Tartrate-resistant Acid Phosphatase Levels Research Articles

Cedrol alleviates postmenopausal osteoporosis in rats through inhibiting the activation of the NF-κB signaling pathway.

Pharmacological studies have shown that Cedrol (CE) exhibits extensive biological activities, including anti-inflammatory and analgesic. Moreover, it can inhibit the NF-κB pathway and the expression of various associated proteins. This study aimed to investigate the role of CE in postmenopausal osteoporosis. The results showed that intragastric administration of CE (10 and 20mg/kg) significantly improved the bone microstructure damage and increased bone mineral density, trabecular bone volume, and bone trabecular thickness in ovariectomized (OVX) rats (p < 0.05). CE treatment additionally made a well-organized arrangement of bone trabeculae and improved its thickness and density. Compared with the OVX group, the levels of tartrate-resistant acid phosphatase from 5b and C-terminal telopeptide of type I collagen were significantly reduced by 42.75% and 49.27% in the OVX + CE rats (p < 0.05). TRAP staining visually showed that the number of osteoclasts in the femur tissue of CE-treated rats was less than that of the OVX group. The expressions of nuclear factor of activated T-cells, cytoplasmic 1, acid phosphatase 5, and cathepsin K in OVX + CE rats were significantly decreased by 51.61%, 46.07%, and 50.34% compared to the OVX group (p < 0.01). In addition, CE intervention effectively reduced the phosphorylation levels of P65 and IκBα and inhibited the NF-κB signaling pathway. Meanwhile, CE diminished the number of multinucleated osteoclasts induced by receptor activator for nuclear factor-κB ligand and hindered cell fusion as well as nuclear translocation of osteoclast precursor cells P65. In conclusion, CE inhibits osteoclastogenesis by suppressing the activation of the NF-κB signaling pathway, thereby alleviating postmenopausal osteoporosis.

Dietary supplementation with calcitriol or quercetin improved eggshell and bone quality by modulating calcium metabolism

This study was aimed to investigate the effects of dietary calcitriol or quercetin supplementation on eggshell and bone quality of laying hens. In trial 1, 72 Hy-Line Brown layers (80-week-old) with weak-shelled strength (25 to 30 N) were assigned into 4 dietary treatments with 6 replicates of 3 birds and fed a basal diet (4% calcium level) or basal diets supplemented with 0.5% calcium, 5 μg/kg calcitriol or 500 mg/kg quercetin for 4 weeks. In trial 2, 360 Hy-Line Brown layers (60-week-old) were divided into 3 groups with 8 replicates of 15 birds: control group (basal diet), calcitriol group (basal diet+5 μg/kg calcitriol), and quercetin group (basal diet+500 mg/kg quercetin). This trial lasted for 12 weeks. The results showed that dietary calcitriol or quercetin improved eggshell quality in both trials (P<0.05). In trial 2, compared with the control group, both calcitriol and quercetin supplementations improved femoral bone quality, calcium retention of hens and calcium content in uterine fluid at 18.5 h post-oviposition (PO) (P<0.05), along with enhancing uterine morphology. Compared to the control group, supplemental calcitriol or quercetin up-regulated the relative mRNA expression levels of uterine transient receptor potential cation channel, subfamily V, member 6 (TRPV6) at 8.5 h PO and plasma membrane calcium-ATPase (PMCA), vitamin D receptor (VDR), estrogen receptor alpha (ERα) at 18.5 h PO (P<0.05), but down-regulated the uterine caspase 3 (CASP3) relative mRNA expression level at 8.5 h PO (P<0.05). Meanwhile, the femoral relative mRNA expression levels of tartrate-resistant acid phosphatase (TRAP) (up-regulated at 8.5 and 18.5 h PO) and alkaline phosphatase (ALP) (up-regulated at 8.5 h PO but down-regulated at 18.5 h PO) were also affected by calcitriol or quercetin supplementation (P<0.05). Compared to the calcitriol, quercetin increased hen-day egg production and femoral medullary bone volume/bone tissue volume but reduced femoral stiffness (P<0.05), which were accompanied by increased relative mRNA expression levels of uterine TRPV6, estrogen receptor beta (ERβ) at 18.5 h PO (P<0.05). Overall, both dietary calcitriol and quercetin could improve eggshell and bone quality by modulating calcium metabolism of aged layers. Compared to calcitriol, dietary quercetin up-regulated the expression of uterine calcium transporters, without affecting eggshell quality.

Icariin promotes bone marrow mesenchymal stem cells osteogenic differentiation via the mTOR/autophagy pathway to improve ketogenic diet-associated osteoporosis

BackgroundIcariin, a traditional Chinese medicine, has demonstrated anti-osteoporotic properties in ovariectomized mice. However, its effectiveness in preventing bone loss induced by ketogenic diet (KD), which mimics osteoporosis in human, remains unexplored. This study aims to investigate icariin’s impact on KD-induced bone loss in mice.MethodsThirty mice were divided into: sham, KD, and KD + icariin groups. Post a 12-week intervention, evaluation including bone microstructures, serum concentrations of tartrate-resistant acid phosphatase (TRAP) and bone-specific alkaline phosphatase (ALP), and femoral tissue expression levels of osteocalcin (OCN) and TRAP. The expression levels of mammalian target of rapamycin (mTOR), ALP, peroxisome proliferator-activated receptor gamma (PPAR-γ), phosphorylated mTOR (p-mTOR), and the autophagy adaptor protein (p62) were also analyzed. Alizarin granule deposition and cellular ALP levels were measured following the induction of bone marrow mesenchymal stem cells (BMSCs) into osteogenesis.ResultsThe study found that KD significantly impaired BMSCs' osteogenic differentiation, leading to bone loss. Icariin notably increased bone mass, stimulated osteogenesis, and reduced cancellous bone loss. In the KD + icariin group, measures such as bone tissue density (TMD), bone volume fraction (BV/TV), trabecular number (Tb.N), and trabecular thickness (Tb.Th) were significantly higher than in the KD group. Additionally, bone trabecular separation (Tb.Sp) was markedly lower in the KD + icariin group. Moreover, icariin increased OCN and ALP levels while suppressing PPAR-γ, TRAP, p62, and p-mTOR. In cellular studies, icariin encouraged osteogenic development in BMSCs under KD conditions.ConclusionsIcariin effectively counteracts bone thinning and improves bone microstructure. Its mechanism likely involves stimulating BMSCs osteogenic differentiation and inhibiting bone resorption, potentially through mTOR downregulation. These findings suggest icariin's potential as an alternative treatment for KD-induced bone loss.

Understanding the mechanisms and implications of acacetin in mitigating diabetic osteoporosis: Insights from a zebrafish model

Osteoporosis, a condition characterised by reduced bone mineral density is more prevalent in individuals with diabetes. Acacetin, a natural flavonoid, possesses anti-inflammatory and antioxidant properties. However, its protective role in diabetic osteoporosis remains unexplored. Hence, the present study aimed to investigate the antihyperglycemic, antioxidant, and osteoprotective effects of acacetin using streptozotocin (STZ)-induced diabetic osteoporosis models in larval and adult zebrafish. The larvae were subjected to assays such as embryotoxicity, detection of oxidative stress markers (reactive oxygen species (ROS), superoxide levels, lipid peroxidation), apoptosis, and gene expression related to diabetes and osteoporosis. In adult zebrafish, body weight, blood glucose level, alkaline phosphatase, and tartrate resistant acid phosphatase (TRAP) activity were estimated, along with assays like alizarin red staining, histopathology, and gene expression. Interaction of acacetin and RANK/RANKL/OPG pathway was detected using molecular docking analysis. Acacetin exhibited antihyperglycemic effects in the zebrafish model, as shown by reduced blood glucose levels and increased body weight. Acacetin also demonstrated antioxidant properties by decreasing ROS, superoxide radicals, and lipid peroxidation levels. Additionally, acacetin treatment resulted in reduction of apoptosis and tartrate-resistant acid phosphatase levels and improved bone histology with increased bone density and enhanced alkaline phosphatase activity. In gene expression analysis, acacetin treatment has upregulated osteoprotegerin gene and downregulated RANKL gene. In conclusion, acacetin holds potential as a therapeutic agent for diabetic osteoporosis by targeting hyperglycemia, oxidative stress, apoptosis, and the RANK/RANKL/OPG signalling pathway.

Azilsartan inhibits inflammation-triggered bone resorption and osteoclastogenesis in vivo via suppression of TNF-α expression in macrophages.

Hypertension is a major risk factor for cardiovascular disease (CVD) and is associated with increased bone loss due to excessive activity of the local renin-angiotensin system (RAS). Angiotensinogen/Angiotensin (ANG) II/Angiotensin II type 1 receptor (AT1R) axis is considered as the core axis regulating RAS activity. Azilsartan is an FDA-approved selective AT1R antagonist that is used to treat hypertension. This study aimed to determine whether azilsartan affects formation of osteoclast, resorption of bone, and the expression of cytokines linked with osteoclastogenesis during lipopolysaccharide (LPS)-triggered inflammation in vivo. In vivo, following a 5-day supracalvarial injection of LPS or tumor necrosis factor-alpha (TNF-α) with or without azilsartan, the proportion of bone resorption and the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells, which are identified as osteoclasts on mice calvariae were counted. The mRNA expression levels of TRAP, cathepsin K, receptor activator of NF-κB ligand (RANKL), and TNF-α were also evaluated. In vitro, the effect of azilsartan (0, 0.01, 0.1, 1, and 10 μM) on RANKL and TNF-α-triggered osteoclastogenesis were investigated. Also, whether azilsartan restrains LPS-triggered TNF-α mRNA and protein expression in macrophages and RANKL expression in osteoblasts were assessed. Furthermore, western blotting for analysis of mitogen-activated protein kinases (MAPKs) signaling was conducted. Azilsartan-treated calvariae exhibited significantly lower bone resorption and osteoclastogenesis than those treated with LPS alone. In vivo, LPS with azilsartan administration resulted in lower levels of receptor activator of RANKL and TNF-α mRNA expression than LPS administration alone. Nevertheless, azilsartan did not show inhibitory effect on RANKL- and TNF-α-triggered osteoclastogenesis in vitro. Compared to macrophages treated with LPS, TNF-α mRNA and protein levels were lower in macrophages treated by LPS with azilsartan. In contrast, RANKL mRNA and protein expression levels in osteoblasts were the same in cells co-treated with azilsartan and LPS and those exposed to LPS only. Furthermore, azilsartan suppressed LPS-triggered MAPKs signaling pathway in macrophages. After 5-day supracalvarial injection, there is no difference between TNF-α injection group and TNF-α with azilsartan injection group. These findings imply that azilsartan prevents LPS-triggered TNF-α production in macrophages, which in turn prevents LPS-Triggered osteoclast formation and bone resorption in vivo.

The mechanism of metformin combined with total flavonoids of Rhizoma Drynariae on ovariectomy-induced osteoporotic rats

The present study evaluated the in vitro effect of metformin (Met) and total flavonoids of Rhizoma Drynariae (TFRD) on osteoclasts, osteocytes, and osteoblasts at different stages. We also assessed the effect and mechanism of treatment with Met combined with TFRD on ovariectomy (OVX)-induced osteoporosis in rats. The results showed that Met combined with TFRD significantly promoted the migration of osteoprogenitor cells and stimulated the differentiation and maturation of osteoblast precursor cells. Furthermore, Met combined with TFRD treatment significantly inhibited the osteogenic inhibitor sclerostin (SOST)/dickkopf 1 (DKK1) protein expression and the osteoclast differentiation factor receptor activator of nuclear factor-κB ligand (RANKL)/osteoprotegerin (OPG) ratio in osteocytes. In the in vivo study, Met combined with TFRD effectively reduced bone resorption markers levels, including type-I collagen carboxy-terminal peptide (CTX-1) and tartrate-resistant acid phosphatase (TRAP), and remarkably increased the bone formation marker propeptide of type I procollagen (PINP) level in the serum of rats with osteoporosis. Met combined with TFRD treatment improved bone mineral density (BMD), trabecular microstructure, and mechanical properties of osteoporotic rats. Mechanistically, Met combined with TFRD downregulated SOST and DKK1 levels, and upregulated Wnt10b, β-catenin, runt-related transcription factor 2 (Runx2) et al. Meanwhile, Met combined with TFRD treatment reduced the RANKL/OPG ratio, and reduced the receptor activator of nuclear factor-κB (RANK), nuclear factor of activated T cells c1 (NFATC1), and TRAP levels. In conclusion, Met combined with TFRD ameliorated bone mass in osteoporotic rats through regulating Wnt/β-catenin signaling pathway and OPG/RANKL/RANK axis.

Modulatory influence of melatonin on apical periodontitis in Wistar rats fed a high-fat diet

ObjectiveThis study aimed to evaluate the influence of high-fat diet (HFD) and melatonin (MEL) treatment on the progression of inflammation and alveolar bone resorption (ABR) in rats with AP. DesignForty male Wistar rats were divided into four groups: apical periodontitis (AP), HFDAP, APMEL and HFDAPMEL. The animals were fed an HFD or standard diet for 107 days. On the 7th day, the rats were subjected to AP, and after 70 days, the rats in the MEL groups were treated with MEL for 30 days. Post treatment, the animals were euthanized, and their jaws were retrieved for evaluation of bone resorption, intensity of the inflammatory response, and immunohistochemical analysis including tartrate-resistant acid phosphatase (TRAP) and interleukin-1β (IL-1β) levels and tumor necrosis factor (TNF)-α expression. ResultsThe APMEL group showed reduction in the inflammatory infiltrate and IL-1β expression relation to HFDAP, while the TNF-α levels did not differ among the groups. The HFDAP group showed an increase in the ABR. MEL reduced the TRAP levels in the APMEL and HFDAPMEL groups. Conclusionswhile MEL could reduce TRAP levels in the APMEL and HFDAPMEL groups, the reduction in the HFDAPMEL group was smaller than that in the APMEL group, demonstrating that the interaction between AP and HFD decreased the anti-resorptive effects of MEL.

Total lignans from Vitex negundo seeds attenuate osteoarthritis and their main component vitedoin A alleviates osteoclast differentiation by suppressing ERK/NFATc1 signaling.

The seeds of Vitex negundo have been used for inflammation-related disease treatment in traditional medicine. This study focused on the anti-osteoarthritis (OA) effects of the total lignans of V. negundo seeds (TOV) in monosodium iodoacetate-induced osteoarthritis rats and its pharmacokinetic properties, as well as the effects and potential mechanism of its main components VN1 (6-hydroxy-4-(4-hydroxy-3-methoxy-phenyl)-3-hydro-xymethyl-7-methoxy-3,4-dihydro-2-naphthaldehydeb) and VN2 (vitedoin A) on receptor activator of NF-κB ligand (RANKL)-induced osteoclast differentiation in bone marrow macrophages (BMMs). TOV significantly attenuated osteoarthritis, leading to an increase in pain thresholds, improvement of knee articular cartilages and chondrocytes loss, and decreased total joint scores and serum levels of TNF-α, interleukin-1β (IL-1β), and prostaglandin E2 (PGE2) in osteoarthritis rats. The half-time (T1/2 ) was 2.82 h and 1.33 h, and the bioavailability was 15.34%-21.89% and 16.29%-22.11%, for VN1 and VN2, respectively. VN2, rather than VN1, remarkably inhibited tartrate-resistant acid phosphatase (TRAP) activity, reduced the number of TRAP-positive multinuclear cells, diminished the formation of actin ring, and decreased mRNA levels of cathepsin K (CTSK), TRAP, nuclear factor of activated T cell 1 (NFATc1), and osteoclast-associated receptor, as well as downregulated protein levels of p-ERK (phosphorylated extracellular signal-regulated kinase), TRAP, CTSK and NFATc1 in BMMs. These findings suggest TOV has promising therapeutic potential for OA treatment and VN2, in particular, attenuates osteoclast differentiation by suppressing ERK/NFATc1 signaling and actin ring, mainly accounting for the anti-OA efficacy of TOV.

Icariin inhibits thioacetamide-induced osteoclast differentiation through RANKL-p38/ERK-NFAT pathway

This study aims to investigate the therapeutic effect of icariin(ICA) on thioacetamide(TAA)-induced femoral osteolysis in rats. RAW264.7 cells were treated with TAA and ICA. Cell counting kit-8(CCK-8) assay was used to detect cell proliferation, and tartrate-resistant acid phosphatase(TRAP) staining to examine the formation of osteoclasts. The expression of TRAP, cathepsin K, c-FOS, and NFATc1 in RAW264.7 cells was determined by Western blot and immunofluorescence method. Thirty-two SD rats were randomized into the control group, TAA group(intraperitoneal injection of TAA at 300 mg·kg~(-1)), ICA group(gavage of ICA at 600 mg·kg~(-1)) and TAA + ICA group(intraperitoneal injection of TAA at 300 mg·kg~(-1) and gavage of ICA at 600 mg·kg~(-1)). Administration was performed every other day for 6 weeks. Body weight and length of femur were recorded at execution. Pathological injury and osteoclast differentiation of femur were observed based on hematoxylin-eosin(HE) staining and TRAP staining, and the changes of bone metabolism-related indexes alkaline phosphatase(ALP), calcium(Ca), phosphorus(P), magnesium(Mg), and cross-linked N-telopeptide of type Ⅰ collagen(NTX-Ⅰ) in serum were detected. Three-point bending test and micro-CT were applied to evaluate the quality of femur, and Western blot to detect the levels of osteoclast-related proteins TRAP, cathepsin K, RANK, RANKL, p38, p-p38, ERK, p-ERK, JNK, p-JNK, c-Fos, and NFATc1. The results showed ICA could inhibit TAA-induced production of TRAP-positive cells, the expression of osteoclast-related proteins, and nuclear translocation of NFATc1. ICA alleviated the weight loss, reduction of femur length, and growth inhibition induced by TAA in SD rats. ICA ameliorated the decline of femur elastic modulus caused by TAA and significantly restored trabecular bone mineral density(BMD), trabecular pattern factor(Tb.Pf), trabecular number(Tb.N), trabecular thickness(Tb.Th), and structure model index(SMI), thus improving bone structure. Western blot results showed ICA suppressed femoral osteoclast differentiation induced by TAA through RANKL-p38/ERK-NFATc1 signaling pathway. ICA inhibits osteoclast differentiation and prevents TAA-induced osteolysis by down-regulating RANKL-p38/ERK-NFAT signaling pathway.

Lithium chloride stimulates bone formation in extraction socket repair in rats.

Previous evidence shows that lithium chloride (LiCl), a suppressor of glycogen synthase kinase-3β (GSK-3β), may enhance bone formation in several medical and dental conditions. Thus, the purpose of the current study was to assess the effects of LiCl on extraction socket repair in rats. Thirty rats were randomly assigned into a control group (administration of water; n = 15) or a LiCl group (administration of 150mg/kg of LiCl; n = 15). LiCl and water were given every other day, starting at 7days before the extraction of upper first molars until the end of each experiment period. Histological sections from five rats per group were obtained at 10, 20, and 30days post-extractions. Histometrical analysis of newly formed bone (NB) and the levels of tartrate-resistant acid phosphatase (TRAP)-stained cells were evaluated at 10, 20, and 30days post-extractions. Immunohistochemical staining for receptor activator of nuclear factor kappa-Β ligand (RANKL), osteoprotegerin (OPG), bone sialoprotein (BSP), osteocalcin (OCN), and osteopontin (OPN) was assessed at 10days post-extractions. The LiCl group had a greater proportion of NB than the control group at 20days (P < 0.05). At 30days, the rate of TRAP-stained cells was lower in the LiCl group than in the control group (P < 0.05). At 10days, the LiCl group presented stronger staining for OPG, BSP, OPN, and OCN, when compared to the control group (P < 0.05). Systemic LiCl enhanced extraction socket repair, stimulated an overall increase in bone formation markers, and restricted the levels of TRAP in rats.

Effects of rearing systems on the eggshell quality, bone parameters and expression of genes related to bone remodeling in aged laying hens.

Public concerns regarding animal welfare are changing the selection of rearing systems in laying hens. This study investigated the effects of rearing systems on eggshell quality, bone parameters and relative expression levels of genes related to bone remodeling in aged laying hens. A total of 2,952 55-day-old Jing Tint Six pullets were randomly assigned to place in the conventional caging system (CCS) or aviary system (AVS) and kept until 95 weeks of age. The AVS group delayed the decrease of eggshell quality and alleviated the symptoms of osteoporosis in the humerus rather than in the femur. Eggshell breaking strength, thickness, weight, weight ratio, stiffness and fracture toughness were decreased linearly with age (from 55 to 95 weeks of age, p < 0.05). The AVS group had higher eggshell breaking strength, stiffness and fracture toughness than the CCS group (p < 0.05). Higher total calcium and phosphorus per egg were presented in the AVS group at 95 weeks of age (p < 0.05). At 95 weeks of age, the AVS group had a humerus with higher weight, volume, length, midpoint perimeter, cortical index, fat-free dry weight, ash content, total calcium per bone, total phosphorus per bone, average bone mineral density, strength, stiffness and work to fracture compared to the CCS group (p < 0.05). Such differences did not appear in the femur. The relative expression levels of alkaline phosphatase (ALP) and osteocalcin (OCN) genes in the femur and hormone receptors (vitamin D receptor (VDR), estrogen receptor alpha (ERα) and fibroblast growth factor 23 (FGF23)) genes in the humerus were significantly upregulated (p < 0.05) in the AVS group. The level of tartrate-resistant acid phosphatase (TRAP) transcripts was also increased (p < 0.05) in the femur of the AVS group. Overall, compared with the CCS, the AVS alleviated the deterioration of eggshell and bone qualities of aged laying hens, which may be related to the changes in the expression of genes associated with bone remodeling.

Novel peptides extracted from Muraenesox cinereus bone promote calcium transport, osteoblast differentiation, and calcium absorption

Extracting peptides from fish bones as calcium supplements is of great significance for patients with osteoporosis. Novel peptides were obtained from Muraenesox cinereus bone (MCBP) by enzymatic hydrolysis and membrane separation. Molecular dynamics simulation and electron microscope analysis shown MCBP could chelate with calcium ions. A Caco-2 cell monolayer model was used to investigate the effect of peptide–calcium chelates on promoting calcium transport. MC3T3-E1 osteoblasts were treated with different concentrations of MCBP (0, 0.25, 0.5, or 1 mg/mL) to analyze the effects of MCBP on calcium absorption and cell differentiation factors. The bone formation of zebrafish evaluate the therapeutic effect of MCBP on the zebrafish OP model. The results showed that there were at least 13 kinds of calcium chelation modes in the mixed peptide, and peptide–calcium chelate could significantly improve calcium transport in the Caco-2 cell monolayer. After 14 days of culture with different concentrations of MCBP, the calcium deposition in MC3T3-E1 cells increased significantly. The type I collagen (COL-I), osteocalcin (OCN) and run-related transcription factor 2 (RUNX2), and activities of alkaline phosphatase (ALP) were dose-dependent with MCBP. MCBP can increase bone mineral density in an OP model of zebrafish. After being treated with MCBP, the expression levels of ALP, OCN, osteoprotegerin (OPG) and transforming growth factor-β (TGF-β) in the model group increased, while the expression levels of tartrate-resistant acid phosphatase (TRAP) decreased. The results indicated that MCBP may act as a nutritional food and provide an effective means for dietary supplementation in OP patients.

DJ-1 controls T cell differentiation and osteoclastogenesis in rheumatoid arthritis

Herein, we investigated the effect of DJ-1 on helper T cell differentiation, fibroblast-like synoviocyte (FLS) activation, and osteoclastogenesis in rheumatoid arthritis (RA). Serum and synovial fluid (SF) of RA and osteoarthritis (OA) patients were collected, and DJ-1 and H2O2 levels were investigated. CD4+ cells from peripheral blood mononuclear cells (PBMCs) were cultured under type 17 helper T cell (Th17) polarization conditions, and CD4+ T cell differentiation, pro-inflammatory cytokine levels, and soluble receptor activator of nuclear factor kappa-Β ligand (RANKL) were assessed. RA-FLSs were stimulated with 50 μM H2O2, and DJ-1 (10, 50, 100 ng/mL) to evaluate MMP-9, VEGF, TNF-α, and sRANKL production, while RANKL+ FLSs were assessed using flow cytometry. Monocytes were cultured with RANKL or IL-17A with or without DJ-1 and H2O2-pretreated RA-FLS, and tartrate-resistant acid phosphatase (TRAP) staining and RT-qPCR of osteoclast-related genes were performed. The levels of DJ-1 and H2O2 in serum and SF of RA patients were higher than those of OA patients. Under Th17-polarizing conditions, CD4+RANKL+ and CD4+CCR4+CCR6+CXCR3- T cells decreased, whereas CD4+CD25highFoxp3+ T cell increased after DJ-1 administration. Additionally, IL-17A, TNF-α, and sRANKL levels decreased in DJ-1-treated groups. DJ-1 lowered MMP-9, VEGF, TNF-α, and sRANKL levels, and RANKL+ FLS in ROS-stimulated RA-FLS. Both RANKL and IL-17A stimulated osteoclast differentiation, DJ-1 decreased TRAP+ cell count, and the expression levels of TRAP, ATP6v0d2, NFATc1, and CTSK. These findings were also observed in in vitro osteoclastogenesis with DJ-1 pretreated RA-FLS. As DJ-1 regulates Th17/Treg imbalance, pro-inflammatory cytokine production, RA-FLS activation, and osteoclastogenesis, it holds potential for RA therapy.

Clinical, radiological and pathochemical substantiation of osteosubstitution by hydroxyapatite ceramics with β-tricalcium phosphate and platelet-enriched autofibrin in fragmentary bone fractures in dogs

Due to the similarity of its composition to bone tissue, bioactive ceramics based on hydroxyapatite are considered the most promising for osteosubstitution of fragmentary fractures, and topical application of pharmacological agents and autologous agents bioactive substances, in particular, platelet-rich fibrin (PRF). The work aims to clinically and radiologically, and pathochemically substantiate osteosubstitution by hydroxyapatite ceramics with β-tricalcium phosphate and platelet-enriched fibrin for fragmentary bone fractures in dogs. A control and two experimental groups of animals were formed, including patients (n = 10) with humerus and forearm fractures. After general and local anesthesia and extracortical osteosynthesis, the first experimental group was injected with platelet-rich autofibrin between bone fragments. The second group was osteosubstituted with a combination of PRF and hydroxyapatite ceramics; in the control group, the defects remained without replacement. For biochemical studies, blood samples were taken after injury no later than 24 hours and on the 3rd, seventh, 14th, 21st, and 42nd day after osteosynthesis. In the postoperative period, clinical and radiological studies were performed. The activity of bone isoenzyme alkaline phosphatase (BAP), tartrate-resistant acid phosphatase (TrAP), the level of total calcium (Ca), inorganic phosphorus (P), and total protein were determined in blood serum. It was found that complete recovery of limb function in the experimental groups was 1.2–1.4 times faster (P &lt; 0.001) compared with the control group and radiologically confirmed earlier consolidation of fractures. In both experimental groups, the maximum activity of BAP was observed on the 14th day, in the absence of a significant difference between these groups, while in control – it peaked on the 21st day. In the case of bone injury, the level of TrAP decreased by 1.2 times (P &lt; 0.05) and then gradually increased, which was significant in the control group on the 42nd day, in the first experimental group on the 21st, and the second on the 14th day. On the third day after osteosynthesis, the concentration of P was still lower in the control (P &lt; 0.05) and the first experimental (P &lt; 0.01) group compared with clinically healthy animals. The concentration of Ca on the third day was reduced in all groups by 1.2 times (P &lt; 0.001) and then gradually returned to normal. The use of PRF with calcium-phosphate ceramics for osteosubstitution helps to optimize regeneration processes in the area of bone injury by activating osteoblastic cells and reducing the intensity of the osteoresorption reaction with an earlier reaction of bone regeneration remodeling.

Effects of long term diabetogenic high fat diet on bone in ovariectomized female rats

ABSTRACT A diabetogenic high fat diet (HFD) can be used to induce insulin resistance and obesity in animal models; however, its effects on bone are unknown. We investigated the effects of long term HFD on bone in ovariectomized (OVX) female rats. We used 12-week-old female rats divided randomly into four groups: sham operation (sham), sham operation with HFD (SHFD), OVX and OVX with HFD (OVX + HFD). Ovaries were removed in the OVX and OVX + HFD groups and the SHFD and OVX + HFD groups were fed a HFD for 28 weeks. Serum estrogen, testosterone, lipid, adiponectin, leptin, tartrate-resistant acid phosphatase (TRAP) and N-mid fragment of osteocalcin (N-MID-OT) levels were measured. Structure, apoptosis and specific transcription factors in bone were evaluated using pathologic, densitometric and immunohistochemical analysis. Body weight, serum leptin, TRAP and testosterone levels were increased, while serum N-MID-OT, estrogen and adiponectin levels were decreased in the SHFD, OVX and OVX + HFD groups. Expression of BCL2-associated X protein, caspase-3, matrix metalloproteinase-9 and calcitonin was increased, while bone mineral density (BMD) and content (BMC) in femurs and lumbar spine, and expression of B cell lymphoma 2, type 1 collagen and osteocalcin were decreased in the bones of the SHFD, OVX and OVX + HFD groups. All indices were greatest in the OVX + HFD group and HFD produced a detrimental effect on bone in both normal and OVX rats, which may be due to increased apoptosis in bone and increased leptin and decreased adiponectin levels in serum. The effects of HFD and OVX may be synergistic.

PHYSICAL THERAPY OF PATIENTS WITH POST-TRAUMATIC GONARTHROSIS WITH IMBALANCE OF OSTEO-ASSOCIATED MACRONUTRIENTS

According to modern concepts, bone remodeling disorders that are directly dependent on the imbalanceof osteo-associated trace elements play a key role in the pathogenesis of OA. Therefore, the purpose of ourstudy is to develop the program of physical rehabilitation of patients with post-traumatic gonarthrosisand to investigate its effect on the composition of osteoassociated macronutrients.To determine the metabolic function of bone tissue there were the results of the study of mineralhomeostasis by the level in the peripheral blood of phosphorus and magnesium, total calcium and itsionized form, as well as the amount of urinary calcium excretion. The processes of bone formation wereevaluated by the content of bone isoenzyme of alkaline phosphatase, bone resorption was evaluated bythe level of tartrate-resistant acid phosphatase. 42 patients with post-traumatic osteoarthritis of the kneejoints of I–II stages according to Kellgren-Lowrence were observed.The complex of physical therapy included the educational program, medical gymnastics and massage.The theoretical part of the program included 6 topics covering the main issues related to the risk factorsand prognosis for the disease, the main mechanisms of its progression, as well as the basics of differentphysical therapy use. The practical part of the program was carried out in a small-group way and includedthe gradual application of static, dynamic, active and proprioceptive neuromuscular stretching usingthe exercises with elastic bands and metered loading. As for the massage technology a combinationof segmental massage with Swedish technique was used in the program. The total duration of the complexprogram was 9 months, after which patients were advised to perform independent classes. The applicationof the developed program of physical rehabilitation had a positive effect on the processes of boneremodeling, and allowed to reduce the number of exacerbations during the year.

Effects of High-Fructose Corn Syrup on Bone Health and Gastrointestinal Microbiota in Growing Male Mice.

Here, we explored the correlation between gut microbiota and bone health and the effects of high-fructose corn syrup (HFCS) on both. Sixteen 3-week-old male C57BL/6J mice were randomly divided into two groups and given purified water (control group) or 30% HFCS in water (HFCS group) for 16 weeks. The effects of HFCS were assessed via enzyme-linked immunosorbent assays, histopathological assays of colon and bone, and 16S rDNA sequence analysis of gut microbiota. The serum of HFCS group mice had lower levels of bone alkaline phosphatase (BALP), bone Gla protein (BGP), insulin-like growth factor 1 (IGF-1), and testosterone, and higher levels of type I collagen carboxyl-terminal telopeptide (ICTP) and tartrate-resistant acid phosphatase (TRAP) than that of the control group. HFCS caused trabecular bone damage by decreasing trabecular number and thickness and increasing trabecular separation. The HFCS group colons were shorter than the control group colons. The HFCS-fed mice showed mild, localized shedding of epithelial cells in the mucosal layer, focal lymphocytic infiltration of the lamina propria, mild submucosal edema, and loosely arranged connective tissue. The HFCS group displayed lower abundance and altered composition of gut microbiota. The abundance of Defluviitaleaceae UCG-011, Erysipelatoclostridium, Ruminococcaceae UCG-009, Lactobacillus, Blautia, and Parasutterella increased, positively correlating with BALP, BGP, IGF-1, and testosterone levels, and negatively correlating with ICTP and TRAP levels. Our study revealed a potential diet-gut microbiota-bone health axis.

The effect of osteosubstitution by platelet-rich autofibrin and hydroxyapatite ceramic with ß-tricalcium phosphate on biochemical parameters of blood in rabbits

The paper presents the results of a rabbit blood biochemical study after the substitution of simulated defects in spongy and compact bone tissue by platelet-rich fibrin and its combination with hydroxyapatite granules with ß-tricalcium phosphate. The aim of the work is to establish the biochemical features of reparative osteogenesis in the time of osteosubstitution by platelet-enriched fibrin and its combination with hydroxyapatite and ß-tricalcium phosphate. Bone defects (d = 3 mm) were modeled in rabbits from control and experimental groups (n = 12) under general anaesthesia. In the control group, a replacement was performed with autologous fibrin enriched with platelets (PRF), and in the experimental group, it was combined with hydroxyapatite ceramics. Content of biochemical substances was determined: soluble fibrin, the activity of antithrombin-III in blood plasma, nitric oxide (NO), haptoglobin, and markers of bone metabolism (bone isoenzyme alkaline phosphatase (BAP) and tartrate-resistant acid phosphatase (TrAP) in serum. Measurements were performed with a spectrophotometer Stat Fax 4500. It is established that the dynamics of changes in biochemical parameters reflect certain features of osteosubstitution by different materials. In particular, the inflammatory-resorptive stage of reparative osteogenesis in the period up to the 7th day is characterized by an increase in blood concentrations of NO, haptoglobin, and soluble fibrin. However, the intensity of the acute phase in the case of combined osteosubstitution is significantly lower. The next peak of NO concentration on the 14th day indicates early angiogenesis with combined osteosubstitution. The two-phase degree of osteoresorption at the level of TrAP is significantly lower than combined osteosubstitution, and the intensity of osteogenic processes at the level of BAP, on the contrary, earlier. Also used biochemical parameters can be prognostic criteria for the analysis of fracture consolidation in conditions of osteosubstitution with different materials.

Effects of gold nanoparticles combined with human \u03b2-defensin 3 on the alveolar bone loss of periodontitis in rat

BackgroundNanomaterials of biomedicine and tissue engineering have been proposed for the treatment of periodontitis in recent years. This study aimed to investigate the effects of gold nanoparticles (AuNPs) combined with human β-defensin 3 (hBD3) on the repair of the alveolar bones of experimental periodontitis in rats.MethodsA model of experimental periodontitis was established by ligation of the maxillary second molars with silk thread in rats, which were treated with or without AuNPs combined with hBD3. Micro‐computerized tomography (micro-CT) scanning, enzyme-linked immunosorbent assay, and histological and immunohistochemical staining, including alkaline phosphatase (ALP), osteoprotegerin (OPG), tartrate-resistant acid phosphatase (TRAP), and receptor activator of NF-κB ligand (RANKL), were used to analyze the samples.ResultsMicro-CT demonstrated that the alveolar bone resorption was significantly reduced after the treatment with AuNPs combined with hBD3. Levels of TNF-α and IL-6 were decreased markedly compared with the ligation group. H&E and Masson staining showed that AuNPs combined with hBD3 group had less inflammatory cell infiltration, collagen fibrosis and fracture, but higher calcification in the new bone tissue. Moreover, the administration of AuNPs combined with hBD3 increased the expression levels of ALP and OPG (related to bone formation) while decreasing the expression levels of TRAP and RANKL (related to bone resorption) expression.ConclusionsAuNPs combined with hBD3 had a protective effect on the progression of experimental periodontitis in rats and played a certain role in suppressing osteoclastogenesis and alleviating the inflammatory destruction of periodontitis along with the promotion of bone repair.

Abnormal Bone Metabolism May Be a Primary Causative Factor of Keel Bone Fractures in Laying Hens.

Simple SummaryKeel is an essential structural bone, providing anchorage for the attachment of large breast muscles in birds, allowing them to flap wings and provide proper ventilation for their lungs during flight. Previous studies reported that keel bone damage (especially fractures) negatively affects the welfare, health, production performance, eggshell quality, and mobility of laying hens contained in different housing systems. Furthermore, various factors affect keel bone damage, including nutrition, age, housing systems, and strains of laying hens. However, studies on the effects of abnormal bone metabolism and development on keel bone damage in laying hens are limited. Therefore, this study aimed to investigate the impacts of bone metabolism and development status on keel bone damage by determining the levels of serum bone turnover markers in laying hens. The results showed that laying hens with impaired keel bone had significantly altered levels of serum Ca and P metabolism-related and osteoblast and osteoclast activity-related markers compared to those in laying hens with normal keel bone. Thus, these results indicated that abnormal bone metabolism before keel bone damage reflected by varying levels of serum bone turnover markers might be a pivotal factor causing keel bone damage in laying hens. Our results also provide new insights into the occurrence of keel bone damage in laying hens.Keel bone damage negatively affects the welfare, production performance, egg quality, and mobility of laying hens. This study aimed to investigate whether abnormal bone metabolism causes keel bone damage in laying hens. Eighty Hy-line Brown laying hens were housed in eight furnished cages with 10 birds per cage and studied from 18 to 29 weeks of age (WOA). Accordingly, keel bone status was assessed at 18, 22, 25, and 29 WOA using the X-ray method, and the serum samples of laying hens with normal keel (NK), deviated keel (DK), and fractured keel (FK) that occurred at 29 WOA were collected across all the time-points. Subsequently, the serum samples were used to measure markers related to the metabolism of Ca and P and activities of osteoblast and osteoclast. The results showed that FK laying hens had lighter bodyweight than NK and DK birds throughout the trial (p < 0.05), while the keel bone length and weight were not different in NK, DK, and FK hens at 29 WOA (p > 0.05). Moreover, bone hematoxylin and eosin (H&E) staining and tartrate-resistant acid phosphatase (TRAP) staining indicated that damaged keel bone had evident pathological changes. In the FK hens, serum P level was reduced but serum 1,25-dihydroxy-vitamin D3 (1,25-(OH)2D3) and 25-hydroxyvitamin D3 (25-OHD3) levels were elevated compared to NK hens (p < 0.05). Additionally, DK hens had higher levels of serum 1,25-(OH)2D3, parathyroid hormone (PTH) and calcitonin (CT), and lower level of serum 25-OHD3 than the NK birds (p < 0.05). Furthermore, serum alkaline phosphatase (ALP), osteocalcin (OC), osteoprotegerin (OPG), TRAP, and corticosterone (CORT) levels were elevated in DK and FK hens compared to NK hens (p < 0.05). The levels of serum Ca, P, PTH, ALP, TRAP, OPG, OC, and CORT in laying hens fluctuated with the age of the birds. Generally, the results of this study indicate that keel bone damage, especially fractures, could be associated with abnormal bone metabolism in laying hens.