The Experts below are selected from a list of 255 Experts worldwide ranked by ideXlab platform
Ying Yang - One of the best experts on this subject based on the ideXlab platform.
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Tracking calcification in tissue-engineered Bone using synchrotron micro-FTIR and SEM
Analytical and Bioanalytical Chemistry, 2015Co-Authors: Anthony J. Deegan, Gianfelice Cinque, Katia Wehbe, Sandeep Konduru, Ying YangAbstract:One novel tissue engineering approach to mimic in vivo Bone formation is the use of aggregate or micromass cultures. Various qualitative and quantitative techniques, such as histochemical staining, protein assay kits and RT-PCR, have been used previously on Cellular aggregate studies to investigate how these intricate arrangements lead to mature Bone tissue. However, these techniques struggle to reveal spatial and temporal distribution of proliferation and mineralization simultaneously. Synchrotron-based Fourier transform infrared microspectroscopy (micro-FTIR) offers a unique insight at the molecular scale by coupling high IR sensitivity to organic matter with the high spatial resolution allowed by diffraction limited SR microbeam. This study is set to investigate the effects of culture duration and aggregate size on the dynamics and spatial distribution of calcification in engineered Bone aggregates by a combination of micro-FTIR and scanning electron microscopy (SEM)/energy-dispersive X-ray spectroscopy (EDX). A murine Bone Cell Line has been used, and small/large Bone aggregates have been induced using different chemically treated culture substrates. Our findings suggest that Bone Cell aggregate culturing can greatly increase levels of mineralization over short culture periods. The size of the aggregates influences mineralisation rates with larger aggregates mineralizing at a faster rate than their smaller counterparts. The micro-FTIR mapping has demonstrated that mineralization in the larger aggregates initiated from the periphery and spread to the centre, whilst the smaller aggregates have more minerals in the centre at the early stage and deposited more in the periphery after further culturing, implying that aggregate size influences calcification distribution and development over time. SEM/EDX data correlates well with the micro-FTIR results for the total mineral content. Thus, synchrotron-based micro-FTIR can accurately track mineralization process/mechanism in the engineered Bone. Graphical Abstract FTIR mapping images of PO_4 regions showing the big intensity and distribution difference between small and large aggregates cultured for 72 h.
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Tracking calcification in tissue-engineered Bone using synchrotron micro-FTIR and SEM
Analytical and bioanalytical chemistry, 2014Co-Authors: Anthony J. Deegan, Gianfelice Cinque, Katia Wehbe, Sandeep Konduru, Ying YangAbstract:One novel tissue engineering approach to mimic in vivo Bone formation is the use of aggregate or micromass cultures. Various qualitative and quantitative techniques, such as histochemical staining, protein assay kits and RT-PCR, have been used previously on Cellular aggregate studies to investigate how these intricate arrangements lead to mature Bone tissue. However, these techniques struggle to reveal spatial and temporal distribution of proliferation and mineralization simultaneously. Synchrotron-based Fourier transform infrared microspectroscopy (micro-FTIR) offers a unique insight at the molecular scale by coupling high IR sensitivity to organic matter with the high spatial resolution allowed by diffraction limited SR microbeam. This study is set to investigate the effects of culture duration and aggregate size on the dynamics and spatial distribution of calcification in engineered Bone aggregates by a combination of micro-FTIR and scanning electron microscopy (SEM)/energy-dispersive X-ray spectroscopy (EDX). A murine Bone Cell Line has been used, and small/large Bone aggregates have been induced using different chemically treated culture substrates. Our findings suggest that Bone Cell aggregate culturing can greatly increase levels of mineralization over short culture periods. The size of the aggregates influences mineralisation rates with larger aggregates mineralizing at a faster rate than their smaller counterparts. The micro-FTIR mapping has demonstrated that mineralization in the larger aggregates initiated from the periphery and spread to the centre, whilst the smaller aggregates have more minerals in the centre at the early stage and deposited more in the periphery after further culturing, implying that aggregate size influences calcification distribution and development over time. SEM/EDX data correlates well with the micro-FTIR results for the total mineral content. Thus, synchrotron-based micro-FTIR can accurately track mineralization process/mechanism in the engineered Bone.
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Development of a ‘mechano-active’ scaffold for tissue engineering
Biomaterials, 2002Co-Authors: Ying Yang, L. M. Cooling, Julia L. Magnay, Alicia J. El HajAbstract:Abstract In this study, we investigate the potential for manipulating Bone Cell mechanotransducers in tissue engineering. Membrane ion channels such as voltage operated calcium channels (VOCC) have been shown to be a critical component of the Bone Cell transduction pathway with agonists and inhibitors of this pathway having profound effects on the load signal. By encapsulating a calcium channel agonist with slow release within a poly( l -lactide) (PLLA) scaffold, we can generate a ‘mechano-active’ scaffold for use in skeletal tissue engineering. PLLA scaffolds with and without a calcium channel agonist, BAY K8644, were seeded with primary human Bone Cells or the human MG63 Bone Cell Line and cultured for 1–3 weeks followed by mechanical stimulation with a four-point bending model. Our results show that addition of the agonist for slow release is sufficient to enhance the load-related responses in Bone Cells within the scaffolds. Specifically, collagen type I expression and the ratio of alkaLine phosphatase to protein are elevated in response to cyclical mechanical stimulation of approximately 1000 μstr which is then further enhanced in the ‘mechano-active’ scaffolds. As the agonists only act when the calcium channels are open by attenuating the calcium flux, the stimulation is specifically targeted to scaffolds subjected to load either in vitro or ultimately in vivo. Our results suggest that manipulating the VOCC and attenuating the opening of the calcium channels may be an effective technique to amplify matrix production via mechanical stimulation which may be applied to Bone tissue engineering and potentially engineering of other load-bearing connective tissues.
Anthony J. Deegan - One of the best experts on this subject based on the ideXlab platform.
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Tracking calcification in tissue-engineered Bone using synchrotron micro-FTIR and SEM
Analytical and Bioanalytical Chemistry, 2015Co-Authors: Anthony J. Deegan, Gianfelice Cinque, Katia Wehbe, Sandeep Konduru, Ying YangAbstract:One novel tissue engineering approach to mimic in vivo Bone formation is the use of aggregate or micromass cultures. Various qualitative and quantitative techniques, such as histochemical staining, protein assay kits and RT-PCR, have been used previously on Cellular aggregate studies to investigate how these intricate arrangements lead to mature Bone tissue. However, these techniques struggle to reveal spatial and temporal distribution of proliferation and mineralization simultaneously. Synchrotron-based Fourier transform infrared microspectroscopy (micro-FTIR) offers a unique insight at the molecular scale by coupling high IR sensitivity to organic matter with the high spatial resolution allowed by diffraction limited SR microbeam. This study is set to investigate the effects of culture duration and aggregate size on the dynamics and spatial distribution of calcification in engineered Bone aggregates by a combination of micro-FTIR and scanning electron microscopy (SEM)/energy-dispersive X-ray spectroscopy (EDX). A murine Bone Cell Line has been used, and small/large Bone aggregates have been induced using different chemically treated culture substrates. Our findings suggest that Bone Cell aggregate culturing can greatly increase levels of mineralization over short culture periods. The size of the aggregates influences mineralisation rates with larger aggregates mineralizing at a faster rate than their smaller counterparts. The micro-FTIR mapping has demonstrated that mineralization in the larger aggregates initiated from the periphery and spread to the centre, whilst the smaller aggregates have more minerals in the centre at the early stage and deposited more in the periphery after further culturing, implying that aggregate size influences calcification distribution and development over time. SEM/EDX data correlates well with the micro-FTIR results for the total mineral content. Thus, synchrotron-based micro-FTIR can accurately track mineralization process/mechanism in the engineered Bone. Graphical Abstract FTIR mapping images of PO_4 regions showing the big intensity and distribution difference between small and large aggregates cultured for 72 h.
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Tracking calcification in tissue-engineered Bone using synchrotron micro-FTIR and SEM
Analytical and bioanalytical chemistry, 2014Co-Authors: Anthony J. Deegan, Gianfelice Cinque, Katia Wehbe, Sandeep Konduru, Ying YangAbstract:One novel tissue engineering approach to mimic in vivo Bone formation is the use of aggregate or micromass cultures. Various qualitative and quantitative techniques, such as histochemical staining, protein assay kits and RT-PCR, have been used previously on Cellular aggregate studies to investigate how these intricate arrangements lead to mature Bone tissue. However, these techniques struggle to reveal spatial and temporal distribution of proliferation and mineralization simultaneously. Synchrotron-based Fourier transform infrared microspectroscopy (micro-FTIR) offers a unique insight at the molecular scale by coupling high IR sensitivity to organic matter with the high spatial resolution allowed by diffraction limited SR microbeam. This study is set to investigate the effects of culture duration and aggregate size on the dynamics and spatial distribution of calcification in engineered Bone aggregates by a combination of micro-FTIR and scanning electron microscopy (SEM)/energy-dispersive X-ray spectroscopy (EDX). A murine Bone Cell Line has been used, and small/large Bone aggregates have been induced using different chemically treated culture substrates. Our findings suggest that Bone Cell aggregate culturing can greatly increase levels of mineralization over short culture periods. The size of the aggregates influences mineralisation rates with larger aggregates mineralizing at a faster rate than their smaller counterparts. The micro-FTIR mapping has demonstrated that mineralization in the larger aggregates initiated from the periphery and spread to the centre, whilst the smaller aggregates have more minerals in the centre at the early stage and deposited more in the periphery after further culturing, implying that aggregate size influences calcification distribution and development over time. SEM/EDX data correlates well with the micro-FTIR results for the total mineral content. Thus, synchrotron-based micro-FTIR can accurately track mineralization process/mechanism in the engineered Bone.
Naftali Stern - One of the best experts on this subject based on the ideXlab platform.
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New vitamin D less-calcemic analog affect human Bone Cell Line and cultured vascular smooth muscle Cells similar to other less-calcemic analogs.
The Journal of steroid biochemistry and molecular biology, 2013Co-Authors: Dalia Somjen, Urszula Kulesza, Orly Sharon, Esther Knoll, Naftali SternAbstract:Abstract Primary cultures of human Bone and vascular Cells respond to vitamin D treatment by modulation of Cell proliferation measured by DNA synthesis (DNA) and energy metabolism measured by creatine kinase specific activity (CK) via binding to vitamin D receptors (VDR) which are expressed in these Cells. Vitamin D compounds also modulate the response to estradiol-17β (E 2 ) and the expression mRNAs of estrogen receptors (ERα and ERβ), VDR, 25-hydroxy vitamin D 3 1-α hydroxylase (1OHase) and lipoxygenases (12LO and 15LO). We now compared our newly synthesized analog: 1α,25-dihydroxy-9-methylene-19-norvitamin D 3 JK152 (JK), on Bone and vascular Cells compared to other analogs. Human Bone Cell Line SaOS 2 respond to JK by increased DNA and stimulated CK dose-dependently, similar to the less-calcemic analogs CB 1093 (CB) and EB 1089 (EB). JK also up-regulated the response to E 2 in terms of DNA and CK. JK inhibited DNA synthesis and increased CK in primary human vascular smooth muscle Cells (VSMC) dose-dependently similar to EB and CB. JK up regulated the response to E 2 in terms of CK with no effect on DNA. JK similar to CB and EB stimulated mRNA expression of VDR and ERα, 12LO and 15LO, with no effect on ERβ and 1OHase mRNA expression in SaOS 2 measured by real time PCR. Similar treatments of VSMC with JK, CB and EB stimulated 12LO and 15LO, VDR and ERα mRNA expression with no effect on ERβ and 1OHase mRNA expression. The results presented here demonstrate that the new vitamin D less-calcemic analog JK is similar to other analogs in its effects on human cultured Cells and therefore may be used in combined hormone replacement treatment (HRT) both in vitro and in vivo .
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Estrogens and Hyperglycemic Modulation of mRNAs Expressions Involved in Bone Metabolism: An Overshadowed Association?
Connective tissue research, 2013Co-Authors: Dalia Somjen, Esther Knoll, Sara Katzburg, Orli Sharon, Naftali SternAbstract:AbstractHuman Bone Cell Line (SaOS2) express different mRNAs involved in Bone biology and physiology such as estrogen receptor α (ERα), estrogen receptor β (ERβ), vitamin D receptor (VDR), 1α, 25 hydroxy vitamin D3 hydroxylase (1OHase) as well as 12 and 15 lipoxygenases (12LO and 15LO). These mRNAs are modulated by estrogenic compounds. Since the skeletal protective effects of estrogens are not discernible in diabetic women, we tested whether the expression of the parameters measured here and their modulations by estrogens, in SaOS2 Cells grown in growth medium containing high glucose (HG; 9.0 g/L; 44 mM) compared to normal glucose (NG; 4.5 g/L; 22 mM). High Glucose (HG) significantly increased DNA synthesis and creatine kinase (CK) specific activity in SaOS2 Cells. Stimulations of DNA but not of CK by E2, by 4, 4′, 4′′-[4-propyl-(1H)-pyrazol-1, 3, 5- triyl] tris-phenol (PPT, ERα specific agonist), or by 2, 3-bis (4-hydroxyphenyl)-propionitrile (DPN, ERβ specific agonist), were abolished by HG. HG itself ...
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the effects of estrogen receptors α and β specific agonists and antagonists on Cell proliferation and energy metabolism in human Bone Cell Line
Journal of Cellular Biochemistry, 2011Co-Authors: Dalia Somjen, Orly Sharon, Esther Knoll, Sara Katzburg, Meital Graficohen, Naftali SternAbstract:In cultured human osteoblasts estradiol-17β (E2) modulated DNA synthesis, the specific activity of creatine kinase BB (CK), 12 and 15 lipoxygenase (LO) mRNA expression and formation of 12- and 15-hydroxyeicosatetraenoic acid (HETE). We now investigate the response of human Bone Cell Line (SaOS2) to phytoestrogens and estrogen receptors (ER)-specific agonists and antagonists. Treatment of SaSO2 with E2, 2,3-bis (4-hydroxyphenyl)-propionitrile (DPN; ERβ-specific agonist), 4,4′,4″-[4-propyl-(1H)-pyrazol-1,3,5-triyl] tris-phenol (PPT; ERα-specific agonist), biochainin A (BA), daidzein (D), genistein (G) and raloxifene (Ral) showed increased DNA synthesis and CK. Ral inhibited completely all stimulations except DPN and to some extent D. The ERα-specific antagonist methyl-piperidino-pyrazole (MPP) and the ERβ-specific antagonist 4-[2-phenyl-5,7-bis (tri-fluoro-methyl) pyrazolo [1,5-a]pyrimidin-3-yl] phenol (PTHPP) inhibited DNA synthesis, CK and reactive oxygen species (ROS) formation induced by estrogens according to their receptors affinity. The LO inhibitor baicaleine inhibited only E2, DPN and G's effects. E2 and Ral unlike all other compounds had no effect on ERα mRNA expression, while ERβ mRNA expression was stimulated by all compounds. All compounds modulated the expression of 12LO and 15LO mRNA, except E2, PPT and Ral for 12LO, and 12- and 15-HETE productions and stimulated ROS formation which was inhibited by NADPH oxidase inhibitors diphenyleneiodonium chloride (DPI) and N-acetyl cysteine and the estrogen inhibitor ICI. DPI did not affect hormonal-induced DNA and CK. In conclusion, we provide evidence for the separation of mediation via ERα and ERβ pathways in the effects of estrogenic compounds on osteoblasts, but the role of LO/HETE/ROS is unclear. J. Cell. Biochem. 112: 625–632, 2011. © 2010 Wiley-Liss, Inc.
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The effects of estrogen receptors α‐ and β‐specific agonists and antagonists on Cell proliferation and energy metabolism in human Bone Cell Line
Journal of cellular biochemistry, 2011Co-Authors: Dalia Somjen, Orly Sharon, Esther Knoll, Sara Katzburg, Meital Grafi-cohen, Naftali SternAbstract:In cultured human osteoblasts estradiol-17β (E2) modulated DNA synthesis, the specific activity of creatine kinase BB (CK), 12 and 15 lipoxygenase (LO) mRNA expression and formation of 12- and 15-hydroxyeicosatetraenoic acid (HETE). We now investigate the response of human Bone Cell Line (SaOS2) to phytoestrogens and estrogen receptors (ER)-specific agonists and antagonists. Treatment of SaSO2 with E2, 2,3-bis (4-hydroxyphenyl)-propionitrile (DPN; ERβ-specific agonist), 4,4′,4″-[4-propyl-(1H)-pyrazol-1,3,5-triyl] tris-phenol (PPT; ERα-specific agonist), biochainin A (BA), daidzein (D), genistein (G) and raloxifene (Ral) showed increased DNA synthesis and CK. Ral inhibited completely all stimulations except DPN and to some extent D. The ERα-specific antagonist methyl-piperidino-pyrazole (MPP) and the ERβ-specific antagonist 4-[2-phenyl-5,7-bis (tri-fluoro-methyl) pyrazolo [1,5-a]pyrimidin-3-yl] phenol (PTHPP) inhibited DNA synthesis, CK and reactive oxygen species (ROS) formation induced by estrogens according to their receptors affinity. The LO inhibitor baicaleine inhibited only E2, DPN and G's effects. E2 and Ral unlike all other compounds had no effect on ERα mRNA expression, while ERβ mRNA expression was stimulated by all compounds. All compounds modulated the expression of 12LO and 15LO mRNA, except E2, PPT and Ral for 12LO, and 12- and 15-HETE productions and stimulated ROS formation which was inhibited by NADPH oxidase inhibitors diphenyleneiodonium chloride (DPI) and N-acetyl cysteine and the estrogen inhibitor ICI. DPI did not affect hormonal-induced DNA and CK. In conclusion, we provide evidence for the separation of mediation via ERα and ERβ pathways in the effects of estrogenic compounds on osteoblasts, but the role of LO/HETE/ROS is unclear. J. Cell. Biochem. 112: 625–632, 2011. © 2010 Wiley-Liss, Inc.
Dalia Somjen - One of the best experts on this subject based on the ideXlab platform.
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The Effects of Lipophilic and Hydrophilic Statins on Bone Tissue Mineralization in Saos2 Human Bone Cell Line– In vitro Comparative Study
Pharmaceutica Analytica Acta, 2015Co-Authors: Oleg Dolkart, Dalia Somjen, Moshe Salai, Pritsch T, Zachary T. Sharfman, Maman EAbstract:We analyzed the effects of of commonly used statins that belong to lipophilic and hydrophilic groups on human osteoblastic Cell activity in vitro, specifically proliferation and tissue mineralization. Proliferation and mineralisation assays were performed on the following drugs: rosuvastatin, atorvastatin, pravastatin, simvastatin and mevastatin. Cells were exposed to the drugs for 24 h and analyzed for DNA synthesis. Mineralization was analyzed after 21 days of drugs treatment. Rosuvastatin, atorvastatin, pravastatin and simvastatin stimulated DNA synthesis to different extents while mevastatin had no effect. The most effective drugs in terms of proliferation were rosuvastatin (8 μg/ml by 219+25%)> pravastatin (10 μg/ml by 185+16%)> atorvastatin (10 μg/ml by 171+6%)> simvastatin (30 μg/ml by 152+10%). Rosuvastatin inhibited mineralization by 57+3% and pravastatin stimulated it by 127+5%, while all other compounds totally destroyed Cells. Our results indicate that particular statins increase Bone proliferation and Bone mineralization in Cell Lines, suggesting a potential for these compounds to be beneficial in patients with established osteoporosis and and may enhance a fracture healing process. However, other statins may inhibit the mineralization process, and even induce Cell death.
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New vitamin D less-calcemic analog affect human Bone Cell Line and cultured vascular smooth muscle Cells similar to other less-calcemic analogs.
The Journal of steroid biochemistry and molecular biology, 2013Co-Authors: Dalia Somjen, Urszula Kulesza, Orly Sharon, Esther Knoll, Naftali SternAbstract:Abstract Primary cultures of human Bone and vascular Cells respond to vitamin D treatment by modulation of Cell proliferation measured by DNA synthesis (DNA) and energy metabolism measured by creatine kinase specific activity (CK) via binding to vitamin D receptors (VDR) which are expressed in these Cells. Vitamin D compounds also modulate the response to estradiol-17β (E 2 ) and the expression mRNAs of estrogen receptors (ERα and ERβ), VDR, 25-hydroxy vitamin D 3 1-α hydroxylase (1OHase) and lipoxygenases (12LO and 15LO). We now compared our newly synthesized analog: 1α,25-dihydroxy-9-methylene-19-norvitamin D 3 JK152 (JK), on Bone and vascular Cells compared to other analogs. Human Bone Cell Line SaOS 2 respond to JK by increased DNA and stimulated CK dose-dependently, similar to the less-calcemic analogs CB 1093 (CB) and EB 1089 (EB). JK also up-regulated the response to E 2 in terms of DNA and CK. JK inhibited DNA synthesis and increased CK in primary human vascular smooth muscle Cells (VSMC) dose-dependently similar to EB and CB. JK up regulated the response to E 2 in terms of CK with no effect on DNA. JK similar to CB and EB stimulated mRNA expression of VDR and ERα, 12LO and 15LO, with no effect on ERβ and 1OHase mRNA expression in SaOS 2 measured by real time PCR. Similar treatments of VSMC with JK, CB and EB stimulated 12LO and 15LO, VDR and ERα mRNA expression with no effect on ERβ and 1OHase mRNA expression. The results presented here demonstrate that the new vitamin D less-calcemic analog JK is similar to other analogs in its effects on human cultured Cells and therefore may be used in combined hormone replacement treatment (HRT) both in vitro and in vivo .
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Estrogens and Hyperglycemic Modulation of mRNAs Expressions Involved in Bone Metabolism: An Overshadowed Association?
Connective tissue research, 2013Co-Authors: Dalia Somjen, Esther Knoll, Sara Katzburg, Orli Sharon, Naftali SternAbstract:AbstractHuman Bone Cell Line (SaOS2) express different mRNAs involved in Bone biology and physiology such as estrogen receptor α (ERα), estrogen receptor β (ERβ), vitamin D receptor (VDR), 1α, 25 hydroxy vitamin D3 hydroxylase (1OHase) as well as 12 and 15 lipoxygenases (12LO and 15LO). These mRNAs are modulated by estrogenic compounds. Since the skeletal protective effects of estrogens are not discernible in diabetic women, we tested whether the expression of the parameters measured here and their modulations by estrogens, in SaOS2 Cells grown in growth medium containing high glucose (HG; 9.0 g/L; 44 mM) compared to normal glucose (NG; 4.5 g/L; 22 mM). High Glucose (HG) significantly increased DNA synthesis and creatine kinase (CK) specific activity in SaOS2 Cells. Stimulations of DNA but not of CK by E2, by 4, 4′, 4′′-[4-propyl-(1H)-pyrazol-1, 3, 5- triyl] tris-phenol (PPT, ERα specific agonist), or by 2, 3-bis (4-hydroxyphenyl)-propionitrile (DPN, ERβ specific agonist), were abolished by HG. HG itself ...
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Effects of commonly used medications on Bone tissue mineralisation in SaOS-2 human Bone Cell Line: An in vitro study
The bone & joint journal, 2013Co-Authors: Moshe Salai, Dalia Somjen, Sara Katzburg, Roy Gigi, O. Yakobson, Oleg DolkartAbstract:We analysed the effects of commonly used medications on human osteoblastic Cell activity in vitro, specifically proliferation and tissue mineralisation. A list of medications was retrieved from the records of patients aged > 65 years filed in the database of the largest health maintenance organisation in our country (> two million members). Proliferation and mineralisation assays were performed on the following drugs: rosuvastatin (statin), metformin (antidiabetic), metoprolol (β-blocker), citalopram (selective serotonin reuptake inhibitor [SSRI]), and omeprazole (proton pump inhibitor (PPI)). All tested drugs significantly stimulated DNA synthesis to varying degrees, with rosuvastatin 5 µg/ml being the most effective among them (mean 225% (sd 20)), compared with metformin 10 µg/ml (185% (sd 10)), metoprolol 0.25 µg/ml (190% (sd 20)), citalopram 0.05 µg/ml (150% (sd 10)) and omeprazole 0.001 µg/ml (145% (sd 5)). Metformin and metoprolol (to a small extent) and rosuvastatin (to a much higher extent) inhibited Cell mineralisation (85% (sd 5)). Our results indicate the need to evaluate the medications prescribed to patients in terms of their potential action on osteoblasts. Appropriate evaluation and prophylactic treatment (when necessary) might lower the incidence and costs associated with potential medication-induced osteoporosis. Cite this article: Bone Joint J 2013;95-B:1575–80.
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the effects of estrogen receptors α and β specific agonists and antagonists on Cell proliferation and energy metabolism in human Bone Cell Line
Journal of Cellular Biochemistry, 2011Co-Authors: Dalia Somjen, Orly Sharon, Esther Knoll, Sara Katzburg, Meital Graficohen, Naftali SternAbstract:In cultured human osteoblasts estradiol-17β (E2) modulated DNA synthesis, the specific activity of creatine kinase BB (CK), 12 and 15 lipoxygenase (LO) mRNA expression and formation of 12- and 15-hydroxyeicosatetraenoic acid (HETE). We now investigate the response of human Bone Cell Line (SaOS2) to phytoestrogens and estrogen receptors (ER)-specific agonists and antagonists. Treatment of SaSO2 with E2, 2,3-bis (4-hydroxyphenyl)-propionitrile (DPN; ERβ-specific agonist), 4,4′,4″-[4-propyl-(1H)-pyrazol-1,3,5-triyl] tris-phenol (PPT; ERα-specific agonist), biochainin A (BA), daidzein (D), genistein (G) and raloxifene (Ral) showed increased DNA synthesis and CK. Ral inhibited completely all stimulations except DPN and to some extent D. The ERα-specific antagonist methyl-piperidino-pyrazole (MPP) and the ERβ-specific antagonist 4-[2-phenyl-5,7-bis (tri-fluoro-methyl) pyrazolo [1,5-a]pyrimidin-3-yl] phenol (PTHPP) inhibited DNA synthesis, CK and reactive oxygen species (ROS) formation induced by estrogens according to their receptors affinity. The LO inhibitor baicaleine inhibited only E2, DPN and G's effects. E2 and Ral unlike all other compounds had no effect on ERα mRNA expression, while ERβ mRNA expression was stimulated by all compounds. All compounds modulated the expression of 12LO and 15LO mRNA, except E2, PPT and Ral for 12LO, and 12- and 15-HETE productions and stimulated ROS formation which was inhibited by NADPH oxidase inhibitors diphenyleneiodonium chloride (DPI) and N-acetyl cysteine and the estrogen inhibitor ICI. DPI did not affect hormonal-induced DNA and CK. In conclusion, we provide evidence for the separation of mediation via ERα and ERβ pathways in the effects of estrogenic compounds on osteoblasts, but the role of LO/HETE/ROS is unclear. J. Cell. Biochem. 112: 625–632, 2011. © 2010 Wiley-Liss, Inc.
Adnan A. Bekhit - One of the best experts on this subject based on the ideXlab platform.
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PHNQ from Evechinus chloroticus Sea Urchin Supplemented with Calcium Promotes Mineralization in Saos-2 Human Bone Cell Line.
Marine drugs, 2020Co-Authors: Yakun Hou, Alan Carne, Michelle Mcconnell, Sonya Mros, Elena A. Vasileva, Natalia P. Mishchenko, Keegan Burrow, Ke Wang, Adnan A. BekhitAbstract:Polyhydroxylated naphthoquinones (PHNQs), known as spinochromes that can be extracted from sea urchins, are bioactive compounds reported to have medicinal properties and antioxidant activity. The MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) Cell viability assay showed that pure echinochrome A exhibited a cytotoxic effect on Saos-2 Cells in a dose-dependent manner within the test concentration range (15.625–65.5 µg/mL). The PHNQ extract from New Zealand sea urchin Evechinus chloroticus did not induce any cytotoxicity within the same concentration range after 21 days of incubation. Adding calcium chloride (CaCl2) with echinochrome A increased the number of viable Cells, but when CaCl2 was added with the PHNQs, Cell viability decreased. The effect of PHNQs extracted on mineralized nodule formation in Saos-2 Cells was investigated using xylenol orange and von Kossa staining methods. Echinochrome A decreased the mineralized nodule formation significantly (p < 0.05), while nodule formation was not affected in the PHNQ treatment group. A significant (p < 0.05) increase in mineralization was observed in the presence of PHNQs (62.5 µg/mL) supplemented with 1.5 mM CaCl2. In conclusion, the results indicate that PHNQs have the potential to improve the formation of Bone mineral phase in vitro, and future research in an animal model is warranted.