Serum Tartrate-resistant Acid Phosphatase Levels Research Articles

Monitoring of bone matrix acidification by TRAP and ERK biomarkers in the chronic hypercholesterolemia male rats.

Hypercholesterolemia is frequently linked to an elevated risk of cardiovascular diseases, including heart attacks and strokes. Additionally, it could be connected to a higher susceptibility to osteoporosis. Hypercholesterolemia can stimulate the differentiation and activity of osteoclasts, leading to enhanced bone reabsorption and a subsequent net loss of bone tissue. The purpose of this study was to examine the influence of a high-cholesterol diet on osteoporosis in male rats with differences in biological and oxidative indicators in the hypercholesterolemia diet in male rats. The samples in this study were twenty male rats, ranging between 1.5 and 2 months, were separated into two groups. In one group, 10 rats were fed a regular diet, while in another group, 10 rats were fed a high-cholesterol diet (2%) over the course of 8 weeks. Samples of blood were obtained at the last stage of the experiment. To calculate physiological and biological markers including extracellular signal-regulated kinase (ERK), tartrate-resistant acid phosphatase (TRAP), hormones, malondialdehyde (MDA), and glutathione (GSH). The results of this study demonstrated a decrease in GSH levels, an increase in ERKs, no significant change in serum TRAP levels, an increase in MDA levels in the blood, and elevated levels of parathyroid hormone, calcitonin, and vitamin D in the cholesterol group. Increased oxidative stress, altered signaling, and disruptions in calcium/bone metabolism associated with cholesterol-related conditions and monitoring biomarker ERK can provide valuable information about disease progression.

Evaluation of Gingival Crevicular Fluid and Serum Tartrate-resistant Acid Phosphatase Levels in Subjects with Clinically Healthy Periodontium and Chronic Periodontitis - A Clinico-biochemical Study.

Background:Periodontitis is a chronic inflammatory disease with conglomerate etiology making it difficult to diagnose at the early stages. Potential biomarkers in gingival crevicular fluid (GCF) would determine the presence of the current disease activity, predict sites vulnerable for future breakdown, and assess the response to periodontal interventions. Merely elevated levels of inflammatory soft-tissue markers do not indicate bone destruction. Since there is no single ideal biomarker established, bone-related biomarkers such as telopeptide of type I collagen, osteocalcin, calprotectin, osteopontin, and tartrate-resistant acid phosphatase (TRAP) seem to hold great promise as predictive markers to determine bone destruction and active phases in the disease progression. The present study is intended to explore the biologic plausibility of the levels of TRAP in health and chronic periodontitis.Materials and Methods:The present cross-sectional clinico-biochemical study comprised 30 systemically healthy subjects with 15 periodontally healthy and 15 chronic periodontitis subjects who were age and gender matched. GCF and blood samples were collected from all the patients. TRAP estimation was done in both the samples using an enzyme-linked immunosorbent assay kit. The data were analyzed using independent t-test and Pearson correlation test.Results:Serum and GCF TRAP levels in chronic periodontitis subjects were significantly higher when compared to the periodontally healthy group. There were no significant correlations found among serum and GCF TRAP levels with increasing age and gender in both the groups. An increase in disease severity, i.e., increase in probing pocket depth and clinical attachment level, did not show correlation with the GCF and serum TRAP levels in the chronic periodontitis group.Conclusion:Based on the findings of the present study, increased GCF TRAP levels in chronic periodontitis seem to be a potential marker for identifying ongoing periodontal destruction.

Asiatic Acid Inhibits OVX-Induced Osteoporosis and Osteoclastogenesis Via Regulating RANKL-Mediated NF-κb and Nfatc1 Signaling Pathways.

Asiatic acid is a triterpenoid compound extracted from a medicinal plant Centella asiatica. It has been used as a highly efficient compound for the treatment of cancer and hyperlipidemia, as well as possessing potential antiinflammatory properties. However, its effects on bone metabolism and osteoporosis haven’t been reported. The purpose of our research were to reveal the biomolecular effects of asiatic acid on osteoclasts, and its underlying molecular mechanisms regulating its effects on receptor activator of NF-κB ligand (RANKL)-induced signaling pathways. We found that asiatic acid inhibited multinucleated tartrate-resistant acid phosphatase (TRAcP)-positive osteoclast differentiation and osteoclast induced bone loss. Real time PCR showed that asiatic acid reduced the expression of down-cascade target genes including Ctsk, Nfatc1, Calcr, and Atp6v0d2. Western blot and luciferase reporter gene assays revealed that asiatic acid inhibits RANKL mediated NF-κB and NFATc1 signalings. Further, in vivo study demonstrated asiatic acid attenuates estrogen deficiency-induced bone loss in ovariectomized mice. MicroCT and histology analyses revealed that osteoclast numbers were significantly suppressed in asiatic acid treated groups. Furthermore, serum levels of TRAcP and CTX-1 were downregulated in treated groups. Taken together, our data show that asiatic acid can inhibit osteoclastic formation and reduce OVX-induced bone resorption through RANKL-activated NF-κB or NFATc1 signaling, suggesting that asiatic acid may be a potential and effective natural compound for the therapy of excessive RANKL-related osteolytic diseases.

Sialoglycoproteins isolated from the eggs of Carassius auratus prevents osteoporosis by suppressing the activation of osteoclastogenesis related NF-κB and MAPK pathways

A sialoglycoprotein was isolated from the eggs of Carassius auratus (Ca-SGP) and its anti-osteoporotic activities were first investigated in in vitro and in vivo systems. In vitro experiments using osteoclast precursors showed that Ca-SGP significantly decreased the formation of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated osteoclasts, bone resorption pits and the expression of osteoclastic marker genes, indicating its inhibitory effect on RANKL-induced osteoclastogenesis. qRT-PCR and western blotting results suggested that Ca-SGP significantly decreased mRNA and protein expressions of NFATc1 and c-Fos, two key transcription factors of osteoclastogenesis regulated by NF-κB and MAPK pathways. Further investigation revealed that Ca-SGP suppressed the activation of NF-κB and MAPK signaling pathways by diminishing the mRNA expression and phosphorylation of p38, ERK, JNK, IκB, NF-κB. In vivo experiment using ovariectomized (OVX)-induced osteoporotic model rats indicated that Ca-SGP prominently declined the levels of serum TRAP, CSTK, MMP-9, urine DPD and ameliorated the damaged trabecular. These results suggest that Ca-SGP can improve osteoporosis by inhibiting bone resorption via suppressing the activation of osteoclastogenesis related NF-κB and MAPK pathways.

F-Spondin Deficient Mice Have a High Bone Mass Phenotype

F-spondin is a pericellular matrix protein upregulated in developing growth plate cartilage and articular cartilage during osteoarthritis. To address its function in bone and cartilage in vivo, we generated mice that were deficient for the F-spondin gene, Spon1. Spon1 − /− mice were viable and developed normally to adulthood with no major skeletal abnormalities. At 6 months, femurs and tibiae of Spon1 − /− mice exhibited increased bone mass, evidenced by histological staining and micro CT analyses, which persisted up to 12 months. In contrast, no major abnormalities were observed in articular cartilage at any age group. Immunohistochemical staining of femurs and tibiae revealed increased levels of periostin, alkaline phosphate and tartrate resistant acid phosphatase (TRAP) activity in the growth plate region of Spon1 − /− mice, suggesting elevated bone synthesis and turnover. However, there were no differences in serum levels of TRAP, the bone resorption marker, CTX-1, or osteoclast differentiation potential between genotypes. Knockout mice also exhibited reduced levels of TGF-β1 in serum and cultured costal chondrocytes relative to wild type. This was accompanied by increased levels of the BMP-regulatory SMADs, P-SMAD1/5 in tibiae and chondrocytes. Our findings indicate a previously unrecognized role for Spon1 as a negative regulator of bone mass. We speculate that Spon1 deletion leads to a local and systemic reduction of TGF-β levels resulting in increased BMP signaling and increased bone deposition in adult mice.

Preservation of bone structure and function by Lithothamnion sp. derived minerals.

Progressive bone mineral loss and increasing bone fragility are hallmarks of osteoporosis. A combination of minerals isolated from the red marine algae, Lithothamnion sp. was examined for ability to inhibit bone mineral loss in female mice maintained on either a standard rodent chow (control) diet or a high-fat western diet (HFWD) for 5, 12, and 18 months. At each time point, femora were subjected to μ-CT analysis and biomechanical testing. A subset of caudal vertebrae was also analyzed. Following this, individual elements were assessed in bones. Serum levels of the 5b isoform of tartrate-resistant acid phosphatase (TRAP) and procollagen type I propeptide (P1NP) were also measured. Trabecular bone loss occurred in both diets (evident as early as 5 months). Cortical bone increased through month 5 and then declined. Cortical bone loss was primarily in mice on the HFWD. Inclusion of the minerals in the diet reduced bone mineral loss in both diets and improved bone strength. Bone mineral density was also enhanced by these minerals. Of several cationic minerals known to be important to bone health, only strontium was significantly increased in bone tissue from animals fed the mineral diets, but the increase was large (5-10 fold). Serum levels of TRAP were consistently higher in mice receiving the minerals, but levels of P1NP were not. These data suggest that trace minerals derived from marine red algae may be used to prevent progressive bone mineral loss in conjunction with calcium. Mineral supplementation could find use as part of an osteoporosis-prevention strategy.

Bone turnover markers predict changes in bone mineral density after parathyroidectomy in patients with renal hyperparathyroidism

Patients on long-term dialysis may develop secondary hyperparathyroidism (SHPT), which causes varying degrees of bone mass loss. This condition is treated with parathyroidectomy (PTX). We investigated whether serial serum bone turnover markers could predict changes in bone mineral density (BMD) after PTX. Renal patients on maintenance haemodialysis who received PTX for refractory SHPT (n = 26, male/female: 13/13; mean age: 48·6 ± 10·7 year) and control subjects without SHPT (n = 25) were prospectively followed for 1 year at two tertiary hospitals in Taiwan. Serum intact parathyroid hormone (iPTH), bone-specific alkaline phosphatase (BAP) and type 5b tartrate-resistant acid phosphatase (TRAP) were measured serially. Additionally, femoral neck (FN) and lumbar spine (LS) BMD were measured before and 1 year after PTX. After PTX, iPTH levels decreased markedly and persistently. BMDs increased in both the FN and LS, but particularly in the LS. Serum BAP progressively increased to a peak at 2 weeks after PTX. Serum TRAP levels progressively decreased over 6 months after PTX. In univariate correlation analyses, baseline iPTH correlated positively with T-score changes in FN (r = 0·45, P = 0·021) and LS (r = 0·48, P = 0·013). In multivariate regression models, changes in FN T-scores were negatively predicted by baseline BAP levels (r = -0·615, P = 0·005) and baseline FN T-scores (r = -0·563, P = 0·012), and they were positively predicted by baseline TRAP(r = 0·6, P = 0·007). Changes in LS T-scores were positively predicted by baseline TRAP values (r = 0·528, P = 0·01) and negatively predicted by the percentage change in BAP after 2 weeks (r = -0·501, P = 0·015). Parathyroidectomy provided marked, sustained improvements in BMD for up to 1 year. Furthermore, markers of bone turnover predicted 1-year changes in FN and LS BMDs after PTX.

Parathyroidectomy leads to decreased blood lead levels in patients with refractory secondary hyperparathyroidism

ObjectiveSecondary hyperparathyroidism (SHPT) is characterized by high bone turnover and may result in increased release of lead (Pb) from bone stores. Parathyroidectomy (PTX) drastically changes bone remodeling. This study investigated the effects of PTX on the levels of blood lead (blood Pb) in patients with refractory SHPT. MethodsThe study included 30 patients on long-term hemodialysis (HD) who underwent PTX in the nephrology units of two Taiwanese hospitals. Changes in serum levels of intact parathyroid hormone (iPTH), bone-specific alkaline phosphatase (BAP), type 5b tartrate-resistant acid phosphatase (TRAP), total calcium (tCa), and blood Pb were analyzed. ResultsAfter PTX, serum iPTH was markedly decreased while serum BAP was progressively increased and peaked 2weeks after PTX. Serum TRAP levels were progressively decreased during the 4week follow-up period. Serum tCa and blood Pb levels decreased sharply immediately after PTX. There was a positive correlation between the percentage of decrease in tCa and blood Pb at one day after PTX. Further analysis indicated a significant positive correlation between levels of blood Pb and serum iPTH (r=0.378, p<0.001), blood Pb and serum TRAP (r=0.180, p<0.05), and a negative correlation between blood Pb and serum BAP (r=−0.205, p<0.05). ConclusionPTX effectively suppressed the elevated levels of blood Pb and serum bone remodeling markers, which are common features of SHPT. In addition to decreased Pb release from bone, an increased store of Pb in bone may play a role in decreasing serum blood Pb levels. These findings suggest that patients undergoing PTX for refractory SHPT should strictly avoid environmental exposure to Pb.

Synergistic effects of green tea polyphenols and alphacalcidol on chronic inflammation-induced bone loss in female rats

Studies suggest that green tea polyphenols (GTP) or alphacalcidol is promising agent for preventing bone loss. Findings that GTP supplementation plus alphacalcidol administration increased bone mass via a decrease of oxidative stress and inflammation suggest a significant role of GTP plus alphacalcidol in bone health of patients with chronic inflammation. Studies have suggested that green tea polyphenols (GTP) or alphacalcidol are promising dietary supplements for preventing bone loss in women. However, the mechanism(s) related to the possible osteo-protective role of GTP plus D(3) in chronic inflammation-induced bone loss is not well understood. This study evaluated bioavailability, efficacy, and related mechanisms of GTP in combination with alphacalcidol in conserving bone loss in rats with chronic inflammation. A 12-week study of 2 (no GTP vs. 0.5% GTP in drinking water) × 2 (no alphacalcidol vs. 0.05 μg/kg alphacalcidol, 5×/week) factorial design in lipopolysaccharide-administered female rats was performed. In addition, a group receiving placebo administration was used to compare with a group receiving lipopolysaccharide administration only to evaluate the effect of lipopolysaccharide. Lipopolysaccharide administration resulted in lower values for bone mass, but higher values for serum tartrate-resistant acid phosphatase (TRAP), urinary 8-hydroxy-2'-deoxyguanosine, and mRNA expression of tumor necrosis factor-α and cyclooxygenase-2 in spleen. GTP supplementation increased urinary epigallocatechin and epicatechin concentrations. Both GTP supplementation and alphacalcidol administration resulted in a significant increase in bone mass, but a significant decrease in serum TRAP levels, urinary 8-hydroxydeoxyguanosine levels, and mRNA expression of tumor necrosis factor-α and cyclooxygenase-2 in spleen. A synergistic effect of GTP and alphacalcidol was observed in these parameters. Neither GTP nor alphacalcidol affected femoral bone area or serum osteocalcin. We conclude that a bone-protective role of GTP plus alphacalcidol during chronic inflammation bone loss may be due to a reduction of oxidative stress damage and inflammation.

High-fat diet decreases cancellous bone mass but has no effect on cortical bone mass in the tibia in mice

Body mass has a positive effect on bone health. Whether mass derived from an obesity condition or excessive fat accumulation is beneficial to bone has not been established; neither have the mechanisms by which obesity affects bone metabolism. The aim of this study was to examine the effects of obesity on bone structure and osteoblastic expression of key markers involved in bone formation and resorption in a diet-induced obesity mouse model. Six-wk-old male C57BL/6 mice ( n = 21) were assigned to two groups and fed either a control (10 kcal% energy as fat) or high-fat diet (HFD, 45 kcal% energy as fat) for 14 weeks. Bone marrow stromal/osteoblastic cells (BMSC) were cultured. Osteoprogenitor activity [alkaline phosphatase (ALP) positive colonies] and mineralization (calcium nodule formation) were determined. Gene expression was measured using quantitative real-time PCR. Bone structure of proximal and midshaft tibia was evaluated by micro-computed tomography. Mice fed the HFD were 31% heavier ( P < 0.01) than those fed the control diet. There were more ALP positive colony forming units at d 14 and calcium nodules at d 28 of culture by BMSC from HFD mice than from control mice ( P < 0.01). Receptor activator of NF-κB ligand (RANKL) mRNA levels and the ratio of RANKL to osteoprotegerin expression in HFD animals was higher ( P < 0.01) than in control diet animals. Serum tartrate-resistant acid phosphatase levels were higher in HFD fed mice when compared to control diet fed mice ( P < 0.05). There were no significant differences in tibial fat-free weight, length, and cortical parameters of midshaft between the two groups. Compared with control mice, tibial trabecular bone volume was reduced, and trabecular separation was increased in HFD mice. Trabecular number was lower ( P < 0.05) and connectivity density tended to be less ( P = 0.07) in HFD mice than in control mice. In conclusion, our data indicate that obesity induced by a high-fat diet decreases cancellous bone mass but has no effect on cortical bone mass in the tibia in mice.

Development of immunoassays for type-5 tartrate-resistant acid phosphatase in human serum

Background: Tartrate-resistant acid phosphatase (TRAP) is known as a marker of bone resorption. The purpose of this study was the development of a sensitive and specific immunoassay for TRAP. Methods: We have developed two types of immunoassays, enzyme-linked immunosorbent assay (ELISA) and immunoselective enzyme immunoassay (ISEA) using monoclonal antibodies to recombinant TRAP, for determination of TRAP in human serum. To evaluate assay performance, recovery and dilution tests were performed. Further, we determined serum TRAP levels of patients with secondary hyperparathyroidism at different pH conditions. Results: The detected ranges of ELISA and ISEA were between 0.08 and 5 μg/l and between 0.063 and 4 U/l. Different concentrations of TRAP added were recovered on average at 98.0% in ELISA and 102.9% in ISEA. In the serial dilution test, serum TRAP levels were on average at 101.6% and 109.6% of the expected values in ELISA and ISEA, respectively. The serum TRAP levels of patients with secondary hyperparathyroidism were significantly higher than those of normal controls in ELISA and ISEA. Similar TRAP levels were obtained in the conditions at pH 5.5 and 6.1 in ISEA. Conclusion: The present findings suggest that our assay methods are applicable for clinical tests, and strengthen the idea that serum TRAP is useful as a marker for bone resorption.

Modulation of bone mass and turnover in growing rats by voluntary weight-bearing exercise and glucose supplementation.

Female Sprague-Dawley rats, 9 weeks of age, were assigned to four groups: Group 0 (n = 8) was dissected for base-line control, and the other three groups were fed for 3 mo: Group 1 (n = 9), sedentary controls; Group 2 (n = 6), running rats housed in a cage with a treadmill and pair-fed with Group 1; and Group 3 (n = 7), running rats, pair-fed and allowed free access to additional glucose. The distances of voluntary running did not significantly differ between Groups 2 and 3. Menstrual cycles in these rats were apparently maintained as observed from daily running distances. The amount of glucose taken by rats in Group 3 was 3.5 +/- 0.4 (mean and SE) g/d. Body weight (BW) at the end of the experiment for Groups 1, 2, and 3 were 295.0 +/- 7.9, 211.7 +/- 5.4 (p < 0.001 vs. Group 1), and 259.0 +/- 3.5 g (p < 0.01 vs. Group 2), respectively. The parameters of bone mass such as ash weights of the femur and bone mineral content of the lumbar spine and the tibia in Groups 1 and 2 did not differ, but the values were significantly greater in Group 3 than in Group 2. However, these parameter values corrected for BW were significantly greater in Group 2 than in Group 1 and did not significantly differ between Groups 2 and 3. The parameters of bone formation, such as serum bone alkaline phosphatase activity levels and trabecular bone formation rates corrected for BW, were significantly greater in Group 2 than in Group 1 but did not differ between Group 2 and 3. However, the parameters of bone resorption, such as serum tartrate resistant acid-phosphatase levels, were significantly less in Group 3 than in Group 2. These results suggest that voluntary running augments the age-dependent increase in bone mass by modulating the bone turnover when an adequate energy source is supplied under conditions of normal menstruation, and an adequate supply of energy could be necessary to enhance the age-dependent increase in bone mass.

Purified Pasteurella multocida protein toxin reduces acid phosphatase-positive osteoclasts in the ventral nasal concha of gnotobiotic pigs.

To study the in vivo response of conchal (turbinate) osteoclasts to Pasteurella multocida toxin, four gnotobiotic pigs (7 days of age) were inoculated subcutaneously with 0.2 microgram/kg of purified toxin. One toxin-treated pig along with one control pig were necropsied at 2, 5, 9, and 14 days postinoculation. The entire length of nasal concha from the nasal planum toi ethmoid region was removed, blocked by transverse cuts into five areas, decalcified, sectioned, and then stained with tartrate-resistant acid phosphatase (TRAP) to identify osteoclasts. In each section, total area of concha, total osteoclast cytoplasmic area, and number of osteoclasts were determined using an image analysis morphometric unit. Also collected from pigs were blood and serum for complete blood counts, electrolyte levels, liver enzymes, and TRAP levels. Conchal atrophy increased in severity with time after 2 days postinoculation. In general, the ventral conchae from toxin-treated pigs at 9 and 14 days postinoculation had decreased surface area, osteoclast cytoplasmic area, and numbers of osteoclasts. Serum levels of TRAP were mildly elevated when compared with age-matched controls. No other significant alterations in blood cells or chemistries occurred and no lesions were present histologically in tissues (liver, kidney, lung, heart, and spleen) other than concha. This study shows that the P. multocida toxin induces rapid bone resorption and increases serum levels of acid phosphatase but leads to diminished acid phosphatase expression and presumably, numbers of osteoclasts.

Tartrate-resistant acid phosphatase as a biological marker of bone modelling and turnover in women in relation to their gonadal state

SummarySerum tartrate-resistant acid phosphatase (TRAP) is considered a biological marker of bone resorption as it is synthesised by osteoclasts. Oestrogens inhibit bone resorption, and this may justify the influence of this hormone on osteoclastic activity. Thus, we have quantified serum TRAP levels in women to define their biological profile and observe any changes in relation to their hormone status.The TRAP level of 18 girls under 5 years of age was 8–1 ± 1–7 u/1. This value decreased (P< 0–001) in girls aged 5 to 12 years (n = 20, mean 6–3 ± 1–3 u/1). Following the menarche, another decrease was observed in 12 to 15 year old girls (n = 25, mean 5.5 ± 0–6u/l, P < 0–01). The mean TRAP value continued to decrease during the childbearing years, 16 to 48 (n = 40, 3.8 ± 0–3 u/1, P < 0–001). Finally, the mean level increased again at the menopause (n = 26, 4–5 ±0–4 u/1, P < 0–001). In women, serum TRAP changes reflect the dependence of bone modelling and turnover on hormone status.

Clinical usefulness of serum tartrate-resistant acid phosphatase in paget's disease of bone: Correlation with other biochemical markers of bone demodelling

Tartrate resistant acid phosphatase (TRAP) has been proposed as a new biochemical marker for bone resorption. We have compared this new marker, TRAP, with the classical biochemical markers of bone remodelling, serum alkaline phosphatase (sAP), serum osteocalcin (sBGP), and with the urinary hydroxyproline/creatinine ratio (uOHProl/creatinine), a routine marker of bone resorption. Serum TRAP was significantly higher in pagetic patients (n = 43) than in control subjects (n = 12) (13.02 +/- 4.7 vs 5.48 +/- 1.31 IU/L, P less than 0.001) and a significantly positive linear correlation was found between the sTRAP and uOHProl/creatinine ratio (y = 0.0051x - 0.0069, r = 0.82, P less than 0.001), between sTRAP and sAP (y = 19.3x - 85.0, r = 0.71, P less than 0.001) and also between sTRAP and sBGP (y = 0.02x + 2.23, r = 0.52, P less than 0.01). Serum TRAP levels were higher than the upper limit of normality in all our pagetic patients except for two, whose uOHProl/creatinine levels were in the normal range. We conclude that (1) sTRAP could be a parameter as sensitive as uOHProl/creatinine in the diagnosis of Paget's disease; (2) sTRAP and uOHProl/creatinine are both good markers of bone resorption; (3) the correlation found between sTRAP and formation markers (sAP and sBGP) makes sTRAP a marker of disease activity in Paget's disease of bone; (4) the assay of sTRAP is easier, faster, and of lower cost than the urinary hydroxyproline determination. We suggest that sTRAP determination could be used as a routine marker of bone resorption in Paget's disease of bone, as is the case with uOHProl determination.