scholarly journals CLEC-2 suppresses calcification in cultured osteoblasts

2019 ◽  
Author(s):  
Takenori Kanai ◽  
Yoshihiko Sawa ◽  
Kenyo Takara ◽  
Koichiro Kajiwara ◽  
Takahiro Fujita ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Collagen Type ◽  
Collagen Type I ◽  
Medullary Cavity ◽  
Type I ◽  
Alizarin Red ◽  
Phosphatase Activities

AbstractPodoplanin is the only counter-receptor of platelet CLEC-2 and is expressing on mature osteoblast, but there is no report on the role of podoplanin and CLEC-2 in calcification. This study aimed to investigate the role of podoplanin binding to CLEC-2 in the calcification of osteoblasts carrying homozygously deleted Pdpn alleles (PdpnΔ/Δ) by heterozygously expressing collagen type I alpha 1 promoter (Col1a)-driven Cre recombinase. There were no macroscopic abnormalities in the bone and dentin of Col1a11-Cre;PdpnΔ/Δ mice but the coccygeal bone medullary cavity was very narrow. In the quantitative analysis for alizarin red-stained products and alkaline phosphatase activities on the cultured calvarial osteoblasts, the amounts of calcified products and alkaline phosphatase activity of calvarial osteoblasts of both Pdpnfl/fl and Col1a11-Cre;PdpnΔ/Δ mice were significantly higher in the calcification medium than in the α-mem. Both the amounts of calcified products and alkaline phosphatase activity of calvarial osteoblasts from Pdpnfl/fl mice were significantly lower in the calcification medium with CLEC-2 than without CLEC-2 while there were no significant differences in the amounts of calcified products and alkaline phosphatase activities of calvarial osteoblasts from Col1a11-Cre;PdpnΔ/Δ mice with CLEC-2. Platelet CLEC-2 may play a role in regulating the calcification via binding to podoplanin on mature osteoblasts expressing podoplanin in the medullary cavity of a part of the bone.

The inhibitory effects of vancomycin on rat bone marrow–derived mesenchymal stem cell differentiation

2021 ◽  
pp. 1-5
Author(s):  
Kari Hanson ◽  
Carly Isder ◽  
Kristen Shogren ◽  
Anthony L. Mikula ◽  
Lichun Lu ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Alkaline Phosphatase ◽  
Osteogenic Differentiation ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
High Dose ◽  
Inhibitory Effects ◽  
Alizarin Red ◽  
Osteogenic Factors

OBJECTIVE The use of intrawound vancomycin powder in spine surgery has been shown to decrease the rate of surgical site infections; however, the optimal dose is unknown. High-dose vancomycin inhibits osteoblast proliferation in vitro and may decrease the rate of solid arthrodesis. Bone marrow–derived mesenchymal stem cells (BMSCs) are multipotent cells that are a source of osteogenesis in spine fusions. The purpose of this study was to determine the effects of vancomycin on rat BMSC viability and differentiation in vitro. METHODS BMSCs were isolated from the femurs of immature female rats, cultured, and then split into two equal groups; half were treated to stimulate osteoblastic differentiation and half were not. Osteogenesis was stimulated by the addition of 50 µg/mL l-ascorbic acid, 10 mM β-glycerol phosphate, and 0.1 µM dexamethasone. Vancomycin was added to cell culture medium at concentrations of 0, 0.04, 0.4, or 4 mg/mL. Early differentiation was determined by alkaline phosphatase activity (4 days posttreatment) and late differentiation by alizarin red staining for mineralization (9 days posttreatment). Cell viability was determined at both the early and late time points by measurement of formazan colorimetric product. RESULTS Viability within the first 4 days decreased with high-dose vancomycin treatment, with cells receiving 4 mg/mL vancomycin having 40%–60% viability compared to the control. A gradual decrease in alizarin red staining and nodule formation was observed with increasing vancomycin doses. In the presence of the osteogenic factors, vancomycin did not have deleterious effects on alkaline phosphatase activity, whereas a trend toward reduced activity was seen in the absence of osteogenic factors when compared to osteogenically treated cells. CONCLUSIONS Vancomycin reduced BMSC viability and impaired late osteogenic differentiation with high-dose treatment. Therefore, the inhibitory effects of high-dose vancomycin on spinal fusion may result from both reduced BMSC viability and some impairment of osteogenic differentiation.

scholarly journals WNT-5A regulates TGF-β-related activities in liver fibrosis

2017 ◽  
Vol 312 (3) ◽  
pp. G219-G227 ◽  
Author(s):  
Leonie Beljaars ◽  
Sara Daliri ◽  
Christa Dijkhuizen ◽  
Klaas Poelstra ◽  
Reinoud Gosens
Keyword(s):  
Liver Fibrosis ◽  
Functional Role ◽  
Collagen Type ◽  
Collagen Type I ◽  
Matrix Proteins ◽  
Type I ◽  
Human Livers

WNT-5A is a secreted growth factor that belongs to the noncanonical members of the Wingless-related MMTV-integration family. Previous studies pointed to a connection between WNT-5A and the fibrogenic factor TGF-β warranting further studies into the functional role of WNT-5A in liver fibrosis. Therefore, we studied WNT-5A expressions in mouse and human fibrotic livers and examined the relation between WNT-5A and various fibrosis-associated growth factors, cytokines, and extracellular matrix proteins. WNT-5A gene and protein expressions were significantly increased in fibrotic mouse and human livers compared with healthy livers. Regression or therapeutic intervention in mice resulted in decreased hepatic WNT-5A levels paralleled by lower collagen levels. Immunohistochemical analysis showed WNT-5A staining in fibrotic septa colocalizing with desmin staining indicating WNT-5A expression in myofibroblasts. In vitro studies confirmed WNT-5A expression in this cell type and showed that TGF-β significantly enhanced WNT-5A expression in contrast to PDGF-BB and proinflammatory cytokines IL-1β and TNF-α. Additionally, TGF-β induces the expression of the WNT receptors FZD2 and FZD8. After silencing of WNT-5A, reduced levels of collagen type I, vimentin, and fibronectin in TGF-β-stimulated myofibroblasts were measured compared with nonsilencing siRNA-treated controls. Interestingly, the antifibrotic cytokine IFNγ suppressed WNT-5A in vitro and in vivo. IFNγ-treated fibrotic mice showed significantly less WNT-5A expression compared with untreated fibrotic mice. In conclusion, WNT-5A paralleled collagen I levels in fibrotic mouse and human livers. WNT-5A expression in myofibroblasts is induced by the profibrotic factor TGF-β and plays an important role in TGF-β-induced regulation of fibrotic matrix proteins, whereas its expression can be reversed upon treatment, both in vitro and in vivo. NEW & NOTEWORTHY This study describes the localization and functional role of WNT-5A in human and mouse fibrotic livers. Hepatic WNT-5A expression parallels collagen type I expression. In vivo and in vitro, the myofibroblasts were identified as the key hepatic cells producing WNT-5A. WNT-5A is under control of TGF-β and its activities are primarily profibrotic.

Acidosis inhibits osteoblastic and stimulates osteoclastic activity in vitro

1992 ◽  
Vol 262 (3) ◽  
pp. F442-F448 ◽  
Author(s):  
N. S. Krieger ◽  
N. E. Sessler ◽  
D. A. Bushinsky
Keyword(s):  
Alkaline Phosphatase ◽  
Metabolic Acidosis ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Collagen Synthesis ◽  
Calcium Flux ◽  
Osteoclastic Activity ◽  
Glucuronidase Activity

Metabolic acidosis induces net calcium flux (JCa) from cultured neonatal mouse calvariae through physicochemical and cell-mediated mechanisms. To determine the role of osteoblasts in acid-induced JCa, collagen synthesis and alkaline phosphatase activity were assessed in calvariae incubated in reduced pH and bicarbonate medium, a model of metabolic acidosis (Met), and compared with controls (Ctl). Collagen synthesis fell from 30.5 +/- 1.1 in Ctl to 25.1 +/- 0.4% with Met, and alkaline phosphatase decreased from 403 +/- 25 in Ctl to 298 +/- 21 nmol Pi.min-1.mg protein-1 with Met. During acidosis JCa was correlated inversely with percent collagen synthesis (r = -0.743, n = 11, P = 0.009) and with alkaline phosphatase activity (r = -0.453, n = 22, P = 0.034). To determine the role of osteoclasts in acid-induced JCa, osteoclastic beta-glucuronidase activity was determined in Ctl and Met in the absence or presence of the osteoclastic inhibitor calcitonin (CT, 3 x 10(-9) M). Met increased beta-glucuronidase (5.9 +/- 0.2) compared with Ctl (4.6 +/- 0.3 micrograms phenolphthalein released.bone-1.h-1), whereas CT inhibited beta-glucuronidase in both Ctl and Met (3.1 +/- 0.2 and 3.5 +/- 0.3, respectively). During acidosis JCa was correlated directly with beta-glucuronidase activity (r = 0.683, n = 42, P less than 0.001). Thus the cell-mediated component of JCa during acidosis in vitro appears to result from a combination of inhibited osteoblastic and stimulated osteoclastic activity.

RICKETS, DEFICIENCY OF "ALKALINE" PHOSPHATASE ACTIVITY AND PREMATURE LOSS OF TEETH IN CHILDHOOD

PEDIATRICS ◽  
1953 ◽  
Vol 11 (4) ◽  
pp. 309-322
Author(s):  
EDNA H. SOBEL ◽  
LELAND C. CLARK ◽  
R. PHYLLIS FOX ◽  
MEINHARD ROBINOW
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Enzyme Treatment ◽  
Deciduous Teeth ◽  
Local Factor ◽  
Plasma Alkaline Phosphatase ◽  
Serum Inorganic Phosphate ◽  
Genetically Determined

A child, studied between the ages of 1½ and 3½ years, presented an abnormally low plasma alkaline phosphatase activity (0.8-1.64 Bessey-Lowry u.), a deformed skeleton and the loss of most of her deciduous teeth. The serum Ca was normal; the serum inorganic phosphate remained at the normal relatively high levels of infancy as the child grew older Roentgenograms demonstrated deficient mineralization of the skeleton and teeth. Biopsies of the liver and the costochondral junction displayed a deficiency of tissue alkaline phosphatase activity. The architecture of the rib was consistent with rickets. There was no evidence for the presence of an inhibitor of alkaline phosphatase, such as beryllium, or for an excessive excretion of the enzyme. Treatment with purified growth hormone, ascorbic acid and thiamin chloride had no effect, while vitamin D 500 thousand u. caused little change in the enzyme activity in a 10 day period. The father had low plasma alkaline phosphatase activity and a number of similar patients are mentioned, for whom there was also evidence that the deficiency in alkaline phosphatase activity may be genetically determined. While the precise role of alkaline phosphatase activity in the metabolism of bone is not clear, the findings in this patient suggest that growing bone may require the presence of alkaline phosphatase for normal calcification, and that the skeletal disorder, which could not be distinguished from rickets, may be related to a disturbance in the local factor.

scholarly journals Cytocompatibility evaluation of hydroxyapatite/collagen composites doped with Zn+2

2007 ◽  
Vol 12 (2) ◽  
pp. 307-312 ◽  
Author(s):  
Maria Helena Santos ◽  
Ana Paula M. Shaimberg ◽  
Patricia Valerio ◽  
Alfredo M. Goes ◽  
Maria de Fátima Leite ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Orthophosphoric Acid ◽  
Type I Collagen ◽  
Enzymatic Digestion ◽  
Osteoblastic Cells ◽  
Type I ◽  
Post Test ◽  
Synthetic Hydroxyapatite

The cytocompatibility of synthetic hydroxyapatite/collagen composites alone or doped with Zn+2 was tested by using primary culture of osteoblasts. The hydroxyapatite (HAP) was synthesized having calcium hydroxide and orthophosphoric acid as precursors. A new HAP composite was developed adding 1.05 w% of Zn(NO3)2.6H2O forming HAPZn. The pure type I collagen (COL) was obtained from bovine pericardium by enzymatic digestion method. The HAP/COL and HAPZn/COL composites were developed and characterized by SEM/EDS. The cell viability and alkaline phosphatase activity in the presence of composites were evaluated by MTT assay and NBT-BCIP assay, respectively, and compared to osteoblastic cells of the control. Three individual experiments were accomplished in triplicates and submitted to the variance analysis and Bonferroni’s post-test with statistically significant at p<0.05. The HAPZn/COL composite did not stimulate the proliferation and increasing of alkaline phosphatase activity of the osteoblastic cells. The tested composites did not alter the cellular viability neither caused alterations in the cellular morphology in 72 h showing adequate properties for biological applications.

Alkaline phosphatase activity in the plasma of children and adolescents.

1977 ◽  
Vol 23 (3) ◽  
pp. 469-472 ◽  
Author(s):  
G A Fleisher ◽  
E S Eickelberg ◽  
L R Elveback
Keyword(s):  
Alkaline Phosphatase ◽  
Public Schools ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Serum Alkaline Phosphatase ◽  
Intraclass Correlation ◽  
Serum Alkaline Phosphatase Activity ◽  
Upper Limits ◽  
Prepubertal Girls

Abstract We determined plasma (serum alkaline phosphatase activity in 854 healthy students of the Rochester, Minnesota, public schools. Prepubertal girls had somewhat greater upper limits than did boys, and there was a low trend of increasing activity in both sexes. At the beginning of adolescence increasing activities were observed, which peaked at ages 11 to 12 years in girls and at ages 13 to 14 in boys. Adult values were not reached until six to eight years later. In 180 pairs of siblings, a significant intraclass correlation was noted. A possible role of alkaline phosphatase in the regulation of protein synthesis is suggested.

Role of catechin on collagen type I stability upon oxidation: a NMR approach

2019 ◽  
Vol 34 (1) ◽  
pp. 53-62 ◽  
Author(s):  
Massimo Lucarini ◽  
Fabio Sciubba ◽  
Donatella Capitani ◽  
Maria Enrica Di Cocco ◽  
Laura D’Evoli ◽  
...  
Keyword(s):  
Collagen Type ◽  
Collagen Type I ◽  
Type I

scholarly journals The Effects of Morinda citrifolia (Noni) on the Cellular Viability and Osteogenesis of Stem Cell Spheroids

Medicina ◽  
2020 ◽  
Vol 56 (8) ◽  
pp. 389
Author(s):  
Sae Kyung Min ◽  
Jaekwen Oh ◽  
Jun-Beom Park
Keyword(s):  
Alkaline Phosphatase ◽  
Stem Cell ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Morinda Citrifolia ◽  
Herbal Remedies ◽  
Cellular Viability ◽  
Alizarin Red ◽  
Anthraquinone Dye ◽  
Cell Spheroids

Background and objectives: Morinda citrifolia (Noni) has been widely used in herbal remedies to treat and prevent various kinds of diseases. We conducted this study to evaluate the effects of Noni extract on the maintenance of morphology, the improvement of cellular viability, and the enhancement of osteogenesis of stem cell spheroids. Materials and Methods: We cultured stem cell spheroids made with gingiva-derived stem cells in the presence of Noni extract at concentrations of 10, 100 and 200 ng/mL. We performed analysis of the cell morphology and changes in the cellular viability. We conducted alkaline phosphatase activity assays using a kit, and mineralization assays using an anthraquinone dye to evaluate the osteogenesis of stem cell spheroids with the addition of Noni extract. Results: The applied cells formed spheroids well, and the addition of Noni at 10, 100 and 200 ng/mL concentrations did not produce significant morphological changes. The quantitative values for cellular viability on Day 3 showed that the absorbance values at 450 nm were 0.314 ± 0.013, 0.318 ± 0.008, 0.304 ± 0.000 and 0.300 ± 0.011 for Noni at 0, 10, 100 and 200 ng/mL concentrations, respectively. The results of alkaline phosphatase activity with absorbance values at 405 nm were 0.189 ± 0.019, 0.174 ± 0.023, 0.192 ± 0.014 and 0.210 ± 0.062 for Noni at 0, 10, 100 and 200 ng/mL concentrations, respectively, on Day 4. There were significantly higher values of Alizarin Red S staining for Noni in the 10, 100 and 200 ng/mL groups, with the highest value at 100 ng/mL when compared with the unloaded control on Day 14. Conclusions: Based on these findings, we concluded that Noni extract might be applied for the enhanced osteogenic differentiation of stem cell spheroids.

Circulating levels of 1,25 dihydroxyvitamin D, alkaline phosphatase, hydroxyproline, and osteocalcin associated with antler growth in white-tailed deer

1988 ◽  
Vol 118 (3) ◽  
pp. 407-414 ◽  
Author(s):  
Karen L. Van Der Eems ◽  
Robert D. Brown ◽  
Caren M. Gundberg
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Serum Alkaline Phosphatase ◽  
Serum Samples ◽  
Serum Alkaline Phosphatase Activity ◽  
Dihydroxyvitamin D ◽  
Antler Growth ◽  
Antler Cycle

Abstract. Alkaline phosphatase, hydroxyproline, osteocalcin, and 1,25(OH)2D were measured in biweekly serum samples obtained from 6 adult (> 4 years), 4 juvenile (1–4 years) and 4 fawn (< 1 year) male white-tailed deer from Oct. 1983 to Oct. 1984. Antler length, from the pedicle to the tip, was measured at the time of serum sampling. Serum alkaline phosphatase activity and levels of hydroxyproline and osteocalcin were higher (P < 0.05) in fawns compared with juveniles and adults reflecting increased bone metabolism in the younger deer. In adult deer serum alkaline phosphatase activity and hydroxyproline levels were elevated (P < 0.05) during the period of antler growth, whereas serum osteocalcin and 1,25(OH)2D increased (P < 0.05) during antler mineralization. Similar but less pronounced trends occurred in juvenile deer, possibly a reflection of skeletal growth in the younger animals. The data lend support for utilization of the deer antler cycle as a model for studies of bone disorders. Further work is needed to help clarify the role of hydroxyproline, osteocalcin, and 1,25(OH)2D in the antler cycle.

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