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Index / Bioassays / QuantiChrom™ Alkaline Phosphatase Assay Kit, Quantitative determination of alkaline phosphatase ALP activity using stable p-nitrophenol phosphate substrate at 405nm. Kit size 250 tests. Detection lim / Product Detail : DALP-250 QuantiChrom™ Alkaline Phosphatase Assay Kit, Quantitative determination of alkaline phosphatase ALP activity using stable p-nitrophenol phosphate substrate at 405nm. Kit size 250 tests. Detection lim
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Product name : QuantiChrom™ Alkaline Phosphatase Assay Kit, Quantitative determination of alkaline phosphatase ALP activity using stable p-nitrophenol phosphate substrate at 405nm. Kit size 250 tests. Detection lim

Catalog number : DALP-250

Quantity: 250tests

Availability: Yes

Supplier name : Bioassays

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QuantiChrom™ Alkaline Phosphatase Assay Kit, Quantitative determination of alkaline phosphatase ALP activity using stable p-nitrophenol phosphate substrate at 405nm. Kit size 250 tests. Detection lim antibody storage GENTAUR recommends for long therm storage to freeze at -24 C. For short time storage up to 30 days we suggest fridge storage at 1 to 10 C. Prevent multiple freeze taw cycles of QuantiChrom™ Alkaline Phosphatase Assay Kit, Quantitative determination of alkaline phosphatase ALP activity using stable p-nitrophenol phosphate substrate at 405nm. Kit size 250 tests. Detection lim.

More Details about

For quantitative determination of alkaline phosphatase (ALP) activity using stable p-nitrophenol phosphate substrate.
Method: OD405nm.
Samples: serum, plasma etc.
Species: all.
Procedure: 4 min.
Size: 250 tests.
Detection limit: 2 U/L.

Alkaline phosphatase (ALP) catalyzes the hydrolysis of phosphate esters in
an alkaline environment, resulting in the formation of an organic radical and
inorganic phosphate. In mammals, this enzyme is found mainly in the liver
and bones. Marked increase in serum ALP levels, a disease known as
hyperalkalinephosphatasemia, has been associated with malignant biliary
obstruction, primary biliary cirrhosis, primary sclerosing cholangitis, hepatic
lymphoma and sarcoidosis.
Simple, direct and automation-ready procedures for measuring ALP activity
in serum are becoming popular in Research and Drug Discovery. BioAssay
Systems' QuantiChromTM Alkaline Phosphatase Assay Kit is designed to
measure ALP activity directly in biological samples without pretreatment.
The improved method utilizes p-nitrophenyl phosphate that is hydrolyzed by
ALP into a yellow colored product (maximal absorbance at 405nm). The
rate of the reaction is directly proportional to the enzyme activity.
p-Nitrophenyl phosphate p-nitrophenol + phosphate
High sensitivity and wide linear range. Use 5 μL serum or plasma
sample. The detection limit is 2 U/L, linear up to 800 U/L.
Homogeneous and simple procedure. Simple “mix-and-measure”
procedure allows reliable quantitation of ALP activity within 5 minutes.
Robust and amenable to HTS. All reagents are compatible with highthroughput
liquid handling instruments.
Direct Assays: ALP activity in serum, plasma and other sources.
Characterization and Quality Control for ALP production.
Drug Discovery: high-throughput screen for ALP inhibitors and evaluation
of ALP inhibitors.
KIT CONTENTS (250 tests in 96-well plates)
Assay Buffer: 50 mL, pH 10.5 Mg Acetate: 1.5 mL 0.2 M
pNPP Liquid: 600 μL 1 M Calibrator: 10 mL Tartrazine
Storage conditions. The kit is shipped at room temperature. Store pNPP
Liquid at -20°C and other components at 4°C. Shelf life of 12 months after
Precautions: reagents are for research use only. Normal precautions for
laboratory reagents should be exercised while using the reagents.

Pipeting devices and accessories (e.g. multi-channel pipettor).
Procedure using 96-well plate:
Clear bottom 96-well plates (e.g. Corning Costar) and plate reader.
Procedure using cuvette:
Spectrophotometer and cuvets for measuring OD 405nm.

Sample Type: serum

Species:  mouse


References: Wan, Y et al (2007). PPAR-g regulates osteoclastogenesis in mice. Nature Med. 13(12): 1496-1503.

Pubmed ID: 18059282

Pubmed link:

Abstract: Osteoclasts are bone-resorbing cells derived from hematopoietic precursors of the monocyte-macrophage lineage. Regulation of osteoclast function is central to the understanding of bone diseases such as osteoporosis, rheumatoid arthritis and osteopetrosis. Although peroxisome proliferator-activated receptor-gamma (PPAR-gamma) has been shown to inhibit osteoblast differentiation, its role, if any, in osteoclasts is unknown. This is a clinically crucial question because PPAR-gamma agonists, "such as thiazolidinediones-" a class of insulin-sensitizing drugs, have been reported to cause a higher rate of fractures in human patients. Here we have uncovered a pro-osteoclastogenic effect of PPAR-gamma by using a Tie2Cre/flox mouse model in which PPAR-gamma is deleted in osteoclasts but not in osteoblasts. These mice develop osteopetrosis characterized by increased bone mass, reduced medullary cavity space and extramedullary hematopoiesis in the spleen. These defects are the result of impaired osteoclast differentiation and compromised receptor activator of nuclear factor-kappaB ligand signaling and can be rescued by bone marrow transplantation. Moreover, ligand activation of PPAR-gamma by rosiglitazone exacerbates osteoclast differentiation in a receptor-dependent manner. Our examination of the underlying mechanisms suggested that PPAR-gamma functions as a direct regulator of c-fos expression, an essential mediator of osteoclastogenesis. Therefore, PPAR-gamma and its ligands have a previously unrecognized role in promoting osteoclast differentiation and bone resorption.

 [PubMed - indexed for MEDLINE]


References: Kim, HJ et al (2006). Glucocorticoids suppress bone formation via the osteoclast. J Clin Invest. 116(8):2152-60

Pubmed ID: 16878176

Pubmed link:

Abstract: The pathogenesis of glucocorticoid-induced (GC-induced) bone loss is unclear. For example, osteoblast apoptosis is enhanced by GCs in vivo, but they stimulate bone formation in vitro. This conundrum suggests that an intermediary cell transmits a component of the bone-suppressive effects of GCs to osteoblasts in the intact animal. Bone remodeling is characterized by tethering of the activities of osteoclasts and osteoblasts. Hence, the osteoclast is a potential modulator of the effect of GCs on osteoblasts. To define the direct impact of GCs on bone-resorptive cells, we compared the effects of dexamethasone (DEX) on WT osteoclasts with those derived from mice with disruption of the GC receptor in osteoclast lineage cells (GRoc-/- mice). While the steroid prolonged longevity of osteoclasts, their bone-degrading capacity was suppressed. The inhibitory effect of DEX on bone resorption reflects failure of osteoclasts to organize their cytoskeleton in response to M-CSF. DEX specifically arrested M-CSF activation of RhoA, Rac, and Vav3, each of which regulate the osteoclast cytoskeleton. In all circumstances GRoc-/- mice were spared the impact of DEX on osteoclasts and their precursors. Consistent with osteoclasts modulating the osteoblast-suppressive effect of DEX, GRoc-/- mice are protected from the steroid's inhibition of bone formation.

[PubMed - indexed for MEDLINE] PMCID: PMC1518793


References: Bhattacharya, A et al (2006). Effect of fish oil on bone mineral density in aging C57BL/6 female mice. J. Nutr. Biochem 18(6):372-379.

Pubmed ID: 16963250

Pubmed link:

Abstract: Life expectancy has increased considerably over the last century in the United States. It is expected that this longevity will be accompanied by an increase in the prevalence of osteoporosis and accompanying complications in the elderly population. Age-related loss of bone mass and bone fragility are major risk factors for osteoporosis, leading to an increased risk of fractures. Therefore, nutritional strategies and lifestyle changes that prevent age-related osteoporosis and improve the quality of life for the elderly population are urgently needed. Hence, the present study compared the effects of corn oil (CO; n-6 fatty acids; commonly present in Western diets) and fish oil (FO; n-3 fatty acids) on bone mineral density (BMD) in aging C57BL/6 female mice. After 6 months of dietary treatment, we found that 18-month-old FO-fed mice maintained higher BMD in different bone regions compared to CO-fed mice. These findings were accompanied by a decreased activity of pro-inflammatory cytokines, tumor necrosis factor-alpha and interleukin-6 in stimulated splenocytes; a nonsignificant but greater increase in bone formation markers alkaline phosphatase and osteocalcin in the serum; and lower osteoclast generation in bone marrow cell cultures in FO-fed mice. In conclusion, these findings suggest that providing n-3 fatty acids may have a beneficial effect on bone mass during aging by modulating bone formation and bone resorption factors.

 [PubMed - indexed for MEDLINE]


Sample Type: plasma

Species:  mouse


References: Sekiya, S, Suzuki, A (2011). Direct conversion of mouse fibroblasts to hepatocyte-like cells by defined factors. Nature 475(7356):390-3

Pubmed ID: 21716291

Pubmed link:

Abstract: The location and timing of cellular differentiation must be stringently controlled for proper organ formation. Normally, hepatocytes differentiate from hepatic progenitor cells to form the liver during development. However, previous studies have shown that the hepatic program can also be activated in non-hepatic lineage cells after exposure to particular stimuli or fusion with hepatocytes. These unexpected findings suggest that factors critical to hepatocyte differentiation exist and become activated to induce hepatocyte-specific properties in different cell types. Here, by screening the effects of twelve candidate factors, we identify three specific combinations of two transcription factors, comprising Hnf4α plus Foxa1, Foxa2 or Foxa3, that can convert mouse embryonic and adult fibroblasts into cells that closely resemble hepatocytes in vitro. The induced hepatocyte-like (iHep) cells have multiple hepatocyte-specific features and reconstitute damaged hepatic tissues after transplantation. The generation of iHep cells may provide insights into the molecular nature of hepatocyte differentiation and potential therapies for liver diseases.

[PubMed - indexed for MEDLINE]


Sample Type: hydrogels

Species:  human cells


References: Keskar, V et al (2009). In vitro evaluation of macroporous hydrogels to facilitate stem cell infiltration, growth, and mineralization. Tissue Eng Part A. 15(7):1695-707.

Pubmed ID: 19119921

Pubmed link:

Abstract: Hydrogels have gained acceptance as biomaterials in a wide range of applications, including pharmaceutical formulations, drug delivery, and tissue sealants. However, exploiting the potential of hydrogels as scaffolds for cell transplantation, tissue engineering, and regenerative medicine still remains a challenge due to, in part, scaffold design limitations. Here, we describe a highly interconnected, macroporous poly(ethylene glycol) diacrylate hydrogel scaffold, with pores ranging from 100 to 600 microm. The scaffold exhibits rapid cell uptake and cell seeding without the need of any external force or device with high incorporation efficiency. When human mesenchymal stem cells are seeded within the porous scaffolds, the scaffolds were found to promote long-term stem cell viability, and on exposure to osteogenic medium, elicit an mineralization response as evaluated by an increased alkaline phosphatase activity (per cell) and calcium and phosphate content within the constructs. The atomic composition of the mineralized matrix was further determined by energy dispersive spectroscopy and found to be similar to calcium-deficient hydroxyapatite, the amorphous biological precursor of bone. The macroporous design of the hydrogel appears advantageous over similar porous hydrogel scaffolds with respect to ease of synthesis, ease of stem cell seeding, and its ability to support long-term stem cell survival and possible differentiation.

[PubMed - indexed for MEDLINE] PMCID: PMC2810413


Sample Type: cells

Species:  rat


References: Panizo, S et al (2009). RANKL increases vascular smooth muscle cell calcification through a RANK-BMP4-dependent pathway. Circ Res. 104(9):1041-8.

Pubmed ID: 19325147

Pubmed link:

Abstract: Vascular calcification commonly associated with several pathologies and it has been suggested to be similar to bone mineralization. The axis RANKL-OPG (receptor activator of nuclear factor kappaB ligand-osteoprotegerin) finely controls bone turnover. RANKL has been suggested to increase vascular calcification, but direct evidence is missing. Thus, in the present work, we assess the effect of RANKL in vascular smooth muscle cell (VSMC) calcification. VSMCs incubated with RANKL showed a dose-dependent increase in calcification, which was abolished by coincubation with OPG. To test whether the effect was mediated by signaling to its receptor, knockdown of RANK was accomplished by short hairpin (sh)RNA. Indeed, cells lacking RANK showed no increases in vascular calcification when incubated with RANKL. To further elucidate the mechanism by which RANK activation increases calcification, we blocked both nuclear factor (NF)-kappaB activation pathways. Only IKKalpha inactivation inhibited calcification, pointing to an involvement of the alternative NF-kappaB activation pathway. Furthermore, RANKL addition increased bone morphogenetic protein (BMP)4 expression in VSMCs, and that increase disappeared in cells lacking RANK or IKKalpha. The increase in calcification was also blunted by Noggin, pointing to a mediation of BMP4 in the calcification induced by RANKL. Furthermore, in an in vivo model, the increase in vascular calcium content was parallel to an increase in RANKL and BMP4 expression, which was localized in calcified areas. However, blood levels of the ratio RANKL/OPG did not change. We conclude that RANKL increases vascular smooth muscle cell calcification by binding to RANK and increasing BMP4 production through activation of the alternative NF-kappaB pathway.

Comment in Circ Res. 2009 May 8;104(9):1032-4.

 [PubMed - indexed for MEDLINE]


Sample Type:  primary osteoblast cells

Species:  mouse


References: Wang, Y et al (2011). Small interfering RNA knocks down the molecular target of alendronate, farnesyl pyrophosphate synthase, in osteoclast and osteoblast cultures. Mol Pharm. 8(4):1016-24.

Pubmed ID: 21186792

Pubmed link:

Abstract: Farnesyl pyrophosphate synthase (FPPS), an enzyme in the mevalonate pathway, is the inhibition target of alendronate, a potent FDA-approved nitrogen-containing bisphosphonate (N-BP) drug, at the molecular level. Alendronate not only inhibits osteoclasts but also has been reported to positively affect osteoblasts. This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures. Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate. Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability. FPPS siRNA is more potent than 10 μM alendronate, but less potent than 50 μM alendronate on reducing osteoclast viability. Despite FPPS knockdown, no significant changes were observed in osteoblast proliferation. FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition. However, compared with 50 μM alendronate dosing, FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation. Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization. Overall, results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways.

 [PubMed - in process] PMCID: PMC3084905[Available on 2012/8/1]


Sample Type:  culture

Species:  cells


References: Lian, Q et al (2006). Establishing clonal cell lines with endothelial-like potential from CD9(hi), SSEA-1(-) cells in embryonic stem cell-derived embryoid bodies. PLoS One 1:e6.

Pubmed ID: 17183690

Pubmed link:


BACKGROUND: Differentiation of embryonic stem cells (ESCs) into specific cell types with minimal risk of teratoma formation could be efficiently directed by first reducing the differentiation potential of ESCs through the generation of clonal, self-renewing lineage-restricted stem cell lines. Efforts to isolate these stem cells are, however, mired in an impasse where the lack of purified lineage-restricted stem cells has hindered the identification of defining markers for these rare stem cells and, in turn, their isolation.

METHODOLOGY/PRINCIPAL FINDINGS: We describe here a method for the isolation of clonal lineage-restricted cell lines with endothelial potential from ESCs through a combination of empirical and rational evidence-based methods. Using an empirical protocol that we have previously developed to generate embryo-derived RoSH lines with endothelial potential, we first generated E-RoSH lines from mouse ESC-derived embryoid bodies (EBs). Despite originating from different mouse strains, RoSH and E- RoSH lines have similar gene expression profiles (r(2) = 0.93) while that between E-RoSH and ESCs was 0.83. In silico gene expression analysis predicted that like RoSH cells, E-RoSH cells have an increased propensity to differentiate into vasculature. Unlike their parental ESCs, E-RoSH cells did not form teratomas and differentiate efficiently into endothelial-like cells in vivo and in vitro. Gene expression and FACS analysis revealed that RoSH and E-RoSH cells are CD9(hi), SSEA-1(-) while ESCs are CD9(lo), SSEA-1(+). Isolation of CD9(hi), SSEA-1(-) cells that constituted 1%-10% of EB-derived cultures generated an E-RoSH-like culture with an identical E-RoSH-like gene expression profile (r(2) = 0.95) and a propensity to differentiate into endothelial-like cells.

CONCLUSIONS: By combining empirical and rational evidence-based methods, we identified definitive selectable surface antigens for the isolation and propagation of lineage-restricted stem cells with endothelial-like potential from mouse ESCs.

 [PubMed - indexed for MEDLINE] PMCID: PMC1762397


Sample Type:  mesenchymal stem cells

Species:  equine


References: Stewart, AA et al (2008). Effect of dexamethasone supplementation on chondrogenesis of equine mesenchymal stem cells. Am J Vet Res. 69(8):1013-21.

Pubmed ID: 18672964

Pubmed link:


OBJECTIVE: To determine whether expansion of equine mesenchymal stem cells (MSCs) by use of fibroblast growth factor-2 (FGF-2) prior to supplementation with dexamethasone during the chondrogenic pellet culture phase would increase chondrocytic matrix markers without stimulating a hypertrophic chondrocytic phenotype.

SAMPLE POPULATION: MSCs obtained from 5 young horses.

PROCEDURES: First-passage equine monolayer MSCs were supplemented with medium containing FGF-2 (0 or 100 ng/mL). Confluent MSCs were transferred to pellet cultures and maintained in chondrogenic medium containing 0 or 10(7)M dexamethasone. Pellets were collected after 1, 7, and 14 days and analyzed for collagen type II protein content; total glycosaminoglycan content; total DNA content; alkaline phosphatase (ALP) activity; and mRNA of aggrecan, collagen type II, ALP, and elongation factor-1alpha.

RESULTS: Treatment with FGF-2, dexamethasone, or both increased pellet collagen type II content, total glycosaminoglycan content, and mRNA expression of aggrecan. The DNA content of the MSC control pellets decreased over time. Treatment with FGF-2, dexamethasone, or both prevented the loss in pellet DNA content over time. Pellet ALP activity and mRNA were increased in MSCs treated with dexamethasone and FGF-2-dexamethasone. After pellet protein data were standardized on the basis of DNA content, only ALP activity of MSCs treated with FGF-2-dexamethasone remained significantly increased.

CONCLUSIONS AND CLINICAL RELEVANCE: Dexamethasone and FGF-2 enhanced chondrogenic differentiation of MSCs, primarily through an increase in MSC numbers. Treatment with dexamethasone stimulated ALP activity and ALP mRNA, consistent with the progression of cartilage toward bone. This may be important for MSC-based repair of articular cartilage.

 [PubMed - indexed for MEDLINE]


Sample Type:  blood

Species:  human


References: Ponnapakkam, T et al (2010). A treatment trial of vitamin D supplementation in breast-fed infants: universal supplementation is not necessary for rickets prevention in Southern Louisiana. Clin Pediatr (Phila). 49(11):1053-60.

Pubmed ID: 20724336

Pubmed link:

Abstract: This study was conducted to determine if vitamin D supplementation is required to prevent rickets in breast-fed infants. Breast-feeding rates are increasing, and there are concerns about whether the vitamin D content of breast milk is sufficient. There are a few treatment trials of vitamin D supplementation in breast-fed infants; these were conducted in northern climates. The authors therefore performed a prospective clinical trial comparing vitamin D supplementation with placebo as control in southern Louisiana. Blood samples and questionnaires were collected at birth, 2, 4, and 6 months of age. There were no cases of rickets observed, and no differences in alkaline phosphatase levels between groups. Thus, there was no evidence that vitamin D supplementation reduced rickets risk in the authors' study population. This suggests that the current recommendations for universal vitamin D supplementation of breast-fed infants throughout the United States may need to be revised.

 [PubMed - indexed for MEDLINE]


Sample Type:  osteoblasts

Species:  human


References: Kauther, MD et al (2010). Alpha-calcitonin gene-related peptide can reverse the catabolic influence of UHMWPE particles on RANKL expression in primary human osteoblasts. Int J Biol Sci.6(6):525-36.

Pubmed ID: 20877694

Pubmed link:


BACKGROUND AND PURPOSE: A linkage between the neurotransmitter alpha-calcitonin gene-related peptide (alpha-CGRP) and particle-induced osteolysis has been shown previously. The suggested osteoprotective influence of alpha-CGRP on the catabolic effects of ultra-high molecular weight polyethylene (UHMWPE) particles is analyzed in this study in primary human osteoblasts.

METHODS: Primary human osteoblasts were stimulated by UHMWPE particles (cell/particle ratios 1:100 and 1:500) and different doses of alpha-CGRP (10(-7 )M, 10(-9 )M, 10(-11 )M). Receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG) mRNA expression and protein levels were measured by RT-PCR and Western blot.

RESULTS: Particle stimulation leads to a significant dose-dependent increase of RANKL mRNA in both cell-particle ratios and a significant down-regulation of OPG mRNA in cell-particle concentrations of 1:500. A significant depression of alkaline phosphatase was found due to particle stimulation. Alpha-CGRP in all tested concentrations showed a significant depressive effect on the expression of RANKL mRNA in primary human osteoblasts under particle stimulation. Comparable reactions of RANKL protein levels due to particles and alpha-CGRP were found by Western blot analysis. In cell-particle ratios of 1:100 after 24 hours the osteoprotective influence of alpha-CGRP reversed the catabolic effects of particles on the RANKL expression.

INTERPRETATION: The in-vivo use of alpha-CGRP, which leads to down-regulated RANKL in-vitro, might inhibit the catabolic effect of particles in conditions of particle induced osteolysis.

 [PubMed - indexed for MEDLINE] PMCID: PMC2945923


Sample Type:  osteoblast cells

Species:  mouse

References: Hutchins, HL et al (2011). Eicosapentaenoic acid decreases expression of anandamide synthesis enzyme and cannabinoid receptor 2 in osteoblast-like cells. J Nutr Biochem. 22(2):195-200.

Pubmed ID: 20951563

Pubmed link:

Abstract: Anandamide (AEA) is an endogenous agonist for the cannabinoid receptor 2 (CB2) which is expressed in osteoblasts. Arachidonic acid (AA) is the precursor for AEA and dietary n-3 polyunsaturated fatty acids (PUFA) are known to reduce the concentrations of AA in tissues and cells. Therefore, we hypothesized that n-3 PUFA, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), which reduce AA in cells, could lower AEA in osteoblasts by altering enzyme expression of the endocannabinoid (EC) system. MC3T3-E1 osteoblast-like cells were grown for 6, 10, 15, 20, 25 or 30 days in osteogenic medium. Osteoblasts were treated with 10 μM of AA, EPA, DHA, oleic acid (OA) or EPA+DHA (5 μM each) for 72 h prior to their collection for measurement of mRNA and alkaline phosphatase (ALP) activity. Compared to vehicle control, osteoblasts treated with AA had higher levels of AA and n-6 PUFA while those treated with EPA and DHA had lower n-6 but higher n-3 PUFA. Independent of the fatty acid treatments, osteoblasts matured normally as evidenced by ALP activity. N-acyl phosphatidylethanolamine-selective phospholipase D (NAPE-PLD), fatty acid amide hydrolase (FAAH) and CB2 mRNA expression were higher at 20 days compared to 10 days. NAPE-PLD and CB2 mRNA was lower in osteoblasts treated with EPA compared to all other groups. Thus, mRNA expression for NAPE-PLD, FAAH, and CB2 increased during osteoblast maturation and EPA reduced mRNA for NAPE-PLD and CB2 receptor. In conclusion, EPA lowered mRNA levels for proteins of the EC system and mRNA for AEA synthesis/degradation is reported in osteoblasts.

Copyright © 2011 Elsevier Inc. All rights reserved.

 [PubMed - indexed for MEDLINE]


Sample Type:  small intestine

Species:  chicken


References: Fasina, YO, Thanissery, RR (2011). Comparative efficacy of a yeast product and bacitracin methylene disalicylate in enhancing early growth and intestinal maturation in broiler chicks from breeder hens of different ages. Poult Sci. 90(5):1067-73.

Pubmed ID: 21489956

Pubmed link:

Abstract: The intestine of the newly hatched chick is immature at hatch. Yeast contains nucleotides and β-glucans that enhance intestinal development and chick growth. Accordingly, a 14-d experiment was conducted to evaluate the efficacy of a novel yeast product and bacitracin methylene disalicylate in enhancing early growth and intestinal maturation in chicks obtained from young (26-27 wk old) and old (58 to 59 wk old) breeder hens. Chicks (384) were randomly assigned to 8 dietary treatments. Treatment 1 (YH) consisted of chicks, from young hens, fed corn-soybean meal (SBM) diet alone. Treatment 2 (YHB) consisted of chicks, from young hens, fed corn-SBM basal into which BMD was added at 0.055 g/kg. Treatment 3 (YHE) consisted of chicks, from young hens, fed corn-SBM basal into which yeast extract (YE) was added at 0.075% level. Treatment 4 (YHED) consisted of chicks, from young hens, fed corn-SBM basal into which YE was added at 0.15% level. Treatments 5 (OH), 6 (OHB), 7 (OHE), and 8 (OHED) consisted of chicks from old hens fed diets similar to those given to YH in treatments 1, 2, 3, and 4, respectively. Growth performance (body weight gain and feed conversion ratio) was evaluated on d 7 and 14. Intestinal tissue samples were also analyzed for alkaline phosphatase (ALP) activity as an indicator of intestinal maturation on d 4 and 13 of experiment. Results showed that by d 14 of experiment, only BMD treatments (YHB and OHB) improved body weight gain (P < 0.05). However, the body weight gains of chicks in the yeast-supplemented treatments (YHE, YHED, OHE, and OHED) were statistically similar (P > 0.05) to those of the BMD treatments. Ileal ALP activity was consistently enhanced by BMD and yeast product supplemented at 0.075% of the diet. It was concluded that antibiotic BMD and our novel yeast product supplemented at 0.075% of the diet improved early chick growth and maturation of the ileal segment of the small intestine.

 [PubMed - indexed for MEDLINE]


Sample Type:  bone marrow stromal cells

Species:  rat


References: Henderson, JA et al (2008). Concurrent differentiation of marrow stromal cells to osteogenic and vasculogenic lineages. Macromol Biosci. 8(6):499-507.

Pubmed ID: 17941111

Pubmed link:

Abstract: When rat bone marrow stromal (BMS) cells were seeded on aligned type I collagen scaffolds and cultured in osteogenic media, they underwent simultaneous maturation and differentiation into osteogenic and vascular cell lineages. In addition, these cells produced mineralized matricellular deposits. BMS cells were seeded in Petri dish or the collagen scaffold, cultured in osteogenic media for 3, 6, and 9 d and subsequently processed for immunohistochemical and cytochemical analysis. Immunolocalization of lineage-specific proteins were visualized using confocal microscopy and mRNA transcript analysis was performed by real-time quantitative polymerase chain reaction (RT-qPCR). The alkaline phosphatase activity and calcium content significantly increased over the observed period of time in an osteogenic medium. Sheets of abundant Pecam (CD31), Flk-1 (VEGFR-2), tomato lectin (TL/LEL), and alpha-smooth muscle actin (alpha-SMA) positive cells were observed in the collagen scaffolds. Nascent capillary-like vessels were also seen amidst the osteoblasts in osteogenic culture, augmenting the maturation and differentiation of BMS cells into osteoblasts. In our in vitro study, concurrent differentiation of BMS cells, a heterogeneous cell population with multilineage differentiation potential, to osteogenic and vascular lineages demonstrated that the substrates (three-dimensional (3-D), collagen type I, aligned fibrils) had a profound effect on guiding the differentiation pathway of BMS cells.

 [PubMed - indexed for MEDLINE]


Sample Type:  serum

Species:  rat


References: Milne, TJ et al (2009). Induction of osteopenia during experimental tooth movement in the rat: alveolar bone remodelling and the mechanostat theory. Eur J Orthod. 31(3):221-31.

Pubmed ID: 19458288

Pubmed link:

Abstract: Increases in bone strains above a certain threshold have a positive effect on bone mass, whereas reductions in strain magnitude lead to bone loss and osteopenia; the term 'mechanostat' has been introduced to describe this tissue-level negative feedback mechanism. The mechanobiology of bone and particularly alveolar bone is poorly understood, and whether the mechanostat theory has any relevance to explaining the osseous changes that occur during orthodontic tooth movement remains unclear. To investigate the relationship further, an expansile force of 0.2 N was applied to the maxillary molars of 36, 6-week-old Wistar rats by helical coil springs. The animals were sacrificed at 1, 2, 4, and 8 days and the tissue response analyzed by histological, biochemical, and finite element (FE) methods. Differences between groups were determined by Student's t-test (two-tailed). The appliance produced an increase in the intermolar width averaging 0.5 mm after 8 days. Tetracycline uptake in the control rats suggested a rapid turnover of bone in both the interradicular domain and the bone-periodontal ligament interface. In the experimental group, however, incorporation of tetracycline into the interradicular domain was reduced and conventional histology revealed evidence of bone loss and osteopenia, in both the experimental and a group of sham-treated positive controls (with inactive, annealed springs). Serum alkaline phosphatase declined significantly in both experimental and sham-treated groups over the 8-day time course, indicating decreased bone formation. Serum acid phosphatase also declined, suggesting a concomitant decrease in bone resorption. Three-dimensional FE analysis of the stresses generated in the bone following occlusal (2 N) and orthodontic loading showed that the orthodontic force created a constant loading condition shielding some areas of bone from mechanical stress. Areas of low mechanical stimulation were coincident with sites of bone loss observed histologically, while bone mass was preserved in areas with higher levels of loading. These findings suggest that (1) the osteopenia resulted from stress shielding of the interradicular bone by the appliance, and a consequent reduction in occlusal loading below the critical threshold required for maintaining normal osseous architecture and (2) the mechanostat model can be employed to explain, at least in part, the response of the bone to orthodontic loading.

 [PubMed - indexed for MEDLINE]


Sample Type:  human cell line

Species:  cells


References: Oldinski, RA et al (2010). Synthesis and characterization of a Hyaluronan-polyethylene copolymer for biomedical applications. J Biomed Mater Res B Appl Biomater. 94(2):441-6.

Pubmed ID: 20583303

Pubmed link:

Abstract: Hyaluronan (HA)-based biomaterials are of interest for bone and cartilage tissue engineering because HA plays an important role in orthopedic tissue development, function, and repair. The goal of this project was to develop a biomaterial that incorporated the constituents of both a hydrogel and a hydrophobic polymer for biomedical applications. A series of amphiphilic graft copolymers consisting of HA, a glycosaminoglycan, and high-density polyethylene (HDPE), that is, HA-co-HDPE, were fabricated. The chemical characteristics, physical and viscoelastic properties, and cytocompatibility of novel HA-co-HDPE materials were characterized via Fourier Transform infrared (FTIR) spectroscopy, solid state nuclear magnetic resonance (ssNMR) spectroscopy, differential scanning calorimetry (DSC), dynamic shear testing, and an in vitro human osteoblast cell study. The esterification reaction between HA and functionalized HDPE resulted in semicrystalline, insoluble powder. The dynamic shear properties of HA-co-HDPE concentrated solutions were more like natural proteoglycans than the HA control. HA-co-HDPE was successfully compression molded into disks that swelled upon hydration. Osteoblasts were viable and expressed the osteoblast phenotype after 7 days of culture on HA-co-HDPE materials. These HA-co-HDPE materials may have several biomaterial applications in saline suspension or molded form, including orthopedic tissue repair.

 [PubMed - indexed for MEDLINE]


Sample Type:  culture

Species:  cells


References: Gaharwar, AK et al (2011). Assessment of using laponite cross-linked poly(ethylene oxide) for controlled cell adhesion and mineralization. Acta Biomater. 7(2):568-77.

Pubmed ID: 20854941

Pubmed link:

Abstract: The in vitro cytocompatibility of silicate (Laponite clay) cross-linked poly(ethylene oxide) (PEO) nanocomposite films using MC3T3-E1 mouse preosteoblast cells was investigated while cell adhesion, spreading, proliferation and mineralization were assessed as a function of film composition. By combining the advantageous characteristics of PEO polymer (hydrophilic, prevents protein and cell adhesion) with those of a synthetic and layered silicate (charged, degradable and potentially bioactive) some of the physical and chemical properties of the resulting polymer nanocomposites could be controlled. Hydration, dissolution and mechanical properties were examined and related to cell adhesion. Overall, this feasibility study demonstrates the ability of using model Laponite cross-linked PEO nanocomposites to create bioactive scaffolds.

Copyright © 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

 [PubMed - indexed for MEDLINE]


Sample Type:  leukocyte

Species:  fish

References: Palić, D et al (2011). Use of rapid cytochemical staining to characterize fish blood granulocytes in species of special concern and determine potential for function testing. Fish Shellfish Immunol. 30(2):646-52.

Pubmed ID: 21199672

Pubmed link:

Abstract: Studies of innate immunity in fish species of special concern are essential for better understanding of their health status during hatchery rearing conditions. The cytochemical and morphological characterizations of blood granulocytes have been used to provide information about phylogenetic differences and determine the potential use of neutrophil function assays. Rapid, simple, cytochemical staining kits used routinely for staining mammalian granulocytes have been used to characterize granulocytes from blood of four fish species: Arctic grayling, cutthroat trout, June sucker, and shovelnose sturgeon. Blood smears were stained with Peroxidase 391 (myeloperoxidase, MPO), alkaline phosphatase (AP), Periodic Acid Schiff (PAS) and Diff-quick stain; examined using bright field and differential interference contrast microscopy. Granulocytes on blood smears were evaluated based on the cell morphology, and presence or absence of the specific chromogen. Presence of lymphocytes, monocytes, platelets/thrombocytes and granulocytes was determined in all fish species. Arctic grayling, June sucker, and cutthroat trout had MPO positive granulocytes, while shovelnose sturgeon heterophils had positive reaction for leukocyte AP, but not MPO. Presence of MPO indicated potential to measure oxidative burst and degranulation of neutrophil primary granules in Arctic grayling, cutthroat trout and June sucker. Absence of MPO in shovelnose sturgeon suggested use of different enzyme marker (AP) in degranulation assay for this species. Standardization of cytochemical techniques allowed for rapid screening of leukocyte types, reducing the number of fish, time and effort to select adequate neutrophil function assays to be used in studies of health status in species of special concern.

Copyright © 2010 Elsevier Ltd. All rights reserved.

 [PubMed - indexed for MEDLINE]


Sample Type:  serum

Species:  human


References: Kučukalić-Selimović, E et al (2011). Evaluation of bone remodelling parameters after one year treatment with alendronate in postmenopausal women with osteoporosis. Bosn J Basic Med Sci. 11(1):41-5

Pubmed ID: 21342141

Pubmed link:

Abstract: Antiresorptive agents are widely used to treat osteoporosis. Both reduction in bone turnover and increase in BMD may be necessary to decrease the fracture risk. The aim of the study was to evaluate the effects of aledronate on bone turnover markers and bone mineral density in postmenopausal women with osteoporosis. The study involved a group of 56 postmenopausal women with osteoporosis treated with alendronate (70 mg) weekly at the Institute of Nuclear Medicine Clinical Center University of Sarajevo during a 12-months period. Bone mineral density (BMD) at lumbar spine and proximal femur and bone turnover markers (serum β-CrossLaps, urinary N-telopeptides of type I collagen (NTx), total serum alkaline phosphatase (AP) and serum osteocalcin) were measured at baseline and after 12 months of the treatment with aledronate. BMD values significantly increased both at lumbar spine by 13.46% and proximal femur by 21.96% during the study period (-3.12±0.24 vs. -2.7±0.19 and -2.55±0.2 vs. -1.99±0.19 respectively; p<0.001). Bone turnover markers significantly decreased during the study period; C-terminal telopeptides of type I collagen fragment (β-CrossLaps) 49.0% (0.51±0.05 vs.0.26±0.028 ng/mL), NTX 33.4% (48.3±4.9 vs.32.15±3.25 nMBCE/mM Cr), AP 24.3% (81.1±5.2 to 61.43±5.2 IU/L) and serum osteocalcin by 29.7% (34.3±2.65 to 24.1±1.36 ng/mL)(p<0.001). Alendronate treatment increased BMD and reduced the level of bone turnover markers. Therefore, the treatment with aledronate during 12 months period can be recommended in postmenopausal women with osteoporosis.

 [PubMed - indexed for MEDLINE]


Sample Type:  culture

Species:  Stem cells


References: Zhi, L et al (2011). Synergistic effect of recombinant human bone morphogenic protein-7 and osteogenic differentiation medium on human bone-marrow-derived mesenchymal stem cells in vitro. Int Orthop. 2011 Apr 13.

Pubmed ID: 21487672

Pubmed link:


PURPOSE: The purpose of this study was to investigate the effect of recombinant human bone morphogenetic protein-7 (rhBMP-7) with or without osteogenic differentiation medium (ODM) on osteogenic differentiation of primary human bone-marrow-derived mesenchymal stem cells (hBMSCs) in vitro.

METHOD: The hBMSCs were isolated from medullary reaming tissue. At 80% confluence, hBMSCs were treated with different concentrations of rhBMP-7 with and without ODM. Alkaline phosphatase (ALP) activity, calcium deposition and messenger RNA (mRNA) expression of osteocalcin (OC) and osteopontin (OPN) were examined.

RESULTS: ALP activity and calcium deposits in hBMSC culture were significantly increased by rhBMP-7 at 0.1 μg/ml (0.23 ± 0.07 IU and 28.9 ± 4.2 mg/dl) and 1.0 μg/ml (0.32 ± 0.03 IU and 38.7 ± 3.0 mg/dl), respectively, in the presence of ODM, showing a clearly dose-dependent osteoblastic differentiation. However, the same dose of 0.1 μg/ml rhBMP-7 without ODM and ODM alone induced low level of ALP and calcium deposits, indicating a synergistic effect of rhBMP-7 and ODM on committed osteogenic differentiation. Quantitative real-time reverse-transcriptase polymerase chain reaction (RT-PCR) analysis showed up-regulated OC and OPN mRNA levels, corroborating the synergistic effect of rhBMP-7 and ODM.

CONCLUSION: Our study showed that rhBMP-7 with ODM created a synergistic effect on up-regulation of osteogenic genes as well as osteogenic differentiation of primary hBMSCs in vitro. In the presence of ODM, the lowest concentration of rhBMP-7 needed to induce significant osteogenic differentiation of hBMSCs was 0.1 μg/ml.

 [PubMed - as supplied by publisher]


Sample Type:  cell lysate

Species:  B16 cells


References: Katuru, R et al (2011). Mevalonate depletion mediates the suppressive impact of geranylgeraniol on murine B16 melanoma cells. Exp Biol Med (Maywood) 236(5):604-13.

Pubmed ID: 21540247

Pubmed link:

Abstract: The diterpene geranylgeraniol (all trans-3,7,11,15-tetramethyl-2,6,10,14-hexadecatetraen-1-ol) suppresses the growth of human liver, lung, ovary, pancreas, colon, stomach and blood tumors with undefined mechanisms. We evaluated the growth-suppressive activity of geranylgeraniol in murine B16 melanoma cells. Geranylgeraniol induced dose-dependent suppression of B16 cell growth (IC(50) = 55 ± 13 µmol/L) following a 48-h incubation in 96-well plates. Cell cycle arrest at the G1 phase, manifested by a geranylgeraniol-induced increase in the G1/S ratio and decreased expression of cyclin D1 and cyclin-dependent kinase 4, apoptosis detected by Guava Nexin™ assay and fluorescence microscopy following acridine orange and ethidium bromide dual staining, and cell differentiation shown by increased alkaline phosphatase activity, contributed to the growth suppression. Murine 3T3-L1 fibroblasts were 10-fold more resistant than B16 cells to geranylgeraniol-mediated growth suppression. Geranylgeraniol at near IC(50) concentration (60 µmol/L) suppressed the mRNA level of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase by 50%. The impact of geranylgeraniol on B16 cell growth, cell cycle arrest and apoptosis were attenuated by supplemental mevalonate, the product of HMG-CoA reductase that is essential for cell growth. Geranylgeraniol and d-δ-tocotrienol, a down-regulator of HMG-CoA reductase, additively suppressed the growth of B16 cells. These results support our hypothesis that mevalonate depletion mediates the tumor-specific growth-suppressive impact of geranylgeraniol. Geranylgeraniol may have potential in cancer chemoprevention and/or therapy.

 [PubMed - indexed for MEDLINE]


Sample Type:  lysate

Species:  cells


References: Ahmed I.M. Abu Obid Alla. Ph.D. Thesis. Effects of Strontium on Human Osteoblasts Extracted from a Cleidocranial Dysplasia Patient – In Vitro Study. Unviersity of Regensburg 2010

Pubmed ID: n/a

Pubmed link: n/a

Abstract: n/a


Sample Type:  Alkaline phosphatase

Species:  Enzyme

References: Maskiewicz, R (2009). SUBLIMABLE SUSTAINED RELEASE DELIVERY SYSTEM AND METHOD OF MAKING SAME.  US Patent application number: 20090220602.

Pubmed ID: n/a

Pubmed link: n/a

Abstract: n/a

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