β-CTX Concentrations Research Articles

Hydrolyzed collagen supplementation prior to resistance exercise augments collagen synthesis in a dose-response manner in resistance-trained, middle-aged men.

Resistance exercise (RE) increases collagen synthesis in young and older men, while hydrolyzed collagen (HC) ingestion improves this response to RE in a dose-response manner in young men. However, the collagen synthesis response to RE with and without HC in middle-aged men is unknown. Eight resistance-trained men (age: 49±8 years; height: 1.78±0.02m; mass: 90±4kg) took part in this double-blind, crossover design study, and undertook 4×10 repetitions of lower-limb RE at maximum load, after consuming 0g, 15g, or 30g vitamin C-enriched HC. We analyzed venous blood samples for N-terminal propeptide of type 1 pro-collagen (PINP), β-isomerized C-terminal telopeptide of type 1 collagen (β-CTx) and 18 collagen amino acids throughout all three interventions. The serum PINP concentration×time area-under-the-curve (AUC) was higher following 30g (169±28 µg/mL×h) than 15g (134±23 µg/mL×h, P<0.05) HC ingestion, and both 15g and 30g were higher than 0g HC (96±23 µg/mL×h, P<0.05). RE with 0g HC showed no change in serum PINP concentration. The AUCs for glycine, proline, hydroxyproline, alanine, arginine, lysine, serine, leucine, valine and isoleucine were greater with 30g than 15g and 0g HC ingestion (P<0.05), and greater with 15g than 0g HC ingestion (P<0.05). Plasma β-CTx concentration decreased after RE independently of HC dose. Our study suggests connective tissue anabolic resistance to RE in middle-aged men but ingesting 15g HC rescues the collagen synthesis response, and 30g augments that response further. This dose-response is associated with the increased bioavailability of collagen amino acids in the blood, which stimulate collagen synthesis.

Effects of resistance exercise, collagen ingestion and circulating oestrogen concentration on collagen synthesis in a female athlete: A case report.

We investigated the effects of resistance exercise (RE), hydrolysed collagen (HC) ingestion and circulating oestrogen concentration on collagen synthesis in a naturally menstruating female CrossFit athlete. In a double-blind, randomised cross-over design, the participant (36years; height 1.61m; mass 82.6kg) consumed 0 or 30g HC prior to performing back-squat RE when endogenous circulating oestrogen concentration was low (onset of menses, OM) and high (late follicular phase, LF) during two consecutive menstrual cycles. Ten 5-mL blood samples were collected during each of the four interventions to analyse concentrations of serum 17β-oestradiol, and biomarkers of type I collagen turnover, that is serum procollagen type I N-terminal propeptide (PINP, a biomarker of collagen synthesis) and plasma β-isomerised C-terminal telopeptide of type I collagen (β-CTX, a biomarker of collagen breakdown), as well as the serum concentration of 18 collagen amino acids. 17β-Oestradiol concentration was 5-fold higher at LF (891±116pmolL-1) than OM (180±13pmolL-1). The PINP concentration × time area under the curve (AUC) was higher in the 30g HC OM intervention (201μgL-1h) than the 30g HC LF (144μgL-1h), 0g HC OM (151μgL-1h) and 0g HC LF (122μgL-1h) interventions. β-CTX concentration decreased 1.4-fold from pre-RE to 6h post-RE in all interventions. Thus, high circulating oestrogen concentration was associated with lower collagen synthesis following RE in this female athlete. Ingesting 30g HC, however, augmented the collagen synthesis response at LF and particularly at OM. HIGHLIGHTS: What is the central question of this study? Does resistance exercise-induced collagen synthesis vary according to circulating oestrogen concentration in a naturally menstruating female athlete, and if so, does hydrolysed collagen ingestion have any impact? What is the main finding and its importance? Exercise-induced collagen synthesis was low when circulating oestrogenconcentration was high and vice versa. However, ingesting 30g hydrolysedcollagen prior to exercise reduced the negative effect of oestrogen oncollagen synthesis. As high circulating oestrogen has been associated withgreater injury risk in females, supplementing exercise with hydrolysedcollagen may help protect these tissues from injury.

Effects of antidiabetic drugs on bone metabolism.

The prevalence of diabetes mellitus type 2 (DMT2) is increasing exponentially worldwide. DMT2 patients have been found to be at a higher risk for bone fractures than the healthy population. Hence, improving ourunderstanding of the impact of antidiabetic drugs on bone metabolism is crucial. A descriptive, retrospective study involving 106 patients receiving six groups of antidiabetic drugs: insulin; dipeptidylpeptidase four inhibitors (DPP4i); glucagon-like peptide type 1 receptor agonists (GLP1ra); sulfonylureas; sodium-glucose cotransporter two inhibitors (SGLT2i); and pioglitazone, in which osteocalcin (OC), bone alkaline phosphatase (BAP) and C-terminal telopeptide of collagen type 1 or beta-crosslaps (β-CTx) were determined. β-CTx concentrations were higher in the patients treated with pioglitazone, as compared to patients treated with DPP4i (p=0.035), SGLT2i (p=0.020) or GLP1ra (p<0.001). The lowest β-CTx concentrations were observed in the patients treated with GLP1ra. Bone remodeling is influenced by the type of antidiabetic drug administered to DMT2 patients. In our study, the patients who received pioglitazone showed higher β-CTx concentrations, as compared to patients treated with other types of antidiabetic drugs. This finding highlights theconvenience of avoiding these drugs, especially in postmenopausal women with DMT2. GLP1ra drugs were associated with the lowest β-CTx concentrations, which suggests that these agents could exert beneficial effects on bone metabolism.

Effect of acute resistance exercise on bone turnover in young adults before and after concurrent resistance and interval training.

Weight-bearing physical activity can stimulate bone adaptation. This investigation explored the effect of an acute bout of resistance exercise before and after resistance+interval training on circulating biomarkers of bone metabolism and muscle-bone crosstalk. Healthy young male and female participants (n = 21 male, 28 ± 4 years; n = 17 female, 27 ± 5 years) performed a 6 × 10 squat test (75% 1RM) before and after a 12-week resistance+interval training program. Before and after completion of the training program, blood samples were collected at rest, immediately postexercise, and 2 h postexercise. Blood samples were analyzed for βCTX, P1NP, sclerostin, osteocalcin, IGF-1, and irisin. Significant effects of acute exercise (main effect of time) were observed as increases in concentrations of IGF-1, irisin, osteocalcin, and P1NP from rest to postexercise. A sex*time interaction indicated a greater decline in βCTX concentration from rest to 2 h postexercise and a greater increase in sclerostin concentration from rest to immediately postexercise in male compared with female participants. Sex differences (main effect of sex) were also observed for irisin and P1NP concentrations. In summary, changes in concentrations of biochemical markers of bone metabolism and muscle-bone crosstalk were observed in males and females after an acute bout of resistance exercise and following 12 weeks of resistance+interval training.

Bone turnover biomarkers predict one-year all-cause mortality and walking ability in geriatric hip fracture patients.

To investigate the utility of serum C-terminal cross-linking telopeptides (β-CTX) and procollagen type I N propeptide (PINP) for predicting one-year mortality and walking ability in Chinese geriatric hip fracture patients who underwent surgical interventions. Elderly patients (≥ 60 years) who underwent surgical interventions for unilateral low-energy hip fracture from 2015 to 2020 in our center were included. Demographic data was retrospectively retrieved from the electronic medical database. The PINP and β-CTX concentrations were measured before the surgery. The patients were divided into two groups according to the outcome of mortality and walking ability after hip surgery, respectively. β-CTX and PINP were divided into four grades based on quartiles [Quartile(Q)1-4] for further analysis. All the variables with p < 0.1 in univariable analysis were included in a multivariable model. In univariable analysis, the levels of serum β-CTX (p = 0.007) and PINP (p = 0.025) was associated with one-year mortality, while the association between levels of serum β-CTX (p = 0.072) or PINP (p = 0.055) with one-year disability was marginally significant. After adjustment for confounders, the relative risk [OR (95 % CI), Q4 v sQ1, p-value] of one-year mortality and one-year disability were 7.28 (2.08-29.78, p = 0.003) and 3.97 (1.44-11.69, p = 0.009) for β-CTX and 5.87 (1.70-23.80, p = 0.008) and 3.48 (1.30-9.93, p = 0.016) for PINP, respectively. The coefficient of determination, AUC and bias-corrected C-index of predictive models based on previously reported predictors were significantly improved after integrating β-CTX or PINP. Higher serum β-CTX and PINP are independently associated with an increased risk of one-year mortality and disability in patients with hip fractures. The application of BTMs improves the performance of currently available predictive models.

The Collagen Synthesis Response to an Acute Bout of Resistance Exercise Is Greater when Ingesting 30 g Hydrolyzed Collagen Compared with 15 g and 0 g in Resistance-Trained Young Men

BackgroundResistance exercise (RE) stimulates collagen synthesis in skeletal muscle and tendon but there is limited and equivocal evidence regarding an effect of collagen supplementation and exercise on collagen synthesis. Furthermore, it is not known if a dose–response exists regarding the effect of hydrolyzed collagen (HC) ingestion and RE on collagen synthesis. ObjectiveTo determine the HC dose–response effect on collagen synthesis after high-intensity RE in resistance-trained young men. MethodsUsing a double-blind, randomized crossover design, 10 resistance-trained males (age: 26 ± 3 y; height: 1.77 ± 0.04 m; mass: 79.7 ± 7.0 kg) ingested 0 g, 15 g, or 30 g HC with 50 mg vitamin C 1 h before performing 4 sets’ barbell back squat RE at 10-repetition maximum load, after which they rested for 6 h. Blood samples were collected throughout each of the 3 interventions to analyze procollagen type Ⅰ N-terminal propeptide (PINP) and β-isomerized C-terminal telopeptide of type I collagen (β-CTX) concentration, and the concentration of 18 collagen amino acids. ResultsThe serum PINP concentration × time area under the curve (AUC) was greater for 30 g (267 ± 79 μg·L−1·h) than for 15 g (235 ± 70 μg·L−1·h, P = 0.013) and 0 g HC (219 ± 88 μg·L−1·h, P = 0.002) but there was no difference between 0 and 15 g HC (P = 0.225). The AUCs of glycine and proline were greater for 30 g than for 15 and 0 g HC (P < 0.05). Plasma β-CTX concentration decreased from −1 to +6 h (P < 0.05), with no differences between interventions. ConclusionsIngesting 30 g HC before high-intensity RE augments whole-body collagen synthesis more than 15 g and 0 g HC in resistance-trained young males.

The relationship between coronary artery calcification and bone metabolic markers in maintenance hemodialysis patients

BackgroundTo study the influencing factors for coronary artery calcification (CAC) in maintenance hemodialysis (MHD) patients and the relationship between CAC and bone metabolism markers and to attempt to find a reliable marker linking vascular calcification and bone metabolism in MHD patients.MethodsA total of 123 patients were enrolled. CAC was assessed by multislice spiral computed tomography (MSCT), and the CAC score (CACS) was evaluated using the Agaston method. Routine laboratory parameters, including triglycerides (TG), total cholesterol (TC), glucose (Glu), calcium (Ca), phosphorus (P), magnesium (Mg), etc., were measured. Serum markers of bone metabolism, such as alkaline phosphatase(ALP), calcitonin (CT), 25-hydroxy vitamin D [25-(OH)D], intact parathyroid hormone (iPTH), total type I procollagen amino-terminal peptide (tPINP), N-terminal mid-fragment of osteocalcin (N-MID OC), and β-type I collagen crosslinked carboxyl-terminal peptide (β-CTX), were also measured.ResultsAmong 123 MHD patients, 37 patients (30.08%) did not have CAC, and 86 patients (69.92%) had CAC, including 41 patients (47.67%) with mild calcification and 45 patients (52.33%) with moderate to severe calcification. Age, Body Mass Index(BMI), the prevalence of hypertension and diabetes mellitus, TC, Glu, P, and Ca×P in the calcification group were higher than those in the noncalcification group, whereas Mg, iPTH, tPINP, N-MID OC, and β-CTX were lower than those in the noncalcified group (P < 0.05). Compared with the mild calcification group (0<CACS<400), P and Ca×P levels were higher in the moderate to severe calcification group (CACS ≥ 400), and ALP, iPTH, N-MID OC, tPINP, and β-CTX concentrations were lower (P < 0.05). Correlation analysis showed that the CACS was positively correlated with TC, LDL-C, P, and Ca×P (P < 0.05) and negatively correlated with N-MID OC and β-CTX (P < 0.05). There was no significant correlation between the CACS and other parameters (P > 0.05). A logistic regression model was used to evaluate the influencing factors for CAC. The results showed that age, BMI, TC, Glu, P, and Ca×P were risk factors for CAC and its severity in MHD patients, whereas diabetes mellitus, Mg, and N-MID OC were protective factors for CAC in MHD patients. In addition, N-MID OC was a protective factor for the severity of CAC. After adjusting for the corresponding confounding factors, the results of the risk factors were consistent, and N-MID OC was still an independent protective factor for CAC and its severity.ConclusionsElevated serum P and Ca×P were independent risk factors for CAC in MHD patients, and serum Mg may be an independent protective factor for CAC. CAC was closely related to abnormal bone metabolism and bone metabolic markers in MHD patients. Relatively low bone turnover can promote the occurrence and development of CAC. N-MID OC may be a reliable bone metabolic marker linking vascular calcification and bone metabolism in MHD patients.

Abstract 3208: Bone-derived TGF-ß release is associated with cardiac dysfunction due to oxidation of SR calcium gatekeepers, Ca2+ leak and mitochondrial dysfunction

Abstract Cancer patients with bone metastases are associated with cardiac dysfunction. Heart attack was observed in 3.8% of patients within 30 days of diagnosis. Here, we report that bone turnover marker β-CTX (P=0.002) and platelet-poor plasma TGF-ß (P=0.036) from patients with bone metastasis were significantly higher than those without bone metastasis. In patients, serum β-CTX concentration was positively correlated with TGF-ß (R2=0.67, P&amp;lt;0.0001). Excess TGF-ß release caused skeletal muscle weakness by oxidation of the sarcoplasmic reticulum (SR) Ca2+ ryanodine receptor (RyR), induction of SR and oxidative stress in breast cancer patients with bone metastasis. Thus, we hypothesized that the systemic effects of TGF-ß could have similar effects on cardiac muscle to induce heart failure. Mechanistically, we showed that TGF-β1 increased transcription of NADPH oxidases, aberrant Ca2+ signaling, SR stress and ROS production in adult cardiomyocytes. We tested this in a transgenic mouse model of Camurati-Engelmann Disease (CED) in which mutant TGF-ß1 (H222D) is overexpressed in osteoblasts and causes accelerated bone destruction. We found that CED mice had increased cardiac size, dilated left ventricle and reduced ejection fraction, by echocardiography, compared to littermate controls. This was associated with increased phosphorylation of Smad2/3, myocardial necrosis and fibrosis in cardiac tissues. Similarly, SR Ca2+ channel RyR2 and SERCA2 showed a significant increase in oxidation and nitrosylation and reduced binding of the stabilizing subunits as well as upregulated iNOS and NADPH oxidases consistent with depressed SR Ca2+ re-uptake, RyR-mediated Ca2+ leak and oxidative stress in CED mice left ventricular. Mitochondria isolated from ventricular samples showed increased mitochondrial Ca2+ overload, ROS production, release of apoptotic proteins, reduced mitochondrial DNA and ATP reproduction in CED mice. Thus, a feed-forward cycle of pathological Ca2+ handling is created between SR mitochondria, which underlies TGF-ß-driven cardiac muscle dysfunction. These findings are consistent with mitochondrial calcium toxicity induced by SR Ca2+ re-uptake and leak, and heart failure with reduced ATP production. The data suggest that excess TGF-ß release by pathological bone destruction could have significant impact on cardiac muscle to induce oxidative stress, SR Ca2+ leak and heart failure and could explain heart disease observed clinically with bone metastasis. Targeting this pathway may improve survival in patients with bone metastases. Citation Format: Lei Shi, Truc Kuo, Jade Martinez, Sukanya Suresh, Trupti Trivedi, Gabriel M. Pagnotti, Leah D. Guerra, Xu Cao, Theresa A. Guise. Bone-derived TGF-ß release is associated with cardiac dysfunction due to oxidation of SR calcium gatekeepers, Ca2+ leak and mitochondrial dysfunction [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3208.

Serum Sclerostin and Its Association with Bone Turnover Marker in Metabolic Bone Diseases.

Sclerostin is a secreted inhibitor of Wnt/β-catenin signaling that is mainly produced by osteocytes and is an important regulator of bone remodeling. Some studies have evaluated serum sclerostin levels in metabolic bone diseases, but the results have been contradictory. The profile of serum sclerostin levels in patients with osteogenesis imperfecta (OI), X-linked hypophosphatemia (XLH), and Paget's disease of bone (PDB) was obtained to determine their association with bone turnover marker. Serum sclerostin levels, biochemical parameters, and the bone turnover marker, β-CrossLaps of type 1 collagen containing cross-linked C-telopeptide (β-CTX), were measured in 278 individuals, comprising 71 patients with OI, 51 patients with XLH, 17 patients with PDB, and 139 age- and sex-matched healthy controls. A correlation analysis was performed between sclerostin and β-CTX concentration. The univariate logistic regression analysis was used to analyze factors associated with OI, XLH, and PDB. Patients with PDB (11 male 6 female), aged 44.47 ± 14.75 years; XLH (17 male, 34 female), aged 19.29 ± 15.65 years; and OI (43 male, 28 female), aged 19.57 ± 16.45 years, had higher sclerostin level than age- and sex-matched healthy controls [median(interquartile range): 291.60 (153.42, 357.35) vs. 38.00 (27.06, 68.52) pmol/L, 163.40 (125.10, 238.20) vs. 31.13 (20.37, 45.84) pmol/L, and 130.50 (96.12, 160.80) vs. 119.00 (98.89, 194.80) pmol/L, respectively; P < 0.001]. Patients with PDB had the highest level of serum sclerostin, followed by those with XLH and OI (P < 0.05). Sclerostin was positively correlated with β-CTX in OI and XLH (r = 0.541 and r = 0.661, respectively; P < 0.001). Higher β-CTX and sclerostin levels were associated with a higher risk of OI, XLH, and PBD. Sclerostin may be a biomarker of OI, XLH, and PDB. Whether sclerostin inhibitors can be used in these patients requires further analysis using additional cohorts.

Уровни маркеров метаболизма костной ткани (N-терминального пропептида проколлагена I типа и β-терминального телопептида коллагена I типа) у жителей Украины в зависимости от пола и возраста

Целью исследования является определение уровня N-терминального пропептида проколлагена I типа (PINP) и карбокситерминального телопептида коллагена I типа (β-CTX) и их взаимосвязи в сыворотке крови у здорового населения Украины разного возраста и пола. Материалы и методы. В исследование было включено 3850 человек в возрасте 20–89 лет (3351 женщина (средний возраст 59,05 ± 13,89 года) и 499 мужчин (49,17 ± 15,8 года)). Всех участников распределили по возрасту в соответствии с геронтологической классификацией на 4 группы: молодой возраст — 20–44 года, средний — 45–59 лет, пожилой — 60–74 года и старческий возраст — 75–89 лет, а также на 7 групп по десятилетиям: от 20 до 89 лет. Концентрацию β-CTX и PINP в сыворотке крови определяли методом электрохемилюминесцентного иммуно­анализа ECLIA на анализаторе cobas e 411. Результаты. Выявлено достоверное влияние возраста на вариабельность β-CTX и PINP у мужчин (F = 6,64, р &lt; 0,001, и F = 4,93, р &lt; 0,001) и женщин (F = 16,17, р &lt; 0,001, и F = 10,46, р &lt; 0,001). У женщин по критерию Шеффе обнаружили достоверно более высокий уровень β-CTX в группе 80–89 лет по сравнению с группами 20–29 лет (р = 0,01), 30–39 лет (р &lt; 0,001), 40–49 лет (р &lt; 0,001), 50–59 лет (0,52 ± 0,27, р = 0,002), 60–69 лет (р = 0,001) и 70–79 лет (р = 0,04). Показатели PINP в группе 80–89 лет также были достоверно выше в сравнении с группами 30–39 лет (р &lt; 0,001), 40–49 лет (р &lt; 0,001), 50–59 лет (р = 0,009), 60–69 лет (р = 0,002) и 70–79 лет (р = 0,02). У мужчин обнаружили достоверное снижение концентрации β-CTX в группах: 30–39 лет (р = 0,02), 40–49 лет (р &lt; 0,001), 50–59 лет (р = 0,01) и 60–69 лет (р = 0,001) в сравнении с группой 20–29 лет. Уровень PINP был достоверно ниже в группах 40–49 лет (р &lt; 0,001) и 50–59 лет (р = 0,03) в сравнении с группой 20–29 лет. По результатам исследования, коэффициент корреляции между показателями концентрации PINP и β-CTX изменяется в зависимости от возраста и пола. Самые высокие коэффициенты корреляции среди обоих полов наблюдались в возрастных группах 40–49 (женщины: r = 0,74; мужчины: r = 0,64) и 50–59 (женщины: r = 0,74; мужчины: r = 0,66) лет; низкие коэффициенты корреляции у женщин наблюдались в возрастной группе 20–29 (r = 0,63) и 70–79 (r = 0,65) лет; у мужчин (r = 0,53) в возрасте 20–29 и 70–79 лет. Выводы. Выявлено достоверное влияние возраста на вариабельность β-CTX и PINP в сыворотке крови у мужчин и женщин. Наиболее высокая корреляционная связь между концентрациями β-CTX и PINP наблюдалась в группах 40–49 и 50–59 лет среди обоих полов. Полученные результаты могут быть использованы у практически здоровых мужчин и женщин Украины как референтные, а в будущем будут необходимыми для изучения метаболизма костной ткани при вторичном остеопорозе и его осложнениях.

Bone metabolic marker concentrations across the menstrual cycle and phases of combined oral contraceptive use.

There is a need to further understand the impact of the menstrual cycle and phase of combined oral contraceptive (COC) use on the pre-analytical variability of markers of bone metabolism in order to improve standardisation procedures for clinical practice and research. The aim of this study was to assess bone metabolism marker concentrations across the menstrual cycle and phases of COC use. Carboxy-terminal cross-linking telopeptide of type I collagen (β-CTX), procollagen type 1N propeptide (P1NP) and Bone alkaline phosphatase (Bone ALP) concentrations were assessed in eumenorrheic women (n=14) during the early follicular, ovulatory and mid-luteal phases of the menstrual cycle and in COC (Microgynon®) (n=14) users on day 2-3 of pill consumption (PC1), day 15-16 pill consumption (PC2) and day 3-4 of the pill free interval (PFI). β-CTX was significantly (-16%) lower at PC2 compared to PC1 (P=0.015) in COC users and was not affected by menstrual cycle phase (P>0.05). P1NP and Bone ALP were not significantly different across either menstrual cycle phase or phase of COC use (all P>0.05). There was no difference in pooled bone marker concentrations between eumenorrheic women and COC users (P>0.05). In contrast to some previous studies, this study showed that bone marker concentrations do not significantly fluctuate across the menstrual cycle. Furthermore, bone resorption markers are significantly affected by phase of COC use, although bone formation markers do not significantly vary by COC phase. Therefore, the phase of COC use should be considered in clinical practice and research when assessing markers of bone metabolism as this can impact circulating concentrations of bone metabolic markers yet is not currently considered in existing guidelines for best practice.

Association of Diabetic Kidney Disease Markers and Urinary Beta-crossLaps in Type 2 Diabetes

Introduction: Diabetic Kidney Disease (DKD) is a chronic complication in Type 2 diabetes. The Chronic Kidney Disease (CKD273) peptide classifier has been found to predict development of DKD even before microalbuminuria develops. Seventy four percent of peptides in the CKD273 classifier are Collagen degradation fragments. The Beta-CrossLaps (β-CTx) Enzyme Llinked Immunosorbent Assay (ELISA) assay detects the specific collagen degradation product, C terminal telopeptide of Type 1 collagen. In light of the Capillary Electrophoresis/Mass Spectrometry (CE-MS) findings, linking collagen degradation fragments excretion to early detection of DKD, the significance of urinary β-CTx levels as a DKD biomarker needs to be evaluated. Aim: To study the urinary excretion of β-CTx in type 2 diabetes patients and to evaluate its relation to Microalbuminuria status and estimated Glomerular Filtration Rate (eGFR) of the patients. Materials and Methods: This descriptive cross-sectional study was undertaken at a tertiary care hospital, with enrollment of 82 type 2 diabetes patients from the diabetes Out Patient Department (OPD) Participants were divided into groups based on their Urinary Albumin Creatinine Ratio (UACR) and eGFR levels. The study participants were tested for Urinary β-CTx level, UACR and eGFR. Mean or median was calculated for the parameters with normal and non-normal distribution, respectively. All statistical testing was performed on online calculators available at the site; https://www.socscistatistics.com/. Results: The median urinary β-CTx level observed was 100.6 ng/mmol of creatinine. Among the 82 participants, 15 participants had urinary β-CTx level 15pg/mL, the sensitivity of the kit. Among the remaining 67 participants, the minimum Urinary Beta-CrossLaps: Creatinine ratio observed was 2.6 ng/mmol and the maximum value observed was 2071 ng/mmol (i.e., 2.1 μg/mmol). The median urinary β-CTx level was highest (100.6 ng/mmol creatinine) in the patient group with eGFR in the normal range. The urinary β-CTx level was found to decline with decline in eGFR, with median urinary β-CTx 65.5 ng/mmol creatinine in the patient group with mildly decreased eGFR and 7.2 ng/mmol creatinine in the patient group with moderately decreased eGFR. Conclusion: The Urinary β-CTx concentration in type 2 diabetes patients is dispersed over a wide range. The Urinary β-CTx concentration correlates with the eGFR of the patient and is not influenced by age, gender or duration of diabetes. This parameter is a potential early DKD biomarker.

Applied Value of Serum Bone Turnover Markers in Patients with Myeloma Bone Disease

To explore the clinical value of serum bone turnover markers including β-CrossLaps (β-CTx), N-MID Osteocalcin (Osteocalcin), and total procollagen type 1 amino-terminal propeptide (TP1NP) in patients with myeloma bone disease (MBD). A total of 55 MBD patients(MBD group) and 20 healthy volunteers(control group) were selected, and the serum was collected for detecting β-CTx, Osteocalcin and TP1NP by Roche analyzer of automated electrochemiluminescence. Meanwhile, the imaging techniques such as MRI and CT were used for evaluation of bone destruction and the damage extent in MBD patients. Measurement data is expressed as median (P25, P75) according to distribution characteristics of data. The detection results showed that concentrations of β-CTx in MBD patients and control group were 0.72(0.48, 1.28) ng/mL and 0.53(0.34, 0.61) ng/mL respectively, the β-CTx concentration in MBD patients was significantly higher than that in control group(P=0.002). The ratio β-CTx to TP1NP (%) in MBD patients and control group was 1.50 (1.05, 3.36) and 1.25 (0.86, 1.35) respectively, the ratio in MBD patients was significantly higher than that in control group (P=0.007). The serum β-CTx concentrations in MBD patients of localized bone destruction type and extensive bone destruction type were 0.41(0.31, 0.66) ng/mL and 1.14(0.72, 1.81) ng/mL respectively, the β-CTx concentration in MBD patients of extensive bone destruction type was significantly higher than that in MBD patients of localized bone destruction type (P＜0.001). The ratio of β-CTx to TP1NP(%) in MBD patients of localized and extensive bone destruction type was 1.30 (0.90, 2.49) and 1.98 (1.18, 3.76) respectively, the ratio in the MBD patients of extensive bone destruction type was significantly higher than that in MBD patients of localized bone destruction type (P=0.019). The serum osteocalcin concentrations in MBD patients of localized and extensive bone destruction type were 14.31 (8.82, 19.39) ng/mL and 21.52 (14.42, 47.76) ng/mL respectively, the osteocalcin concentration in MBD patients of extensive bone destruction type was higher than that in MBD patients of localized bone destruction type (P=0.008). The AUC of β-CTx was 0.88 (95% CI: 0.78-0.98)(P＜0.001), and the cut-off value was 0.69 ng/ml in the diagnosis of extensive bone injury for MBD patients, and the sensitivity and specificity were 80.65% and 83.33% respectively. The MBD patients show bone resorption hyperthyroidism and high bone turnover. The β-CTx and osteocalcin in serum bone turnover markers can effectively reflect the extent of bone damage in MBD patients, especially the β-CTx is more efficient for the diagnosis of MBD patients of extensive bone destruction type. However, β-CTx, osteocalcin and TP1NP are not relate with the MM disease progression.

Association between bile acid metabolism and bone mineral density in postmenopausal women

OBJECTIVES:Previous studies have not shown any correlation between bile acid metabolism and bone mineral density (BMD) in women with postmenopausal osteoporosis. Thus, the current study evaluated the association between bile acid levels as well as BMD and bone turnover marker levels in this group of women.METHODS:This single-center cross-sectional study included 150 postmenopausal Chinese women. According to BMD, the participants were divided into three groups: osteoporosis group, osteopenia group, and healthy control group. Serum bile acid, fibroblast growth factor 19 (FGF19), and bone turnover biomarker levels were assessed. Moreover, the concentrations of parathyroid hormone, 25-hydroxy vitamin D [25(OH)D], procollagen type I N-peptide (P1NP), and beta-CrossLaps of type I collagen containing cross-linked C-terminal telopeptide (β-CTX) were evaluated. The BMD of the lumbar spine and proximal femur were examined via dual-energy X-ray absorptiometry.RESULTS:The serum total bile acid levels in the osteoporosis and osteopenia groups (5.28±1.56 and 5.31±1.56 umol/L, respectively) were significantly lower than that in the healthy control group (6.33±2.04 umol/L; p=0.002 and 0.018, respectively). Serum bile acid level was positively associated with the BMD of the lumbar spine, femoral neck, and total hip. However, it negatively correlated with β-CTX concentration. Moreover, no correlation was observed between bile acid and P1NP levels, and the levels of the other biomarkers that were measured did not differ between the groups.CONCLUSION:Serum bile acid was positively correlated with BMD and negatively correlated with bone turnover biomarkers reflecting bone absorption in postmenopausal women. Thus, bile acid may play an important role in bone metabolism.

Bone formation and resorption markers at 7 years of age: Relations with growth and bone mineralization.

PurposeWe aimed to describe bone formation and resorption markers in generally healthy prepubertal children using total alkaline phosphatase (tALP), osteocalcin (OC) and β-isomerized C-terminal telopeptides of type I collagen (β-CTx) serum concentrations and to estimate markers’ correlations with anthropometric growth (height, weight, body mass index and trajectories of weight gain) as well as bone mineral content (BMC) and areal density (aBMD).MethodsWe assessed 395 7-year-old children from the Generation XXI cohort with tALP, OC and β-CTx concentrations determined from a fasting venous blood sample and BMC/aBMD measured by dual-energy X-ray absorptiometry. Gender-specific reference intervals for tALP, OC and β-CTx in 7-year-old children were established by calculating the 2.5th and 97.5th percentiles. Pearson and partial correlation coefficients (controlling for sex, age, body size and season) between bone markers and growth measures were computed.ResultstALP increased with height (rpartial controlled for sex = 0.26, 95%CI: 0.17, 0.35), was higher in overweight than in healthy weight children, and in children who gained weight above average during infancy. No correlations were found between OC or β-CTx and growth. In girls, OC was slightly correlated with subtotal BMC (rpartial = 0.22, 95%CI: 0.08, 0.35), subtotal aBMD (rpartial = 0.20, 95%CI: 0.06, 0.33) and lumbar spine aBMD (rpartial = 0.23, 95%CI: 0.09, 0.36). tALP and β-CTx were not correlated with any of the DXA-derived bone measures.ConclusionThis study contributed to the description of bone turnover at 7 years of age and suggested that bone metabolism markers measured in a single point in time have limited ability to describe anthropometric growth and overall bone status in generally healthy prepubertal children.

Study on diagnostic value of P1NP and β-CTX in bone metastasis of patients with breast cancer and the correlation between them.

This study aimed to investigate the diagnostic value of the total amino-terminal propeptide of type 1 procollagen (P1NP) and C-terminal telopeptide of β-I collagen (β-CTX) in bone metastasis of patients with breast cancer and the correlation between them. The medical records of 73 patients were retrospectively analyzed. These patients with breast cancer were treated in Oncology, General Surgery, and Orthopedic Departments in The Third People's Hospital of Qingdao from March 2014 to April 2017, including 40 patients with bone metastasis (bone metastasis group) and 33 patients with no bone metastasis (non-bone metastasis group). Other 40 healthy people who received physical examination in the same period were selected as the control group. The expression of P1NP and β-CTX in plasma were detected by the Enzyme-linked immunosorbent assay, and the correlation between them was analyzed. There were significant differences in P1NP and β-CTX concentrations among the three groups (p<0.05). The concentrations of P1NP in the control group and the non-bone metastasis group were significantly lower than that in the bone metastasis group (p<0.05); the concentrations of β-CTX in the control group and the non-bone metastasis group were significantly lower than that in the bone metastasis group (p<0.05). P1NP: AUC=0.852, sensitivity: 72.5%, specificity: 93.9%, CUT OFF=66.44. β-CTX: AUC=0.883, sensitivity: 85.0%, specificity: 84.8%, CUT OFF=69.8. Joint detection: AUC=0.952, sensitivity: 84.8%, specificity: 99.5%, CUT OFF=99.5. The results of the concentrations of P1NP and β-CTX in the bone metastasis group detected by the Pearson correlation analysis showed that their concentrations were positively correlated in the bone metastasis group (r=0.764, p<0.05). P1NP and β-CTX in plasma have a high diagnostic value for bone metastasis of breast cancer and have important significance in the diagnosis of bone metastasis and disease monitoring.

LncRNA SNHG1 modulates p38 MAPK pathway through Nedd4 and thus inhibits osteogenic differentiation of bone marrow mesenchymal stem cells

AimsCurrent study aimed to investigate the effects of lncRNA SNHG1 on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and explore the underlying mechanisms. Main methodsMouse model of osteoporosis was created by ovariectomy (OVX). The BMD of mice spine, the serum level of β-CTX and the ALP activity were measured. The expression of SNHG1, JNK, p-JNK, p-38, p-p38 and Osterix were examined by qRT-PCR and Western blot. Co-IP assay was used to verify the effect of SNHG1 on the interaction between p-p38 and Nedd4. Ubiquitination assay was used to evaluate the roles of SNHG1 in ubiquitination of p-p38. Key findingsIn the mice with osteoporosis, BMD was decreased and β-CTX concentration and SNHG1 expression were increased. ALP activity and p-p38 protein level were elevated and SNHG1 expression was down-regulated in BMSCs stimulated by osteogenic inducer, while the effects were reversed by SNHG1 over-expression. SNHG1 over-expression enhanced the interaction between Nedd4 and p-p38, disrupted protein stability of p-p38, and promoted the ubiquitination of p-p38. In addition, pcDNA-SNHG1 down-regulated p-p38 protein level, and sh-Nedd4 removed the trend. Nedd4 silencing elevated ALP activity and Osterix protein level, while p-38 inhibitor abrogated the effects. In vivo, SNHG1 silence increased BMD and Osterix protein level, and decreased endogenous SNHG1 expression in mice with OVX. SignificanceThis study proved the regulation mechanism that lncRNA SNHG1 negatively regulates p38 MAPK signal pathway through ubiquitination mediated by Nedd4, and thus inhibits osteogenic differentiation of BMSCs.

Bone metabolic responses to low energy availability achieved by diet or exercise in active eumenorrheic women.

We aimed to explore the effects of low energy availability (EA)[15 kcal·kg lean body mass (LBM)-1·d-1] achieved by diet or exercise on bone turnover markers in active, eumenorrheic women. By using a crossover design, ten eumenorrheic women (VO2 peak: 48.1 ± 3.3 ml·kg-1·min-1) completed all three, 3-day conditions in a randomised order: controlled EA (CON; 45 kcal·kgLBM-1·d-1), low EA through dietary energy restriction (D-RES; 15 kcal·kgLBM-1·d-1) and low EA through increasing exercise energy expenditure (E-RES; 15 kcal·kgLBM-1·d-1), during the follicular phase of three menstrual cycles. In CON, D-RES and E-RES, participants consumed diets providing 45, 15 and 45 kcal·kgLBM-1·d-1. In E-RES only, participants completed supervised running sessions (129 ± 10 min·d-1) at 70% of their VO2 peak that resulted in an exercise energy expenditure of 30 kcal·kg LBM-1·d-1. Blood samples were collected at baseline (BASE) and at the end of the 3-day period (D6) and analysed for bone turnover markers (β-CTX and P1NP), markers of calcium metabolism (PTH, albumin-adjusted Ca, Mg and PO4) and hormones (IGF-1, T3, insulin, leptin and 17β-oestradiol). In D-RES, P1NP concentrations at D6 decreased by 17% (BASE: 54.8 ± 12.7 μg·L-1, D6: 45.2 ± 9.3 μg·L-1, P < 0.001, d = 0.91) and were lower than D6 concentrations in CON (D6: 52.5 ± 11.9 μg·L-1, P = 0.001). P1NP did not change significantly in E-RES (BASE: 55.3 ± 14.4 μg·L-1, D6: 50.9 ± 15.8 μg·L-1, P = 0.14). β-CTX concentrations did not change following D-RES (BASE: 0.48 ± 0.18 μg·L-1, D6: 0.55 ± 0.17 μg·L-1) or E-RES (BASE: 0.47 ± 0.24 μg·L-1, D6: 0.49 ± 0.18 μg·L-1) (condition × time interaction effect, P = 0.17). There were no significant differences in P1NP (P = 0.25) or β-CTX (P = 0.13) responses between D-RES and E-RES. Both conditions resulted in reductions in IGF-1 (-13% and - 23% from BASE in D-RES and E-RES, both P < 0.01) and leptin (-59% and - 61% from BASE in D-RES and E-RES, both P < 0.001); T3 decreased in D-RES only (-15% from BASE, P = 0.002) and PO4 concentrations decreased in E-RES only (-9%, P = 0.03). Low EA achieved through dietary energy restriction resulted in a significant decrease in bone formation but no change in bone resorption, whereas low EA achieved through exercise energy expenditure did not significantly influence bone metabolism. Both low EA conditions elicited significant and similar changes in hormone concentrations.

Definition of reference ranges for β-isomerized carboxy-terminal telopeptide collagen type I for children and adolescents.

Bone metabolism involves many complex pathways that are disturbed by several bone diseases. The literature shows some limitations concerning pediatric reference intervals to bone markers, mainly because of the low number of patients included in the studies, the heterogeneity of methods, beyond the fact that it is time-consuming and expensive. The aim of this study was to determine reference values for β-isomerized carboxy-terminal telopeptides collagen type I (β-CTX), a marker of bone resorption, for children and adolescents. Blood samples from 246 patients were collected and β-CTX was measured using an electrochemiluminescence immunoassay (ECLI). We propose reference ranges for β-CTX concentration from the 2.5 percentile and 97.5 percentile for each age group. The reference values obtained, concerning children and adolescents, might be useful in the evaluation of diseases such as osteosarcoma and anorexia in both childhood as adolescence.

The Effect of Postexercise Carbohydrate and Protein Ingestion on Bone Metabolism

ABSTRACT This study aimed to investigate the effect of feeding carbohydrate and protein (CHO + PRO), immediately or 2 h after an exhaustive run, on the bone turnover response in endurance runners. Ten men (age = 28 ± 5 yr, height = 1.74 ± 0.05 m, body mass [BM] = 69.7 ± 6.3 kg) performed treadmill running at 75% V˙O2max, until exhaustion, on three occasions. Blood was collected before and immediately, 1, 2, 3, 4, and 24 h postexercise, for measurement of β-CTX, P1NP, parathyroid hormone, PO4, ACa, and Ca2+. This was a randomized, counterbalanced, placebo-controlled, and single-blinded crossover study. The three trials were (i) placebo (PLA), where the PLA solution was ingested immediately and 2 h postexercise; (ii) immediate feeding (IF), where CHO + PRO (1.5 g·kg−1 BM dextrose and 0.5 g·kg−1 BM whey) was ingested immediately postexercise and PLA 2 h postexercise; and (iii) delayed feeding (DF), where PLA was ingested immediately postexercise and CHO + PRO solution 2 h postexercise. Data were analyzed using repeated-measures ANOVA and Tukey's HSD post hoc test. At 1 and 2 h postexercise, β-CTX concentrations were lower in the IF trial compared with the DF and PLA trials (P ≤ 0.001). At 3 h postexercise, β-CTX concentrations were higher in the PLA trial compared with the IF (P ≤ 0.001) and DF trials (P = 0.026). At 4 h postexercise, β-CTX concentrations were lower in the DF trial compared with the IF (P = 0.003) and PLA trials (P ≤ 0.001). At 4 h postexercise, P1NP was higher in the IF trial compared with the DF (P = 0.026) and PLA trials (P = 0.001). At 3 h postexercise, parathyroid hormone was higher in the IF trial compared with the DF trial (P ≤ 0.001). After exhaustive running, immediate ingestion of CHO + PRO may be beneficial, as it decreases bone resorption marker concentrations and increases bone formation marker concentrations, creating a more positive bone turnover balance.