Search Results (35)

Background: The juvenile-pubertal period is a critical window for linear growth and bone mass accumulation. This study investigated the joint effects of folic acid (FA) and colostrum basic protein (CBP)-fortified milk powder on growth, bone health, and metabolic safety in juvenile mice. Methods: Three-week-old C57BL/6J mice (n = 120) were acclimatized for 1 week and then randomly assigned to three isocaloric diet groups for an 8-week intervention starting at 4 weeks of age: Control (AIN-93M), Milk (AIN-93M + FA/CBP-fortified milk powder), and Positive Control (AIN-93G). Body length and weight were measured twice weekly. Bone microarchitecture was assessed by micro-computed tomography, and bone remodeling was evaluated through histology and serum biomarkers. The GH–IGF-1 axis and related metabolic parameters were also assessed. Results: FA–CBP–fortified milk powder significantly accelerated linear growth at intervention week 2, with body length higher in the Milk group than in the Control group (p < 0.01). After 8 weeks, the Milk group showed improved trabecular bone mass and microarchitecture compared with Control, especially in males (p < 0.01). Bone remodeling was transiently elevated at intervention week 4, as indicated by higher serum osteocalcin and CTX-I, and by increased osteoclast and cartilage matrix formation versus Control (p < 0.05). The GH–IGF-1 axis was also temporarily activated at week 4, with elevated serum GH and IGF-1/IGFBP-3 ratio compared with Control (p < 0.05). These skeletal benefits occurred without excess weight gain or adverse metabolic effects compared with Control (all p > 0.05). Conclusions: FA-CBP-fortified milk significantly enhanced linear growth during puberty and improved bone mass and microstructure in early adulthood. These skeletal benefits are consistent with the transient activation of the GH–IGF-1 axis. Importantly, no adverse metabolic effects were detected from early intervention through adulthood, supporting its potential application in growth-promoting nutritional strategies. Full article

Disuse-induced bone loss during stall confinement and immobilization is a major concern in horses because it impairs recovery and increases susceptibility to further injury. Experimental models are needed to evaluate therapeutic options, but most available equine models rely on cast immobilization, which is technically demanding and may be associated with complications. This study aimed to assess a simpler and less restrictive model to induce a quantifiable decrease in bone density in horses. Six French Standardbred horses underwent eight weeks of stall confinement, with a wooden wedge fitted to one front foot to elevate the heels during the last four weeks. Bone density was assessed using computed tomography (CT) examinations of both forelimbs performed at the beginning (M0) and after the confinement period (M2). Serum markers of bone metabolism (CTX-I, osteocalcin, bone-specific alkaline phosphatase, and hydroxyproline) were analyzed monthly from baseline to 2 months post confinement. Statistical analysis used Wilcoxon signed-rank tests and mixed models as appropriate. Computed tomography revealed a significant decrease in bone density after confinement (p < 0.05), more pronounced distally in the wedge limb. CTX-I levels varied with physical activity. This model provides a practical and reproducible alternative to cast immobilization for inducing equine bone demineralization. Full article

Background: Periodontitis is a chronic disease triggered by disturbed oral microbiota. We have previously reported that hepatocyte growth factor (HGF) could mitigate early-stage experimental periodontitis but exacerbate the condition in its late stage. Here, we investigated the impact of HGF on the periodontal microbiome during periodontitis progression. Methods: We established ligation-induced periodontitis in wild-type (WT) mice and HGF high-expression transgenic (HGF-Tg) mice. We quantified the levels of IL-6 and TNF-α in periodontal tissues, as well as the serum concentrations of CTXI and PINP. Ligatures were collected on days 0, 7, and 28 after ligation for 16S rRNA sequencing and microbial analysis. Results: HGF significantly altered the diversity of ligatures during periodontitis. Interestingly, specific microbial genera, such as Lactobacillus, exhibited opposing trends between the two disease stages of HGF-Tg mice, aligning with the different effects of HGF on periodontitis progression. We also identified some taxa, such as Sphingomonas, associated with IL-6, TNF-α, CTXI, and PINP. The predicted inflammatory pathways (e.g., IL-17 signaling pathways) were enriched in HGF-Tg mice on day 28 but decreased on day 7. Conclusions: HGF exerted different influences on the microbiota of ligatures during early and late stages of periodontitis, which may account for the divergent effects of HGF on periodontitis progression. Full article

Laying hens usually have 16 h of light and 8 h of darkness during egg laying, with eggshell formation primarily occurring during darkness when dietary calcium is lacking, leading to bone calcium resorption and osteoporosis. This study examined how interrupting the dark phase affects bone health in 396 Hy-line W36 hens assigned to control (C) or treatment groups (W1 and W2). All hens received 16 h of light and 8 h of darkness daily in different variations of scotophase interruption. Blood samples were taken at weeks 20, 30, 50, and 70, serum calcium was measured during darkness at two timepoints (SRT and END), and bone demineralization markers were examined using enzyme concentrations (TRACP-5b and CTX-I). Across weeks, tibias were CT-scanned for density (mg/cm³) and area (mm²), then used for breakage strength analysis (N) and ash%. No SRT Ca level differences emerged, but C hens had lower END Ca levels compared to W1 and W2 hens across all weeks, while W1 and W2 hens showed no significant differences. C hens displayed higher TRACP-5b and CTX-I concentrations across all weeks compared to W1 and W2 (all p ≤ 0.05). At week 70, C hens had the lowest cortical bone cross-sectional area and mineral density compared to W1 and W2 (all p ≤ 0.05). Tibiotarsi bone breakage strength was lower in C hens compared to W1 and W2. C hens had significantly lower ash% than treatment birds. Interrupting the scotophase period improved overall bone health in Hy-line W36 laying hens. Full article

Hemophilia, which is a rare disease, results from congenital deficiencies of coagulation factors VIII and IX, respectively, leading to spontaneous bleeding into joints, resulting in hemophilic arthropathy (HA). HA involves complex processes, including synovial proliferation, angiogenesis, and tissue remodeling. Despite ongoing research, factors contributing to HA progression, especially in adults with severe HA experiencing joint pain, remain unclear. Blood markers, particularly collagen-related ones, have been explored to assess joint health in hemophilia. For example, markers like CTX-I and CTX-II reflect bone and cartilage turnover, respectively. Studies indicate elevated levels of certain markers post-bleeding episodes, suggesting joint health changes. However, longitudinal studies on collagen turnover and basement membrane or endothelial cell markers in relation to joint outcomes, particularly during painful episodes, are scarce. Given the role of the CX3CL1/CX3XR1 axis in arthritis, other studies investigate its involvement in HA. The importance of different inflammatory and bone damage biomarkers should be assessed, alongside articular cartilage and synovial membrane morphology, aiming to enhance understanding of hemophilic arthropathy progression. Full article

High dose bolus cholecalciferol supplementation has been associated with falls and fracture, and this does not appear to be due to hypercalcaemia. The primary aim of this study was to determine the change in free vitamin D and metabolites after high dose bolus supplementation. This was a single centre, double-blinded, randomised, controlled trial of three different oral bolus doses of vitamin D₃ (50,000 IU, 150,000 IU, and 500,000 IU) in otherwise healthy, vitamin D deficient (total 25-hydroxylated vitamin 25(OH)D < 30 nmol/L) postmenopausal women. Thirty-three women were randomized to one of the three treatment groups. Twenty-seven vitamin D sufficient (25(OH)D > 50 nmol/L) postmenopausal women were recruited as a concurrent control group. Participants attended five study visits over three months. We measured total 25(OH)D₃ and free 25(OH)D, total and free 1,25(OH)₂D, parathyroid hormone, fibroblast-growth factor-23, serum calcium, ionised calcium, urinary calcium excretion, and bone turnover markers (procollagen I N-propeptide (PINP), serum C-telopeptides of type I collagen (CTX-I) and Osteocalcin (OC)). We assessed muscle strength and function with grip strength and a short physical performance battery. Postural blood pressure and aldosterone:renin ratio (ARR) was also measured. Total 25(OH)D₃ and free 25(OH)D increased in response to dose, and there were proportionate increases in total and free metabolites. Treatment did not affect serum calcium, postural blood pressure, ARR, or physical function. Bone turnover markers increased transiently one week after administration of 500,000 IU. High dose bolus cholecalciferol supplementation does not cause disproportionate increases in free vitamin D or metabolites. We did not identify any effect on blood pressure regulation or physical function that would explain increased falls after high dose treatment. A transient increase in bone turnover markers one week after a 500,000 IU bolus suggests that very high doses can have acute effects on bone metabolism, but the clinical significance of this transient increase is uncertain. Full article

Despite therapy with growth hormone (GH) in children with Prader–Willi syndrome (PWS), low bone mineral density and various orthopedic deformities have been observed often. Therefore, this study aimed to analyze bone markers, with an emphasis on vitamin K-dependent proteins (VKDPs), in normal-weight children with PWS undergoing GH therapy and a low-energy dietary intervention. Twenty-four children with PWS and 30 healthy children of the same age were included. Serum concentrations of bone alkaline phosphatase (BALP), osteocalcin (OC), carboxylated-OC (Gla-OC), undercarboxylated-OC (Glu-OC), periostin, osteopontin, osteoprotegerin (OPG), sclerostin, C-terminal telopeptide of type I collagen (CTX-I), and insulin-like growth factor-I (IGF-I) were determined using immunoenzymatic methods. OC levels and the OC/CTX-I ratios were lower in children with PWS than in healthy children (p = 0.011, p = 0.006, respectively). Glu-OC concentrations were lower (p = 0.002), but Gla-OC and periostin concentrations were higher in patients with PWS compared with the controls (p = 0.005, p < 0.001, respectively). The relationships between IGF-I and OC (p = 0.013), Gla-OC (p = 0.042), and the OC/CTX-I ratio (p = 0.017) were significant after adjusting for age in children with PWS. Bone turnover disorders in children with PWS may result from impaired bone formation due to the lower concentrations of OC and the OC/CTX-I ratio. The altered profile of OC forms with elevated periostin concentrations may indicate more intensive carboxylation processes of VKDPs in these patients. The detailed relationships between the GH/IGF-I axis and bone metabolism markers, particularly VKDPs, in children with PWS requires further research. Full article

Osteoarthritis (OA) is a chronic degenerative disease leading to articular cartilage destruction. Menopausal and postmenopausal women are susceptible to both OA and osteoporosis. S-equol, a soy isoflavone-derived molecule, is known to reduce osteoporosis in estrogen-deficient mice, but its role in OA remains unknown. This study aimed to explore the effect of S-equol on different degrees of menopausal OA in female Sprague–Dawley (SD) rats induced by estrogen deficiency caused by bilateral ovariectomy (OVX) combined with intra-articular injection of mono-iodoacetate (MIA). Knee joint histopathological change; serum biomarkers of bone turnover, including N-terminal propeptide of type I procollagen (PINP), C-terminal telopeptide of type I collagen (CTX-I) and N-terminal telopeptide of type I collagen (NTX-I); the cartilage degradation biomarkers hyaluronic acid (HA) and N-terminal propeptide of type II procollagen (PIINP); and the matrix-degrading enzymes matrix metalloproteinases (MMP)-1, MMP-3 and MMP-13, as well as the oxidative stress-inducing molecules nitric oxide (NO) and hydrogen peroxide (H₂O₂), were assessed for evaluation of OA progression after S-equol supplementation for 8 weeks. The results showed that OVX without or with MIA injection induced various severity levels of menopausal OA by increasing pathological damage, oxidative stress, and cartilage matrix degradation to various degrees. Moreover, S-equol supplementation could significantly reduce these increased biomarkers in different severity levels of OA. This indicates that S-equol can lessen menopausal OA progression by reducing oxidative stress and the matrix-degrading enzymes involved in cartilage degradation. Full article

Background: Significant improvement in neonatal care has enabled increasing survival of preterm infants. Metabolic bone disease of prematurity is often overlooked due to other comorbidities of preterm birth. The best approach is screening and prevention of the disease in high-risk infants such as preterm infants. Aim: We followed up the clinical, radiological, and serum biochemical markers of metabolic bone disease in extremely preterm infants (<28 weeks of gestation). The clinical applicability and validation of C-terminal telopeptide of type I collagen (CTX-I) as a novel bone turnover marker were assessed. Standard and novel biochemical bone turnover markers and quantitative ultrasound were compared. Method: Patients’ data were collected from medical records. Assessments of calcium, phosphate, alkaline phosphatase, bone-alkaline phosphatase, CTX-I, and quantitative ultrasound were prospectively performed twice in 42 extremely preterm infants at postmenstrual ages of 30–32 weeks and 36–40 weeks. Bone mineral density was measured by quantitative ultrasound. Conclusion: Phosphate, alkaline phosphatase, bone alkaline phosphatase, calcium, or ionized calcium are not related to gestational age, but bone mineral density, measured by quantitative ultrasound, is related. There is no correlation between standard and novel biochemical markers and quantitative ultrasound for the identification of metabolic bone disease. Full article

The role of bone and muscle as endocrine organs may be important contributing factors for children’s growth and development. Myokines, secreted by muscle cells, play a role in regulating bone metabolism, either directly or indirectly. Conversely, markers of bone metabolism, reflecting the balance between bone formation and bone resorption, can also influence myokine secretion. This study investigated a panel of serum myokines and their relationships with bone metabolism markers in children following vegetarian and omnivorous diets. A cohort of sixty-eight healthy prepubertal children, comprising 44 vegetarians and 24 omnivores, participated in this study. Anthropometric measurements, dietary assessments, and biochemical analyses were conducted. To evaluate the serum concentrations of bone markers and myokines, an enzyme-linked immunosorbent assay (ELISA) was used. The studied children did not differ regarding their serum myokine levels, except for a higher concentration of decorin in the vegetarian group (p = 0.020). The vegetarians demonstrated distinct pattern of bone metabolism markers compared to the omnivores, with lower levels of N-terminal propeptide of type I procollagen (P1NP) (p = 0.001) and elevated levels of C-terminal telopeptide of type I collagen (CTX-I) (p = 0.018). Consequently, the P1NP/CTX-I ratio was significantly decreased in the vegetarians. The children following a vegetarian diet showed impaired bone metabolism with reduced bone formation and increased bone resorption. Higher levels of decorin, a myokine involved in collagen fibrillogenesis and essential for tissue structure and function, may suggest a potential compensatory mechanism contributing to maintaining bone homeostasis in vegetarians. The observed significant positive correlations between myostatin and bone metabolism markers, including P1NP and soluble receptor activator of nuclear factor kappa-B ligand (sRANKL), suggest an interplay between muscle and bone metabolism, potentially through the RANK/RANKL/OPG signaling pathway. Full article

Laying hens can experience a progressive increase in bone fragility due to the ongoing mobilization of calcium from bones for eggshell formation. Over time, this escalates their susceptibility to bone fracture, which can reduce their mobility and cause pain. The provision of perches as an exercise opportunity could potentially enhance bone strength, but the timing of exposure to perches during the birds’ development may modulate its impact. The objective of this study was to investigate the enduring impacts of perch provision timing on the musculoskeletal health of laying hens. A total of 812 pullets were kept in different housing conditions (seven pens/treatment, 29 birds/pen) with either continuous access to multi-tier perches from 0 to 40 weeks of age (CP), no access to perches (NP), early access to perches during the rearing phase from 0 to 17 weeks of age (EP), or solely during the laying phase from 17 to 40 weeks of age (LP). At weeks 24, 36, and 40 of age (n = 84 birds/week), three birds per pen were monitored for individual activity level, and blood samples were collected from a separate set of three birds per pen to analyze serum levels of tartrate-resistant acid phosphatase 5b (TRACP-5b) and C-terminal telopeptide of type I collagen (CTX-I) as markers of bone demineralization. At 40 weeks of age, three birds per pen (n = 84) were euthanized for computed tomography scans to obtain tibial bone mineral density (BMD) and cross-sectional area (CSA) with further analysis including muscle deposition, tibial breaking strength, and tibial ash percent. During week 24, hens from CP, EP, and LP pens had the highest overall activity compared to hens from NP pens (p < 0.05) with no differences between treatments for overall activity level during weeks 36 or 40 (p > 0.05). During weeks 24, 36, and 40, hens from CP and LP pens showed greater vertical and less horizontal activity compared to hens from EP and NP pens (p < 0.05). TRACP-5b and CTX-I concentrations did not differ between treatments at week 24 of age (p > 0.05). Hens from CP pens had the lowest TRACP-5b and CTX-I concentrations at 36 weeks of age with EP and LP hens showing intermediate responses and NP hens having the highest concentration (p < 0.05). At 40 weeks of age, CP hens had the lowest TRACP-5b and CTX-I concentrations compared to NP hens (p < 0.05). Total bone CSA did not differ between treatments (p > 0.05), but CP had greater total BMD than NP (p < 0.05) with no differences between EP and LP treatments. CP and LP hens had larger biceps brachii, pectoralis major, and leg muscle groups as well as greater tibial breaking strengths than EP and NP treatments (p < 0.05). CP hens had higher tibial ash percentages compared to EP, LP, and NP (p < 0.05). Our results indicate that providing continuous perch access improves the musculoskeletal health and activity of laying hens at 40 weeks of age compared to no access and that late access to perches has a beneficial impact on activity, muscle deposition, and bone strength. Full article

The castration of stallions is traditionally performed after puberty, at around the age of 2 years old. No studies have focused on the effects of early castration on osteoarticular metabolism. Thus, we aimed to compare early castration (3 days after birth) with traditional castration (18 months of age) in horses. Testosterone and estradiol levels were monitored from birth to 33 months in both groups. We quantified the levels of biomarkers of cartilage and bone anabolism (CPII and N-MID) and catabolism (CTX-I and CTX-II), as well as of osteoarthritis (HA and COMP) and inflammation (IL-6 and PGE₂). We observed a lack of parallelism between testosterone and estradiol synthesis after birth and during puberty in both groups. The extra-gonadal synthesis of steroids was observed around the 28-month mark, regardless of the castration age. We found the expression of estrogen receptor (ESR1) in cartilage and bone, whereas androgen receptor (AR) expression appeared to be restricted to bone. Nevertheless, with respect to osteoarticular metabolism, steroid hormone deprivation resulting from early castration had no discernable impact on the levels of biomarkers related to bone and cartilage metabolism, nor on those associated with OA and inflammation. Consequently, our research demonstrated that early castration does not disrupt bone and cartilage homeostasis. Full article

Nitisinone has been approved for treatment of alkaptonuria (AKU). Non-invasive biomarkers of joint tissue remodelling could aid in understanding the molecular changes in AKU pathogenesis and how these can be affected by treatment. Serological and urinary biomarkers of type I collagen and II collagen in AKU were investigated in patients enrolled in the randomized SONIA 2 (NCT01916382) clinical study at baseline and yearly until the end of the study (Year 4). The trajectories of the biomarkers over time were observed. After treatment with nitisinone, the biomarkers of type I collagen remodelling increased at Year 1 (19% and 40% increase in CTX-I and PRO-C1, respectively), which was potentially reflected in the higher degree of mobility seen following treatment. The biomarkers of type II collagen remodelling decreased over time in the nitisinone group: C2M showed a 9.7% decline at Year 1, and levels then remained stable over the following visits; CTX-II showed a 26% decline at Year 3 and 4 in the nitisinone-treated patients. Nitisinone treatment induced changes in biomarkers of bone and cartilage remodelling. These biomarkers can aid patient management and deepen our knowledge of the molecular mechanisms of this rare disease. Full article

The extension of human life makes it more and more important to prevent and treat diseases of the elderly, including Alzheimer’s disease (AD) and osteoporosis. Little is known about the effects of drugs used in the treatment of AD on the musculoskeletal system. The aim of the present study was to investigate the effects of donepezil, an acetylcholinesterase inhibitor, on the musculoskeletal system in rats with normal and reduced estrogen levels. The study was carried out on four groups of mature female rats: non-ovariectomized (NOVX) control rats, NOVX rats treated with donepezil, ovariectomized (OVX) control rats and OVX rats treated with donepezil. Donepezil (1 mg/kg p.o.) was administered for four weeks, starting one week after the ovariectomy. The serum concentrations of CTX-I, osteocalcin and other biochemical parameters, bone mass, density, mineralization, histomorphometric parameters and mechanical properties, and skeletal muscle mass and strength were examined. Estrogen deficiency increased bone resorption and formation and worsened cancellous bone mechanical properties and histomorphometric parameters. In NOVX rats, donepezil decreased bone volume to tissue volume ratio in the distal femoral metaphysis, increased the serum phosphorus concentration and tended to decrease skeletal muscle strength. No significant bone effects of donepezil were observed in OVX rats. The results of the present study indicate slightly unfavorable effects of donepezil on the musculoskeletal system in rats with normal estrogen levels. Full article

This study was conducted to evaluate the effects of phytase supplementation in low-phosphorus diets on the production performance, phosphorus–calcium metabolism, and bone metabolism in laying hens from 69 to 78 weeks of age. Hy-Line Brown laying hens (n = 1350) were assigned randomly to six treatments with five replicates of 45 birds. A corn–soybean meal-based diet with no inorganic phosphates was formulated to contain 0.12% non-phytate phosphorus (NPP) and 1470 FTU/kg phytase (Released phytate phosphorus content ≥ 0.1%). Inorganic phosphorus (dicalcium phosphate) was supplemented into the basal diet to construct five test diets (level of NPP supplementation = 0.10%, 0.15%, 0.20%, 0.25%, and 0.30%). The level of calcium carbonate was adjusted to ensure that all six experimental diets contained the same calcium percentage (3.81%). The feeding trial lasted 10 weeks (hens from 69 to 78 weeks of age). Upon supplementation with phytase (1470 FTU/kg), supplemental inorganic phosphates (dicalcium phosphate) had no significant effects (p > 0.05) on the production performance or egg quality. Significant differences in serum levels of calcium, phosphorus, copper, iron, zinc, or manganese were not detected across treatments (p > 0.05). Hens fed NPP (0.15%, 0.20%, 0.25%, and 0.30%) had higher levels (p < 0.0001) of tibial ash, calcium, and phosphorus than those not fed inorganic phosphates. The tibial breaking strength of the group without inorganic phosphates was significantly lower than that of the other groups (p < 0.01). Dietary supplementation with inorganic phosphates had no effect (p > 0.05) on serum levels of calcitonin (CT) and 1,25-dihydroxy-vitamin D3 (1,25-(OH)2D3). Hens that did not receive supplementation with inorganic phosphates had higher serum levels of parathyroid hormone (PTH), osteoprotegerin (OPG), type-I collagen c-telopeptide (CTX-I), and tartrate-resistant acid phosphatase 5b (TRACP-5b) compared with those in the other groups (p < 0.01). Serum levels of CTX-I and TRACP-5b were significantly lower in the NPP-supplementation groups of 0.25% and 0.30% than in the 0.10% NPP-supplementation group (p < 0.01). Dietary supplementation with inorganic phosphates had no effect (p > 0.05) on serum levels of bone-alkaline phosphatase (BAP), osteocalcin (OCN), or osteopontin (OPN). Hens not fed inorganic phosphate had the highest renal expression of phosphorus transporter type IIa Na/Pi cotransporter (NaPi-Ⅱa). Renal expression of NaPi-Ⅱa was increased significantly in NPP-supplementation groups of 0.10–0.20% compared with that in NPP-supplementation groups of 0.25% and 0.30% (p < 0.0001). The results indicated that a reduction in NPP supplementation to 0.15% (dietary NPP level = 0.27%) with phytase inclusion did not have an adverse effect on the production performance or bone health of laying hens from 69 to 78 weeks of age, which might be attributed to renal phosphorus reabsorption and bone resorption. These findings could support the application of low-phosphorus diets in the poultry industry. Full article