal torque and a histomorphometric evaluation of the RBM treated implants with the RBM followed by laser treated implants: an experimental study in rabbits.

BackgroundIn the osseointegration of dental implants, the implant surface properties have been reported to be some of the most important critical factors. The effect of implant’s surfaces created by resorbable blast media (RBM) followed by laser ablation on bone tissue reactions was examined using the removal torque test and histomorphometric analysis.MethodsTwo types of dental implants, RBM-laser implants (experimental group) and RBM implants (control group) (CSM implant system, Daegu, Korea; L=6 mm, diameter=3.75 mm) were placed into the right and left distal femoral metaphysis of 17 adult rabbits. Six weeks after placement, removal torque was measured and histomorphometric analysis was performed.ResultsThe mean removal torque was 24.0±10.2 Ncm and 46.6±16.4 Ncm for the control and test specimens, respectively. The experimental RBM-laser implants had significantly higher removal torque values than the control RBM implants (p=0.013). The mean values of total and cortical bone to implant contact (BIC) were respectively 46.3±10.8% and 65.3±12.5% for the experimental group, and 41.9±18.5% and 57.6±10.6% for the control group. The experimental RBM-laser implants showed a higher degree of total and cortical BIC compared with RBM implants, but there was no statistical significance (p=0.482, 0.225).ConclusionThe removal torque and BIC of the test group were higher than those of the control group. In this study, the surface treatment created by RBM treatment followed by laser ablation appears to have a potential in improving bone tissue reactions of dental implants.

Short-term intermittent PTH 1–34 administration and bone marrow blood vessel ossification in Mature and Middle-Aged C57BL/6 mice

Intermittent parathyroid hormone (PTH) administration augments bone and progressive bone marrow blood vessel (BMBV) ossification occurs with advancing age. Since intermittent PTH administration augments bone, it may also serve to increase BMBV ossification. We assessed the influence of 5- and 10-days of intermittent PTH 1–34 administration on trabecular and cortical bone and BMBV ossification in mature (6–8 mon; n = 30) and middle-aged (10–12 mon; n = 30) male and female C57BL/6 mice. Mice were divided accordingly: control (CON) and 5-days (5dPTH) and 10-days (10dPTH) of PTH. Mice were given PBS (50 μl) or PTH 1–34 (43 μg/kg/d) for 5- and 10-consecutive days. Trabecular bone microarchitecture (i.e., BV/TV [%], Tb.Th [μm], Tb.N [/mm], and Tb.Sp [μm]) was assessed in the distal femoral metaphysis and cortical bone parameters (i.e., Ct.Th [μm] and CSMI [mm4]) at the femoral mid-shaft. BMBV ossification (i.e., ossified vessel volume [OsVV, %] and ossified vessel thickness [OsV.Th, μm]) was assessed in the medullary cavity of the femoral shaft. All parameters were determined by μCT. At this sample size, no gender-related differences were observed so female and male data were pooled. There were no main effects nor interactions for trabecular microarchitecture and Ct.Th. However, CSMI was larger (p < 0.05) in Middle-Age vs. Mature and larger (p < 0.05) in CON and 10dPTH vs. 5dPTH. OsVV tended (p = 0.057) to be higher (0.18 ± 0.04% vs. 0.09 ± 0.02%, respectively) and OsV.Th was higher (p < 0.05; 17.4 ± 1.6 μm vs. 12.1 ± 1.4 μm, respectively) in Middle-Aged vs. Mature mice. OsVV was not altered, but ossified vessels tended (p = 0.08) to be thicker in 10dPTH (17.6 ± 2.0 μm) vs. CON (12.5 ± 1.7 μm). No interactions were observed for OsVV and OsV.Th. In conclusion, this is the first report of ossified BMBV in C57BL/6 mice. The increased OsV.Th in Middle-Aged mice coincides with previous reports of increased OsVV in aged rats. The tendency of augmented OsV.Th in 10dPTH suggests that this treatment may ultimately impair the patency of bone marrow blood vessels.

Tetrahydroxy stilbene glucoside protected against diabetes‐induced osteoporosis in mice with streptozotocin‐induced hyperglycemia

Tetrahydroxy stilbene glucoside (TSG), an active component from medicinal herb Polygonum multiflorum Thunb, could block the activity of the tissue renin-angiotensin system (RAS), which plays a critical role in development of diabetic osteoporosis. This study aimed to determine if TSG therapy could alleviate bone deteriorations in diabetic mouse model induced by streptozotocin. The diabetic mice showed the loss of trabecular bone mass and the changes of trabecular bone microarchitectural parameters as well as the increase in amount of tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts at the distal metaphysis of femur when compared with those of nondiabetic mice. Treatment with TSG significantly elevated calcium content in serum and bone and improved biological parameters of trabecular bone, accompanied by increasing messenger RNA (mRNA) expression of RUNX-2, COL-I, and OCN and protein expression of β-catenin as well as down-regulating protein expression of RAS components including renin and AT1R. In addition, TSG repressed diabetes-induced decrease in ratio of OPG/RANKL expression and increase in sclerostin expression in bone. The similar effects of TSG on osteoblasts-specific genes were found in MC3T3-E1 cells. Taken together, the present study demonstrated the osteopreserve effects of TSG in diabetic mice, and the underlying mechanism might be attributed to its regulation on osteogenesis and osteoclastogenesis.

Effects of a Pasty Bone Cement Containing Brain-Derived Neurotrophic Factor-Functionalized Mesoporous Bioactive Glass Particles on Metaphyseal Healing in a New Murine Osteoporotic Fracture Model.

The development of new and better implant materials adapted to osteoporotic bone is still urgently required. Therefore, osteoporotic muscarinic acetylcholine receptor M3 (M3 mAChR) knockout (KO) and corresponding wild type (WT) mice underwent osteotomy in the distal femoral metaphysis. Fracture gaps were filled with a pasty α-tricalcium phosphate (α-TCP)-based hydroxyapatite (HA)-forming bone cement containing mesoporous bioactive CaP-SiO2 glass particles (cement/MBG composite) with or without Brain-Derived Neurotrophic Factor (BDNF) and healing was analyzed after 35 days. Histologically, bone formation was significantly increased in WT mice that received the BDNF-functionalized cement/MBG composite compared to control WT mice without BDNF. Cement/MBG composite without BDNF increased bone formation in M3 mAChR KO mice compared to equally treated WT mice. Mass spectrometric imaging showed that the BDNF-functionalized cement/MBG composite implanted in M3 mAChR KO mice was infiltrated by newly formed tissue. Leukocyte numbers were significantly lower in M3 mAChR KO mice treated with BDNF-functionalized cement/MBG composite compared to controls without BDNF. C-reactive protein (CRP) concentrations were significantly lower in M3 mAChR KO mice that received the cement/MBG composite without BDNF when compared to WT mice treated the same. Whereas alkaline phosphatase (ALP) concentrations in callus were significantly increased in M3 mAChR KO mice, ALP activity was significantly higher in WT mice. Due to a stronger effect of BDNF in non osteoporotic mice, higher BDNF concentrations might be needed for osteoporotic fracture healing. Nevertheless, the BDNF-functionalized cement/MBG composite promoted fracture healing in non osteoporotic bone.

Prevalence of glucocorticoid induced osteonecrosis in the mouse is not affected by treatments that maintain bone vascularity

ObjectiveDetermine if LLP2A-Ale or PTH (1–34) affects the prevalence of glucocorticoid-induced osteonecrosis (ON) in a mouse model. MethodsEight-week-old young adult male BALB/cJ mice were weight-randomized into Control (Con), glucocorticoid (GC)-only, or concurrent treatments with GC and LLP2A-Ale (250 μg/kg or 500 μg/kg, IV, Days 1, 14, 28) or parathyroid hormone hPTH (1–34) (40 μg/kg, 5×/week). Mice were necropsied after 45 days for qualitative evaluation of prevalent ON and quantitative evaluation of vascularity in the distal femoral epiphysis (DFE); and quantitative evaluation of bone mass, microarchitecture, and strength in the distal femoral metaphysis and lumbar vertebral body. ResultsThe prevalence of ON was 14% in the Con group and 36% in the GC-only group (P = 0.07). The prevalence of ON did not differ among GC-only, GC + LLP2A-Ale, and GC + PTH groups. GC-only mice had significantly lower trabecular and cortical bone strength than Con, while GC + LLP2A-Ale (500 μg/kg) and GC + PTH (1–34) groups had significantly greater trabecular bone strength than the GC-only group. GC + LLP2A-Ale (250 μg/kg and 500 μg/kg) and GC + PTH had significantly higher trabecular bone volume than GC-only mice at the vertebrae, distal femoral epiphyses and distal femoral metaphyses. DFE vascularity was lower in GC-only mice than in all other groups. ConclusionNeither LLP2A-Ale nor hPTH (1–34) reduced the prevalence of GC-induced ON, compared to GC-only mice. However, GC-treated mice given LLP2A-Ale or hPTH (1–34) had better bone mass, microarchitecture, and strength in trabecular-rich regions, and higher levels of vascularity than GC-only mice.

RESPON TULANG FEMUR TIKUS OSTEOPOROSIS YANG MENGKONSUMSI CALCITRIOL

Calcitriol supplementation in ovariectomized rats decreased bone resorption and increased bone formation, however, it depend on dose. The objective of the research was to study the response of femur bone in osteoporosis rats consuming calcitriol. Thirty female Wistar rats at 8 weeks of age were randomly divided into six groups (sham operated rats as normal control rats/group N and NK, ovariectomized control rats/group Ov and OvK, ovariectomized rats/group OvDand OvE) of five each. All rats were fed standard diet for 8 weeks. At 16 weeks of age, group N and Ov were euthanized, the right femur were taken for histopathological examination. Group NK and OvK were fed a standard diet, group OvD was fed a standard diet +40ng calcitriol; and group OvE was fed a standard diet+25µg ethynil ethyl estradiol. Treatments were done for six weeks. At the end of study, blood samples were taken from plexus orbitalis medialis for estrogen analysis. All rats were euthanized using ketamine10% and xylazine 2%. Right femur was taken for histopathological examination using hematoxylin and eosin stain, and immunohistochemistry using monoclonal antibody anti TRAP5b which was detected with streptavidin-biotin. The results showed that estradiol level of the rats in group OvD was not significantly different compared with the rats in OvK group, however, it was significanly lower compared to the rats in group OvE. Histopathologic figure of right distal femur metaphysis in group OvD was shown lesser adipocyte in the bone marrow and more trabecular bone speculum compared to group OvK, however, there was more adipocyte in the bone marrow and lesser trabecular bone speculum compared to group OvE. Immunohistochemistry of distal femur metaphysis in group OvD and OvE were revealed tartrate resistant alkaline phosphatase 5b (TRAP5b) expression in trabecular bone, which was located in bone marrow space and trabecular speculum surface as well. Based on the results, it can be concluded that calcitriol 40ng/day supplementation in osteoporosis rats for 6 weeks decreased bone resorption and increased bone formation distal femur metaphysis.

Bone Mineral Density Around the Knee Joint: Correlation With Central Bone Mineral Density and Associated Factors

Introduction: The aims of this study were to (1) assess the bone mineral density (BMD) around the knee joint, (2) determine the correlation between central and knee BMDs, and (3) investigate the factors associated with BMD around the knee joint in patients with knee osteoarthritis (OA). Methodology: This cross-sectional study included 122 patients who underwent total knee arthroplasty. Central and knee dual-energy X-ray absorptiometry was performed preoperatively. BMD at 6 regions of interest (ROIs) around the knee joint were measured, and their correlations with central BMD were determined using Spearman's correlation analysis. Lower limb alignment, severity of OA, body mass index (BMI), preoperative functional and pain scores were assessed to elucidate the factors associated with knee BMD using linear regression analysis. Results: Around the knee joint, BMD was the lowest at the distal femoral metaphysis and lateral tibial condyle. Knee BMD was significantly correlated with central BMD. However, the correlation coefficients varied by the ROI. Additionally, multivariate analysis revealed different associations with respect to the regions around the knee joint. Varus alignment of the lower limb was associated with increased BMD of the medial condyles and decreased BMD of lateral condyles. High grade OA was a protective factor; it was associated with increased BMD at the lateral condyles of the femur and tibia. Higher BMI was an independent protective factor in all ROIs around the knee joint except the lateral femoral condyles. Lower functional level was not associated with decreased BMD, whereas a higher pain score was significantly associated with lower BMD at the proximal tibial metaphysis. Conclusions: Knee BMD was significantly correlated with central BMD. However, the correlations varied with the regions around the knee joint probably due to their independent association with the alignment of the lower limb, severity of OA, BMI, and preoperative pain level.

Whole body vibration with rest days could improve bone quality of distal femoral metaphysis by regulating trabecular arrangement.

Low-magnitude, high-frequency vibration (LMHFV) with rest days (particularly seven rest days) was considerably effective in improving the morphological and mechanical properties of rat proximal femur. However, current knowledge is limited regarding the possible benefit of this mechanical regimen to other bone sites and whether the optimal rest days are the same. This study followed our previous experiment on LMHFV loading with rest days for three-month-old male Wistar rats. The experiment involved seven groups, namely, vibrational loading for X day followed with X day rest (X=1, 3, 5, 7), daily vibrational loading, tail suspension and baseline control. Micro-computed tomography (micro-CT) scanning was used to evaluate the microarchitecture of the distal femoral trabecular bone. Micro-CT image-based microfinite element analysis was performed for each distal femoral metaphysis. LMHFV with rest days substantially changed the trabecular arrangement from remarkably plate-like to rod-like. Vibrational loading with 1 day rest was substantially effective in improving the architecture and apparent- and tissuelevel mechanical properties of the rat distal femoral metaphysis. This study may provide an improved understanding of the sitespecific responses of bone tissue to LMHFV with rest days for a substantially effective therapy of a targeted bone site.