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BACKGROUND: Many studies have found that the pathological progression of steroid-induced osteonecrosis of the femoral head is related to abnormal proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells, in which a disintegrin and metalloproteases 10 (ADAM10) is highly involved, but the specific mechanism remains unclear. OBJECTIVE: To investigate the effects of ADAM10 on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells derived from steroidinduced osteonecrosis of the femoral head patients and its possible mechanism. METHODS: Bone marrow was harvested from proximal femur in 23 patients with steroid-induced osteonecrosis of the femoral head and 17 patients with femoral neck fracture under sterile conditions. Bone marrow mesenchymal stem cells from steroid-induced osteonecrosis of the femoral head (S-BMSCs) and femoral neck fracture (F-BMSCs) were isolated by density gradient centrifugation, and cultured to the third generation in vitro. The proliferation activity of cells in the two groups was detected by CCK-8 assay. Alizarin red staining and alkaline phosphatase staining were used to observe the osteogenic differentiation ability of cells in the two groups. qRT-PCR and western blot assay were used to detect the levels of ADAM10 mRNA and protein in cells of the two groups. S-BMSCs were infected with ADAM10 overexpression lentivirus vector, and then treated with Notch signaling pathway inhibitor DAPT. The proliferation activity of these cells was detected by CCK-8 assay. Alizarin red staining and alkaline phosphatase staining were used to observe the osteogenic differentiation of cells. qRT-PCR was used to detect the expression levels of osteogenic markers alkaline phosphatase, Runt-related transcription factor 2, and osteocalcin mRNA. The expression levels of Notch signaling pathway related proteins Notch1, NICD, and Hes1 were detected by western blot assay. RESULTS AND CONCLUSION: (1) The proliferation activity, osteogenic differentiation ability and the expression level of ADAM10 in S-BMSCs were significantly lower than those in F-BMSCs (P < 0.01). (2) After ADAM10 overexpression, the proliferation activity, osteogenic differentiation ability, the expression levels of alkaline phosphatase, Runt-related transcription factor 2, and osteocalcin mRNA and Notch1, NICD and Hes1 protein of S-BMSCs were significantly increased (P < 0.05). However, combined intervention with DAPT significantly inhibited the promoting effects of ADAM10 overexpression on proliferation activity and osteogenic differentiation ability of S-BMSCs, and inhibited the activation of Notch signaling pathway. (3) The results show that ADAM10 can be underexpressed in S-BMSCs, and its overexpression can enhance the proliferation and osteogenic differentiation ability of S-BMSCs, and its mechanism may be related to the activation of Notch signaling pathway. [ABSTRACT FROM AUTHOR]

Introduction: Osteocalcin (OC) and its effect on insulin resistance in animals have been studied and demonstrated. But, the same is yet to be established in humans. Moreover, the association of Gestational Diabetes Mellitus (GDM) with serum Osteocalcin (OC) levels hasn't been studied much and hence, is an area of scope for research. This study is aimed at assessing the difference between serum OC levels in non-diabetic pregnant women (Group I - Controls) and women diagnosed with GDM (Group II - Cases) and determine whether they can be used in early prediction of GDM. Methods: A thorough and systematic search of PUBMED and SCOPUS databases was done to identify relevant articles, using the keywords Osteocalcin, pregnancy, diabetes and gestational diabetes. Studies which had included atleast 10 study participants per group and had measured serum OC levels in various forms viz., undercarboxylated Osteocalcin (ucOC), N terminal Mid fragment Osteocalcin (N MID OC) and total Osteocalcin (tOC) were considered relevant. Out of 45 studies, 9 studies were selected for the present study. Results: There was no significant difference in the mean ucOC and N MID OC levels between the GDM and nondiabetic controls, whereas, a significant difference was found in the tOC levels between the two groups. Conclusion: Thus, this meta-analysis shows that tOC may be used as potential marker for GDM rather than ucOC and N MID OC. [ABSTRACT FROM AUTHOR]

Uncarboxylated osteocalcin (GluOC), a small-molecule protein specifically synthesized and secreted by osteoblasts, is important in the regulation of energy metabolism. In our previous study, GluOC was shown to be effective in ameliorating dyslipidemia and hepatic steatosis in KKAy mice. However, the underlying mechanism of GluOC action on hepatocytes has not been well validated. In this study, oleic acid/palmitic acid (OA/PA)-induced HepG2 and NCTC 1469 cells were used as non-alcoholic fatty liver disease (NAFLD) cell models, and triacylglycerol (TG) levels were measured by oil red O staining, Nile Red staining, and ELISA. The fatty acid synthesis-related protein expression was detected by real-time quantitative polymerase chain reaction, Western blotting, and immunofluorescence. The results show that GluOC reduced triglyceride levels, and decreased the expression of sterol regulatory element-binding protein-1c (SREBP-1c) and stearyl-coenzyme A desaturase 1 (SCD1). si-SCD1 mimicked the lipid accumulation-reducing effect of GluOC, while overexpression of SCD1 attenuated the effect of GluOC. In addition, GluOC activated AMP-activated protein kinase (AMPK) phosphorylation to affect lipid metabolism in hepatocytes. Overall, the results of this study suggest that GluOC decreases SCD1 by activating AMPK to alleviate hepatocyte lipid accumulation, which provides a new target for improving NAFLD in further research. [ABSTRACT FROM AUTHOR]

This work describes the design, preparation, and deep investigation of "intelligent nanobiomaterials" that fulfill the safety rules and aim to serve as "signal deliverers" for osteogenesis, harboring a specific peptide that promotes and enhances osteogenesis at the end of their hydrogel fibers. The de novo synthesized protein fibers, besides their mechanical properties owed to their protein constituents from elastin, silk fibroin and mussel-foot adhesive protein-1 as well as to cell-attachment peptides from extracellular matrix glycoproteins, incorporate the Bone Morphogenetic Protein-2 (BMP2) peptide (AISMLYLDEN) that, according to our studies, serves as "signal deliverer" for osteogenesis. The osteogenetic capacity of the biomaterial has been evidenced by investigating the osteogenic marker genes ALP, RUNX2, Osteocalcin, COL1A1, BMPR1A, and BMPR2, which were increased drastically in cells cultured on scaffold-BMP2 for 21 days, even in the absence of osteogenesis medium. In addition, the induction of phosphorylation of intracellular Smad-1/5 and Erk-1/2 proteins clearly supported the osteogenetic capacity of the biomaterial. [ABSTRACT FROM AUTHOR]

The clinical application of biochemical indicators of bone metabolism provides molecular biological basis for the diagnosis， differential diagnosis， prediction of fracture risk and evaluation of therapeutic effect of anti-osteoporosis treatment of osteoporosis， and has important significance in the epidemiological study of osteoporosis， pathogenesis， and development and research of osteoporosis drugs. Because of the high specificity and sensitivity of bone metabolism biochemical indicators， its application is increasingly widespread. This paper has searched a large number of Chinese and foreign literatures， compiled and reviewed the Expert Consensus on Clinical Application of Bone Metabolism Biochemical Indicators （2023 Revision）， and discussed the classification of bone metabolism biochemical indicators， the methodology and biological significance of bone metabolism biochemical indicators， and the detection and variation of bone metabolism indicators. [ABSTRACT FROM AUTHOR]

Background: The aim of this study was to investigate and compare the cytotoxicity and gene expression of Bio-C Repair, Mineral Trioxide Aggregate (MTA) HP Repair, and Biodentine on stem cells derived from exfoliated deciduous teeth. Materials and Methods: In this in vitro study MTT assay was used to assess the cellular viability at three different dilutions. The gene expression of Runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), osteocalcin [OCN], and dentin matrix protein-1 (DMP-1) was measured with real-time polymerase chain reaction after 7 days, 14 days, and 21 days of incubation. One-way analysis of variance and Bonferroni posttest were used for statistical analysis (p=o.o5). Results: After 72 h of incubation at dilution 1:4, stem cells derived from human exfoliated deciduous teeth (SHEDs) cultivated in Biodentine, followed by Bio-C Repair and MTA Repair HP reported with highest cellular viability. The highest mRNA expression of Runx2, ALP, OCN, and DMP-1 was reported in SHEDs cultured in Biodentine (after 21 days of incubation). Conclusion: Bio-C Repair and MTA HP Repair are biocompatible and capable of odontogenic differentiation similar to Biodentine when cultured in stem cells derived from exfoliated primary teeth. [ABSTRACT FROM AUTHOR]

The study aims to investigate the quality of dentin barriers and pulp reaction to EndoSequence Root Repair Material (ERRM) combined with low‐level laser application. In eight dogs, pulps were exposed via class V, half of the samples received low‐level diode laser at 870 nm. Thereafter, cavities were capped with fast‐set or regular‐set ERRM. The specimens were processed for histomorphological and immunohistochemical examination after 2 weeks and 2 months. Dentin bridges were observed in all samples, and 87.5% were complete. The low‐level laser group had significantly more reparative dentin area than the non‐lased group (p < 0.05). The dentin bridges were found to have an unprecedented tubularity of 43%–89%. Tiny dentin island formation was observed within the material particles. Initial mild‐to‐moderate inflammatory reactions were observed, which subsided after 2 months. RUNX2 and osteocalcin staining were evident for all samples at both time intervals. Low‐level laser combined with bioactive ERRM is effective in inducing reparative dentin formation. [ABSTRACT FROM AUTHOR]

Background and purpose: Chronic liver diseases are associated with increased bone fracture risk, mostly in end-stage disease and cirrhosis; besides, data in non-alcoholic fatty liver disease (NAFLD) are limited. Aim of this study was to investigate bone mineralization and microstructure in obese individuals with NAFLD in relation to the estimated liver fibrosis. Methods: For this cross-sectional investigation, we analyzed data from 1872 obese individuals (44.6 ± 14.1 years, M/F: 389/1483; BMI: 38.3 ± 5.3 kg/m2) referring to the Endocrinology outpatient clinics of Sapienza University, Rome, Italy. Participants underwent clinical work-up, Dual-Energy X-ray Absorptiometry for assessing bone mineral density (BMD) and microarchitecture (trabecular bone score, TBS). Liver fibrosis was estimated by Fibrosis Score 4 (FIB-4). Serum parathyroid hormone (PTH), 25(OH) vitamin D, osteocalcin and IGF-1 levels were measured. Results: Obese individuals with osteopenia/osteoporosis had greater FIB-4 than those with normal BMD (p < 0.001). FIB-4 progressively increased in presence of degraded bone microarchitecture (p < 0.001) and negatively correlated with the serum osteocalcin (p < 0.001) and IGF-1 (p < 0.001), which were both reduced in presence of osteopenia/osteoporosis. FIB-4 predicted IGF-1 reduction in multivariable regression models adjusted for confounders (β: − 0.18, p < 0.001). Higher FIB-4 predicted bone fragility with OR 3.8 (95%C.I:1.5–9.3); this association persisted significant after adjustment for sex, age, BMI, diabetes, smoking status and PTH at the multivariable logistic regression analysis (OR 1.91 (95%C.I:1.15–3.17), p < 0.01), with AUROC = 0.842 (95%C.I:0.795–0.890; p < 0.001). Conclusion: Our data indicate the presence of a tight relation between NAFLD-related liver fibrosis, lower bone mineral density and degraded microarchitecture in obese individuals, suggesting potential common pathways underlying liver and bone involvement in obesity and insulin resistance-associated disorders. [ABSTRACT FROM AUTHOR]

This systematic review and meta-analysis was performed to compare the alterations in bone turnover markers between SG and RYGB. A literature search was conducted in PubMed, Medline, Scopus, Web of Science, and the Cochrane Central Register of Controlled Trials (CENTRAL) databases to find the studies. There was significant less increment in osteocalcin [WMD = − 5.98, 95% CI (− 9.30, − 2.47) P < 0.01] and parathyroid hormone (PTH) [WMD = − 9.59, 95% CI (− 15.02, − 4.16) P < 0.01] in the SG group compared to the RYGB group. No significant differences were seen in change of C-terminal telopeptide of type I collagen (CTX), N-terminal propeptide of type I collagen (PINP), Ca, and 25(OH)-D between SG and RYGB groups. According to our meta-analysis, bone formation markers appear to have more increment following RYGB than SG. This observation is accompanied by a larger increase in PTH after RYGB patients compared to SG patients. PROSPERO: CRD42022308985. [ABSTRACT FROM AUTHOR]

Introduction: The bone matrix protein osteocalcin (OC), secreted by osteoblasts, displays endocrine effects. We tested the hypothesis that OC modulates parathyroid tumor cell function. Methods: Primary cell cultures derived from parathyroid adenomas (PAds) and HEK293 cells transiently transfected with the putative OC receptor GPRC6A or the calcium sensing receptor (CASR) were used as experimental models to investigate g-carboxylated OC (GlaOC) or uncarboxylated OC (GluOC) modulation of intracellular signaling. Results: In primary cell cultures derived from PAds, incubation with GlaOC or GluOC modulated intracellular signaling, inhibiting pERK/ERK and increasing active b-catenin levels. GlaOC increased the expression of PTH, CCND1 and CASR, and reduced CDKN1B/p27 and TP73. GluOC stimulated transcription of PTH, and inhibited MEN1 expression. Moreover, GlaOC and GluOC reduced staurosporin-induced caspase 3/7 activity. The putative OC receptor GPRC6A was detected in normal and tumor parathyroids at membrane or cytoplasmic level in cells scattered throughout the parenchyma. In PAds, the membrane expression levels of GPRC6A and its closest homolog CASR positively correlated; GPRC6A protein levels positively correlated with circulating ionized and total calcium, and PTH levels of the patients harboring the analyzed PAds. Using HEK293A transiently transfected with either GPRC6A or CASR, and PAds-derived cells silenced for CASR, we showed that GlaOC and GluOC modulated pERK/ERK and active b-catenin mainly through CASR activation. Conclusion: Parathyroid gland emerges as a novel target of the bone secreted hormone osteocalcin, which may modulate tumor parathyroid CASR sensitivity and parathyroid cell apoptosis. [ABSTRACT FROM AUTHOR]

Osteocalcin (Ocn), also known as bone Gla protein, is synthesized by osteoblasts and thought to regulate energy metabolism, testosterone synthesis and brain development. However, its function in bone is not fully understood. Mice have three Ocn genes: Bglap, Bglap2 and Bglap3. Due to the long span of these genes in the mouse genome and the low expression of Bglap3 in bone, researchers commonly use Bglap and Bglap2 knockout mice to investigate the function of Ocn. However, it is unclear whether Bglap3 has any compensatory mechanisms when Bglap and Bglap2 are knocked out. Considering the controversy surrounding the role of Ocn in bone, we constructed an Ocn-deficient mouse model by knocking out all three genes (Ocn-/-) and analyzed bone quality by Raman spectroscopy (RS), Scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and MicroCT (μCT). The RS test showed that the alignment of hydroxyapatite crystals and collagen fibers was significantly poorer in Ocn-/- mice than in wild-type (WT) mice. Ocn deficiency resulted in a looser surface structure of bone particles and a larger gap area proportion. FTIR analysis showed few differences in bone mineral index between WT and Ocn-/- mice, while μCT analysis showed no significant difference in cortical and trabecular regions. However, under tail-suspension simulating bone loss condition, the disorder of hydroxyapatite and collagen fiber alignment in Ocn-/- mice led to more obvious changes in bone mineral composition. Collectively, our results revealed that Ocn is necessary for regulating the alignment of minerals parallel to collagen fibrils. [ABSTRACT FROM AUTHOR]

Conventional intake of drugs and active substances is most often based on oral intake of an appropriate dose to achieve the desired effect in the affected area or source of pain. In this case, controlling their distribution in the body is difficult, as the substance also reaches other tissues. This phenomenon results in the occurrence of side effects and the need to increase the concentration of the therapeutic substance to ensure it has the desired effect. The scientific field of tissue engineering proposes a solution to this problem, which creates the possibility of designing intelligent systems for delivering active substances precisely to the site of disease conversion. The following review discusses significant current research strategies as well as examples of polymeric and composite carriers for protein and non-protein biomolecules designed for bone tissue regeneration. [ABSTRACT FROM AUTHOR]

Background: Serum osteocalcin levels are closely related to metabolic syndrome and cardiovascular disease. This study aimed to investigate the relationship between serum osteocalcin levels and cardiometabolic risk factors in patients with type 2 diabetes (T2D) according to age and sex. Methods: This cross-sectional study included 1500 patients with T2D (991 men and 509 women) aged ≥ 18 years old. The age- and sex-specific disparities in glycemic and lipid control, as well as cardiometabolic risk factors were evaluated. Results: The levels of serum osteocalcin were significantly higher in women aged > 50 years compared with women aged ≤ 50 years (15.6 ± 6.5 ng/mL vs. 11.3 ± 4.5 ng/mL, p < 0.0001). However, this was lower in men aged > 50 years than men aged ≤ 50 years (12.2 ± 4.2 ng/mL vs. 12.9 ± 4.3 ng/mL, p = 0.0081). We performed correlation analyses of serum osteocalcin and cardiometabolic parameters. Serum osteocalcin concentrations were negative associated with FBG and HbA1c levels in women and men ≤ 50 years old, but not in men aged > 50 years old. Serum osteocalcin were negatively correlated with TG and positively correlated with HDL-C and LDL-C only in men aged ≤ 50 years. In binary logistic regression analysis, serum osteocalcin levels were associated with multiple cardiovascular risk factors, as follows: overweight/obese (odds ratio [OR], 0.944; 95% confidence interval [CI], 0.9–0.991, p = 0.02) in men aged > 50 years; high HbA1C and high FBG in women and men aged ≤ 50 years, but not in men aged > 50 years; after adjustment for confounding factors, high TG (OR, 0.905; 95% CI 0.865–0.947, p < 0.0001), metabolic syndrome (OR, 0.914; 95% CI 0.874–0.956, p < 0.0001), and low high-density lipoprotein cholesterol (OR, 0.933; 95% CI, 0.893–0.975, p = 0.002) were seen in men aged ≤ 50 years only. Conclusions: Serum osteocalcin level has significant relationships with cardiometabolic risk factors and several age- and sex-related differences in patients with T2D. Decreased serum osteocalcin levels are associated with a worse cardiometabolic risk profile. [ABSTRACT FROM AUTHOR]

Control of tissue metabolism and growth involves interactions between organs, tissues, and cell types, mediated by cytokines or direct communication through cellular exchanges. Indeed, over the past decades, many peptides produced by adipose tissue, skeletal muscle and bone named adipokines, myokines and osteokines respectively, have been identified in mammals playing key roles in organ/tissue development and function. Some of them are released into the circulation acting as classical hormones, but they can also act locally showing autocrine/paracrine effects. In recent years, some of these cytokines have been identified in fish models of biomedical or agronomic interest. In this review, we will present their state of the art focusing on local actions and inter-tissue effects. Adipokines reported in fish adipocytes include adiponectin and leptin among others. We will focus on their structure characteristics, gene expression, receptors, and effects, in the adipose tissue itself, mainly regulating cell differentiation and metabolism, but in muscle and bone as target tissues too. Moreover, lipid metabolites, named lipokines, can also act as signaling molecules regulating metabolic homeostasis. Regarding myokines, the best documented in fish are myostatin and the insulin-like growth factors. This review summarizes their characteristics at a molecular level, and describes both, autocrine effects and interactions with adipose tissue and bone. Nonetheless, our understanding of the functions and mechanisms of action of many of these cytokines is still largely incomplete in fish, especially concerning osteokines (i.e., osteocalcin), whose potential cross talking roles remain to be elucidated. Furthermore, by using selective breeding or genetic tools, the formation of a specific tissue can be altered, highlighting the consequences on other tissues, and allowing the identification of communication signals. The specific effects of identified cytokines validated through in vitro models or in vivo trials will be described. Moreover, future scientific fronts (i.e., exosomes) and tools (i.e., co-cultures, organoids) for a better understanding of inter-organ crosstalk in fish will also be presented. As a final consideration, further identification of molecules involved in inter-tissue communication will open new avenues of knowledge in the control of fish homeostasis, as well as possible strategies to be applied in aquaculture or biomedicine. [ABSTRACT FROM AUTHOR]

I previously reported that F-box/leucine-rich repeat protein 14 (FBXL14) expressed in periosteum-derived cells, and F-box and WD-40 domain-containing protein 2 (FBXW2) in the periosteum form a fiber-like structure. Here, two culture medium conditions, that is, media with and without ascorbic acid, were compared during explant culture. In the absence of ascorbic acid, the expression patterns of osteocalcin, FBXW2, and elastin were compared using fluorescent immunostaining during weeks 3–5. By observing the periosteum, cambium layer and bone, I demonstrated FBXL14 expression in micro-vessels and bone lacuna. Fluorescent immunostaining revealed that, without ascorbic acid, the FBXL14 layer was thin. Conversely, in the presence of ascorbic acid, FBXL14 formed a thick membrane-like structure inside the periosteum, and the multilayer of periosteum-derived cells (PDCs) was strong. The expression patterns of osteocalcin and FBXW2 were similar. Elastin retained its fiber structure for up to five weeks. Although osteocalcin and FBXW2 were expressed in regions similar to elastin, they could not retain their fiber structures. In conclusion, FBXL14 appears to play a role in preparing a native scaffold for forming a multilayered sheet of PDCs inside the periosteum. FBXW2 and osteocalcin appear to separate from elastic fibers during calcification. [ABSTRACT FROM AUTHOR]

Elastic fibers form vessel walls, and elastic fiber calcification causes serious vascular diseases. Elastin is a well-known elastic fiber component; however, the insoluble nature of elastic fibers renders elastic fiber component analysis difficult. A previous study investigated F-box and WD-40 domain-containing protein 2 (FBXW2) in the cambium layer of bovine periosteum and hypothesized that fiber structures of FBXW2 are coated with osteocalcin during explant culture. Here, FBXW2 was expressed around some endothelial cells but not in all microvessels of the bovine periosteum. The author hypothesized that FBXW2 is expressed only in blood vessels with elastic fibers. Immunostaining and Elastica van Gieson staining indicated that FBXW2 was expressed in the same regions as elastic fibers and elastin in the cambium layer of the periosteum. Alpha-smooth muscle actin (αSMA) was expressed in microvessels and periosteum-derived cells. Immunostaining and observation of microvessels with serial sections revealed that osteocalcin was not expressed around blood vessels at 6 and 7 weeks. However, blood vessels and periosteum connoted elastic fibers, FBXW2, and αSMA. These findings are expected to clarify the processes involved in the calcification of elastic fibers in blood vessels. [ABSTRACT FROM AUTHOR]

Background: The interplay between vitamin K (vitK) (as carboxylation cofactor, partially produced by the gut microbiota) and short-chain fatty acids (SCFAs), the end-product of fiber fermentation in the gut, has never been assessed in mother–newborn pairs, although newborns are considered vitK deficient and with sterile gut. Methods: We collected venous blood from 45 healthy mothers with uncomplicated term pregnancies and umbilical cord blood from their newborns at birth. The concentrations of total SCFAs and hepatic/extra-hepatic vitK-dependent proteins (VKDPs), as proxies of vitK status were assayed: undercarboxylated and total matrix Gla protein (ucMGP and tMGP), undercarboxylated osteocalcin (ucOC), undercarboxylated Gla-rich protein (ucGRP), and protein induced by vitK absence II (PIVKA-II). Results: We found significantly higher ucOC (18.6-fold), ucMGP (9.2-fold), and PIVKA-II (5.6-fold) levels in newborns, while tMGP (5.1-fold) and SCFAs (2.4-fold) were higher in mothers, and ucGRP was insignificantly modified. In mother–newborn pairs, only ucGRP (r = 0.746, p < 0.01) and SCFAs (r = 0.428, p = 0.01) levels were correlated. Conclusions: We report for the first time the presence of SCFAs in humans at birth, probably transferred through the placenta to the fetus. The increased circulating undercarboxylated VKDPSs in newborns revealed a higher vitamin K deficiency at the extrahepatic level compared to liver VKDPs. [ABSTRACT FROM AUTHOR]

Osteocalcin (OC), an abundant non-collagenous protein in bone extracellular matrix, plays a vital role in both its biological and mechanical function. OC undergoes post-translational modification, such as glycation; however, it remains unknown whether glycation of OC affects bone's resistance to fracture. Here, for the first time, we demonstrate the formation of pentosidine, an advanced glycation end-product (AGE) cross-link on mouse OC analyzed by ultra-performance liquid chromatography. Next, we establish that the presence of OC in mouse bone matrix is associated with lower interlamellar separation (distance) and thicker bridges spanning the lamellae, both of which are critical for maintaining bone's structural integrity. Furthermore, to determine the impact of modification of OC by glycation on bone toughness, we glycated bone samples in vitro from wild-type (WT) and osteocalcin deficient (Oc−/−) mice, and compared the differences in total fluorescent AGEs and fracture toughness between the Oc−/− glycated and control mouse bones and the WT glycated and control mouse bones. We determined that glycation resulted in significantly higher AGEs in WT compared to Oc−/− mouse bones (delta-WT > delta-OC, p = 0.025). This observed change corresponded to a significant decrease in fracture toughness between WT and Oc−/− mice (delta-WT vs delta-OC, p = 0.018). Thus, we propose a molecular deformation and fracture mechanics model that corroborates our experimental findings and provides evidence to support a 37%–90% loss in energy dissipation of OC due to formation of pentosidine cross-link by glycation. We anticipate that our study will aid in elucidating the effects of a major non-collagenous bone matrix protein, osteocalcin, and its modifications on bone fragility and help identify potential therapeutic targets for maintaining skeletal health. [ABSTRACT FROM AUTHOR]

Objective: o study the serum insulin-like growth factor-binding protein 3 (IGFBP-3), serum N-terminal osteocalcin (N-MID), 25-hydroxyvitamin D (25(OH)D), bone mineral density in osteoporosis patients (BMD) detection effect. Methods: Data were collected from 80 patients with osteoporosis who were admitted to our hospital from April 2021 to May 2022. Results: The levels of IGFBP-3, 25(OH)D, N-MID, and BMD in the research group were lower than those in the reference group (P<0.05). 25(OH)D, N-MID, and BMD levels were positively correlated with each other (P<0.05). Multiple regression analysis showed that among the main factors affecting BMD, IGFBP-3, 25(OH) D and N-MID are both protective factors for osteoporosis. Conclusion: The detection of IGFBP-3, 25(OH)D and N-MID in patients with osteoporosis can effectively observe the disease and provide reference data for disease treatment, which is beneficial to improve the prognosis and has a significant effect. [ABSTRACT FROM AUTHOR]

Osteocalcin (OC) is a type of multiple peptide hormone formed and secreted by osteoblasts, which is closely related to bone formation and is clinically used as a marker of bone formation. Recently, it has been found that OC regulates many philological and developmental activities, including brain development, nervous construction, and cognitive function, through its active form, non-carboxylated form. It is known that non-carboxylated OC may penetrate blood-brain barrier and concentrated in the brainstem, thalamus, and hypothalamus. After specific binding with serotoninergic neuron in the post raphe nuclei and raphe nuclei, OC stimulates seronine formation, calcium flow, and frequency of action potential. Meanwhile, exercise improves cognitive function and is involved in the prevention of dementia through OC. Different type of exercise may cause different change of OC. In this review, we summarize the advances in the effect and regulation of OC on cognitive function, aiming to provide a basis for further exploration of exercise in promotion of brain health. [ABSTRACT FROM AUTHOR]

Background/purpose: To investigate the surface characteristics of titanium (Ti) implant materials, which were coated with different thicknesses of nanoscale tin oxide (SnO2) using the atomic layer deposition technique, and evaluated its biological performance on human embryonic palatal mesenchyme (HEPM) cells.Methods: The thickness of the coating layer on Ti was 0 (Ti0), 20 nm (Ti20), 50 nm (Ti50), and 100 nm (Ti100), respectively. The surface morphology was observed with an SEM and AFM. The root mean square roughness of micron-scale (mRq) and nanoroughness (nRq) of Ti discs' surface were measured. The Alamar blue (AB) assay and F-actin fluorescence staining were used to evaluate the biocompatibility, and the osteocalcin (OCN) was measured to clarify the differentiation of HEPM cells on materials.Results: In the coating groups, the mRq was decreased, but the nRq was increased. The spreading and polygonal morphology of HEPMs was apparent in coating groups. On Day 4, the survival rate of HEPM cells on Ti0 was higher than on Ti20 and Ti50. There was no significant difference on Day 7, Day 10, and Day 14. The OCN was significantly higher on Day 14 in all the coating groups than Ti0.Conclusion: The results showed that the cell growth was intensified with rough surfaces. However, the OCN and morphology change was prominent when the nanoroughness was increased, which meant the increased nanoroughness might enhance OCN production and improve the tendency of osseointegration. The nanoscale SnO2 coating could increase the ability of bone formation but not cell growth. [ABSTRACT FROM AUTHOR]

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Background: The goal was to investigate the correlations of peripheral blood Omentin-1 and leptin (LEP) levels with bone metabolism and plasma glucose in patients with type 2 diabetes mellitus (T2DM) complicated with osteoporosis (OP). Methods: One hundred patients with T2DM admitted from September 2019 to September 2021 were divided into group A (n = 36, OP with T-score ≤ -2.5), group B (n = 50, osteopenia with T-score between -1 and -2.5), and group C (n = 14, non-OP with T-score > -1) according to the values of bone mineral density (BMD). Thirty healthy adults physically examined in the same period were selected as group D. The levels of peripheral blood Omentin-1 and LEP, bone metabolism, and plasma glucose were compared among the four groups. The correlations of peripheral blood Omentin-1 and LEP levels with bone metabolism and plasma glucose were explored by Pearson's analysis. Results: In group A, the levels of Omentin-1 and LEP in peripheral blood were lowest, the serum levels of beta Cterminal cross-linked telopeptides of type I collagen (β-CTX) and osteocalcin (OCN) were highest, the serum level of total N-terminal propeptide of type I procollagen (tPINP) was lowest, and the levels of fasting plasma glucose (FPG), 2-hours postprandial plasma glucose (2hPG) and glycosylated hemoglobin A1c (HbA1c) were highest, sequentially followed by those of group B, group C, and group D (p < 0.05). Omentin-1 and LEP in peripheral blood were negatively correlated with β-CTX, OCN, 2hPG, and HbA1c and positively correlated with tPINP and FPG (p < 0.05). Conclusions: The expressions of Omentin-1 and LEP in peripheral blood have correlations with bone metabolism and plasma glucose in patients with T2DM complicated with OP. [ABSTRACT FROM AUTHOR]

Hormones are important signaling molecules controlling physiological homeostasis. ELISA kits are commonly used to measure hormones; however, few ELISA kits are multiplex, not all species-specific ELISA kits are commercially available, and ELISA kits typically require a significant volume of biological fluids. Pigs resemble humans in digestive physiology, making them an excellent model in preclinical research of nutrition and metabolism. In this study, we developed and validated a simple liquid–liquid extraction procedure and LC-MS/MS method for the simultaneous quantification of insulin, cortisol, glucagon-like peptide-1 (GLP-1) (7-37) and (7-36), acyl and des-acyl ghrelin, and carboxylated osteocalcin in pig serum. The proposed method is specific, highly sensitive (LOQ in ng/mL and pg/mL), reasonably accurate (more than 76.2% of all quality control samples within 20% error from nominal values), and precise (intra-day CV ≤ 10% and inter-day CV ≤ 23.1%). The recoveries of all analytes and corresponding internal standards ranged from 83.7 to 116.0%. The method also requires a low serum volume of 50–100 μL, which is invaluable when sample volume is limited. These methods could be easily extended for use in other mammalian species.

The aim of this study was to investigate the levels of bone alkaline phosphatase (BALP) and osteocalcin (OC) in the saliva of growing patients of different maturation levels. The sample consisted of 55 patients (34 females and 21 males of 7–16 years old). Two milliliters of saliva were collected and BALP and OC levels were assessed. Skeletal age was estimated using the cervical vertebral maturation method (CVM). The relationship between the biomarkers’ concentration in saliva and skeletal age was examined with the Spearman’s coefficient “ρ” (rho). Correlations between skeletal age groups and BALP and OC concentrations were assessed with the Kruskal–Wallis or the Mann–Whitney tests. No statistically significant differences in the levels of BALP (p = 0.568) and OC (p = 0.996) in saliva were identified according to the patient’s skeletal age. The use of BALP and OC levels in saliva seems to be dubious for skeletal growth assessment. However, slightly differentiated levels of those biomarkers, especially of BALP, through the different maturation stages, with higher concentrations at the pubertal phase, have been noticed. More studies are needed to clarify the exact potential role of these biomarkers as predictors of pubertal onset.

Purpose: The possibility that the sympathetic nervous system (SNS) controls bone remodeling has been raised; however, the actual function of the SNS in osseointegration is still unknown. This study aimed to investigate the effect of chemical sympathectomy on peri-implant osseointegration in adult mice. Materials and Methods: Forty C57BL/6J mice (8-week-old) were divided into two groups: a sympathectomy group and a control group, which were administered 6-hydroxydopamine and saline, respectively, by intraperitoneal injection for 5 days. Then, the mice were exposed to implant surgery. Analyses of serum chemistry, microcomputed tomography, biomechanical test, and bone histomorphometry were employed at 2 and 4 weeks. Results: Compared with the control, the chemical sympathectomy group had a higher serum level of C-terminal collagen I cross-links but lower serum osteocalcin. After 4 weeks, peri-implant trabecular microstructure, including trabecular volume, trabecular thickness, the percentage of osseointegration, and bone-to-implant contact, was lower; however, the trabecular separation was higher in the sympathectomy group mice in comparison with the control group. In addition, the strength of bone-titanium integration measured by the biomechanical resistance test was lower. Furthermore, histomorphologic evidence revealed that the osteoclast counts were higher in the sympathectomy group, while the mineral apposition rate and the bone formation rate per bone surface were significantly lower. Conclusion: Within the limitations of this experimental study, the data showed that chemical sympathectomy has a negative effect in peri-implant osseointegration, suggesting that the SNS may need to be taken into consideration in terms of peri-implant bone healing. [ABSTRACT FROM AUTHOR]

The relationships of osteocalcin (OC) and C-telopeptide of type I collagen (CTX) with long-term incidence of hip fracture were examined in 1680 post-menopausal women from a population-based study. CTX, but not OC, levels were associated with incident hip fracture in these participants, a relationship characterized by an inverted U-shape.IntroductionWe sought to investigate the relationships of OC, a marker of bone formation, and CTX, a marker of bone resorption, with long-term incidence of hip fracture in older women.MethodsWe included 1680 women from the population-based Cardiovascular Health Study (mean [SD] age 74.5 [5.0] years). The longitudinal association of both markers with incidence of hip fracture was examined using multivariable Cox models.ResultsDuring a median follow-up of 12.3 years, 288 incident hip fractures occurred. Linear spline analysis did not demonstrate an association between OC levels and incident hip fracture. By contrast, increasing levels of CTX up to the middle-upper range were associated with a significantly greater risk of hip fracture (HR = 1.52 per SD increment, 95% CI = 1.10-2.09), while further increases were associated with a marginally non-significant lower risk (HR = 0.80 per SD increment, 95% CI = 0.63-1.01), after full adjustment for potential confounders. In analyses of quartiles, CTX exhibited a similar inverted U-shaped relationship with incident fracture after adjustment, with a significant association observed only for the comparison of quartile 3 to quartile 1 (HR = 1.63, 95% CI = 1.10-2.43). In a subset with available measures, both OC and CTX were inversely associated with bone mineral density of the hip.ConclusionCTX, but not OC, levels were associated with incident hip fracture in post-menopausal women, a relationship characterized by an inverted U-shape. These findings highlight the complex relationship of bone turnover markers with hip fracture risk.

Bone has long been considered as a silent organ that provides a reservoir of calcium and phosphorus, traditionally. Recently, further study of bone has revealed additional functions as an endocrine organ connecting systemic organs of the whole body. Communication between bone and other organs participates in most physiological and pathological events and is responsible for the maintenance of homeostasis. Here, we present an overview of the crosstalk between bone and other organs. Furthermore, we describe the factors mediating the crosstalk and review the mechanisms in the development of potential associated diseases. These connections shed new light on the pathogenesis of systemic diseases and provide novel potential targets for the treatment of systemic diseases.

Abstract Background Osteocalcin, a protein secreted mainly by mature osteoblasts, has been shown to be involved in glucose metabolism through various pathways. However, few studies has explored the association between osteocalcin and Time in range (TIR). Continuous glucose monitoring (CGM) -derived metrics, such as TIR and other indexes have been gradually and widely used in clinical practice to assess glucose fluctuations. The main purpose of this study was to investigate the correlation between osteocalcin and indexes from CGM in patients with type 2 diabetes mellitus (T2DM). Method The total number of 376 patients with T2D were enrolled, all of them performed three consecutive days of monitoring. They were divided into four groups on account of the quartile of osteocalcin. Time in range, Time below range (TBR), Time above range(TAR) and measures of glycemic variability (GV) were assessed for analysing. After a 100 g standard steamed bread meal, blood glucose (Glu0h Glu0.5 h, Glu1h, Glu2h, GLu3h), C-peptide (Cp0h, Cp0.5 h, Cp1h, Cp2h, Cp3h), serum insulin (INS0h, INS0.5 h, INS1h, INS2h, INS3h) concentrations at different time points were obtained. HOMA-IS, HOMA-βwas calculated to evaluate insulin sensitivity and insulin secreting of the participants. Results Patients with higher osteocalcin level had higher TIR (P 0.05). Multiple stepwise regression showed that osteocalcin was an independent contributor to TIR, TAR and HOMA-IS. Conclusion Circulating osteocalcin is positively correlated with TIR and negatively correlated with MODD, ADDR, and MAGE. Osteocalcin may have a beneficial impact on glucose homeostasis in T2DM patients.

Abstract Objectives Osteoporosis is a common skeletal disorder attributed to age and is defined as a systematic degradation of bone mass and the microarchitecture leading to bone fractures. Exosomes have been reported in almost all biological fluids and during the failure of bone remodeling. 20 ml of blood samples were obtained from osteoporotic and non-osteoporotic postmenopausal women. After the isolation of peripheral blood mononuclear cells (PBMCs), T cells were separated via the magnetic-activated cell sorting (MACS) technique. Exosomes were driven from T cells of non-osteoporotic and osteoporotic volunteers. Subsequently, normal osteoblasts were treated with obtained T cell exosomes to assess osteoblastic function and gene expression. Results Runx2, type I collagen, osteopontin, and osteocalcin expression decreased in osteoblasts treated by osteoporotic T cell exosomes. In contrast, an increased expression of the mentioned genes was observed following non-osteoporotic T cell exosome treatment. Additionally, osteoblast alkaline phosphatase (ALP) activity treated with non-osteoporotic T cell exosomes increased. However, this activity decreased in another group. Our data demonstrated that T cell exosomes obtained from osteoporotic and non-osteoporotic individuals could alter the osteoblastic function and gene expression by affecting the genes essential for bone remodeling.

Abstract Osteocalcin (OCN) is a bone‐derived and vitamin K dependent hormone that affects energy metabolism and vascular calcification. The relationship between serum OCN and vascular function in patients with chronic kidney disease (CKD) is uncertain. This study investigated the association between serum OCN and vascular function as expressed with reactive hyperemia index (RHI) and augmentation index (AIx) measured by Endo‐PAT 2000 device. This cross‐sectional analysis was based on 256 pre‐dialysis CKD patients who had completed the Endo‐PAT 2000 test and serum OCN at the First Center of Chinese PLA Hospital from November 2017 to December 2019. Based on whether the RHI was less than 1.67, the patients were divided into endothelial dysfunction and normal endothelial function groups. Multiple logistic and linear regression were used to analyze the association between OCN and vascular function. Subgroup analyses were performed to examine the effects of OCN on vascular function in different CKD populations. After multivariate adjustment, CKD with low OCN were more likely to have endothelial dysfunction (OR: 0.794; 95%CI: 0.674‐0.934; P = .006); on the contrary, patients with high OCN had a higher degree of arterial stiffness (standardized β: 0.174; P = .003). Subgroup analyses showed that higher OCN was associated with severe arterial stiffness but a better endothelial function in young (age

Context: Histological quantification of osteoclasts and osteoblasts can evaluate biological responses to orthodontic tooth movement. Histological analysis of bone samples can be technically challenging. Aims: To evaluate the differences between hematoxylin and eosin (HE) staining and immunohistochemistry (IHC) in quantifying osteoblast and osteoclast cells following the application of static orthodontic force. Methods: Orthodontic force was applied using a rubber separator around the maxilla incisor of Cavia porcellus. Tooth samples were taken at 0, 4, 8, 14, 21, and 28 days after applying orthodontic force. HE and IHC staining quantify osteoblast and osteoclast cells in the alveolar bone. IHC staining, i.e., Tartrate-resistant acid phosphatase (TRAP) staining, was used to identify osteoclasts, and osteocalcin (OCN) staining was used to identify osteoblasts. Results: Significantly higher numbers of osteoclasts and osteoblasts were observed with IHC compared to HE staining (p

Abstract Background The association between osteocalcin and mortality has been scantly studied. We aimed to investigate the association between osteocalcin along with its trajectories and mortality based on long-term longitudinal data. Methods We performed a retrospective cohort study of 9413 type 2 diabetic patients with at least three measurements of total serum osteocalcin within 3 years since their first inpatient diagnosis of type 2 diabetes. Baseline, mean values of osteocalcin levels and their trajectories were used as exposures. A multivariable-adjusted Cox proportional hazards model was used to estimate the association of osteocalcin levels and their trajectories with mortality. Results During a mean follow-up of 5.37 years, 1638 patients died, of whom 588 were due to cardiovascular events. Multivariable-adjusted hazard ratios (HRs) across quintiles of baseline osteocalcin levels were 2.88 (95% confidence interval (CI) 2.42–3.42), 1.65 (95% CI 1.37–1.99), 1.17 (95% CI 0.96–1.42), 1.00, and 1.92 (95% CI 1.60–2.30) for all-cause mortality, and 3.52 (95% CI 2.63–4.71), 2.00 (95% CI 1.46–2.73), 1.03 (95% CI 0.72–1.47), 1.00, 1.67 (95% CI 1.21–2.31) for CVD mortality, respectively. When we used the mean values of osteocalcin as the exposure, U-shaped associations were also found. These U-shaped associations were consistent among patients of different baseline characteristics. Patients with a stable or even increasing trajectory of osteocalcin may have a lower risk of both all-cause and CVD mortality. Conclusions A U-shape association between baseline osteocalcin and mortality was observed among patients with type 2 diabetes. Patients with lower levels of serum osteocalcin during follow-ups had higher risks for all-cause and cardiovascular mortality.

IntroductionKetosis is a predominant metabolic problem and a risk factor for several postpartum diseases. This retrospective study aimed to evaluate the complete blood count (CBC), plasma biochemistry, and osteocalcin and identify significant prepartum and early postpartum values expressed in ketotic cows.MethodsIn 135 Holstein Friesian cows, 210 parturitions of 114 primiparous and 96 multiparous cows were examined. According to the plasma concentrations of β-hydroxybutyrate (BHB; ≥ 1.4 mmol/L) or non-esterified fatty acids (NEFA; ≥ 0.7 mmol/L) in the postpartum period, cows were divided into healthy cows (CON) and ketotic cows (KET). Analyses of CBC and biochemistry profiles were performed from −6 to 4 weeks of parturition every 2 weeks (prepartum; BW–5, BW–3, and BW–1, postpartum; BW1 and BW3), and osteocalcin ELISA tests were performed using blood samples from −2 to 2 weeks of parturition (BW–1 and BW1).ResultsIn primiparous KET (n = 114) before parturition, lower lymphocyte (Lym) in BW–5 and BW–3, lower red blood cell (RBC) in BW–5, higher mean corpuscular volume (MCV) in BW–1, and higher NEFA in BW–3 were significant compared with CON. Primiparous KET showed lower carboxylated osteocalcin (cOC) levels and a significant decrease after parturition. In multiparous KET (n = 96) before parturition, lower neutrophil (Neu) in BW–5, higher hemoglobin (HGB) in BW–5, higher MCV in BW–5 and BW–1, higher MCH in BW–5, lower total cholesterol (TC) in BW–5, higher triglyceride (TG) in BW–3, higher NEFA in BW–1, higher glucose (Glu) in BW–3, lower γ-glutamyl transferase (GGT) in BW–5, lower inorganic phosphate (iP) in BW–3, and higher body condition score (BCS) in BW–5 and BW–3 were significant compared with CON. Multiparous KET showed decreased cOC and uncarboxylated osteocalcin (ucOC) after parturition, which was lower than that in the CON group.DiscussionThe blood parameters expressing different values between CON and KET in prepartum or early postpartum periods are presumed to show individual nutrition and health states, liver function, and overweight status. These parameters could be valuable indicators that can be used to prevent the occurrence of ketosis and improve management practices by recognizing these differences in ketotic cows before calving.

This study aimed to investigate the effects and mechanisms of osteocalcin on autophagy in myoblasts, as well as its possible therapeutic effects in aging muscle. Starved murine myoblast C2C12 cells with or without interleukin (IL)-6 siRNA were treated with osteocalcin. Expression of the autophagy protein marker LC3, as well as IL-6 and phosphorylated STAT3 were detected by immunoblotting, immunofluorescence, or immunohistochemistry. Autophagosomes were observed with transmission electron microscopy. Levels of reactive oxygen species (ROS) were detected by flow cytometry. Fasted young mice were injected intraperitoneally with osteocalcin, with or without the JAK inhibitor CP-690550 to inhibit IL-6 signaling. Older mice were treated with osteocalcin and muscle mass, grip strength and muscle structure were assessed. The results revealed that compared to control and serum-starved cells, osteocalcin treatment significantly increased the relative expression of LC3-II/LC3-I protein, the numbers of autophagosomes, and levels of intracellular ROS. Osteocalcin injection in mice also resulted in increased relative LC3-II/LC3-I protein expression and autophagosome numbers. Osteocalcin treatment significantly increased the secretion of IL-6 in muscle cells and tissue, and activated STAT3 signaling. Moreover, knockdown of IL-6 or blocking IL-6 signaling inhibited the phosphorylation of STAT3, and further inhibited autophagy in starved myoblasts and fasting-treated murine muscle tissue. In addition, osteocalcin treatment significantly increased muscle mass and grip strength in both aged mice and aged fasting mice. In conclusion, the inhibition of osteocalcin on muscle cell aging is accompanied by the induction of IL-6-STAT3-dependent autophagy, indicating osteocalcin might be a promising therapeutic candidate for aging-related myopathies.

IntroductionThe bone matrix protein osteocalcin (OC), secreted by osteoblasts, displays endocrine effects. We tested the hypothesis that OC modulates parathyroid tumor cell function.MethodsPrimary cell cultures derived from parathyroid adenomas (PAds) and HEK293 cells transiently transfected with the putative OC receptor GPRC6A or the calcium sensing receptor (CASR) were used as experimental models to investigate γ-carboxylated OC (GlaOC) or uncarboxylated OC (GluOC) modulation of intracellular signaling.ResultsIn primary cell cultures derived from PAds, incubation with GlaOC or GluOC modulated intracellular signaling, inhibiting pERK/ERK and increasing active β-catenin levels. GlaOC increased the expression of PTH, CCND1 and CASR, and reduced CDKN1B/p27 and TP73. GluOC stimulated transcription of PTH, and inhibited MEN1 expression. Moreover, GlaOC and GluOC reduced staurosporin-induced caspase 3/7 activity. The putative OC receptor GPRC6A was detected in normal and tumor parathyroids at membrane or cytoplasmic level in cells scattered throughout the parenchyma. In PAds, the membrane expression levels of GPRC6A and its closest homolog CASR positively correlated; GPRC6A protein levels positively correlated with circulating ionized and total calcium, and PTH levels of the patients harboring the analyzed PAds. Using HEK293A transiently transfected with either GPRC6A or CASR, and PAds-derived cells silenced for CASR, we showed that GlaOC and GluOC modulated pERK/ERK and active β-catenin mainly through CASR activation.ConclusionParathyroid gland emerges as a novel target of the bone secreted hormone osteocalcin, which may modulate tumor parathyroid CASR sensitivity and parathyroid cell apoptosis.

The skeleton is the third most common site of metastatic disease, which causes serious bone complications and short-term prognosis in cancer patients. Prostate and breast cancers are responsible for the majority of bone metastasis, resulting in osteolytic or osteoblastic lesions. The crosstalk between bone cells and their interactions with tumor cells are important in the development of lesions. Recently, both preclinical and clinical studies documented the clinical relevance of bone-derived factors, including osteocalcin (OC) and its undercarboxylated form (ucOC), fibroblast growth factor 23 (FGF23), sclerostin (SCL), and lipocalin 2 (LCN2) as prognostic tumor biomarkers and potential therapeutic targets in bone metastasis. Both OC and ucOC could be useful targets for the prevention of bone metastasis in breast cancer. Moreover, elevated OC level may be a metastatic marker of prostate cancer. FGF23 is particularly important for those forms of cancer that primarily affect bone and/or are characterized by bone metastasis. In other tumor entities, increased FGF23 level is enigmatic. SCL plays a significant role in the pathogenesis of both osteolytic and osteoblastic lesions, as its levels are high in metastatic breast and prostate cancers. Elevated expression levels of LCN2 have been found in aggressive subtypes of cancer. However, its role in antimetastasis varies significantly between different cancer types. Anyway, all aforementioned bone-derived factors can be used as promising tumor biomarkers. As metastatic bone disease is generally not curable, targeting bone factors represents a new trend in the prevention of bone metastasis and patient care. [ABSTRACT FROM AUTHOR]

Background: Calcific aortic valve disease (CAVD) is the second leading cause of adult heart diseases. The purpose of this study is to investigate whether miR-101-3p plays a role in the human aortic valve interstitial cells (HAVICs) calcification and the underlying mechanisms. Methods: Small RNA deep sequencing and qPCR analysis were used to determine changes in microRNA expression in calcified human aortic valves. Results: The data showed that miR-101-3p levels were increased in the calcified human aortic valves. Using cultured primary HAVICs, we demonstrated that the miR-101-3p mimic promoted calcification and upregulated the osteogenesis pathway, while anti-miR-101-3p inhibited osteogenic differentiation and prevented calcification in HAVICs treated with the osteogenic conditioned medium. Mechanistically, miR-101-3p directly targeted cadherin-11 (CDH11) and Sry-related high-mobility-group box 9 (SOX9), key factors in the regulation of chondrogenesis and osteogenesis. Both CDH11 and SOX9 expressions were downregulated in the calcified human HAVICs. Inhibition of miR-101-3p restored expression of CDH11, SOX9 and ASPN and prevented osteogenesis in HAVICs under the calcific condition. Conclusion: miR-101-3p plays an important role in HAVIC calcification through regulation of CDH11/SOX9 expression. The finding is important as it reveals that miR-1013p may be a potential therapeutic target for calcific aortic valve disease. [ABSTRACT FROM AUTHOR]

We present a topological method for the detection and quantification of bone microstructure from non-linear microscopy images. Specifically, we analyse second harmonic generation (SHG) and two photon excited autofluorescence (TPaF) images of bone tissue which capture the distribution of matrix (fibrillar collagen) structure and autofluorescent molecules, respectively. Using persistent homology statistics with a signed Euclidean distance transform filtration on binary patches of images, we are able to quantify the number, size, distribution, and crowding of holes within and across samples imaged at the microscale. We apply our methodology to a previously characterized murine model of skeletal pathology whereby vascular endothelial growth factor expression was deleted in osteocalcin-expressing cells (OcnVEGFKO) presenting increased cortical porosity, compared to wild type (WT) littermate controls. We show significant differences in topological statistics between the OcnVEGFKO and WT groups and, when classifying the males, or females respectively, into OcnVEGFKO or WT groups, we obtain high prediction accuracies of 98.7% (74.2%) and 77.8% (65.8%) respectively for SHG (TPaF) images. The persistence statistics that we use are fully interpretable, can highlight regions of abnormality within an image and identify features at different spatial scales. [ABSTRACT FROM AUTHOR]

Cleidocranial dysplasia (CCD) is a rare, autosomal dominant hereditary disorder characterized by skeletal malformations and dental abnormalities. The purpose of this study was to explore the functional role of a novel mutation in the pathogenesis of CCD. Genomic DNA was extracted from peripheral blood mononuclear cells collected from family members of a Chinese patient with CCD. An analysis of their RUNX Family Transcription Factor 2 (RUNX2) gene sequences was performed by PCR amplification and Sanger sequencing. The function of the mutant RUNX2 was studied by bioinformatics, real‐time PCR, western blotting, and subcellular localization analysis. Sanger sequencing identified a novel single‐base deletion (NM_001024630.4:c.132delG;NP_001019801.3: Val45Trpfs*99) in the RUNX2 gene present in the Chinese patient with CCD. In vitro, functional studies showed altered protein localization and increased expression of mutant RUNX2 mRNA and mutant Runt‐related transcription factor 2 (RUNX2). Luciferase reporter assay demonstrated that the novel RUNX2 mutations significantly increased the transactivation activity of RUNX2 on the osteocalcin gene promoter. In conclusion, we identified a patient with sporadic CCD carrying a novel deletion/frameshift mutation of the RUNX2 gene and performed screening and functional analyses to determine the cause of the CCD phenotype. This study provides new insights into the pathogenesis of CCD.3 [ABSTRACT FROM AUTHOR]

Introduction: Metformin may provide a therapeutic benefit in different types of malignancy. Purpose: We aimed at evaluating the effect of metformin as an adjuvant therapy to letrozole on estradiol and other biomarkers involved in the pathogenesis of breast cancer in overweight and obese postmenopausal women. Methods: Seventy-five postmenopausal stages II–III breast cancer female patients were assessed for eligibility in an open-labeled parallel pilot study. Forty-five patients met the inclusion criteria and were assigned into three arms: the lean arm (n = 15) women who received letrozole 2.5 mg/day, the control arm (n = 15) overweight/obese women who received letrozole 2.5 mg/day, and the metformin arm (n = 15) overweight/obese women who received letrozole 2.5 mg/day plus metformin (2000 ± 500 mg/day). The intervention duration was 6 months. Blood samples were obtained at baseline and 6 months after intervention for the measurement of serum estradiol, leptin, osteocalcin levels, fasting blood glucose concentration, and serum insulin. Results: After the intervention and as compared to the control arm, the metformin arm showed a significantly lower ratio to the baseline (significant reduction) for estradiol (p = 0.0433), leptin (p < 0.0001), fasting blood glucose (p = 0.0128), insulin (p = 0.0360), osteocalcin serum levels (p < 0.0001), and the homeostatic model assessment of insulin resistance "HOMA-IR" value (p = 0.0145). There was a non-significant variation in the lactate ratio to the baseline among the three study arms (p = 0.5298). Conclusion: Metformin may exert anti-cancer activity by decreasing the circulating estradiol, leptin, and insulin. Metformin might represent a safe and promising adjuvant therapy to letrozole in overweight/obese postmenopausal women with breast cancer. Trial registration: ClinicalTrials.gov Identifier: NCT05053841/Registered September 23, 2021 - Retrospectively. [ABSTRACT FROM AUTHOR]

High-dose vitamin D supplementation can increase total osteocalcin concentrations that may reduce insulin resistance in individuals at risk for prediabetes or diabetes mellitus. Magnesium is a cofactor in vitamin D metabolism and activation. The purpose of this study was to determine the combined effect of vitamin D and magnesium supplementation on total osteocalcin concentrations, glycemic indices, and other bone turnover markers after a 12-week intervention in individuals who were overweight and obese, but otherwise healthy. We hypothesized that combined supplementation would improve serum total osteocalcin concentrations and glycemic indices more than vitamin D supplementation alone or a placebo. A total of 78 women and men completed this intervention in 3 groups: a vitamin D and magnesium group (1000 IU vitamin D 3 and 360 mg magnesium glycinate), a vitamin D group (1000 IU vitamin D 3), and a placebo group. Despite a significant increase in serum 25-hydroxyvitamin D concentrations in the vitamin D and magnesium group compared with the placebo group (difference = 5.63; CI, –10.0 to –1.21; P =.001) post-intervention, there were no differences in serum concentrations of total osteocalcin, glucose, insulin, and adiponectin or the homeostatic model assessment of insulin resistance (HOMA-IR) among groups (P >.05 for all). Additionally, total osteocalcin (β = –0.310, P =.081), bone-specific alkaline phosphatase (β = 0.004, P =.986), and C-terminal cross-linked telopeptide (β = 0.426, P =.057), were not significant predictors of HOMA-IR after the intervention. Combined supplementation was not associated with short-term improvements in glycemic indices or bone turnover markers in participants who were overweight and obese in our study. This trial was registered at clinicaltrials.gov (NCT03134417). In a 12-week intervention of individuals who were overweight and obese, despite increasing serum 25(OH)D concentrations, combined vitamin D and magnesium supplementation did not improve serum concentrations of total osteocalcin, glucose, insulin, or adiponectin or affect HOMA-IR. Total osteocalcin, BAP, and CTX were not predictors of HOMA-IR after the intervention. BAP, bone-specific alkaline phosphatase; CTX, carboxy-terminal collagen crosslinks; HOMA-IR, homeostatic model assessment of insulin resistance; 25(OH)D, 25-hydroxyvitamin D. [Display omitted] [ABSTRACT FROM AUTHOR]

Aim: To evaluate the tissue reaction of a tricalcium silicate‐based repair material associated with 30% calcium tungstate (TCS + CaWO4) in comparison to Bio‐C Repair (Bio‐C; Angelus) and to MTA Repair HP (MTA HP; Angelus). Methodology: Polyethylene tubes filled with one of the materials or left empty (control group, CG) were implanted into the subcutaneous tissues of rats for 7, 15, 30 and 60 days (n = 32/group). The capsule thickness, number of inflammatory cells, collagen content, interleukin‐6 (IL‐6), osteocalcin (OCN), von Kossa reaction and analysis under polarized light were evaluated. The data were subjected to generalized linear models for repeated measures, except the OCN. OCN data were submitted to Kruskal–Wallis and Dunn's post hoc test and Friedman followed by Nemenyi's test at significance level of 5%. Results: At all time points, significant differences in the number of inflammatory cells were not observed between TCS + CaWO4 and Bio‐C, whereas, at 15, 30 and 60 days, no significant difference was detected between TCS + CaWO4 and MTA HP. At all periods, significant differences were not detected in the number of fibroblasts in TCS + CaWO4 versus MTA HP, and, at 60 days, no significant difference was demonstrated between these groups and CG. Significant differences in the immunoexpression of IL‐6 were not detected amongst bioceramic materials at all periods. From 7 to 60 days, significant reduction in the number of inflammatory cells, number of IL‐6‐immunopositive cells and in the capsule thickness was accompanied by significant increase in the collagen in all groups. OCN‐immunolabelled cells, von Kossa‐positive structures and amorphous calcite deposits were observed around all materials, whereas, in the CG, these structures were not seen. Conclusions: These findings indicate that the experimental material (TCS + CaWO4) is biocompatible and has a bioactive potential, similar to the MTA HP and Bio‐C Repair, and suggest its use as a root repair material. [ABSTRACT FROM AUTHOR]

Background and objective: Pinctada Maxima which is one of the nacres some of the structures of marine biota whose contents can be used to build tooth structures are damaged due to disease. The aim of this study is to see the effectiveness of pearl shells on osteogenesis of bone graft in periodontal tissue and to determine the effectiveness of pearl shells on periodontal tissue through the formation of osteoblasts so that later it can be considered as an ideal alternative bone graft. Methods: Cavia Porcellus were divided into 3 groups of 30: Bone graft using Hydroxyapatite Pinctada maxima, Bovine Xenograft as positive control and placebo as negative control, then sacrificed on 14th and 21st days. The Real Time-Polymerase Chain Reaction (RTPCR) analysis was performed to see the expression of osteocalcin (OCN) and Scan electron microscope (SEM) examination to see the formation of osteoblasts Results: Data were analyzed with Independent T test and processed in SPSS 24.0. It was shown that the osteoblast increases in the Batan and Placebo groups from day 14 to day 21, in contrast to the Hydroxyapatite Pinctada Maxima test group with SEM analysis. For the OCN increase in all the Batan, Placebo and Hydroxyapatite Pinctada Maxima groups and the most significant increase was in the Hydroxyapatite Pinctada Maxima Test group although it was not significant with the RT-PCR analysis. Coclusion: Hidroxyapatite pinctada maxima has quite good potential as a bone graft. [ABSTRACT FROM AUTHOR]

There exists a marked circadian variation for several bone markers (BM), which is influenced by endogenous as well as exogenous factors including hormones, physical activity, and fasting. Consequently, was the aim of this review to provide an overview of the knowledge of the circadian variation of BM and which factors influence this rhythmicity. A systematic search of PubMed was performed for studies evaluating the circadian variation of BM and which factors influence this rhythmicity. The studies were screened for eligibility by a set of predetermined criteria including a list of relevant BM and a minimum study duration of 24 h with at least 3 blood samples of which two should be at least 6 h apart. In total were 29 papers included. There exists a marked circadian variation for most BM including Carboxy-terminal Cross-Linked Telopeptide of Type I Collagen (CTX) and osteocalcin (OC) with nighttime or early morning peak. Pro-collagen Type I N-terminal Propeptide (PINP) and PTH also showed circadian rhythm but with less amplitude. The inter-osteoblast-osteoclast regulatory markers such as OPG, RANKL, FGF23, and sclerostin showed no circadian rhythm. The markers were differently affected by exogenous factors like fasting, which greatly reduced the circadian variation of CTX but did not affect PINP or OC. The marked circadian variation and the factors which influence the rhythmicity, e.g., fasting are of great consequence when measuring BM. To reduce variation and heighten validity should circadian variation and fasting be kept in mind when measuring BM. [ABSTRACT FROM AUTHOR]

We used an ovariectomy (OVX) rat model to test whether yeast hydrolysate (YH) has therapeutic effects on postmenopausal osteoporosis-induced bone loss. The rats were separated into five treatment groups: the sham group (sham operation); the control group (no treatment after OVX); the estrogen group (estrogen treatment after OVX); YH 0.5% group (drinking water supplementation with 0.5% YH after OVX); and the YH 1% group (drinking water supplementation with 1% YH after OVX). In addition, the YH treatment restored serum testosterone concentration in the OVX rats up to the normal level. Further, YH treatment affected bone markers; a significant increase in serum calcium concentration was observed after adding YH to the diet. The levels of serum alkaline phosphatase, osteocalcin, and cross-linked telopeptides of type I collagen were reduced by YH supplementation, unlike those in the no-treatment control. Although not statistically significant, YH treatment in OVX rats improved trabecular bone microarchitecture parameters. These results show that YH may ameliorate the bone loss caused by postmenopausal osteoporosis because of the normalization of serum testosterone concentration. [ABSTRACT FROM AUTHOR]

Objective: To determine whether resveratrol (Res) can correct osteoporosis induced in a rat model of male hypogonadism. Methods: Thirty-two rats were randomly divided into 4 groups, 8 in each group; 1) a control sham group: underwent a similar surgical procedure for induction of orchiectomy (ORCD) without ligation of any arteries or veins or removal of the testis and epididymis; 2) a control + Res-treated group (Con+Res): underwent sham surgery similar to the control, but was then treated with Res, as described below; 3) an ORCD-induced group: bilateral ORCD surgery as described above, and 4) a ORCD+Res-treated group: bilateral ORCD surgery followed by Res treatment. Res treatment began 4 weeks after ORCD and continued for 12 weeks. After 12 weeks, bone mineral density (BMD) and bone mineral content (BMC) were measured in the tibia and femur of each rat's right hind leg. Blood levels of bone turnover indicators such as deoxypyridinoline (Dpd), N-telopeptide of type I collagen (NTX I), alkaline phosphatase (ALP), and osteocalcin (OC), as well as receptor activator of nuclear factor kappa B (RANK) and osteoprotegerin (OPG) were assessed. Results: ORCD significantly decreased BMD (P<0.01) and significantly increased bone resorption, manifested by increased RANK. In addition, it inhibited serum levels of OPG and OC. Res treatment after ORCD effectively increased serum levels of bone formation markers such as OPG and OC, compared with testisectomized rats (P<0.05). Conclusion: Res could ameliorate bone loss induced by male hypogonadism, possible via restoration of the normal balance between RANK and OPG. [ABSTRACT FROM AUTHOR]

Background and Objective: Teeth overeruption is a problem of clinical significance, but the underlying mechanism how changes in external occlusal force convert to the periodontium remodeling signals has been a largely under explored domain. And recently, periodontal ligament-associated protein-1 (PLAP-1)/ asporin was found to play a pivotal role in maintaining periodontal homeostasis. The aim of this study was to explore the function of PLAP-1 in the periodontally hypofunctional tissue turnover. Methods: After extracting left maxillary molars in mice, the left and right mandibular molars were distributed into hypofunction group (HG) and control group (CG), respectively. Mice were sacrificed for radiographic, histological, and molecular biological analyses after 1, 4 and 12 weeks. In vitro, dynamic compression was applied using Flexcell FX-5000 Compression System to simulate intermittent occlusal force. The expression of PLAP1 in loaded and unloaded human periodontal ligament cells (hP-DLCs) was compared, and its molecular biological effects were further explored by small interfering RNA (siRNA) targeting PLAP1. Results: In vivo, fiber disorder in periodontal ligament (PDL), bone apposition at furcation regions, and bone resorption in alveolar bone were illustrated in the HG compared with the CG. In addition, PLAP-1 positive area decreased significantly in PDL following occlusal unloading. In vitro, the loss of compressive loading relatively down-regulated PLAP1 expression, which was essential to promote collagen I but inhibit osterix and osteocalcin expression in hPDLCs. Conclusions: PLAP-1 presumably plays a pivotal role in occlusal force-regulated periodontal homeostasis by facilitating collagen fiber synthesis in hPDLCs and suppressing excessive osteoblast differentiation, further preventing teeth from overeruption. Further evidence in PLAP-1 conditional knockout mice is needed. [ABSTRACT FROM AUTHOR]