Wnt5a deficiency in osteocalcin-expressing cells could not alleviate the osteoarthritic phenotype in a mouse model of post-traumatic osteoarthritis.

Objectives: Wnt5a, which regulates the activities of osteoblasts and osteoclasts, is reportedly overexpressed in osteoarthritis (OA) tissues. The purpose of this study was to elucidate its role in the development of OA by deleting Wnt5a in osteocalcin (OCN)-expressing cells. Materials and Methods: Knee OA was induced by anterior cruciate ligament transection (ACLT) in OCN-Cre;Wnt5afl/fl knockout (Wnt5a-cKO) mice and control littermates. Eight weeks after surgery, histological changes, cell apoptosis, and matrix metabolism of cartilage were evaluated by toluidine blue, TUNEL staining, and im-immunohistochemistry analyses, respectively. In addition, the subchondral bone microarchitecture of mice was examined by micro-computed tomography (micro-CT). Results: Histological scores show substantial cartilage degeneration occurred in ACLT knees, coupled with decreased collagen type II expression and enhanced matrix metalloproteinase 13 expression, as well as higher proportions of apoptotic cells. Micro-CT results show that ACLT resulted in decreased bone mineral density, bone volume/trabecular volume, trabecular number, and structure model index of subchondral bones in both Wnt5a-cKO and control littermates; although Wnt5a-cKO mice display lower BMD and BV/TV values, no significant difference was observed between Wnt5a-cKO and control mice for any of these values. Conclusion: Our findings indicate that Wnt5a deficiency in OCN-expressing cells could not prevent an osteoarthritic phenotype in a mouse model of post-traumatic OA.

[Effects of fine particulate matter on bone mass and osteogenic differentiation of bone marrow mesenchymal stem cells in mice].

OBJECTIVE: To observe the effects of exposure to fine particulate matter(PM_(2.5)) on bone mass, microstructure, biomechanical properties, and osteogenic differentiation ability of bone marrow mesenchymal stem cells(BMSCs) in mice. METHODS: A total of 16 C57BL/6J mice aged 8 weeks were randomly divided into control group(NS group) and PM_(2.5) exposure group(PM group). NS group was given normal saline, PM group was given 14 mg/kg PM_(2.5) suspension, 50 µL, poisoning every 3 day. After 10 weeks, the lungs of mice were taken for HE staining, and the left tibia was taken for Micro CT detection to analyze parameters related to cancellous and cortical bone. The right tibia was taken for HE staining to observe changes in bone trabeculae. Immunohistochemical staining was used to detect type I collagen(Col I), osteoprotegerin(OPG), and nuclear factor-κB receptor activating factor ligand(RANKL) protein expression, tartrate resistant acid phosphatase(TRAP) staining for detection of osteoclasts. Extract primary BMSCs from bilateral femurs, induce osteogenesis, and then perform alkaline phosphatase(ALP) staining to detect ALP activity, alizarin red staining to detect bone mineralization ability, real-time PCR to detect osteocalcin(OCN), ALP, OPG, and RANKL mRNA expression, and biomechanical testing to test the mechanical properties of the femur. RESULTS: Compared with the NS group, the pulmonary alveolar structure of the PM group mice was disrupted and a large number of inflammatory cells gathered. Prompt for successful PM_(2.5) poisoning operation. Micro CT result showed that the bone mineral density(BMD) and bone volume fraction(BV/TV) of the PM group mice were 276.959±15.152 mg/cm~3 and 0.208%±0.009%, respectively. The NS group had 316.709±28.205 mg/cm~3 and 0.236%±0.019%, respectively. The PM group was lower than the NS group(P<0.05), but the trabecular number(Tb. N) There was no statistically significant difference in parameters such as trabecular thickness(Tb. Th) and trabecular separation(Tb. SP)(P>0.05). The HE staining result of the tibia showed that the trabeculae in the NS group were thick, dense, and uniform. The bone trabeculae in the PM group were slender, with a decrease in number, widened spacing, and sparse arrangement. The expression of Col I(0.023±0.009) and OPG(0.036±0.010) in the PM group increased compared to the NS group(0.079±0.007, 0.059±0.012), while the expression of RANKL(0.036±0.006) decreased compared to the NS group(0.022±0.002)(P<0.05); The number of TRAP positive particles increased in the PM group. The experimental result after osteoinduction of BMSCs in mice showed that compared with the NS group, the PM group had a decrease in the number of ALP positive cells and a decrease in the number of calcium nodules. The relative expression of ALP, OCN, and OPG mRNA in the PM group(0.375±0.021, 0.585±0.088, 0.768±0.112) was significantly reduced compared to the NS group(1.001±0.043, 1.006±0.132, 1.002±0.086), while the relative expression of RANKL mRNA(1.278±0.118) was increased compared to the NS group(1.001±0.057)(P<0.05). The biomechanical experimental result showed that the maximum deflection of the NS group was 0.337±0.031 mm, while the maximum deflection of the PM group was 0.258±0.041 mm. Compared with the NS group, the maximum deflection of the PM group decreased significantly(P<0.05), and the maximum stress and maximum load showed a decreasing trend, but the difference was not statistically significant(P>0.05). CONCLUSION: After 10 weeks of exposure to PM_(2.5), it can affect the bone health of mice, and its mechanism may be related to increased osteoclast activity and inhibition of the osteogenic differentiation ability of BMSCs.

Strontium ranelate retards disc degradation and improves endplate and bone micro-architecture in ovariectomized rats with lumbar fusion induced - Adjacent segment disc degeneration.

Objectives: Adjacent segment disc degeneration (ASDD) is one of the long-term sequelae of spinal fusion, which is more susceptible with osteoporosis. As an anti-osteoporosis drug, strontium ranelate (SR) has been reported to not only regulate bone metabolism but also cartilage matrix formation. However, it is not yet clear whether SR has a reversal or delaying effect on fusion-induced ASDD in a model of osteoporosis. Materials and methods: Fifth three-month-old female Sprague-Dawley rats that underwent L4-L5 posterolateral lumbar fusion (PLF) with spinous-process wire fixation 4 weeks after bilateral ovariectomy (OVX) surgery. Animals were administered vehicle (V) or SR (900 mg/kg/d) orally for 12 weeks post-PLF as follows: Sham+V, OVX + V, PLF + V, OVX + PLF + V, and OVX + PLF + SR. Manual palpation and X-ray were used to evaluate the state of lumbar fusion. Adjacent-segment disc was assessed by histological (VG staining and Scoring), histomorphometry (Disc Height, MVD, Calcification rate and Vascular Bud rate), immunohistochemical (Col-II, Aggrecan, MMP-13, ADAMTS-4 and Caspase-3), and mRNA analysis (Col-I, Col-II, Aggrecan, MMP-13 and ADAMTS-4). Adjacent L6 vertebrae microstructures were evaluated by microcomputed tomography. Results: Manual palpation and radiographs showed clear evidence of the fused segment's immobility. After 12 weeks of PLF surgery, a fusion-induced ASDD model was established. Low bone mass caused by ovariectomy can significantly exacerbate ASDD progression. SR exerted a protective effect on adjacent segment intervertebral disc with the underlying mechanism possibly being associated with preserving bone mass to prevent spinal instability, maintaining the functional integrity of endplate vascular microstructure, and regulating matrix metabolism in the nucleus pulposus and annulus fibrosus. Discussion: Anti-osteoporosis medication SR treatments not only maintain bone mass and prevent fractures, but early intervention could also potentially delay degenerative conditions linked to osteoporosis. Taken together, our results suggested that SR might be a promising approach for the intervention of fusion-induced ASDD with osteoporosis.

Estrogen deficiency impedes fracture healing despite eliminating the excessive absorption of the posterior callus in a semi-fixed distal tibial fracture mouse model.

BACKGROUND: Treatment of distal tibial fractures is a challenge due to their specific anatomical location. However, there is no appropriate mouse model to simulate a clinical distal tibial fracture for basic research. The aim of this investigation was to evaluate the feasibility of simulating a clinical fracture of the distal tibia of mice and to investigate the effect of ovariectomy (OVX)-induced osteoporosis on fracture healing in this model. METHODS: Sixty female 8-week-old C57BL/6 mice were randomly divided into two groups, either sham or OVX. A semi-fixation distal tibia fracture was established in the right tibia after 8 weeks of OVX. The right tibias were collected at 7, 14, 21, and 28 days post fracture. RESULTS: In the semi-fixation distal tibia fracture model, the posterior callus in the sham group showed excessive bone resorption and lower bone mass phenotype compared with the anterior site; a similar trend was not found in the OVX group. At 28 days post fracture, the posterior callus was more mineralized than the anterior callus in the OVX group. Although the fracture healing of the sham group showed a special phenotype in this mode, the progress and quality of fracture healing were still better than those of the OVX group. CONCLUSION: A semi-fixed distal tibial closed fracture mouse model was successfully established. In this model, excess bone resorption of the posterior callus impaired normal fracture healing, but not in OVX-induced osteoporotic bone. Although the stress shielding effect was not observed in the OVX group, impaired bone healing caused by OVX was still present. Our results suggest that this fracture model may have potential for studies on distal tibial fractures and stress shielding.

Impacts and mechanisms of PM2.5 on bone.

Osteoporosis is a metabolic bone disease, which is characterized by a decreased bone mass and deterioration of bone microstructure, resulting in increased bone fragility and a higher risk of fracture. The main pathological process of osteoporosis is the dynamic imbalance between bone absorption and bone formation, which can be caused by various factors such as air pollution. Particulate matter (PM)2.5 refers to the fine particles in the atmosphere, which are small in volume and large in specific surface area. These particles are prone to carrying toxic substances and have negative effects on several extrapulmonary organs, including bones. In this review, we present relevant data from studies, which show that PM2.5 is associated with abnormal bone turnover and osteoporosis. PM2.5 may cause or aggravate bone loss by stimulating an inflammatory response, inducing oxidative damage, reducing estrogen efficiency by competitive binding to estrogen receptors, or endocrine disorder mediated by binding with aromatic hydrocarbon receptors, and affecting the synthesis of vitamin D to reduce calcium absorption. The cellular and molecular mechanisms involved in these processes are also summarized in this review.

Modification of low-salt myofibrillar protein using combined ultrasound pre-treatment and konjac glucomannan for improving gelling properties: Intermolecular interaction and filling effect.

The quality deterioration of low-salt meat products has been gained ongoing focus of researchers. In this study, konjac glucomannan (KGM) was used to alleviate the finiteness of ultrasound treatment on the quality improvement of low-salt myofibrillar protein (MP), and the modification sequence was also investigated. The results revealed that the single and double sequential modification by utilizing KGM and ultrasound significantly influenced the gelation behavior of low-salt MPs. The uniform MP-KGM mixture formed by a single ultrasound treatment had limited protein unfolding, resulting in relatively weak intermolecular forces in the composite gel. Importantly, ultrasound pre-treatment combined with KGM modification promoted the unfolding and moderate thermal aggregation of proteins and remarkably improved the rheological behaviors and gel strength of the composite gel. This result could also be corroborated by the highest percentage of trans-gauche-trans conformation of SS bridges and maximum ß-sheet proportion. Furthermore, molecular dynamic simulation and molecular docking elucidated that the hydrogen bond length between protein and KGM was shortened after ultrasound pre-treatment, which was the molecular basis for the enhanced intermolecular interactions. Therefore, ultrasound pre-treatment combined with KGM can effectively improve the gelling properties of low-salt MPs, providing a practical method for the processing of low-salt meat products.

Liraglutide in Combination with Insulin Has a Superior Therapeutic Effect to Either Alone on Fracture Healing in Diabetic Rats.

Purpose: Fractures in patients with type 2 diabetes mellitus are at a high risk of delayed union or non-union. Previous studies have shown a protective effect of liraglutide on bone. In the present study, we aimed to investigate the effects of a combination of liraglutide and insulin on fracture healing in a rat model of diabetes and the mechanisms involved. Materials and Methods: Closed femoral mid-shaft fractures were established in male Sprague-Dawley rats with or without diabetes mellitus, and the diabetic rats were administered insulin and/or liraglutide. Six weeks after femoral fracture, the femoral callus was evaluated by immunohistochemistry, histology, and micro-computed tomography. Additionally, the effects of liraglutide on high-glucose-stimulated MC3T3-E1 cells were analyzed by Western blotting. Results: Micro-computed tomography and safranin O/fast green staining showed that fracture healing was delayed in the diabetic rats, and this was accompanied by much lower expression of osteogenic markers and greater osteoclast activity. However, treatment with insulin and/or liraglutide prevented these changes. Liraglutide in combination with insulin treatment resulted in lower blood glucose concentrations and significantly higher osteocalcin (OCN) and collagen I (Col I) expression six weeks following fracture. Western blot analysis showed that liraglutide prevented the low expression of the bone morphogenetic protein-2, osterix/SP7, OCN, Col I, and ß-catenin in high-glucose-stimulated MC3T3-E1 cells. Conclusion: These results demonstrate that insulin and/or liraglutide promotes bone fracture healing in the DF model. The combination was more effective than either single treatment, which may be because of the two drugs' additive effects on the osteogenic ability of osteoblast precursors.

Fracture Mode Transition during Assembly of TC4 High-Lock Bolt under Tensile Load: A Combined Experimental Study and Finite Element Analysis.

Fracture during the assembly process is an important failure mode for high-lock bolts used in the aviation industry, which greatly increases the potential of unpredictable accidents during service. In the current study, the underlying reasons for fracture during the assembly of a TC4 high-lock bolt was investigated using a tensile test and finite element analysis (FEA). The microstructure of the as-received bolt consisted of a high proportion of α phase, some ß phase, and a small amount of α' phase formed via martensite phase transformation during the rammer process. The experimental force-displacement curves revealed an average yield load of 55.9 kN and a breaking load of 67.65 kN. The corresponding yield strength was calculated to be 0.9 GPa, which was smaller than the standard value of TC4. This was attributed to the preload-induced stress concentration on the thread surface, leading to obvious strain hardening, which can lead to crack initiation. The effect of preload was further confirmed by the fractographies in which the initial crack was observed on the thread surface. The fractographies suggested that hybrid fracture occurred on the tensile loaded bolt. The initial failure was brittle fracture on the thread surface, transforming into ductile fracture in the screw. The results can contribute to understanding the effect of preload on the load carry capacity of high-lock bolts and provide a strategy to design its assembly specification.

Review of the Effect of Surface Coating Modification on Magnesium Alloy Biocompatibility.

Magnesium alloy, as an absorbable and implantable biomaterial, has been greatly developed in the application field of biomaterials in recent years due to its excellent biocompatibility and biomechanics. However, due to the poor corrosion resistance of magnesium alloy in the physiological environment, the degradation rate will be unbalanced, which seriously affects the clinical use. There are two main ways to improve the corrosion resistance of magnesium alloy: one is by adding alloying elements, the other is by surface modification technology. Compared with adding alloy elements, the surface coating modification has the following advantages: (1) The surface coating modification is carried out without changing the matrix elements of magnesium alloy, avoiding the introduction of other elements; (2) The corrosion resistance of magnesium alloy can be improved by relatively simple physical, chemical, or electrochemical improvement. From the perspective of corrosion resistance and biocompatibility of biomedical magnesium alloy materials, this paper summarizes the application and characteristics of six different surface coating modifications in the biomedical magnesium alloy field, including chemical conversion method, micro-arc oxidation method, sol-gel method, electrophoretic deposition, hydrothermal method, and thermal spraying method. In the last section, it looks forward to the development prospect of surface coating modification and points out that preparing modified coatings on the implant surface combined with various modification post-treatment technologies is the main direction to improve biocompatibility and realize clinical functionalization.

A Distributed Multi-Agent Formation Control Method Based on Deep Q Learning.

Distributed control method plays an important role in the formation of a multi-agent system (MAS), which is the prerequisite for an MAS to complete its missions. However, the lack of considering the collision risk between agents makes many distributed formation control methods lose practicability. In this article, a distributed formation control method that takes collision avoidance into account is proposed. At first, the MAS formation control problem can be divided into pair-wise unit formation problems where each agent moves to the expected position and only needs to avoid one obstacle. Then, a deep Q network (DQN) is applied to model the agent's unit controller for this pair-wise unit formation. The DQN controller is trained by using reshaped reward function and prioritized experience replay. The agents in MAS formation share the same unit DQN controller but get different commands due to various observations. Finally, through the min-max fusion of value functions of the DQN controller, the agent can always respond to the most dangerous avoidance. In this way, we get an easy-to-train multi-agent collision avoidance formation control method. In the end, unit formation simulation and multi-agent formation simulation results are presented to verify our method.

The combined anti-osteoporotic effects of simvastatin and exercise in ovariectomized mice fed a high-fat diet.

BACKGROUND: To evaluate and compare the effects of the combined intervention of simvastatin and exercise on the bone degeneration in a mice model of osteoporosis (OP) induced by obesity and estrogen deficiency. METHODS: 56 female 3-month-old C57BL/6 mice were given a standard diet or a high-fat diet after ovariectomy (OVX) or sham surgery. Drug administration and exercise training were initiated 72 h after surgical operation, which were treated with simvastatin (10 mg/kg/day) or exercise (15 m/min for 30 min/day) or combined with simvastatin and exercise at 72 h for 8 weeks. The pathology of OP was assessed by histomorphology analyses, immunohistochemistry (IHC), micro-computed tomography (Micro-CT), enzyme-linked immunosorbent assay (ELISA) and cell culture. RESULTS: The coexistence of obesity and estrogen deficiency significantly further exacerbated OP pathology, and combined intervention showed a better significant anti-osteoporosis effect than monotherapy. In details, simvastatin combined with exercise ameliorated the abnormal bone mass, microstructure and bone marrow adipocyte differentiation, significantly increased osteoprotegerin (OPG), type 1 collagen (Col-I), RUNX2 and osteocalcin (OCN) expression, decreased the expression of receptor activator of nuclear factor-kappaB ligand (RANKL) and peroxisome proliferator-activated receptor γ (PPARγ). Furthermore, combined intervention markedly improved abnormal metabolic status, reduced the levels of serum glucose, insulin, triglycerides (TG), low-density lipoprotein (LDL), leptin, CTX-1 and IL-1ß, and increased the level of OCN. CONCLUSIONS: The coexistence of obesity and estrogen deficiency further aggravates bone tissue degeneration and abnormal metabolic pathology, which could be better inhibited by the combination with simvastatin and exercise instead of single intervention, suggesting that combined intervention may be a potential candidate for amelioration of the progression of OP.

Combination of metformin and exercise alleviates osteoarthritis in ovariectomized mice fed a high-fat diet.

OBJECTIVE: To evaluate and compare the effects of the combined intervention of metformin and exercise on the degeneration of cartilage and subchondral bone in a mouse model of osteoarthritis (OA) induced by estrogen deficiency and obesity. METHODS: 56 female 3-month-old C57BL/6 mice underwent ovariectomy (OVX) (n = 40) or a sham operation (n = 16) and were randomized into seven groups (n = 8/group): 1) sham-operated mice with a normal diet (Sham), 2) OVX mice with a normal diet (OVX), 3) sham-operated mice with high-fat diet (HFD) (HSVX), 4) OVX mice with HFD (HOVX), 5) OVX mice with HFD + exercise (HOVE), 6) OVX mice with HFD + metformin (HOMX), and 7) OVX mice with HFD + metformin + exercise (HOME). Drug administration and exercise training were initiated 72 h after surgical operation. The pathology of OA was assessed by histomorphology analyses, immunohistochemistry (IHC), tartrate-resistant acid phosphatase (TRAP) staining, micro-computed tomography and enzyme-linked immunosorbent assay (ELISA). RESULTS: Histomorphological analysis revealed that OA was significantly exacerbated by the coexistence of estrogen deficiency and obesity and markedly alleviated by the combined intervention. In details, metformin plus exercise ameliorated the abnormal metabolic status and cartilage lesions, significantly increased aggrecan and collagen-II expression and decreased the expression of ADAMTS-4. Furthermore, combined intervention markedly improved bone degeneration, bone mass and microarchitecture of subchondral bone. And the intervention also increased the concentration of OCN and decreased the serum concentration of IL-1ß and CTX-1 and glucose. CONCLUSIONS: The coexistence of estrogen deficiency and obesity further aggravates abnormal metabolic pathology and articular degeneration, which could be prevented by the combination with metformin and exercise, suggesting that combined intervention may be a potential candidate for amelioration of the progression of OA.

Preparation of medical Mg-Zn alloys and the effect of different zinc contents on the alloy.

In recent years, along with the development and application of magnesium alloys, magnesium alloys have been widely used in automotive, aerospace, medicine, sports, and other fields. In the field of medical materials, magnesium not only has the advantage of light weight, high strength, and a density similar to that of human bone, but also has good biocompatibility and promotes the growth of human bone. However, the mechanical properties and corrosion resistance of magnesium alloys need to be further improved to meet the requirements for human biodegradable implants. In this study, three alloys (mass fractions: Mg-10Zn, Mg-20Zn, and Mg-30Zn (wt.%)) were prepared using powder metallurgy by homogeneously mixing powders of the above materials in a certain amount with magnesium as the substrate through the addition of zinc elements, which also have good biocompatibility. The effect of zinc on the microstructure, mechanical properties, wear performance, and corrosion resistance of magnesium-zinc alloys was studied when the zinc content was different. The results show that compared with the traditional magnesium alloy using powder metallurgy, prepared magnesium alloy has good resistance to compression and bending, its maximum compressive stress can reach up to 318.96 MPa, the maximum bending strength reached 189.41 MPa, and can meet the mechanical properties of the alloy as a human bone-plate requirements. On the polarization curve, the maximum positive shift of corrosion potential of the specimens was 73 mv and the maximum decrease of corrosion-current density was 53.2%. From the comparison of the above properties, it was concluded that the three prepared alloys of which Mg-20% Zn had the best overall performance. Its maximum compressive stress, maximum bending strength, and corrosion-current density reached 318.96 MPa, 189.41 MPa and 2.08 × 10-5 A·cm-2 respectively, which are more suitable for use as human implant bone splints in human-body fluid environment. The mechanical properties of the sintered Mg-Zn alloys were analyzed using powder-metallurgy techniques, and their microstructure, micromotion wear properties, electrochemical corrosion properties and composition of the physical phases were analyzed and discussed.

The Protective Effects of Parathyroid Hormone (1-34) on Cartilage and Subchondral Bone Through Down-Regulating JAK2/STAT3 and WNT5A/ROR2 in a Collagenase-Induced Osteoarthritis Mouse Model.

OBJECTIVE: To assess the effects of PTH (1-34) on bone and cartilage metabolism in a collagenase-induced mouse model of osteoarthritis (OA) and examine whether PTH (1-34) affects the expression of JAK2/STAT3 and WNT5A/ROR2 in this process. METHODS: Eighteen 12-week-old male C57Bl/6 mice were randomly assigned into three groups as follows: sham group (Group A), the collagenase + saline injection group (Group B), and the collagenase + PTH (1-34) treatment group (Group C). Collagenase was injected (intra-articular) into the knee joint of Group B and C. The PTH (1-34)-treatment was started at 6 weeks after the operation and lasted for 6 weeks. Cartilage pathology was evaluated by gross visual, histological, and immunohistochemical assessments. Subchondral bone was evaluated by microcomputed tomography (micro-CT) and immunohistochemical analyses. RESULTS: The OARSI macroscopic and microscopic scores of Group B were higher than those of Group A (P = 0.026; P = 0.002, respectively). Group C showed statistically significant differences in macroscopic and microscopic scores from Group B (P = 0.041; P = 0.008, respectively). The results showed that the Col-II and AGG expression levels in the cartilage tissue were significantly lower in Group B than Group A (P < 0.001; P = 0.008, respectively). The Col-II and AGG expression levels were significantly higher in Group C than Group B (P = 0.009; P = 0.014, respectively). MMP-13, ADAMTS-4, Caspase-3, P53, and Bax expression levels were significantly higher in Group B than the Group A (P < 0.001; P < 0.001; P = 0.04; P < 0.001; P = 0.005, respectively); however, the cartilage tissue in Group C showed significantly less ADAMTS-4, MMP-13, Caspase-3, P53, and Bax expression than Group B (P < 0.001, P < 0.001, P = 0.044; P = 0.002; P = 0.005, respectively). Over-expressed JAK2/STAT3 and WNT5A/ROR2 were observed in both cartilage and subchondral bone in this model; however, these changes were prevented by PTH (1-34) treatment. These parameters (bone mineral density, bone volume ratio, trabecular bone pattern factor, and structure model index) of micro-CT indicated subchondral bone loss and architecture changes in Group B, but improvements in these parameters in Group C. CONCLUSIONS: PTH (1-34) exhibits protective effects on both cartilage and subchondral bone in a collagenase-induced OA mouse model, and it may be involved in down-regulating the expression of JAK2/STAT3 and WNT5A/ROR2.

Denosumab alleviates intervertebral disc degeneration adjacent to lumbar fusion by inhibiting endplate osteochondral remodeling and vertebral osteoporosis in ovariectomized rats.

BACKGROUND: Although adjacent segmental intervertebral disc degeneration (ASDD) is one of the most common complications after lumbar fusion, its exact mechanism remains unclear. As an antibody to RANKL, denosumab (Dmab) effectively reduces bone resorption and stimulates bone formation, which can increase bone mineral density (BMD) and improve osteoporosis. However, it has not been confirmed whether Dmab has a reversing or retarding effect on ASDD. METHODS: Three-month-old female Sprague-Dawley rats that underwent L4-L5 posterolateral lumbar fusion (PLF) with spinous-process wire fixation 4 weeks after bilateral ovariectomy (OVX) surgery were given Dmab 4 weeks after PLF surgery (OVX+PLF+Dmab group). In addition, the following control groups were defined: Sham, OVX, PLF, and OVX+PLF (n=12 each). Next, manual palpation and X-ray were used to evaluate the state of lumbar fusion. The bone microstructure in the lumbar vertebra and endplate as well as the disc height index (DHI) of L5/6 was evaluated by microcomputed tomography (µCT). The characteristic alterations of ASDD were identified via Safranin-O green staining. Osteoclasts were detected using tartrate-resistant acid phosphatase (TRAP) staining, and the biomechanical properties of vertebrae were evaluated. Aggrecan (Agg), metalloproteinase-13 (MMP-13), and a disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS-4) expression in the intervertebral disc were detected by immunohistochemistry and real-time polymerase chain reaction (RT-PCR) analysis. In addition, the expression of CD24 and Sox-9 was assessed by immunohistochemistry. RESULTS: Manual palpation showed clear evidence of the fused segment's immobility. Compared to the OVX+PLF group, more new bone formation was observed by X-ray examination in the OVX+PLF+Dmab group. Dmab significantly alleviated ASDD by retaining disc height index (DHI), decreasing endplate porosity, and increasing vertebral biomechanical properties and BMD. TRAP staining results showed a significantly decreased number of active osteoclasts after Dmab treatment, especially in subchondral bone and cartilaginous endplates. Moreover, the protein and mRNA expression results in discs (IVDs) showed that Dmab not only inhibited matrix degradation by decreasing MMP-13 and ADAMTS-4 but also promoted matrix synthesis by increasing Agg. Dmab maintained the number of notochord cells by increasing CD24 but reducing Sox-9. CONCLUSIONS: These results suggest that Dmab may be a novel therapeutic target for ASDD treatment.

Synthesis and Properties of Mg-Mn-Zn Alloys for Medical Applications.

The magnesium alloys Mg-0.5Mn-2Zn, Mg-1.0Mn-2Zn, and Mg-1.5Mn-2Zn (wt.%) with potential biomedical applications, synthesized by powder metallurgy, were investigated to evaluate the influence of manganese content on their microstructure, mechanical properties, and corrosion resistance. The results show that Mg-Mn-Zn alloys prepared by powder metallurgy reached the maximum compressive stress of 316 MPa and the maximum bending strength of 186 MPa, showing their good resistance to compression and bending, and meeting the mechanical properties required for the human bone plate. With an increase in manganese content, the corrosion resistance improved. In the polarization curve, the maximum positive shift of corrosion potential was 92 mV and the maximum decrease of corrosion current density was 10.2%. It was concluded that, of the alloys tested, Mg-1.0Mn-2.0Zn (wt.%) had the best overall performance, and its maximum compressive stress force and corrosion current density reached 232.42 MPa and 1.32 × 10-5 A·cm-2, respectively, being more suitable for service in human body fluids.

Effects of sodium hexametaphosphate, sodium tripolyphosphate and sodium pyrophosphate on the ultrastructure of beef myofibrillar proteins investigated with atomic force microscopy.

Myofibrillar protein isolated from beef muscles were treated with 3 phosphates (Sodium Hexametaphosphate, sodium tripolyphosphate, sodium pyrophosphate) with different concentrations of 0.3%, 0.6%, 0.9%, 1.2% respectively. Protein solubility, surface hydrophobicity and reactive sulfhydryl group was determined. Atomic force microscopy was used to observe the microscopic protein surface. SDS-PAGE was carried out to determine the proteolysis of myofibrillar protein. The solubility and surface hydrophobic bond of myofibrillar protein was highly increased and the diameter decreased by SHMP, TSPP, STPP. Reactive sulfhydryl groups increased after SHMP addition, but slightly decreased in STPP and TSPP treated MP. TSPP and STPP had the same effect on myofibrillar microstructure and was different from SHMP. Three phosphates all caused MP unfolding. The MP gel complexity was increased, and roughness was decreased after phosphates addition, indicating phosphates helped to construct a more ordered and smoother gel microcosmic surface.

Parathyroid hormone (1-34) ameliorates cartilage degeneration and subchondral bone deterioration in collagenase-induced osteoarthritis model in mice.

AIMS: Parathyroid hormone (PTH) (1-34) exhibits potential in preventing degeneration in both cartilage and subchondral bone in osteoarthritis (OA) development. We assessed the effects of PTH (1-34) at different concentrations on bone and cartilage metabolism in a collagenase-induced mouse model of OA and examined whether PTH (1-34) affects the JAK2/STAT3 signalling pathway in this process. METHODS: Collagenase-induced OA was established in C57Bl/6 mice. Therapy with PTH (1-34) (10 µg/kg/day or 40 µg/kg/day) was initiated immediately after surgery and continued for six weeks. Cartilage pathology was evaluated by gross visual, histology, and immunohistochemical assessments. Cell apoptosis was analyzed by TUNEL staining. Microcomputed tomography (micro-CT) was used to evaluate the bone mass and the microarchitecture in subchondral bone. RESULTS: Enhanced matrix catabolism, increased apoptosis of chondrocytes in cartilage, and overexpressed JAK2/STAT3 and p-JAK2/p-STAT3 were observed in cartilage in this model. All of these changes were prevented by PTH (1-34) treatment, with no significant difference between the low-dose and high-dose groups. Micro-CT analysis indicated that bone mineral density (BMD), bone volume/trabecular volume (BV/TV), and trabecular thickness (Tb.Th) levels were significantly lower in the OA group than those in the Sham, PTH 10 µg, and PTH 40 µg groups, but these parameters were significantly higher in the PTH 40 µg group than in the PTH 10 µg group. CONCLUSION: Intermittent administration of PTH (1-34) exhibits protective effects on both cartilage and subchondral bone in a dose-dependent manner on the latter in a collagenase-induced OA mouse model, which may be involved in regulating the JAK2/STAT3 signalling pathway.Cite this article: Bone Joint Res 2020;9(10):675-688.

Mechanical Characteristics of Cement-Based Grouting Material in High-Geothermal Tunnel.

The cement-based grouting materials used for practical purposes in high-geothermal tunnels are inevitably affected by humidity and high temperature, leading to the deterioration of mechanical properties. Based on the characteristics of changing high temperatures and two typical conditions of hot-humid and hot-dry environments in high-geothermal tunnels, many mechanical strength tests were carried out on the grouting material cured under different environmental conditions. The study results indicated that high temperature and low relative humidity were unfavorable to the development of mechanical characteristics of grouting material, but the coupling effect of two factors could improve the strength at early ages and reduce the degradation of long-term strength. As the curing temperature exceeded 56.3 °C, the humidity effect on strength played a more important role in recovering the strength of grouting material damaged by high temperature. Temperature had more significant impact on the relative peak stress while the relative humidity had greater influence on the relative peak strain. A calculation compressive constitutive model was prospered, which considering both temperature and relative humidity. The study results may provide much valuable experimental data and theoretical supporting for the design of compression constitutive of cement-based grouting material in high-geothermal tunnel.