Bone Surface/bone Volume Research Articles

Long-acting bulleyaconitine A microspheres via intra-articular delivery for multidimensional therapy of rheumatoid arthritis

Bulleyaconitine A (BLA) is a promising candidate for treating rheumatoid arthritis (RA) with diverse pharmacological activities, including anti-inflammatory, analgesic and bone repair. Herein, the long-acting bulleyaconitine A microspheres (BLA-MS) were developed to treat RA comprehensively by forming drug reservoirs in joint cavities. The BLA-MS were prepared by emulsion/solvent evaporation method. The particle size and distribution were assessed by SEM. The crystalline state was investigated by DSC and PXRD. The drug loading (DL), encapsulation efficiency (EE) and cumulative release in vitro were determined by HPLC. The DL and EE were 23.93 ± 0.38 % and 95.73 ± 1.56 % respectively, and the cumulative release was up to 69 days with a stable release curve. The pharmacodynamic results in collagen induced arthritis (CIA) rats showed a noticeable reduction in paw thickness (5.66 ± 0.32 mm), and the decreasing expression level of PGE2, TNF-α and IL-6 which diminished the infiltration of inflammatory cells, thereby alleviating the progression of erosion and repairing the damaged bones (BV/TV (Bone Volume / Total Volume): 81.97 %, BS/BV (Bone Surface / Bone Volume): 6.08 mm−1). In conclusion, intra-articular injection of BLA-MS should have a promising application in the treatment of RA and may achieve clinical transformation in the future.

Preparation and in vivo osteogenesis of acellular dermal matrix/dicalcium phosphate composite scaffold for bone repair

To investigate the physicochemical properties, osteogenic properties, and osteogenic ability in rabbit model of femoral condylar defect of acellular dermal matrix (ADM)/dicalcium phosphate (DCP) composite scaffold. ADM/DCP composite scaffolds were prepared by microfibril technique, and the acellular effect of ADM/DCP composite scaffolds was detected by DNA residue, fat content, and α-1，3-galactosyle (α-Gal) epitopes; the microstructure of scaffolds was characterized by field emission scanning electron microscopy and mercury porosimetry; X-ray diffraction was used to analyze the change of crystal form of scaffold; the solubility of scaffolds was used to detect the pH value and calcium ion content of the solution; the mineralization experiment in vitro was used to observe the surface mineralization. Twelve healthy male New Zealand white rabbits were selected to prepare the femoral condylar defect models, and the left and right defects were implanted with ADM/DCP composite scaffold (experimental group) and skeletal gold ® artificial bone repair material (control group), respectively. Gross observation was performed at 6 and 12 weeks after operation; Micro-CT was used to detect and quantitatively analyze the related indicators [bone volume (BV), bone volume/tissue volume (BV/TV), bone surface/bone volume (BS/BV), trabecular thickness (Tb.Th), trabecular number (Tb.N), trabecular separation (Tb.Sp), bone mineral density (BMD)], and HE staining and Masson staining were performed to observe the repair of bone defects and the maturation of bone matrix. Gross observation showed that the ADM/DCP composite scaffold was a white spongy solid. Compared with ADM, ADM/DCP composite scaffolds showed a significant decrease in DNA residue, fat content, and α-Gal antigen content ( P<0.05). Field emission scanning electron microscopy showed that the ADM/DCP composite scaffold had a porous structure, and DCP particles were attached to the porcine dermal fibers. The porosity of the ADM/DCP composite scaffold was 76.32%±1.63% measured by mercury porosimetry. X-ray diffraction analysis showed that the crystalline phase of DCP in the ADM/DCP composite scaffolds remained intact. Mineralization results in vitro showed that the hydroxyapatite layer of ADM/DCP composite scaffolds was basically mature. The repair experiment of rabbit femoral condyle defect showed that the incision healed completely after operation without callus or osteophyte. Micro-CT showed that bone healing was complete and a large amount of new bone tissue was generated in the defect site of the two groups, and there was no difference in density between the defect site and the surrounding bone tissue, and the osteogenic properties of the two groups were equivalent. There was no significant difference in BV, BV/TV, BS/BV, Tb.Th, Tb.N, and BMD between the two groups ( P>0.05), except that the Tb.Sp in the experimental group was significantly higher than that in the control group ( P<0.05). At 6 and 12 weeks after operation, HE staining and Masson staining showed that the new bone and autogenous bone fused well in both groups, and the bone tissue tended to be mature. The ADM/DCP composite scaffold has good biocompatibility and osteogenic ability similar to the artificial bone material in repairing rabbit femoral condylar defects. It is a new scaffold material with potential in the field of bone repair.

Comparative evaluation of 3D-printed and conventional implants in vivo: a quantitative microcomputed tomographic and histomorphometric analysis

In recent years, 3D-printing technology to fabricate dental implants has garnered widespread attention due to its patient-specific customizability and cost-effectiveness. This preclinical animal study analyzed the radiographic and histomorphometric outcomes of 3D-printed implants (3DIs) placed immediately after extraction and compared them to conventional implants (CIs). 3DIs and CIs of the same dimensions placed immediately were analyzed at 2, 6, and 12 weeks. The micro-computed tomography (micro-CT) analysis revealed statistically significant differences at 2 weeks in favor of 3DIs over the CIs in terms of bone volume/tissue volume (BV/TV), bone surface/bone volume (BS/BV), trabecular bone pattern factor (Tb.Pf), and structure model index (SMI). At 2 weeks, the mean bone-to-implant contact (BIC) of the 3DIs was greater than that of the CIs; the mean bone area fraction occupancy (BAFO) and the number of Haversian canals of the 3DIs showed no statistically significant differences compared to CIs at 2 weeks. At 6 and 12 weeks, there were no statistically significant differences between the 3DIs and CIs in any parameters. Within limitations, in the early stage of extraction socket healing, the 3DIs demonstrated a higher BIC than the CIs, presenting that 3DIs may be a potential option for immediate placement to enhance osseointegration.

Protaetia brevitarsis seulensis larvae ethanol extract inhibits RANKL-stimulated osteoclastogenesis and ameliorates bone loss in ovariectomized mice

Modulation of osteoclast formation could be a therapeutic target for inhibiting pathological bone destruction. The receptor activator of nuclear factor (NF)-κB ligand (RANKL) is known to be an essential factor in osteoclast differentiation and activation inducers. However, whether Protaetia brevitarsis seulensis (P. brevitarsis) larvae—a traditional animal-derived medicine used in many Asian countries—can inhibit RANKL-induced osteoclast formation and prevent ovariectomy (OVX)-induced bone loss has not been evaluated. Here, we aimed to investigate the anti-osteoporotic effects of P. brevitarsis larvae ethanol extract (PBE) in RANKL-stimulated RAW264.7 cells and OVX mice. In vitro, PBE (0.1, 0.5, 1, and 2 mg/mL) decreased RANKL‑induced tartrate-resistant acid phosphatase (TRAP) activity and expression of osteoclastogenesis-associated genes and proteins. Furthermore, PBE (0.1, 0.5, 1, and 2 mg/mL) significantly inhibited the phosphorylation of p38 and NF-κB. Female C3H/HeN mice were divided into five groups (n = 5 per group), namely, sham-operated, OVX, OVX+PBEL (100 mg/kg, oral gavage), OVX+PBEH (200 mg/kg, oral gavage), and OVX+estradiol (0.03 μg/day, subcutaneous injection). High doses of PBE significantly increased femoral bone mineral density (BMD) and bone volume/tissue volume (BV/TV), whereas femoral bone surface/bone volume (BS/BV) and osteoclastogenesis-associated protein expression decreased compared to those in the OVX group. Moreover, PBE (200 mg/kg) significantly increased estradiol and procollagen type I N-terminal propeptide and decreased N-terminal telopeptide of type I collagen and C-terminal telopeptide of type I collagen compared to those in the OVX group. Our results suggest that PBE can be an effective therapeutic candidate for preventing or treating postmenopausal osteoporosis.

Naringin promotes osteogenesis and ameliorates osteoporosis development by targeting JAK2/STAT3 signalling.

Osteoporosis is a systemic bone metabolism disorder, which increases the risk of fractures, and in severe cases it may cause disability or even death. An important factor contributing to osteoporosis is the imbalance between bone formation and resorption. Naringin was reported to promote osteoblast differentiation, thus enhancing bone formation and alleviating osteoporosis development. However, the signalling pathways related to the regulatory mechanism of naringin in osteoporosis development are not clear. Proliferation of bone mesenchymal stem cells (BMSCs) treated with naringin in vitro was detected by CCK-8. An osteogenesis differentiation medium supplemented with naringin was applied to explore the effects of naringin on BMSC osteogenic differentiation, as detected by Alizarin red staining. Ovariectomy (OVX)-induced postmenopausal osteoporosis (PMOP) rats were orally administered with naringin. Dual-energy X-ray absorptiometry (DEXA) and micro-CT were applied to measure bone mineral density (BMD), bone volume/total volume (BV/TV), trabecula thickness (Tb.Th), trabecula number (Tb.N), trabecular separation (Tb.Sp) and bone surface/bone volume (BS/BV). H&E staining was performed to show pathological changes of the femur in PMOP rats after naringin treatment. Bone metabolism indicators were assessed by ELISA. We found that naringin suppressed the activation of the JAK2/STAT3 pathway. Naringin promoted BMSC proliferation and osteogenic differentiation. Furthermore, naringin alleviates bone loss and improves abnormal bone metabolism of PMOP rats. Collectively, naringin promotes BMSC osteogenic differentiation to ameliorate osteoporosis development by targeting JAK2/STAT3signalling.

Combined Effects of Low-Frequency Pulsed Electromagnetic Field and Melatonin on Ovariectomy-Induced Bone Loss in Mice.

Pulsed electromagnetic field (PEMF) therapy and melatonin (MEL) supplementation are expected to be important strategies for the treatment of osteoporosis. The aim of the current study was to investigate the efficacy of PEMF therapy, MEL supplementation, a combination of PEMF therapy, and MEL supplementation (PEMF + MEL) in mice with bilateral ovariectomy (OVX)-induced osteoporosis. Forty 12-week-old female C57/BL mice were randomly assigned to five groups (n = 8/group): OVX, PEMF, MEL, PEMF + MEL, and sham-operation (sham) groups. All mice in the first four groups were subjected to OVX. The mice in the PEMF and PEMF + MEL groups were exposed to PEMF (75 Hz, 1.6 mT, 1 h/day for 12 weeks), while those in the MEL and PEMF + MEL groups were administered MEL (50 mg/kg, i.p.). Body mass, micro-computed tomography, histology, immunohistochemistry, and real-time polymerase chain reaction were performed. PEMF + MEL treatment enhanced bone volume fraction (BV/TV) 2.2-fold over OVX control (P < 0.001) and increased expression levels of collagen type I (COL1) 1.9-fold and bone morphogenetic protein 2 (BMP2) 2.5-fold. PEMF + MEL also reduced the ratio of bone surface/bone volume (BS/BV) by 40% (P < 0.05) and appeared to reduce the number of osteoclasts in the metaphysis area. Preservation of bone value and bone microarchitecture in the combined therapy group were found to be superior to those in the single treatment groups. However, there were no apparent differences between the PEMF and MEL groups. The use of a combination of PEMF therapy and MEL supplementation may be an effective method to treat osteoporosis. © 2021 Bioelectromagnetics Society.

The Embryonic Chick Femur Organotypic Model as a Tool to Analyze the Angiotensin II Axis on Bone Tissue

Activation of renin–angiotensin system (RAS) plays a role in bone deterioration associated with bone metabolic disorders, via increased Angiotensin II (AngII) targeting Angiotensin II type 1 receptor/Angiotensin II type 2 receptor (AT1R/AT2R). Despite the wide data availability, the RAS role remains controversial. This study analyzes the feasibility of using the embryonic chick femur organotypic model to address AngII/AT1R/AT2R axis in bone, which is an application not yet considered. Embryonic day-11 femurs were cultured ex vivo for 11 days in three settings: basal conditions, exposure to AngII, and modulation of AngII effects by prior receptor blockade, i.e., AT1R, AT2R, and AT1R + AT2R. Tissue response was evaluated by combining µCT and histological analysis. Basal-cultured femurs expressed components of RAS, namely ACE, AT1R, AT2R, and MasR (qPCR analysis). Bone formation occurred in the diaphyseal region in all conditions. In basal-cultured femurs, AT1R blocking increased Bone Surface/Bone Volume (BS/BV), whereas Bone Volume/Tissue Volume (BV/TV) decreased with AT2R or AT1R + AT2R blockade. Exposure to AngII greatly decreased BV/TV compared to basal conditions. Receptor blockade prior to AngII addition prevented this effect, i.e., AT1R blockade induced BV/TV, whereas blocking AT2R caused lower BV/TV increase but greater BS/BV; AT1R + AT2R blockade also improved BV/TV. Concluding, the embryonic chick femur model was sensitive to three relevant RAS research setups, proving its usefulness to address AngII/AT1R/AT2R axis in bone both in basal and activated conditions.

Bovine lactoferrin inhibits alveolar bone destruction in an orthodontic rat model with periodontitis

ObjectiveWe aimed to evaluate the effect of bovine lactoferrin (bLF) on alveolar bone destruction and remodelling under orthodontic force (OF) in periodontitis-affected rats. Material and methodsAfter establishing the periodontitis-affected rat model with lipopolysaccharides (LPS), the left maxillary first molars were moved orthodontically under a force of 0.2N. Based on saline or bLF gavage, 54 Sprague-Dawley (SD) rats were randomized into 5 groups: A (blank), P1 (LPS+OF+bLF), P2 (LPS+OF+saline), C1 (OF+bLF), and C2 (OF+saline). Animals were evaluated using micro-computed tomography (micro-CT) followed by haematoxylin and eosin (H&E) and tartrate-resistant acid phosphatase (TRAP) staining, and the LF level was determined using ELISA in the gingival crevicular fluid (GCF) of the experimental teeth. Immunohistochemistry helped to detect expression changes in RANKL, OPG and COX-2. ResultsMicro-CT results indicated that compared with group P2, trabecular number (Tb.N) and trabecular thickness (Tb.Th) in group P1 were higher and bone surface/bone volume (BS/BV) was lower on day 14, while trabecular separation (Tb.Sp) decreased significantly on Day 5 and Day 14 after bLF gavage (P<0.05). This was supported by changes in H&E and TRAP staining. bLF down-regulated RANKL level at both timepoints and up-regulated OPG level on Day 14 in periodontitis rats (P<0.05). The significant changes mentioned above were not observed between group C1 and C2 (P>0.05). No significant change in COX-2 levels were observed in any group (P>0.05). The lactoferrin level in GCF increased significantly after bLF gavage (P<0.05). ConclusionBovine lactoferrin inhibited LPS-induced bone destruction, but the bone healing effect was independent of orthodontic aseptic inflammatory bone remodelling.

AEG-1 deletion promotes cartilage repair and modulates bone remodeling-related cytokines via TLR4/MyD88/NF-κB inhibition in ovariectomized rats with osteoporosis.

BackgroundOsteoporosis is a systemic skeletal disorder that can impact a variety of bones throughout the body. Astrocyte-elevated gene-1 (AEG-1) is involved in multiple pro-tumorigenic functions and participates in various inflammatory reactions. However, whether it has an impact on osteoporosis-related cartilage repair and bone remodeling remains unknown.MethodsWe utilized an ovariectomy mouse model with AEG-1 deletion to investigate the role of AEG-1 in osteoporosis. The mRNA level of AEG-1 was detected by RT-PCR, bone markers, bone volume/total volume (BV/TV), trabecular bone surface/bone volume (BSA/BV) and trabecular bone thickness (Tb. Th) were detected by micro computed tomography (µCT), bone injury was observed by HE and alcian blue staining. The contents of IL-6, IL-17, iNOS and IL-10 in peripheral blood of the three groups were detected by ELISA. The expression of OSX, coi1a1, OC, TLR4, MyD88 and NF-κB were detected by Western Blot.ResultsµCT revealed increased bone volume in the AEG-1 knockout (KO) ovariectomy (OVX) group compared to the wildtype (WT) OVX group 4 weeks after surgery, indicating restored bone formation after AEG-1 deletion. Flow sorting revealed that AEG-1 deletion inhibited the production of inflammatory factors. Western blot demonstrated activation of the TLR4/MyD88/NF-κB pathway after LPS exposure, which was reduced by AEG-1 deletion. AEG-1 deletion also improved lipopolysaccharide (LPS) induced adverse reactions.ConclusionsTaken together, these findings indicate that AEG-1 deletion improves cartilage repair and bone remodeling during osteoporosis, which may partly occur through the inhibition of the TLR4/MyD88/NF-κB signaling pathway.

Pulsed Electromagnetic Field Versus Whole Body Vibration on Cartilage and Subchondral Trabecular Bone in Mice With Knee Osteoarthritis.

Pulsed electromagnetic field (PEMF) and whole body vibration (WBV) interventions are expected to be important strategies for management of osteoarthritis (OA). The aim of the study was to investigate the comparative effectiveness of PEMF versus WBV on cartilage and subchondral trabecular bone in mice with knee OA (KOA) induced by surgical destabilization of the medial meniscus (DMM). Forty 12-week-old male C57/BL mice were randomly divided into four groups (n = 10): Control, OA, PEMF, and WBV. OA was induced (OA, PEMF, and WBV groups) by surgical DMM of right knee joint. Mice in PEMF group received 1 h/day PEMF exposure with 75 Hz, 1.6 mT for 4 weeks, and the WBV group was exposed to WBV for 20 min/day with 5 Hz, 4 mm, 0.3 g peak acceleration for 4 weeks. Micro-computed tomography (micro-CT), histology, and immunohistochemistry analyses were performed to evaluate the changes in cartilage and microstructure of trabecular bone. The bone volume fraction (BV/TV), trabecular thickness (Tb.Th), and trabecular number (Tb.N) increased, and bone surface/bone volume (BS/BV) decreased by micro-CT analysis in PEMF and WBV groups. The Osteoarthritis Research Society International (OARSI) scores in PEMF and WBV groups were significantly lower than in the OA group. Immunohistochemical results showed that PEMF and WBV promoted expressions of Aggrecan, and inhibited expressions of IL-1β, ADAMTS4, and MMP13. Superior results are seen in PEMF group compared with WBV group. Both PEMF and WBV were effective, could delay cartilage degeneration and preserve subchondral trabecular bone microarchitecture, and PEMF was found to be superior to WBV. Bioelectromagnetics. 2020;41:298-307 © 2020 Bioelectromagnetics Society.

Elevated Serum 25-Hydroxyvitamin D Levels Are Associated with Improved Bone Formation and Micro-Structural Measures in Elderly Hip Fracture Patients.

Vitamin D, along with calcium, is generally considered necessary for bone health and reduction of fractures. However, he effects of improving vitamin D status have not always been observed to improve bone mineral density (BMD). We have investigated whether varying vitamin D status in humans, as measured by serum 25(OH)D levels, relate to micro-structural and histomorphetric measures of bone quality and quantity, rather than density. Intertrochanteric trabecular bone biopsies and serum samples were collected from patients undergoing hip arthroplasty (65 females, 38 males, mean age 84.8 ± 8.3 years) at Royal Adelaide Hospital. Estimated GFR, serum ionized calcium, alkaline phosphatase, albumin, supplement and medication intake prior to surgery were taken from patient case records. Serum 25(OH)D, 1,25(OH)2D, and parathyroid hormone (PTH) levels were measured by immunoassays. Trabecular bone structural indices were determined by high-resolution micro-CT. Mean wall thickness (MWT) was measured on toluidine blue-stained histological sections. Bone mRNA levels for vitamin D metabolising enzymes CYP27B1 and CYP24A1 were measured by qRT-PCR. While serum 25(OH)D levels did not associate with bone volume/tissue volume (BV/TV%), serum 25(OH)D levels were strongly and independently associated with MWT (r = 0.81 p < 0.0001) with values significantly greater in patients with higher serum 25(OH)D levels. Furthermore, serum 25(OH)D levels were negatively associated with Bone Surface/Bone Volume (BS/BV) (r = −0.206, p < 0.05) and together with bone CYP27B1 and CYP24A1 mRNA accounted for 10% of the variability of BS/BV (p = 0.001). These data demonstrate that serum 25(OH)D is an independent positive predictor of micro-structural and bone formation measures and may be dependent, in part, on its metabolism within the bone.

A novel device to prevent osteoporosis by promoting bone metabolism using a newly developed double-loading stimulation with vibration and shaking.

In Japan, 13 million people have osteoporosis, including approximately 9 hundred thousand people who are bedridden owing to bone fractures from falls. Preventing osteoporosis is considered to be an important and effective way of preventing fall-related fractures. Thus, we developed a novel method of locomotor stimulation and analyzed its effectiveness in mice. Specifically, we created a double-loading device that combines vibration and shaking stimulation. The device was used to continuously stimulate ovariectomy-induced decreased bone density mouse models 30 minutes daily for 10 weeks. We then collected femur samples, created undecalcified tissue slices, calculated parameters using bone histomorphomtry, and conducted comparative testing. BS/TV (bone surface/tissue volume), N.Oc/ES (osteoclast number/eroded surface), Oc.S/ES (osteoclast osteoid surface/eroded surface), Omt (osteoid maturation time), Tb.N (trabecular number), Mlt (mineralization lag time) < (p < 0.01), N.Ob (osteoblast number), N.Ob/TV (osteoblast number/tissue volume), sLS (single labeled suface), N.Mu.Oc/ES (multinucle osteoclast number/eroded surface), and N.Mo.Oc/ES (mononucle osteoclast number/eroded surface) (p < 0.05) were significantly higher in the stimulation group than in the non-stimulation group. In addition, BS/BV (bone surface/bone volume), Tb.Sp (trabecular separation), MAR (mineral apposition rate), Aj.Ar (adjusted apposition rate) (p < 0.01), ES (eroded surface ), ES/BS (eroded surface/bone surface), and BRs.R (bone resorption rate) (p < 0.05) were significantly lower in the stimulation group than in the non-stimulation group. These results suggest that stimulation activated osteoblasts and osteoclasts, thereby leading to highly active bone remodeling. We anticipate that bone mineralization will subsequently occur, suggesting that this stimulation technique is effective in preventing osteoporosis by alleviating sudden bone density loss.

Pulsed electromagnetic field improves subchondral bone microstructure in knee osteoarthritis rats through a Wnt/β-catenin signaling-associated mechanism.

Pulsed electromagnetic field (PEMF) is often used for management of osteoarthritis (OA). The aim of the study was to determine whether PEMF can successfully improve subchondral bone microstructure through a Wnt/β-catenin signaling-associated pathway in rats with knee OA induced by low-dose monosodium iodoacetate (MIA). Seventy-two 12-week-old male Sprague-Dawley rats were randomly assigned to three groups: OA (n = 24), PEMF (n = 24), and Control (n = 24). OA was induced (OA and PEMF groups) by injecting 0.2 mg MIA in rats' right knee joint. The control rats received a single sterile saline injection in the right knee. Rats in the PEMF group were exposed to daily 2 h PEMF exposure with 75 Hz, 1.6 mT for 4 weeks. After 4 weeks, micro-computed tomography (micro-CT), real-time PCR, and immunohistochemistry staining were performed. The PEMF group increased bone volume fraction (BV/TV), trabecular thickness (Tb.Th), trabecular number (Tb.N), and suppressed bone surface/bone volume (BS/BV) and trabecular separation (Tb.Sp) levels in micro-CT analysis. Real-time PCR analysis showed that PEMF promoted tibial subchondral bone's gene expressions of Wnt3a, β-catenin, and OPG, but did not alter LRP5 and RANKL mRNA levels. Similar results involved tibial subchondral bone's protein expressions that were observed in immunohistochemistry staining. These results suggest that PEMF preserved the structural integrity of subchondral bone in knee OA rats by promoting the activation of Wnt/β-catenin signaling and OPG/RANKL/RANK signaling. Bioelectromagnetics. 39:89-97, 2018. © 2017 Wiley Periodicals, Inc.

Domestic artificial cervical disc interface pressure distribution and effect of bone-implant interface pressure on osseointegration

To analyze the distribution of stress in the upper and lower plates of the prosthesis-bone interface, and the effect of interface pressure on osseointegration. CT scanning was performed on goats at 1 week after artificial cervical disc replacement to establish the finite element model of C 3, 4. The stress distribution of the upper and lower plates of the interface was observed. At 6 and 12 months after replacement, Micro-CT scan and three dimensional reconstruction were performed to measure the bone volume fraction (BVF), trabecular number (Tb. N), trabecular thickness (Tb. Th), trabecular separation (Tb. Sp), bone mineral density (BMD), bone surface/bone volume (BS/BV), and trabecular pattern factor (Tb. Pf). The C 3 lower plate and C 4 upper plate of 4 normal goat were chosen to made the cylinder of the diameter of 2 mm. The gene expressions of receptor activator for nuclear factor κB ligand (RANKL), osteoprotegerin (OPG), transforming growth factor β (TGF-β), and macrophage colony-stimulating factor (M-CSF) were detected by real time fluorescent quantitative PCR at immediate after cutting and at 24 and 48 hours after culture. The samples of appropriate culture time were selected to made mechanical loading, and the gene expressions of RANKL, OPG, M-CSF, and TGF-β were detected by real time fluorescent quantitative PCR; no mechanical loading samples were used as normal controls. Under 25 N axial loading, the stress of the upper plate of C 3, 4 was concentrated to post median region, and the stress of the lower plate to middle-front region and two orbits. According to stress, the plate was divided into 5 regions. The Micro-CT scan showed that BMD, Tb.Th, BVF, and Tb.N significantly increased, and BS/BV, Tb.Sp, and Tb.Pf significantly decreased at 12 months after replacement when compared with ones at 6 months ( P<0.05). At 24 and 48 hours after culture, the gene expressions of RANKL, OPG, and TGF-β were signifi-cantly higher than those at immediate ( P<0.05), but no significant difference was found between at 24 and 48 hours after culture ( P>0.05). The mechanical loading test results at 24 hours after culture showed that the RANKL and OPG gene expressions and OPG/RANKL ratio in C 3 lower plate and C 4 upper plate were significantly up-regulated when compared with controls ( P<0.05), but no significant difference was shown in TGF-β and M-CSF gene expressions ( P>0.05). Domestic artificial cervical disc endplate has different pressure distribution, the stress of lower plate is higher than that of upper plate. Pressure has important effect on local osseointegration; the higher pressure area is, the osseointegration is better. Under the maximum pressure in interface, the osteoblast proliferation will increase, which is advantageous to the local osseointegration.

Full supervised learning for osteoporosis diagnosis using micro‐CT images

Early osteoporosis diagnosis is of important significance for reducing fracture risk. Image analysis provides a new perspective for noninvasive diagnosis in recent years. In this article, we propose a novel method based on machine-learning method performed on micro-CT images to diagnose osteoporosis. The aim of this work is to find a way to more effectively and accurately diagnose osteoporosis on which many methods have been proposed and practiced. In this method, in contrast to the previously proposed methods in which features are analyzed individually, several features are combined to build a classifier for distinguishing osteoporosis group and normal group. Twelve features consisting of two groups are involved in our research, including bone volume/total volume (BV/TV), bone surface/bone volume (BS/BV), trabecular number (Tb.N), obtained from the software of micro-CT, and other four features from volumetric topological analysis (VTA). Support vector machine (SVM) method and k-nearest neighbor (kNN) method are introduced to create classifiers with these features due to their excellent performances on classification. In the experiment, 200 micro-CT images are used in which half are from osteoporosis patients and the rest are from normal people. The performance of the obtained classifiers is evaluated by precision, recall, and F-measure. The best performance with precision of 100%, recall of 100%, and F-measure of 100% is acquired when all the features are included. The satisfying result demonstrates that SVM and kNN are effective for diagnosing osteoporosis with micro-CT images.

The trabecular heterogeneity of femoral head in male osteoporotic fracture

Objective The aim of this study was to investigate the femoral head trabecular heterogeneity in Chinese male patients with osteoporotic fracture and their effects on osteoporotie fracture.Methods Human femoral heads were obtained from 11 male osteoporotie fracture (OP) patients ranged from 51 to 82 years old [average age (65±9 ) years old], and 7 male trauma ( TM ) patients ranged from 46 to 75 years old [average age (61±11 ) years old] who underwent total hip arthroplasty within two hours after either osteoporotic or trauma hip fracture.The OP was defined as having a fragility fracture.After laying femoral head as living body position and locating mark, nine trabecular specimens were obtained from femoral heads, each of 6 mm × 6 mm× 7 mm.The cortical shell was not included in each specimen.One cube was selected as the primary compressive trabecular region and the other 8 specimens as non-primary compressive trabecular region.These cubes were scanned using high-resolution microcomputed tomography scanner (μCT).After scanning, the data of total cubes, primary compressive trabecular region and noncompressive trabecular region were used for analysis by t test.Results In OP group volumetric bone mineral deosity(vBMD) [( 182.15±66.00) mg/mm3 vs (223.97±70.92) mg/mm3, t =3.041], tissue bone mineral density (tBMD) [(538.76±64.72) mg/mm3 vs (580.01±63.86 ) mg/mm3, t = 3.160],bone volume fraction (TV/BV) [(0.22 ± 0.06) % vs (0.26 ± 0.07 ) %, t = 2.821], trabecular thickness (Tb.Th.) [( 161.07 ±42.75 ) μm vs ( 205.47 ± 74.44 ) μm, t = 3.233] were significantly decreased while bone surface/bone volume ( BS/BV ) [( 13.75 ± 2.55 ) mm-1 vs ( 12.28 ± 2.70 ) mm-1, t =-2.777] was significantly increased in the non-primary compressive trabecular region than that in the primary compressive trabecular region ( P < 0.05 ).vBMD [( 182.15 ± 66.00) mg/mm3 vs ( 248.05 ±105.48) mg/mm3, t = - 3.598], tBMD [(538.76 ± 64.72) mg/mm3 vs ( 570.54 ± 100.32) mg/mm3,t=-2.108],TV/BV [(0.22±0.06) % vs (0.28±0.12) %, t= -3.466], Tb.Th.[(161.07±42.75) μm vs (200.31 ±96.63) μm, t= -2.866], trabecular number (Tb.N.)[(1.46±0.23)/mm3 vs ( 1.57 ± 0.29)/mm3, t = - 2.396] were significantly decreased while trabecular separation ( Tb.Sp.) [(780.82 ± 144.85 )μm vs ( 653.09 ± 119.64) μm, t = 5.470], degree of anisotropy (DA) ( 1.57±0.20 vs 1.47±0.18, t = 2.930 ) were significantly increased in OP than in TM in the non-compressive trabecular region( P < 0.05 ).No significant differents were found between OP and TM for any of the parameters measured in the primary compressive trabecular region.Tb.Th.[(199.37±68.22)μm vs (176.33 ±71.21 )μm, t = 2.060,P < 0.05] were significantly increased in the primary compressive trabecular region than that in the non-primary compressive trabecular region and no significant differences were found in the other parameters in the all 18 specimens.Conclusions The femoral head trabeculae had a heterogenic distribution in OP.Bone loss in OP primarily takes place in non-compressive trabecular region.Femoral neck fracture cannot be prevented though the bone microstructure do not loss in the primary compressive trabecular region.Tb.Th.in the femoral head could be an interesting parameter which is closely related to the femoral neck fracture. Key words: Osteoporosis; Femoral head; Tomography,X-ray computed

Effect of rosiglitazone on bone microarchitecture of bilaterally ovariectomized rats

Objective:To study the effect of rosiglitazone on the bone microarchitecture of bilaterally ovariectomized(OVX) rats.Methods: Twenty-one 24-week-old skeletally mature rats were bilaterally ovariectomized,and were randomly divided into three groups(n=7):Control group(C group),Rosiglitazone A group(RA group),and Rosiglitazone B group(RB group).Rats in RA group were given i.g 0.35 mg/d rosiglitazone for four weeks and those in the RB group were given i.g 0.70 mg/d rosiglitazone.Rats in the control group were given normal saline in the same manner.The rats were sacrificed four weeks later.The bone volume/tissue volume(BV/TV),the trabecular number(Tb.N),the trabecular thickness(Tb.Th),the trabecular separation(Th.Sp),and the bone surface/bone volume(BS/BV)of the femur,tibia,and lumbar were determined by micro-computered tomography(micro-CT).Results: Compared with the control group,the Tb.N,Tb.Th,and BS/BV of the femur,tibia,and lumbar all decreased significantly in RA group(except for the BS/BV of tibia)(P0.05,P0.01);the Tb.SP of tibia and lumbar in the RA group were significantly increased(P0.05);and there were no differences in BV/TV between the two groups.Compared with control group,the Tb.N,Tb.Th,and BS/BV of the femur,tibia,and lumbar were decreased significantly in RB group(P0.05,P0.01);the Tb.Sp of tibia and lumbar increased significantly(P0.05);and the BV/TV had no differences between the two groups.There were no significant differences between the RA group and RB group concerning all the parameters.Conclusion: Rosiglitazone can cause injury to the bone microarchitecture,especially to the cancellous part of the bone.

Trabecular bone micro-architecture and bone mineral density in adolescent idiopathic and congenital scoliosis

Objective: To investigate the microstructure of trabecular bone in adolescent idiopathic scoliosis (AIS) and age-matched congenital scoliosis (CS), and to evaluate the bone mineral status of CS patients compared with normal controls and AIS patients. Methods: This study included 15 AIS and 16 CS female patients and 35 healthy female adolescents. Corrective surgery was indicated for the AIS and CS patients, from whom iliac crest biopsies were collected during autograft harvesting, and scanned by micro-computer tomography. Bone mineral status was assessed at the lumbar and hip areas in every patient by dual energy X-ray absorptiometry (DEXA). Results: Significantly lower lumbar and femoral neck bone mineral density (BMD) was found in AIS patient compared with normal controls. All BMD and bone mineral content (BMC) parameters were significantly lower in CS patients compared with age-matched normal controls. Under DEXA assessment significant associations between bone volume/tissue volume (BV/TV) and BMD values were observed. In the 3D model, BV/TV was significantly higher in AIS (19.9% ± 3.4%) than in CS (13.3% ± 3.0%, P < 0.05). Significant differences between AIS and CS were also found in trabecular thickness (Tb.Th) and bone surface/bone volume (BS/BV) (155.5 ± 54.9 µm vs. 108.1 ± 17.4 µm and 16.4% ± 3.3% vs. 22.0% ± 3.4% respectively, P < 0.05 in both). Conclusion: Lower bone mineral status and weak trabecular bone structure observed in AIS and CS justify further investigation of the bone mineral status in scoliosis of various etiologies.

Trabecular bone micro‐architecture and bone mineral density in adolescent idiopathic and congenital scoliosis

To investigate the microstructure of trabecular bone in adolescent idiopathic scoliosis (AIS) and age-matched congenital scoliosis (CS), and to evaluate the bone mineral status of CS patients compared with normal controls and AIS patients. This study included 15 AIS and 16 CS female patients and 35 healthy female adolescents. Corrective surgery was indicated for the AIS and CS patients, from whom iliac crest biopsies were collected during autograft harvesting, and scanned by micro-computer tomography. Bone mineral status was assessed at the lumbar and hip areas in every patient by dual energy X-ray absorptiometry (DEXA). Significantly lower lumbar and femoral neck bone mineral density (BMD) was found in AIS patient compared with normal controls. All BMD and bone mineral content (BMC) parameters were significantly lower in CS patients compared with age-matched normal controls. Under DEXA assessment significant associations between bone volume/tissue volume (BV/TV) and BMD values were observed. In the 3D model, BV/TV was significantly higher in AIS (19.9% ± 3.4%) than in CS (13.3% ± 3.0%, P < 0.05). Significant differences between AIS and CS were also found in trabecular thickness (Tb.Th) and bone surface/bone volume (BS/BV) (155.5 ± 54.9 µm vs. 108.1 ± 17.4 µm and 16.4% ± 3.3% vs. 22.0% ± 3.4% respectively, P < 0.05 in both). Lower bone mineral status and weak trabecular bone structure observed in AIS and CS justify further investigation of the bone mineral status in scoliosis of various etiologies.

Micro-CT evaluation and histological analysis of screw-bone interface of expansive pedicle screw in osteoporotic sheep

To investigate the properties of screw-bone interface of expansive pedicle screw (EPS) in osteoporotic sheep by micro-CT and histological observation. Six female sheep with bilateral ovariectomy-induced osteoporosis were employed in this experiment. After EPS insertion in each femoral condyle, the sheep were randomly divided into two groups: 3 sheep were bred for 3 months (Group A), while the other 3 were bred for 6 months (Group B). After the animals being killed, the femoral condyles with EPS were obtained, which were three-dimensionally-imaged and reconstructed by micro-CT. Histological evaluation was made thereafter. The trabecular microstructure was denser at the screw-bone interface than in the distant parts in expansive section, especially within the spiral marking. In the non-expansive section, however, there was no significant difference between the interface and the distant parts. The regions of interest (ROI) adjacent to EPS were reconstructed and analyzed by micro-CT with the same thresholds. The three-dimensional (3-D) parameters, including tissue mineral density (TMD), bone volume fraction (BVF, BV/TV), bone surface/bone volume (BS/BV) ratio, trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp), were significantly better in expansive sections than non-expansive sections (P less than 0.05). Histologically, newly-formed bony trabeculae crawled along the expansive fissures and into the center of EPS. The newly-formed bones, as well as the bones at the bone-screw interface, closely contacted with the EPS and constructed four compartments. The findings of the current study, based on micro-CT and histological evaluation, suggest that EPS can significantly provide stabilization in osteoporotic cancellous bones.