Strategies for inhibiting advanced glycation endproduct (age) induced vascular calcification in a smooth muscle cell culture model

Research output: Contribution to journalAbstractpeer-review

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Strategies for inhibiting advanced glycation endproduct (age) induced vascular calcification in a smooth muscle cell culture model. / Sidgwick, Gary P; Walling, Peter; Shabbir, Ambreen; Weston, Ria; Schiro, Andrew; Serracino-inglott, Ferdinand; Jones, Alan M; Kamalov, Meder; Brimble, Margaret A; Wilkinson, Fiona L; Yvonne Alexander, M.

In: Heart, Vol. 102, No. Suppl 6, 196, 01.06.2016, p. A132.

Research output: Contribution to journalAbstractpeer-review

Harvard

Sidgwick, GP, Walling, P, Shabbir, A, Weston, R, Schiro, A, Serracino-inglott, F, Jones, AM, Kamalov, M, Brimble, MA, Wilkinson, FL & Yvonne Alexander, M 2016, 'Strategies for inhibiting advanced glycation endproduct (age) induced vascular calcification in a smooth muscle cell culture model', Heart, vol. 102, no. Suppl 6, 196, pp. A132. https://doi.org/10.1136/heartjnl-2016-309890.196

APA

Sidgwick, G. P., Walling, P., Shabbir, A., Weston, R., Schiro, A., Serracino-inglott, F., Jones, A. M., Kamalov, M., Brimble, M. A., Wilkinson, F. L., & Yvonne Alexander, M. (2016). Strategies for inhibiting advanced glycation endproduct (age) induced vascular calcification in a smooth muscle cell culture model. Heart, 102(Suppl 6), A132. [196]. https://doi.org/10.1136/heartjnl-2016-309890.196

Vancouver

Author

Sidgwick, Gary P ; Walling, Peter ; Shabbir, Ambreen ; Weston, Ria ; Schiro, Andrew ; Serracino-inglott, Ferdinand ; Jones, Alan M ; Kamalov, Meder ; Brimble, Margaret A ; Wilkinson, Fiona L ; Yvonne Alexander, M. / Strategies for inhibiting advanced glycation endproduct (age) induced vascular calcification in a smooth muscle cell culture model. In: Heart. 2016 ; Vol. 102, No. Suppl 6. pp. A132.

Bibtex

@article{996cf7fe1bb645ceae270fd5fbd21e92,
title = "Strategies for inhibiting advanced glycation endproduct (age) induced vascular calcification in a smooth muscle cell culture model",
abstract = "Vascular calcification is implicated in a range of cardiovascular disease mechanisms, leading to an associated increase in morbidity and mortality. One such trigger are advanced glycation endproducts (AGEs), the tissue accumulation of which increases with age and is more prevalent in diabetic subjects due to oxidative stress and poor glycaemic control. The aim of this study was to investigate the osteogenic potential of AGEs and elucidate mechanisms of inhibiting these processes in a smooth muscle cell (SMC) culture model.Osteogenic differentiation of SMCs was induced using {\^I}²-glycerophosphate ({\^I}²-GP), carboxymethyllysine (CML), carboxyethyllysine (CEL) methylglyoxal (MGO) and glycated low density lipoprotein (gly-LDL). The cells were subsequently treated with aminoguanidine (AG), an inhibitor of AGE formation, and novel glycomimetic compounds in order to determine their anti-calcification potential in vitro using qPCR, ELISA, Alkaline phosphatase (ALP) activity and Alizarin red staining.Gly-LDL (10 µg/ml) and CML (2.5nM) increased the level of calcification observed compared to the {\^I}²-GP (5 mM) positive control after 21 days (p < 0.05), with gly-LDL induced calcification apparent after 14 days. Both AG (250 µM) and the novel glycomimetic compounds reduced the level of mineralisation observed at 21 days compared with osteogenic treatments (p < 0.05). CEL (2.5 nM) and MGO (0.1 mM) both induced calcification, however mineralization was not as extensive as with {\^I}²-GP. When compared to the structure of CML, the side-chain of CEL contains an extra methyl group, suggesting this group impacts RAGE receptor binding. It was also shown that {\^I}²-GP combined with increased glucose concentration induced more extensive calcification unlike low glucose levels and {\^I}²-GP alone. ALP activity, when stimulated with {\^I}²-GP, CML and gly-LDL was greater at day 4 than at day 7, with AG reducing ALP activity measurements at day 4. Gly-LDL increases gene expression of OCN at day 4 compared with {\^I}²-GP and CML, however this was reduced at day 7, corresponding with an increased expression of OPN and OPG. NOTCH-3 gene expression was also reduced at day 7. Gene expression of OPN, OPG and NOTCH-3 were reduced at both day 4 and day 7 compared with osteogenic treatments ({\^I}²-GP, CML and gly-LDL).In summary, we conclude that gly-LDL and CML are potent inducers of calcification compared with {\^I}²-GP, and that their osteogenic potential can be modulated by both AG and novel glycomimetic compounds.",
author = "Sidgwick, {Gary P} and Peter Walling and Ambreen Shabbir and Ria Weston and Andrew Schiro and Ferdinand Serracino-inglott and Jones, {Alan M} and Meder Kamalov and Brimble, {Margaret A} and Wilkinson, {Fiona L} and {Yvonne Alexander}, M",
year = "2016",
month = jun,
day = "1",
doi = "10.1136/heartjnl-2016-309890.196",
language = "English",
volume = "102",
pages = "A132",
journal = "Heart",
issn = "1355-6037",
publisher = "BMJ Publishing Group",
number = "Suppl 6",

}

RIS

TY - JOUR

T1 - Strategies for inhibiting advanced glycation endproduct (age) induced vascular calcification in a smooth muscle cell culture model

AU - Sidgwick, Gary P

AU - Walling, Peter

AU - Shabbir, Ambreen

AU - Weston, Ria

AU - Schiro, Andrew

AU - Serracino-inglott, Ferdinand

AU - Jones, Alan M

AU - Kamalov, Meder

AU - Brimble, Margaret A

AU - Wilkinson, Fiona L

AU - Yvonne Alexander, M

PY - 2016/6/1

Y1 - 2016/6/1

N2 - Vascular calcification is implicated in a range of cardiovascular disease mechanisms, leading to an associated increase in morbidity and mortality. One such trigger are advanced glycation endproducts (AGEs), the tissue accumulation of which increases with age and is more prevalent in diabetic subjects due to oxidative stress and poor glycaemic control. The aim of this study was to investigate the osteogenic potential of AGEs and elucidate mechanisms of inhibiting these processes in a smooth muscle cell (SMC) culture model.Osteogenic differentiation of SMCs was induced using β-glycerophosphate (β-GP), carboxymethyllysine (CML), carboxyethyllysine (CEL) methylglyoxal (MGO) and glycated low density lipoprotein (gly-LDL). The cells were subsequently treated with aminoguanidine (AG), an inhibitor of AGE formation, and novel glycomimetic compounds in order to determine their anti-calcification potential in vitro using qPCR, ELISA, Alkaline phosphatase (ALP) activity and Alizarin red staining.Gly-LDL (10 µg/ml) and CML (2.5nM) increased the level of calcification observed compared to the β-GP (5 mM) positive control after 21 days (p < 0.05), with gly-LDL induced calcification apparent after 14 days. Both AG (250 µM) and the novel glycomimetic compounds reduced the level of mineralisation observed at 21 days compared with osteogenic treatments (p < 0.05). CEL (2.5 nM) and MGO (0.1 mM) both induced calcification, however mineralization was not as extensive as with β-GP. When compared to the structure of CML, the side-chain of CEL contains an extra methyl group, suggesting this group impacts RAGE receptor binding. It was also shown that β-GP combined with increased glucose concentration induced more extensive calcification unlike low glucose levels and β-GP alone. ALP activity, when stimulated with β-GP, CML and gly-LDL was greater at day 4 than at day 7, with AG reducing ALP activity measurements at day 4. Gly-LDL increases gene expression of OCN at day 4 compared with β-GP and CML, however this was reduced at day 7, corresponding with an increased expression of OPN and OPG. NOTCH-3 gene expression was also reduced at day 7. Gene expression of OPN, OPG and NOTCH-3 were reduced at both day 4 and day 7 compared with osteogenic treatments (β-GP, CML and gly-LDL).In summary, we conclude that gly-LDL and CML are potent inducers of calcification compared with β-GP, and that their osteogenic potential can be modulated by both AG and novel glycomimetic compounds.

AB - Vascular calcification is implicated in a range of cardiovascular disease mechanisms, leading to an associated increase in morbidity and mortality. One such trigger are advanced glycation endproducts (AGEs), the tissue accumulation of which increases with age and is more prevalent in diabetic subjects due to oxidative stress and poor glycaemic control. The aim of this study was to investigate the osteogenic potential of AGEs and elucidate mechanisms of inhibiting these processes in a smooth muscle cell (SMC) culture model.Osteogenic differentiation of SMCs was induced using β-glycerophosphate (β-GP), carboxymethyllysine (CML), carboxyethyllysine (CEL) methylglyoxal (MGO) and glycated low density lipoprotein (gly-LDL). The cells were subsequently treated with aminoguanidine (AG), an inhibitor of AGE formation, and novel glycomimetic compounds in order to determine their anti-calcification potential in vitro using qPCR, ELISA, Alkaline phosphatase (ALP) activity and Alizarin red staining.Gly-LDL (10 µg/ml) and CML (2.5nM) increased the level of calcification observed compared to the β-GP (5 mM) positive control after 21 days (p < 0.05), with gly-LDL induced calcification apparent after 14 days. Both AG (250 µM) and the novel glycomimetic compounds reduced the level of mineralisation observed at 21 days compared with osteogenic treatments (p < 0.05). CEL (2.5 nM) and MGO (0.1 mM) both induced calcification, however mineralization was not as extensive as with β-GP. When compared to the structure of CML, the side-chain of CEL contains an extra methyl group, suggesting this group impacts RAGE receptor binding. It was also shown that β-GP combined with increased glucose concentration induced more extensive calcification unlike low glucose levels and β-GP alone. ALP activity, when stimulated with β-GP, CML and gly-LDL was greater at day 4 than at day 7, with AG reducing ALP activity measurements at day 4. Gly-LDL increases gene expression of OCN at day 4 compared with β-GP and CML, however this was reduced at day 7, corresponding with an increased expression of OPN and OPG. NOTCH-3 gene expression was also reduced at day 7. Gene expression of OPN, OPG and NOTCH-3 were reduced at both day 4 and day 7 compared with osteogenic treatments (β-GP, CML and gly-LDL).In summary, we conclude that gly-LDL and CML are potent inducers of calcification compared with β-GP, and that their osteogenic potential can be modulated by both AG and novel glycomimetic compounds.

U2 - 10.1136/heartjnl-2016-309890.196

DO - 10.1136/heartjnl-2016-309890.196

M3 - Abstract

VL - 102

SP - A132

JO - Heart

JF - Heart

SN - 1355-6037

IS - Suppl 6

M1 - 196

ER -