C-terminal Cross-linking Telopeptide Of Type Research Articles

Associations between the serum levels of selected bone turnover markers and biological traits in nursing home women aged 80+ without inflammation. A pilot study

Bone turnover markers (BTM) reflect the status of bone remodeling processes responsible for bone mineral density. The existing body of evidence that osseous tissue can interact with many other body tissues and organs suggests that the cross-talks can lead to different relationships. The biological traits associated with BTMs have not been thoroughly studied in the elderly despite bone turnover being known to increase with age. To determine whether the C-terminal crosslinking telopeptides of type I collagen (CTXI) and the serum levels of total (TAP) and bone-specific (BAP) alkaline phosphatase are associated with the biological traits in nursing home women aged 80-92years without inflammation and, if so, to indicate the best predictors of these BTM's blood concentrations. A group of 64 female volunteers aged 80years and older, the residents of nursing homes, were screened for the study. Fifty two women were ineligible as they met the exclusion criteria. As a result, the study group consisted of 12 participants (85.1±3.9years; 58.1±8.7kg; 1.52±0.06m), all having blood C-reactive protein (CRP) levels below 3mg/l. Also assessed were the participants' morphology, glucose and insulin levels, lipid profiles, CTXI, TAP, and BAP. Other measured parameters included body composition, resting heart rate and arterial blood pressure, isometric knee extension peak torque (IKEPT), and walking capacity (6-min walk test). The statistical analysis was performed using Pearson's correlation coefficients, the Benjamini-Hochberg procedure, and a stepwise multiple regression analysis with backward elimination. Inverse correlations were found between CTX-I and hemoglobin concentration (HGB) (r=-0.680; p=.015), red blood cells count (RBC) (r=-0.664; p=.019), fat-free mass (r=-0.633; p=.027), body weight (r=-0.589; p=.044), and total cholesterol (r=-0.581; p=.048). The multiple regression analysis of CTX-I showed that body weight was the only independent variable that was statistically significant (r2=0.346; p<.05; SEE=0.347ng/ml). BAP was positively correlated with double product (DP) (r=0.742; p=.006), RBC (r=0.650; p=.022), HGB (r=0.637; p=.026), mean arterial pressure (MAP) (r=0.622; p=.031), diastolic blood pressure (DBP) (r=0.612; p=.034), body height (r=0.603; p=.038), IKEPT (r=0.565; p=.056), and systolic blood pressure (SBP) (r=0.538; p=.071). BAP, a dependent variable, was the most closely correlated with DP (r2=0.550; p<.01; SEE=9.161U/l). TAP was also significantly associated with DP (r=0.775; p=.003), with the association being stronger than between BAP and DP (r2=0.600; p<.01; SEE=1000.5 beats/min*mmHg). In relatively healthy the oldest-old nursing home women without inflammation, total body weight was the best predictor of bone resorption shown by the CTX-I concentration, whereas the rate pressure product (DP) turned out to best predict osteoblastic activity determinable from serum alkaline phosphatase activity. The results of the study suggest that the activity of serum TAP and BAP can be enhanced by different mechanisms.

Effect of Denosumab on Bone Mineral Density of Hematopoietic Stem Cell Transplantation Recipients.

Purpose Denosumab is a monoclonal antibody that prevents the development of osteoclasts. The effect of denosumab in solid organ transplant recipients has been elucidated, but its effect in haematopoietic stem cell transplantation recipients has not been studied yet. The aim of this study was to determine the effectiveness and safety of denosumab in haematopoietic stem cell transplantation recipients. Methods We retrospectively evaluated 33 female patients with osteoporosis (mean age 52.6 ± 9.8 years) following allogeneic haematopoietic stem cell transplantation. Patients were treated with denosumab every 6 months for 12 months. Changes in bone mineral density were evaluated for denosumab-treated patients in a 12-month interval after the first administration of denosumab. Results Significant increases in bone mineral density were observed in all measured skeletal sites including 4.39 ± 6.63% in the lumbar spine (p=0.014), 3.11 ± 7.69% in the femoral neck (p=0.048), and 1.97 ± 6.01% in the total hip (p=0.138). The bone turnover marker serum cross-linked C-terminal telopeptide of type 1 collagen was decreased at 18 months (−51.6 ± 17.6%, p < 0.001). No serious symptomatic hypocalcaemia was observed. Serious adverse drug reactions requiring drug discontinuation were not observed. Conclusion Denosumab improved bone mineral density in haematopoietic stem cell transplantation recipients. The use of denosumab could be a good therapeutic option without causing severe adverse effects in recipients of haematopoietic transplantation.

Biochemical bone turnover markers in hormonal disorders in adults: a narrative review.

Hormonal disorders are often associated with abnormal levels of bone turnover markers (BTMs). N-terminal propeptide of type I procollagen (PINP) and serum C-terminal cross-linking telopeptide of type I collagen (CTX-I) are the reference markers of bone formation and bone resorption, respectively. A comprehensive literature search within the MEDLINE and Web of Science databases was performed. Acromegaly is associated with higher BTM levels, which decrease during the remission after treatment. Adult-onset growth hormone deficiency is often associated with decreased BTM levels. Growth hormone replacement therapy stimulates bone turnover and increases BTM levels. Hypothyroidism is characterized by general slowing of bone metabolism which is reflected by lower BTM levels. The replacement thyroid hormone therapy increases the bone turnover rate and BTM levels increase. Patients with thyroid cancer receive a suppressive dose of thyroid hormones and may have slightly elevated BTM levels. Patients with overt hyperthyroidism had higher BTM levels and anti-thyroid therapy induces a rapid decrease in the BTM levels. Patients with overt primary hyperparathyroidism have higher BTM levels, whereas those with asymptomatic and normocalcemic hyperparathyroidism usually have normal BTM levels. Hypoparathyroidism is characterized by slightly decreased BTM levels. Cushing's syndrome is characterized consistently by markedly decreased osteocalcin concentration, whereas data on other BTMs are discordant. BTMs help us to better understand mechanisms of the impact of hormonal disorders and their treatment on bone metabolism. However, it is unknown whether BTMs may be used to monitor the effect of their treatments on bone in the clinical practice.

Bone metabolism in Chinese patients after laparoscopic Roux-en-Y gastric bypass.

BackgroundLaparoscopic Roux-en-Y gastric bypass is a leading metabolic surgery for obese patients with type 2 diabetes mellitus. However, postoperative nutritional deficiencies and abnormal bone metabolism are possible. In this study, we investigated the mid-term impact on bone metabolism of laparoscopic Roux-en-Y gastric bypass.MethodsThis retrospective study involved 238 patients with type-2 diabetes mellitus and obesity who underwent laparoscopic Roux-en-Y gastric bypass in our institution; patients were followed for 3 years. Patients received a calcium supplement (1,200 mg per day), alfacalcidol (0.5 µg per day), and a multivitamin (1 tablet per day), postoperatively. We recorded and analyzed patients’ body mass index, and blood levels of 25-hydroxy-vitamin D, calcium, and parathyroid hormone. Data for 16 patients 6 months postoperatively were selected randomly to evaluate bone-specific alkaline phosphatase and the bone resorption marker, C-terminal crosslinking telopeptide of type I collagen (CTX-1).ResultsOf the 238 patients, 111 were women, and 127 were men (mean age: 46.91±12.03 years; mean body mass index: 31.37 kg/m2). 25-hydroxy vitamin D3 levels did not change (P>0.05) compared with preoperative levels during the 3-year follow-up. Parathyroid hormone (PTH) levels increased (P<0.05), while serum calcium decreased (P<0.05). Patients’ body mass index decreased sharply 3 months postoperatively (preoperative vs. postoperative mean: 31.37±3.52 vs. 25.60±2.86, respectively; P<0.05). We saw no decrease in 25-hydroxy vitamin D3 levels with supplementation, postoperatively. CTX-1 and bone-specific alkaline phosphatase levels showed no significant changes comparing pre- and postoperative levels at 6-, 12-, and 24 months (P>0.05). However, CTX-1 levels increased in the first 12 months postoperatively, then decreased in the second year; bone alkaline phosphatase levels increased gradually.ConclusionsLaparoscopic Roux-en-Y gastric bypass is effective for weight loss, but also has an impact on bone metabolism, which increases the risk of fracture. Our results highlight the importance of regular follow-ups and full-dose calcium and vitamin supplementation, postoperatively.

Ginsenoside Rg3 Attenuates Aluminum-Induced Osteoporosis Through Regulation of Oxidative Stress and Bone Metabolism in Rats.

Aluminum (Al)-induced bone metabolism disorder is a primary cause of osteoporosis. Ginsenoside Rg3 (Rg3) has demonstrated therapeutic properties in the treatment of osteoporosis. The present study aimed to identify potential bone protection mechanisms of Rg3 against Al-induced osteoporosis in rats. In this study, forty healthy male Sprague-Dawley rats were randomly allocated into groups in which they were treated with AlCl3 (64mg/kg/day) and/or Rg3 (20mg/kg/day). AlCl3 was given orally to rats for 120days, and from the 91st day, treated orally with Rg3 for 30days. Rg3 attenuated AlCl3-induced accumulation of Al by decreasing the bone mineral density in the lumbar spines, femoral metaphysis, and tibia, and inhibited AlCl3-induced oxidative stress in rat bone by decreasing the levels of reactive oxygen species and malondialdehyde, while increasing glutathione peroxidase and superoxide dismutase activity. Rg3 facilitated bone formation by increasing the concentration of calcium, phosphorus, amino-terminal propeptide of type I procollagen, and carboxyl-terminal propeptide of type I procollagen, bone alkaline phosphatase activity in serum, and type I collagen, osteocalcin, and osteopontin protein expressions. Rg3 inhibited bone resorption by decreasing the content of N-terminal cross-linking telopeptide of type I collagen, C-terminal cross-linking telopeptide of type I collagen, and tartrate-resistant acid phosphatase 5b activity in serum. Rg3 promoted the mRNA expression of growth regulation factors by increasing transforming growth factor-β1, bone morphogenetic protein-2, insulin-like growth factor I, and core-binding factor α1. The results demonstrate that Rg3 can significantly attenuate Al accumulation, facilitate bone formation, inhibit bone resorption, resist oxidative stress, and promote the expression of factors that regulate growth. The results indicate that Rg3 is effective in alleviating AlCl3-induced osteoporosis.

Prevalence and contributing factors of osteoporosis in the elderly over 70 years old: an epidemiological study of several community health centers in Shanghai.

OP seriously affects the health status and quality of life of the elderly, even endangers life. This study aimed to investigate the prevalence and contributing factors of osteoporosis (OP) in the elderly over 70 years old of several Community Health Centers in Shanghai. A total of 565 elderly subjects were recruited and received questionnaire survey. The general characteristics, medical history and history of OP treatment were collected. Bone mineral density (BMD) was detected, and the awareness rate, prevalence and treatment rate were determined in all of the subjects. Moreover, activity of daily living and nutritional status were also evaluated. Logistic regression analysis was performed to calculate the OR and its 95% confidence interval. Bone turnover markers (BTMs) [procollagen type 1 N-peptide (P1NP), C-terminal cross-linked telopeptides of type 1 collagen (β-CTX), molecular fragment of N-terminal osteocalcin (N-MID), and 25 hydroxyvitamin D (25OHD)] were detected and their relationship with OP was further evaluated with Pearson correlation analysis. : (I) The awareness rate of OP was 89.7%, the prevalence was 39.5%, and the treatment rate was 59.5% in the elderly over 70 years old of 4 Community Health Center in Shanghai; (II) the male gender and good nutritional status were independent protective factors of OP, and aging and poor activity of daily living were independent risk factors of OP; (III) β-CTX was negatively related to BMD (r=-0.286, P=0.000), but P1NP, N-MID, and 25OHD were positively related to BMD (r=0.254, 0.349 and 0.357, P=0.000, 0.000 and 0.000). Female gender, aging, malnutrition, and inability to take care of themselves are risk factors of OP in people over 70 years old of 4 Community Health Centers in Shanghai. Some BTMs are related to the BMD. BTMs together with BMD may be used to comprehensively evaluate the bone state, identify the old people with OP at early stage, and guide the clinical treatment and early monitoring of OP.

Development of 3-mercaptopropyltrimethoxysilane (MPTS)-modified bone marrow mononuclear cell membrane chromatography for screening anti-osteoporosis components from Scutellariae Radix

Osteoporosis is a bone metabolic disease caused by the imbalance between osteoblasts and osteoclasts due to excess osteoclastogenesis, manifesting in the decrease of bone density and bone strength. Scutellariae Radix shows good anti-osteoporosis activity, but the effective component is still unclear. Cell membrane chromatography (CMC) is a biological affinity chromatography with membrane immobilized on a silica carrier as the stationary phase. It can realize a dynamical simulation of interactions between drugs and receptors on cell membrane, which is suitable for screening active compounds from complex systems. In this study, the components of Scutellariae Radix with potential anti-osteoporosis activity through inhibiting the differentiation from bone marrow mononuclear cells (BMMCs) to osteoclast were screened by a BMMC/CMC analytical system. Firstly, a new 3-mercaptopropyltrimethoxysilane (MPTS)-modified BMMC/CMC stationary phase was developed to realize covalent binding with cell membrane fractions. By investigating the retention time (tR) of the positive drug, the life span of the MPTS-modified CMC columns was significantly improved from 3 to 12 days. Secondly, 6 components of Scutellariae Radix were screened to show affinity to membrane receptors on BMMCs by a two-dimensional BMMC/CMC–TOFMS analytical system. Among them, tectochrysin demonstrated the best anti-osteoporosis effect in vitro, which has never been reported. We found that tectochrysin could inhibit the differentiation of BMMCs into osteoclasts induced by receptor activator of nuclear factor-κΒ ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) in a concentration-dependent manner in vitro. In vivo, it significantly reduced the loss of bone trabeculae in ovariectomized mice, and decreased the level of C-terminal cross-linking telopeptides of type 1 collagen (CTX-1), tartrate-resistant acid phosphatase 5b (TRAP-5b), interleukin 6 (IL-6) in serum. In conclusion, tectochrysin serves as a potential candidate in the treatment of osteoporosis. The proposed two-dimensional MPTS-modified BMMC/CMC-TOFMS analytical system shows the advantages of long-life span and fast recognition ability, which is very suitable for infrequent cell lines.

Can urinary CTX-II be a biomarker for knee osteoarthritis?

BackgroundEarly diagnosis of knee osteoarthritis (OA) remains a diagnostic challenge. Urinary C-terminal cross-linked telopeptide of type II collagen (urinary CTX-II) is one of the potential OA biomarkers. However, conclusive evidence regarding the use of this biomarker as a tool for early diagnosis is still lacking. The purposes of this study were to compare urinary CTX-II levels in patients with knee OA and in healthy controls, to evaluate the correlation between urinary CTX-II levels, radiographic severity of OA, and patient-reported outcomes and to evaluate the effect of age and gender on urinary CTX-II levels in the Asian populations.MethodsTwo groups were studied. The OA group included 78 patients with knee OA aged > 40 years who met the diagnostic criteria for knee OA described by the American College of Rheumatology (ACR). The control group consisted of 51 healthy participants age > 40 years without clinical or radiographic evidence of knee OA. Bilateral knee radiographs were taken and classified according to the Kellgren and Lawrence (KL) grading system. Urinary CTX-II was measured using a competitive ELISA test and Western Ontario and Mcmaster Universities Arthritis Index (WOMAC) was also recorded in all participants.ResultsUrinary CTX-II was significantly higher in the OA group than in the control group (p < 0.001). The severe knee OA group (KL grade 3 and 4) had higher urinary CTX-II levels than mild knee OA group (KL grade 2) but the difference did not reach statistical significance (p = 0.2). There was a moderate correlation between urinary CTX-II levels and KL grades (r = 0.405, p < 0.001) and a weak correlation between urinary CTX-II levels and WOMAC index scores (r = 0.367, p < 0.001). Multiple regression analysis showed that urinary CTX-II was independently associated with KL grades. Whereas age, gender, and WOMAC index had no statistically significant influence on the urinary CTX-II levels.ConclusionsPatients with knee OA had higher urinary CTX-II levels than healthy controls. Moreover, levels of urinary CTX-II were independently correlated with radiographic severity of knee OA. Age, gender, and patient-reported outcomes exerted no effect on the urinary CTX-II levels.Level of evidenceDiagnostic Level III.

A Short-Term Ketogenic Diet Impairs Markers of Bone Health in Response to Exercise.

Objectives: To investigate diet-exercise interactions related to bone markers in elite endurance athletes after a 3.5-week ketogenic low-carbohydrate, high-fat (LCHF) diet and subsequent restoration of carbohydrate (CHO) feeding.Methods: World-class race walkers (25 male, 5 female) completed 3.5-weeks of energy-matched (220 kJ·kg·d−1) high CHO (HCHO; 8.6 g·kg·d−1 CHO, 2.1 g·kg·d−1 protein, 1.2 g·kg·d−1 fat) or LCHF (0.5 g·kg·d−1 CHO, 2.1 g·kg·d−1 protein, 75–80% of energy from fat) diet followed by acute CHO restoration. Serum markers of bone breakdown (cross-linked C-terminal telopeptide of type I collagen, CTX), formation (procollagen 1 N-terminal propeptide, P1NP) and metabolism (osteocalcin, OC) were assessed at rest (fasting and 2 h post meal) and after exercise (0 and 3 h) at Baseline, after the 3.5-week intervention (Adaptation) and after acute CHO feeding (Restoration).Results: After Adaptation, LCHF increased fasting CTX concentrations above Baseline (p = 0.007, Cohen's d = 0.69), while P1NP (p < 0.001, d = 0.99) and OC (p < 0.001, d = 1.39) levels decreased. Post-exercise, LCHF increased CTX concentrations above Baseline (p = 0.001, d = 1.67) and above HCHO (p < 0.001, d = 0.62), while P1NP (p < 0.001, d = 0.85) and OC concentrations decreased (p < 0.001, d = 0.99) during exercise. Exercise-related area under curve (AUC) for CTX was increased by LCHF after Adaptation (p = 0.001, d = 1.52), with decreases in P1NP (p < 0.001, d = 1.27) and OC (p < 0.001, d = 2.0). CHO restoration recovered post-exercise CTX and CTX exercise-related AUC, while concentrations and exercise-related AUC for P1NP and OC remained suppressed for LCHF (p = 1.000 compared to Adaptation).Conclusion: Markers of bone modeling/remodeling were impaired after short-term LCHF diet, and only a marker of resorption recovered after acute CHO restoration. Long-term studies of the effects of LCHF on bone health are warranted.

Higher concentration of serum C-terminal cross-linking telopeptide of type I collagen is positively related with inflammatory factors in postmenopausal women with H-type hypertension and osteoporosis.

ObjectiveTo investigate the changes of inflammatory factors and bone metabolism markers in postmenopausal women with H‐type hypertension and to assess the relationship between them.MethodsPostmenopausal women who were diagnosed with osteoporosis were selected as observation objects. Participants were divided into three groups: only osteoporosis group (osteoporosis group), hypertension combined with osteoporosis group (hypertension group), and H‐type hypertension combined with osteoporosis group (H‐type hypertension group). The changes in bone mineral density and bone metabolic markers (osteocalcin [OC], procollagen type I N‐terminal propeptide (PINP), and C‐terminal cross‐linking telopeptide of type I collagen [CTX]) and inflammatory factors (interleukin‐6 [IL‐6] and tumor necrosis factor‐α [TNF‐α]) were compared among three groups.ResultsIn the hypertension group and the H‐type hypertension group, the bone mineral density of the lumbar spine (0.647 ± 0.038 vs 0.638 ± 0.034 vs 0.668 ± 0.047, P < 0.05) and the femoral neck (0.567 ± 0.047 vs 0.552 ± 0.053 vs 0.618 ± 0.059, P < 0.05) was significantly lower than that in the osteoporosis group. The concentrations of CTX (266.61 ± 64.65 vs 293.09 ± 72.34 vs 235.48 ± 62.85, P < 0.05), IL‐6 (44.36 ± 6.45 vs 48.05 ± 8.04 vs 39.06 ± 7.95, P < 0.05) and TNF‐α (30.53 ± 6.28 vs 34.52 ± 7.15 vs 28.66 ± 6.19, P < 0.01) in the hypertension group and in the H‐type hypertension group were significantly higher than those in the osteoporosis group. The concentrations of OC (30.59 ± 6.43 vs 27.10 ± 6.51, P < 0.05) and PINP (36.36 ± 6.16 vs 33.16 ± 6.77, P < 0.05) in the H‐type hypertension group were increased dramatically. The concentration of CTX was positively correlated with the concentration of IL‐6 (r = 0.587, P < 0.01) and TNF‐α (r = 0.474, P < 0.01) and negatively related with the concentration of OC (r = −0.591, P < 0.01) and PINP (r = −0.646, P < 0.01) and the bone mineral density of the lumbar spine (r = −0.470, P < 0.01) and the femoral neck (r = −0.509, P < 0.01).ConclusionHigher concentration of serum CTX is found in postmenopausal women with H‐type hypertension, which is positively correlated with inflammatory factors. Besides, H‐type hypertension could further enhance the activity of osteoclasts and increase the expressions of inflammatory factors, resulting in the aggravation of osteoporosis.

Effects of maternal intake restriction during early pregnancy on fetal growth and bone metabolism in goats

• Effect of maternal intake restriction on bone metabolism of goats was investigated. • Maternal intake restriction during early gestation decreased the INTP and CTX-1 in fetal umbilical blood. • Nutritional restriction during early gestion regulated the femur metabolic activity of fetal goats. The objective of this study was to investigate the effects of maternal nutritional restriction on fetal growth and bone metabolism in early pregnancy. Twelve twin-bearing goats (Xiangdong black goats) in second pregnancy, with similar weight (31.2 ± 8.1 kg) and age (2.0 ± 0.3 year) were assigned to two dietary treatments in early gestation (26–65 days): control group (CON, 100% feed) and intake restriction group (IR, 60% of CON intake). The umbilical blood, right femur and liver of the fetuses were sampled on day 65 to determine the bone metabolism indicators and expression of genes involved in bone development. Maternal intake restriction did not affect the growth performance of pregnancies ( P > 0.05), increased ( P < 0.05) the body weight, body length, thoracic circumference and umbilical circumference of fetal goats, but decreased ( P < 0.05) the concentrations of cross-linked N-terminal telopeptides of type I collagen and cross-linked C-terminal telopeptides of type I collagen, which involved in bone metabolic activity, in the fetal umbilical blood of the IR. In fetal femurs, the mRNA expression of bone gamma-carboxyglutamate protein, osteoprotegerin and mothers against decapentaplegic homolog 1 both participating in osteoclast differentiation were increased ( P < 0.05), whereas transcript of the low density lipoprotein receptor related protein 5 involved in osteoblast differentiation was decreased ( P < 0.05), compared with the fetuses from the CON. These results indicate that nutrition restriction in early pregnancy regulated the fetal growth and promoted formative activity of femur. The molecular mechanism of maternal intake restriction on bone metabolism needs further investigation.

Study protocol: design and rationale for an exploratory phase II randomized controlled trial to determine optimal vitamin D3 supplementation strategies for acute fracture healing

BackgroundObservational studies have found that 75% of healthy adult fracture patients (ages 18–50) have serum 25-hydroxyvitamin D (25(OH)D) levels < 30 ng/mL. Although lower serum 25(OH)D levels have yet to be correlated to fracture healing complications or poor fracture outcomes, many orthopedic surgeons are routinely prescribing vitamin D supplements to improve fracture healing in healthy non-osteoporotic patients. To address this gap in the literature, we propose a phase II exploratory randomized controlled trial comparing three vitamin D3 dosing regimens for early surrogate treatment response.MethodsWe will conduct a 4-arm blinded exploratory phase II trial in 96 adults aged 18–50 years with a closed or low-grade open (Gustilo type I or II) tibial or femoral shaft fracture. Eligible patients will be randomized in equal allocation ratio of 1:1:1:1 to one of the treatment groups: (1) 150,000 IU loading dose vitamin D3 plus daily dose placebo; (2) loading dose placebo plus 4000 IU vitamin D3 per day, (3) loading dose placebo plus 600 IU vitamin D3 per day, or (4) loading dose placebo plus daily dose placebo. The primary outcome is fracture healing, assessed as follows: (1) clinical fracture healing measured using the Function IndeX for Trauma, (2) radiographic fracture healing measured using the Radiographic Union Score for Tibial fractures, and (3) biological fracture healing measured using serum levels of cross-linked C-terminal telopeptides of type I collagen and amino-terminal procollagen propeptides of collagen type I. The main secondary outcome will be assessed by measuring serum 25(OH)D levels. All outcome analyses will be exploratory and adhere to the intention-to-treat principle. Per-protocol sensitivity analyses will also be conducted.DiscussionStudy results will be disseminated through a publication in an academic journal and presentations at orthopedic conferences. Study results will inform dose selection for a large definitive randomized controlled trial and provide preliminary clinical data on which dose may improve acute fracture healing outcomes in healthy adult patients (18–50 years) at 3 months.Trial registrationVita-Shock (A Blinded Exploratory Randomized Controlled Trial to Determine Optimal Vitamin D3 Supplementation Strategies for Acute Fracture Healing) was registered at ClinicalTrials.gov (identifier NCT02786498) prior to enrollment of participants.

Analysis of bio-markers in osteoarthritis early diagnosis

Through analysis and summary of the biological characteristics of various biomarkers, to explore the reliability of different markers for early diagnosis of osteoarthritis. Cartilage oligomeric matrix protein, N-terminal crosslinked telopeptide of type Ⅰ collagen and C-terminal crosslinked telopeptide of type Ⅱ collagen are the possible effective markers in osteoarthritis early diagnosis. Hyaluronic acid and C-terminal crosslinked telopeptide of type I collagen are more suitable for evaluating the ollurrence and derelopment of osteoarthritis. The efficacy of miRNA and lncRNA in osteoarthritis diagnosis and evaluation remains to be proved. Each marker may has two or more biological effect, this paper will focus on finding out an accurate and stable marker with the analysis and summary of present bio-markers. Key words: Biological markers; Osteoarthritis; Early diagnosis

Impact of Exercise Therapy on Molecular Biomarkers Related to Cartilage and Inflammation in Individuals at Risk of, or With Established, Knee Osteoarthritis: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

To investigate the impact of exercise therapy on molecular biomarkers related to cartilage and inflammation in individuals at risk of, or with established, knee osteoarthritis by conducting a systematic review of randomized controlled trials (RCTs). We conducted a literature search up to September 2017 in 5 major databases with no restriction on publication year or language. Data were extracted from the first available follow-up time point, and we performed a narrative synthesis for the effect of exercise therapy on molecular biomarkers related to cartilage and inflammation. A subset of studies reporting sufficient data was combined in a meta-analysis, using an adjusted random-effects model. Twelve RCTs involving 57 study comparisons at 4 to 24 weeks following an exercise-therapy intervention were included. Exercise therapy decreased molecular biomarkers in 17 study comparisons (30%), had no effect in 36 (63%), and increased molecular biomarkers in 4 study comparisons (7%). Meta-analyses of 9 biomarkers showed that exercise therapy was associated with nonsignificant reductions of the C-reactive protein level, C-terminal crosslinking telopeptide of type II collagen, tumor necrosis factor (TNF), soluble TNF receptors 1 and 2, C2C neoepitope of type II collagen, and cartilage oligomeric matrix protein, compared to nonexercising control groups, and exercise therapy had no effect on interleukin-6 and soluble interleukin-6 receptor. Exercise therapy is not harmful, because it does not increase the concentration of molecular biomarkers related to cartilage turnover and inflammation, implicated in osteoarthritis progression. The overall quality of evidence was downgraded to low because of the limited number of RCTs available.

Guhong Injection promotes fracture healing by activating Wnt/beta-catenin signaling pathway in vivo and in vitro

Guhong Injection (GHI), composed of aceglutamide and safflower aqueous extract, has been applied to the clinical treatment of orthopedic diseases, but the relevant mechanism by which GHI exerts effects on bone remodeling has not been reported. In the present study, we investigated the effects of various concentrations of GHI (2.5, 5 and 10 ml/kg) in accelerating rat tibia healing progress by observing haematoxylin and eosin (HE) stained sections, detecting the activity of bone metabolism biochemical markers such as bone morphogenetic protein-2 (BMP-2), transforming growth factor-beta (TGF-beta), osteocalcin (OC) and C-terminal crosslinking telopeptide of type Ⅰ collagen (CTX-1) in rat serum, as well as measuring the expressions of collagen I (COL-1) and collagen II (COL-2) in rat tibia. Also, we investigated the effects of different concentrations of GHI (30, 60 and 90 μl/ml) on the proliferation and differentiation of osteoblasts (OBs) through proliferating cell nuclear antigen (PCNA), alkaline phosphatase (ALP) and type I collagen (COL-1). At the same time, the expression of important factors of Wnt/beta-catenin signaling pathway including Wnt-3a, beta-catenin, disheveled-1 (Dvl-1), glycogen synthase kinases-3beta (GSK-3beta), lymphoid enhancing factor-1 (LEF-1) and axis inhibition protein-2 (Axin-2) after GHI intervention was detected by quantitative real-time PCR (q-PCR), immunohistochemistry and Western blotting. In vivo, rats of tibia fracture model treated with intraperitoneal injection (ip) of GHI had more mature fibroblasts, as well as shorter period formation of new bone. The levels of BMP-2, TGF-beta and OC in rat serum were significantly up-regulated, while the level of CTX was down-regulated. After 4 weeks of drug treatment, the level of COL-1 in the rat tibia increased, but there was no significant change in the level of COL-2. In vitro, after drug intervention, the number of OBs increased significantly, the activities of PCNA, ALP and COL-1 were enhanced. Treatment with GHI increased the mRNA and protein expression of Wnt-3a, beta-catenin, Dvl-1 and LEF-1, and decreased the expression of mRNA of Axin-2 and GSK-3beta. All results demonstrate that GHI accelerates the proliferation of OBs and shortens the recovery time of bone structure, and the Wnt/beta-catenin signaling pathway is involved in the regulation process.

An exploratory randomized-controlled trial of the efficacy of the Src-kinase inhibitor saracatinib as a novel analgesic for cancer-induced bone pain

Pain is a major symptom of bone metastases from advanced cancer and represents a clinical challenge to treat effectively. Basic neurobiology in preclinical animal models implicates enhanced sensory processing in the central nervous system, acting through N-methyl-D-aspartate (NMDA) glutamate receptors, as an important mechanism underpinning persistent pain. The non-receptor tyrosine kinase Src is thought to act as a hub for regulating NMDA receptor activity and the orally available Src inhibitor saracatinib has shown promise as a potential analgesic in recent animal studies. Here we tested the efficacy of saracatinib as a novel analgesic in an exploratory phase II randomized controlled trial on cancer patients with painful bone metastases. Twelve patients completed the study, with 6 receiving saracatinib 125 mg/day for 28 days and 6 receiving placebo. Pharmacokinetic measurements confirmed appropriate plasma levels of drug in the saracatinib-treated group and Src inhibition was achieved clinically by a significant reduction in the bone resorption biomarker serum cross-linked C-terminal telopeptide of type I collagen. Differences between the saracatinib and placebo groups self-reported pain scores, measured using the short form of the Brief Pain Inventory, were not clinically significant after 4 weeks of treatment. There was also no change in consumption of maintenance analgesia in the saracatinib-treated group and no improvement in Quality-of-Life scores. The data were insufficient to demonstrate saracatinib has efficacy as analgesic, although it may have a role as an anti-bone resorptive agent.

Temporal Change in Biomarkers of Bone Turnover Following Late Evening Ingestion of a Calcium-Fortified, Milk-Based Protein Matrix in Postmenopausal Women with Osteopenia.

The diurnal rhythm of bone remodeling suggests nocturnal dietary intervention to be most effective. This study investigated the effect of bedtime ingestion of a calcium-fortified, milk-derived protein matrix (MBPM) or maltodextrin (CON) on acute (0–4 h) blood and 24-h urinary change in biomarkers of bone remodeling in postmenopausal women with osteopenia. In CON, participants received 804 ± 52 mg calcium, 8.2 ± 3.2 µg vitamin D and 1.3 ± 0.2 g/kg BM protein per day. MBPM increased calcium intake to 1679 ± 196 mg, vitamin D to 9.2 ± 3.1 µg and protein to 1.6 ± 0.2 g/kg BM. Serum C-terminal cross-linked telopeptide of type I collagen (CTX) and procollagen type 1 amino-terminal propeptide (P1NP), and urinary N-telopeptide cross-links of type I collagen (NTX), pyridinoline (PYD) and deoxypyridinoline (DPD) was measured. Analyzed by AUC and compared to CON, a −32% lower CTX (p = 0.011, d = 0.83) and 24% (p = 0.52, d = 0.2) increase in P1NP was observed for MBPM. Mean total 24 h NTX excreted in MBPM was −10% (p = 0.035) lower than CON. Urinary PYD and DPD were unaffected by treatment. This study demonstrates the acute effects of bedtime ingestion of a calcium-fortified, milk-based protein matrix on bone remodeling.

Bone mass density and bone metabolism marker are associated with progression of carotid and cardiac calcified plaque in Chinese elderly population.

Bone mineral density (BMD) and bone metabolism marker may contribute to the progression of carotid and cardiac arterial calcifications. The aim of this study was to investigate whether low bone mass and the change of bone biomarker are associated with the prevalence of calcified atherosclerotic plaque in elderly Chinese. We conducted a five-year prospective study. BMD was measured by dual-energy X-ray absorptiometry scanning. Carotid and cardiac computed tomography angiography (CTA) was conducted using a 64-multidetector row scanner to assess carotid and cardiac arterial plaque at baseline and during follow-up. Of 1571 community residents over 60years of age, 184 (11.7%) subjects developed carotid calcified plaque, 510 (32.5%) subjects developed cardiac calcified plaque and 97 (6.2%) subjects developed co-existence calcified plaques in carotid and cardiac arteries. After adjustment for age and all relevant confounders, Q1, Q2 quartile of BMD, and osteoprotegerin (OPG), osteocalcin (OC), and C-terminal cross-linked telopeptide of type I collagen (CTX) were associated with increased risk of calcified plaques. This study suggested that lower BMD and change of bone metabolism biomarker were associated with a higher risk of carotid and cardiac calcified plaque development.

Evaluation of therapeutic effects of teriparatide in a rat model of zoledronic acid-induced bisphosphonate-related osteonecrosis

Osteonecrosis of the jaw is a complication in patients using bisphosphonate agents. In this study, we aimed to investigate the effects and mechanism of teriparatide (TPTD) in a rat model of bisphosphonate-related osteonecrosis (BRON). Osteonecrosis was induced by administration of zoledronic acid (ZOL). Four weeks after ZOL injection, the rats underwent a surgery in which drilling holes were made. These holes were filled with freeze-dried Aggregatibacter actinomycetemcomitans. After the four-weeks period, TPTD or saline (n = 9) was intermittently administered for four weeks. Later, rats were euthanized, and the mandible and femur bones were examined the amount of necrosis and bone regeneration. Serum receptor activator of NF-κB ligand (RANKL), osteocalcin (OC), and C-terminal crosslinking telopeptide of type I collagen (CTX) were also examined. TPTD administration reduced necrotic bone area of the mandibles and femurs in the BRON rat model and induced new bone formation. In addition, TPTD injection increased the number of osteoclasts. The suggested underlying mechanism is the induction of protein levels of RANKL by TPTD. Furthermore, the serum levels of bone metabolism biomarkers (OC and CTX) were upregulated in the TPTD injection group. In conclusion, ZOL has negative effects on osteoclasts. TPTD was found to be effective in eliminating the negative effects of ZOL. TPTD had positive effects in preventing bone resorption and promoting osteogenesis. In addition, TPTD improved osteoclastogenesis, which in turn led to the improvement of BRON.

LncRNAp21 promotes osteogenic differentiation of mesenchymal stem cells in the rat model of osteoporosis by the Wnt/β-catenin signaling pathway.

The aim of this study was to explore the promoting effect of long non-coding ribonucleic acid p21 (lncRNAp21) on the osteogenic differentiation of mesenchymal stem cells in the rat model of osteoporosis (OP) through the Wnt/β-catenin signaling pathway. A total of 30 healthy female rats were selected and randomly divided into three groups, including the lncRNAp21 group, OP model group (model group) and normal group. Rats in the lncRNAp21 group were given a certain quantity of lncRNAp21 inhibitors for gavage. Rats in the model group were regularly given 0.9% NaCl for gavage every day after the removal of bilateral ovaries. Meanwhile, rats in the normal group were fed normally without any changes. Bone mineral density (BMD) was measured after 12 weeks of modeling. The levels of procollagen type I N-terminal propeptide (PINP), serum estradiol (E2), osteocalcin (OC), bone alkaline phosphatase (BALP), C-terminal cross-linking telopeptide of type I collagen (CTX) and tartrate-resistant acid phosphatase 5b (TRACP-5b) in the bone and serum of rats were measured by enzyme-linked immunosorbent assay (ELISA). Besides, quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blotting were adopted to detect the mRNA and protein expressions of Wnt1 and β-catenin in bone tissues, respectively. Compared with the normal group and lncRNAp21 group, the serum level of E2 in the model group decreased significantly (p<0.05). BMD and phosphorus (P) content in the model group were both markedly lower than those of the normal group and lncRNAp21 group. However, calcium (Ca) content was remarkably higher than that of the normal group and lncRNAp21 group (p<0.05). The serum levels of bone resorption markers (including TRACP-5b and CTX) in the model group were prominently higher than those of the normal group (p<0.05). However, the levels of the two markers in the lncRNAp21 group were significantly lower than the model group (p<0.05). Additionally, bone formation markers (including OC, PINP and BALP) in the serum of rats in the model group were notably higher than those in the normal group and lncRNAp21 group (p<0.05). QRT-PCR and Western blotting results revealed that the mRNA and protein expressions of Wnt1 and β-catenin in bone tissues of the model group were markedly lower than those of the normal group. However, the mRNA and protein expressions of Wnt1 and β-catenin in the lncRNAp21 group were remarkably higher than the model group (p<0.05). Low expression of lncRNAp21 activates the Wnt/β-catenin signaling pathway by increasing E2 secretion, eventually stimulating bone formation and increasing osteogenic differentiation of mesenchymal stem cells in OP model rats.