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Journal of Bone and Mineral Research :... Jun 1994Using immunohistochemical methods we studied the tissue localization of the extracellular matrix proteins osteopontin (OPN), osteocalcin (OC), and dentin sialoprotein...
Using immunohistochemical methods we studied the tissue localization of the extracellular matrix proteins osteopontin (OPN), osteocalcin (OC), and dentin sialoprotein (DSP) during the formation of acellular and cellular cementum in newly born rats. In the layer of acellular cementum of developing incisor and molar teeth we found a very strong staining for OPN but not for DSP or OC. Many cells immediately adjacent to acellular cementum and PDL cells were also positive for OPN but not for DSP or for OC. In contrast, cellular cementum in molar teeth stained strongly for OPN and OC but not for DSP. Consistent with these observations, the cells engaged in the formation of cellular cementum (cementoblasts and cementocytes) reacted strongly for OPN and OC but not for DSP. In advanced stages of dentinogenesis, both crown and root odontoblasts and dentin stained for OPN, OC, and DSP. Cells and matrices of surrounding alveolar bone stained for OPN and OC but not for DSP. We conclude that cementoblasts and cementocytes of cellular cementum produce OPN and OC but not DSP and thus express an osteoblast-like, not an odontoblast-like, phenotype. The cells responsible for the production of acellular cementum are likely cells of the PDL in close contact with the dental root surface. These fibroblast-like cells express OPN but not OC or DSP and accordingly express only a partial osteoblastic phenotype.
Topics: Animals; Animals, Newborn; Cementogenesis; Dental Cementum; Extracellular Matrix Proteins; Immunohistochemistry; Osteocalcin; Osteopontin; Phosphoproteins; Protein Precursors; Rats; Rats, Sprague-Dawley; Sialoglycoproteins; Tooth Root
PubMed: 8079659
DOI: 10.1002/jbmr.5650090609 -
Stem Cells and Development Mar 2012This study aimed to compare clonogenicity, proliferation, stem cell-related marker expression, senescence, and differentiation potential of rat patellar tendon-derived...
This study aimed to compare clonogenicity, proliferation, stem cell-related marker expression, senescence, and differentiation potential of rat patellar tendon-derived stem cells (TDSCs) at early (P5), mid (P10), and late (P20, P30) passages. The clonogenicity of the cells was assessed by colony-forming assay and their proliferative potential was assessed by bromodeoxyuridine assay. The surface expression of CD90 and CD73 was assessed by flow cytometry. The cellular senescence was assessed by β-galactosidase activity. The adipogenic, chondrogenic, and osteogenic differentiation potentials of TDSCs were assessed by standard assays after induction. The mRNA expression of tendon-related markers, scleraxis (Scx) and tenomodulin (Tnmd), was measured by quantitative real-time reverse transcription-polymerase chain reaction. Both the colony numbers and proliferative potential of TDSCs increased with passaging. Concomitantly, there was significant upregulation of β-galactosidase activity with TDSC passaging. The subculture of TDSCs downregulated the expression of CD90 and CD73. Lipid droplets were formed in the early and mid passages of TDSCs upon adipogenic induction, but were absent in the late passages. The expression of peroxisome proliferator activator receptor gamma 2 (PPARγ2) and CCAAT/enhancer binding protein alpha (C/EBPα) in TDSCs after adipogenic induction decreased with passaging. Chondrogenesis, proteoglycan deposition, collagen type II protein expression, collagen type 2A1 (Col2AI), and aggrecan (Acan) mRNA expression were less in pellets formed with later passages of TDSCs after chondrogenic induction. The expression of Scx and Tnmd was lower in the late, compared with early and mid, passages of TDSCs. However, matrix mineralization and expression of alkaline phosphatase (Alpl) and osteocalcin (Bglap) mRNA after osteogenic induction increased with TDSC passaging. Researchers and clinicians should consider the changes of stem cell-related properties of TDSCs when multiplying them in vitro for tissue engineering.
Topics: 5'-Nucleotidase; Adipocytes; Animals; Cell Differentiation; Cell Proliferation; Cells, Cultured; Cellular Senescence; Chondrocytes; Collagen Type II; Colony-Forming Units Assay; Flow Cytometry; Gene Expression; Glycoside Hydrolases; Male; Osteoblasts; Osteocalcin; PPAR gamma; Rats; Rats, Sprague-Dawley; Rats, Transgenic; Reverse Transcriptase Polymerase Chain Reaction; Stem Cells; Tendons; Thy-1 Antigens; Tissue Engineering
PubMed: 21627568
DOI: 10.1089/scd.2011.0160 -
Analytical Sciences : the International... Mar 2021Immunochromatography assay is an easy and rapid on-site detection method. However, conventional sandwich immunochromatographies using two antibodies can only detect...
Immunochromatography assay is an easy and rapid on-site detection method. However, conventional sandwich immunochromatographies using two antibodies can only detect target molecules above a threshold size. Small molecules below 1000 in molecular weight are usually detected using competitive immunoassay. However, competitive immunoassay is not suitable for visual detection of low concentration samples. Based on the principles of open sandwich immunoassay, which detects small molecules via interchain interaction of separated variable region fragments (V and V) from a single antibody, we developed non-competitive open sandwich immunochromatography. Bone Gla protein (BGP)-C7, a peptide containing the seven C-terminal amino acids of human osteocalcin, was selected as the target. By using V fragments fixed on a nitrocellulose membrane, and colored cellulose bead-labeled V fragments, we specifically detected 10 ng/mL of BGP-C7. This is the first report of open sandwich immunochromatography, which is an easy and rapid method for on-site, signal-on detection of small molecules.
Topics: Antigens; Cellulose; Chromatography, Affinity; Osteocalcin
PubMed: 33229828
DOI: 10.2116/analsci.20SCP06 -
Cell Proliferation Oct 2014Amongst the fourth generation of PHAs is bio-plasticpoly3-hydroxybutyrate4-hydroxybutyrate (P34HB); it is thus appropriate to perform novel research on its uses and...
OBJECTIVES
Amongst the fourth generation of PHAs is bio-plasticpoly3-hydroxybutyrate4-hydroxybutyrate (P34HB); it is thus appropriate to perform novel research on its uses and applications. The main objective of this study was to determine whether electrospun P34HB fibres would accommodate viability, growth and differentiation of mouse adipose-derived stem cells (mASCs).
MATERIALS AND METHODS
In the present study, we looked at P34HB in two forms, electrospun P34HB fibres and P34HB film. Morphology of electrospun P34HB fibres and P34HB film were characterized using scanning electron microscopy, fluorescence microscopy and confocal laser scanning microscopy, after cell seeding. Cell adhesion, proliferation and cytotoxicity tests were conducted on both by MTT and CCK-8 assays, respectively. After being cultured with osteogenic induction, expression of adipogenic genes Runx2, OPN and OCN, were examined by real-time PCR.
RESULTS
By scanning electron microscopy, light microscopy and confocal laser scanning microscopy, we observed that the mASCs grew well associated with the P34HB materials. After MTT and CCK-8 assay, we concluded that P34HB would, indeed, be a material suitable for further cell adhesion and proliferation studies. More importantly, we found that the P34HB matrices promoted expression of Runx2, OPN and OCN with osteogenic induction.
CONCLUSIONS
In this investigation, we can confirm that the electrospun P34HB fibres accommodated survival, proliferation and differentiation of mASCs, and we have been able to draw the conclusion that fibre scaffolds produced by the electrospinning process are promising for application of bone tissue engineering.
Topics: Adipose Tissue; Animals; Biocompatible Materials; Cell Adhesion; Cell Differentiation; Cell Proliferation; Cell Survival; Cells, Cultured; Core Binding Factor Alpha 1 Subunit; Green Fluorescent Proteins; Hydroxybutyrates; Mice; Nanofibers; Osteocalcin; Osteopontin; Polyesters; Polyhydroxyalkanoates; Polymers; RNA, Messenger; Real-Time Polymerase Chain Reaction; Stem Cells; Tissue Engineering; Tissue Scaffolds
PubMed: 25124858
DOI: 10.1111/cpr.12122 -
The Journal of Clinical Endocrinology... Jan 2009Bone has recently been described as exhibiting properties of an endocrine organ by producing osteocalcin that increases insulin sensitivity and secretion in animal...
CONTEXT
Bone has recently been described as exhibiting properties of an endocrine organ by producing osteocalcin that increases insulin sensitivity and secretion in animal models.
OBJECTIVE AND DESIGN
We aimed to evaluate circulating osteocalcin in association with insulin sensitivity and insulin secretion in three different studies in nondiabetic subjects: one cross-sectional study in 149 men (using minimal model), and two longitudinal studies in two independent groups (one formed by 26 women, and the other by 9 men and 11 women), after a mean of 7.3 and 16.8% weight loss, and after a mean of 8.7% weight loss plus regular exercise.
RESULTS
In the cross-sectional study, circulating osteocalcin was associated with insulin sensitivity, mainly in lean subjects, and with insulin secretion (only in lean subjects). A mean of 16.8%, but not 7.3% weight loss, led to significant increases in circulating osteocalcin. However, a mean of 8.7% weight loss plus regular exercise led to the more pronounced effects on the serum osteocalcin concentration, which increased in parallel to reduced visceral fat mass, unchanged thigh muscle mass, and increased leg strength and force. The postintervention serum levels of osteocalcin were associated with both insulin sensitivity (r = 0.49; P = 0.03) and fasting triglycerides (r = -0.54; P = 0.01). The change in visceral fat was the parameter that best predicted the change in serum osteocalcin, once age, body mass index, and insulin sensitivity changes were controlled for (P = 0.002).
CONCLUSION
Circulating osteocalcin could mediate the role of bone as an endocrine organ in humans.
Topics: Adult; Aged; Body Mass Index; Cross-Sectional Studies; Energy Intake; Female; Humans; Insulin; Insulin Resistance; Insulin Secretion; Longitudinal Studies; Male; Middle Aged; Osteocalcin; Resistance Training; Weight Loss
PubMed: 18854399
DOI: 10.1210/jc.2008-0270 -
Head & Face Medicine Aug 2023Eggshell peptides (EP) majorly contribute to rapid bone building in chicks, wherefore this paper investigated their potential for stimulating osteogenesis in vitro. In...
Eggshell peptides (EP) majorly contribute to rapid bone building in chicks, wherefore this paper investigated their potential for stimulating osteogenesis in vitro. In this study, the effects of EP, also called putamen ovi peptides and a combination of hyaluronic acid with EP in cell culture medium were tested towards proliferation, differentiation, gene expression and mineralization of bovine osteoprogenitors and primary human osteoblasts. The influence of EP at concentrations of 0.005 g/L, 0.5 g/L and 0.5 g/L with 0.25% hyaluronic acid was analyzed using immunocytochemical staining of bone-specific matrix proteins, namely collagen type I, osteonectin, osteopontin and osteocalcin, to prove osteoblastic differentiation. Additionally, Richardson-staining was performed. All tests revealed a superior osteoblastic differentiation with EP at 0.5 g/L after 5 days of cultivation. Hyaluronic acid alone showed controversial results and partially constrained osteoblastic differentiation in combination with EP to a level as low as for pure EP at 0.005 g/L. Of particular interest is the osteoblast-typical mineralization, as an important indicator of bone formation, which was measured indirectly via the calcium concentration after cultivation over 4 weeks. The mineralization showed an increase by a factor of 286 during the cultivation of primary human osteoblasts with hyaluronic acid and EP. Meanwhile, cell cultures treated with EP (0.5 g/L) only showed an 80-fold increase in calcium concentration.The influence of EP (0.5 g/L) on primary human osteoblasts was investigated by gene expression after 2 weeks of cultivation. Microarray and qRT-PCR analysis showed a strongly increased expression of main important genes in bone formation, bone regeneration and the physiological bone remodelling processes. Namely, BMP 2, osteopontin and the matrix metalloproteinases 1 and 9, were present during in vitro osteoprogenitor culture with EP. By explicitly underlining the potential of eggshell peptides for stimulating osteogenesis, as well as emphasizing complex and controversial interaction with hyaluronan, this manuscript is relevant for developing new functionalized biomaterials for bone regeneration.
Topics: Animals; Cattle; Humans; Osteopontin; Hyaluronic Acid; Calcium; Putamen; Peptides; Osteogenesis; Cell Differentiation; Osteocalcin; Osteoblasts; Cells, Cultured
PubMed: 37553683
DOI: 10.1186/s13005-023-00380-3 -
Low osteocalcin level is a risk factor for impaired glucose metabolism in a Chinese male population.Journal of Diabetes Investigation Jul 2016This study was to assess the association between serum osteocalcin level and glucose metabolism in a Chinese male population.
AIMS/INTRODUCTION
This study was to assess the association between serum osteocalcin level and glucose metabolism in a Chinese male population.
MATERIALS AND METHODS
We carried out a cross-sectional study with a cohort of participants from the Fangchenggang Area Male Health and Examination Survey. The cross-sectional study was carried out among 2,353 men, including 2,139 participants with normal glucose tolerance, 148 with impaired fasting glucose and 66 with type 2 diabetes. A subsample of 1,109 men with measurement of osteocalcin was observed in the cohort. After a 4-year follow-up period, 1,049 non-diabetic and 983 participants with normal glucose tolerance who submitted the available information were enrolled in the cohort. Participants were divided into group-H (≥23.33 ng/mL) and group-L (<23.33 ng/mL) by osteocalcin level.
RESULTS
In the cross-sectional study, osteocalcin levels were highest in participants with normal glucose tolerance, followed by those with impaired fasting glucose and type 2 diabetes (P < 0.001). In partial correlation analysis adjusted for age, serum osteocalcin level was related to glucose level (r = -0.082, P < 0.001), insulin level (r = -0.079, P < 0.001) and insulin resistance (r = -0.065, P = 0.002). Compared with group-H, group-L was associated with an increased risk of type 2 diabetes (odds ratio 2.107, 95% confidence interval 1.123-3.955), impaired fasting glucose (odds ratio 2.106; 95% CI 1.528-2.902), and insulin resistance (odds ratio 1.359, 95% confidence interval 1.080-1.710) adjusted for age, education levels, cigarette smoking and lipid profiles. In the cohort study, the increased risk of impaired fasting glucose was significant in group-L vs group-H (3.3% vs 1.2%, P = 0.026).
CONCLUSIONS
Low serum osteocalcin level was a risk factor for impaired glucose metabolism and subsequent type 2 diabetes.
Topics: Adult; Asian People; China; Cross-Sectional Studies; Glucose; Glucose Metabolism Disorders; Glucose Tolerance Test; Humans; Male; Middle Aged; Osteocalcin; Risk Factors
PubMed: 27181428
DOI: 10.1111/jdi.12439 -
Archives of Endocrinology and Metabolism Aug 2018This study evaluated the effects of combination therapy of curcumin and alendronate on BMD and bone turnover markers in postmenopausal women with osteoporosis. (Randomized Controlled Trial)
Randomized Controlled Trial
OBJECTIVE
This study evaluated the effects of combination therapy of curcumin and alendronate on BMD and bone turnover markers in postmenopausal women with osteoporosis.
SUBJECTS AND METHODS
In a randomized, double-blind trial study, 60 postmenopausal women were divided into three groups: control, alendronate, and alendronate + curcumin. Each group included 20 patients. Total body, total hip, lumbar spine and femoral neck BMDs were measured by dual-energy X-ray absorptiometry (DXA) at baseline and after 12 months of therapy. Bone turnover markers such as bone-specific alkaline phosphatase (BALP), osteocalcin and C-terminal cross-linking telopeptide of type I collagen (CTx) were measured at the outset and 6 months later.
RESULTS
Patients in the control group suffered a significant decrease in BMD and increased bone turnover markers at the end of study. The group treated with only alendronate showed significantly decreased levels of BALP and CTx and increased levels of osteocalcin compared to the control group. The alendronate group also showed significant increases in the total body, total hip, lumbar spine and femoral neck BMDs at the end of study compared to the control group. In the curcumin + alendronate group, BALP and CTx levels decreased and osteocalcin levels increased significantly at the end of study compared to the control and alendronate groups. BMD indexes also increased in four areas significantly at the end of study compared to the control and alendronate groups.
CONCLUSION
The combination of curcumin and alendronate has beneficial effects on BMD and bone turnover markers among postmenopausal women with osteoporosis. Arch Endocrinol Metab. 2018;62(4):438-45.
Topics: Aged; Alendronate; Alkaline Phosphatase; Bone Density; Bone Density Conservation Agents; Bone Remodeling; Collagen Type II; Curcumin; Double-Blind Method; Drug Therapy, Combination; Female; Humans; Middle Aged; Osteocalcin; Osteoporosis, Postmenopausal; Peptide Fragments
PubMed: 30304108
DOI: 10.20945/2359-3997000000060 -
Advances in Nutrition (Bethesda, Md.) Aug 2022Milk contains a number of bone-beneficial nutrients. However, milk, due to the D-galactose content, might have unfavorable effects on bone health. A meta-analysis of... (Meta-Analysis)
Meta-Analysis
Milk contains a number of bone-beneficial nutrients. However, milk, due to the D-galactose content, might have unfavorable effects on bone health. A meta-analysis of randomized controlled trials (RCTs) was performed to clarify the effects of milk supplementation on bone mineral density (BMD), bone turnover markers [N-terminal telopeptide of type I collagen (NTx), C-terminal telopeptide of type 1 collagen (CTx), osteocalcin, bone alkaline phosphatase (BALP), and procollagen type 1 N-propeptide (P1NP)], and hormonal indices related to bone metabolism [parathyroid hormone (PTH), 25-hydroxyvitamin D [25(OH)D], and insulin-like growth factor 1 (IGF-1)] in adults. The PubMed and Web of Science databases were searched. A random-effects model was used to estimate the pooled effect sizes. A total of 20 RCTs were included. The trial duration ranged from 1 mo to 36 mo. Milk supplementation resulted in a small but significant increase in BMD at the hip (+0.004 g/cm2; n = 9 RCTs) and lumbar spine (+0.025 g/cm2; n = 7), but did not significantly affect whole-body BMD (n = 3) and femoral neck BMD (n = 7). Milk supplementation reduced the concentrations of P1NP (-5.20 ng/mL; n = 9), CTx (-0.16 ng/mL; n = 9), and NTx (-8.66 nmol bone collagen equivalents/mmol creatinine; n = 3). The concentrations of osteocalcin (n = 9) and BALP (n = 3) were not affected by milk supplementation. Reduced parathyroid hormone PTH (-1.01 pg/mL; n = 13) concentrations and increased IGF-1 (+1.79 nmol/l; n = 4) concentrations were observed with milk supplementation. 25(OH)D (+3.73 ng/mL; n = 11) concentrations were increased with vitamin-D fortified milk supplementation. The addition of milk to the diet may potentially increase the likelihood of preventing bone loss by restoring bone homeostasis through the modulation of the calcium-vitamin D-PTH axis, bone remodeling rate, and growth hormone/IGF-1 axis.
Topics: Adult; Animals; Biomarkers; Bone Density; Bone Remodeling; Collagen Type I; Dietary Supplements; Humans; Insulin-Like Growth Factor I; Milk; Osteocalcin; Parathyroid Hormone; Randomized Controlled Trials as Topic; Vitamin D
PubMed: 34792092
DOI: 10.1093/advances/nmab136 -
Journal of Trace Elements in Medicine... Jul 2014To study the effects and importance of fluoride on FBs in the development of extraperiosteal calcification and the ossification of skeletal fluorosis, the presence of...
To study the effects and importance of fluoride on FBs in the development of extraperiosteal calcification and the ossification of skeletal fluorosis, the presence of the osteogenic phenotype, which is indicated by the expression of core-binding factor α1 (Cbfa1) and osteocalcin (OCN), in an FB cell line (L929) and in osteoblasts (OBs) exposed to fluoride was determined. Fibroblasts and osteoblasts were exposed to different concentrations of fluoride (0, 0.0001, 0.001, 0.1, 1.0, 10.0 and 20.0 mg/L F(-)). By using RT-PCR and ELISA, the mRNA levels of Cbfa1 and OCN were measured at 48 h, and the protein levels of Cbfa1 and OCN were measured at 2, 4, 24, 48 and 72 h. The data demonstrated the following: (1) The Cbfa1 protein level in fluoride-treated fibroblasts clearly increased at 48 h in the groups treated with 0.0001, 0.001, 0.1, 1.0 and 20.0 mg/L F(-). The Cbfa1 protein level of the group treated with 10 mg/L F(-) at 72 h was higher than that of the control group. The level of Cbfa1 mRNA in the fibroblasts was much higher at 48 h in the group treated with 10.0 mg/L F(-) than in the control group. (2) The OCN protein level in fluoride-treated fibroblasts was significantly higher than that of the control group in the 0.0001, 0.1, 1.0, 10.0 and 20.0 mg/L F(-) groups at 2 h, and in the 0.001 and 0.1 F(-) groups at 4 h. A slightly higher level of OCN mRNA in fluoride-treated fibroblasts was also found in the 1.0 and 20.0 mg/L F(-) groups compared to the control group. (3) The expressions of Cbfa1 and OCN in osteoblasts treated with the same experimental conditions as the fibroblasts were up-regulated by fluoride following the same trend as in the fibroblasts. Our results showed an increase in the expression of Cbfa1 and OCN in fibroblasts and osteoblasts exposed to fluoride and suggested that the osteogenic function of fibroblasts induced by fluoride could play an important role in the development of extraperiosteal ossification during skeletal fluorosis.
Topics: Animals; Cell Line; Cells, Cultured; Core Binding Factor Alpha 1 Subunit; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Fluorides; Immunohistochemistry; Mice; Osteoblasts; Osteocalcin
PubMed: 24680482
DOI: 10.1016/j.jtemb.2014.02.004