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Updated: Jun 26, 2025
Isolation and Characterization of Primary Rat Valve Interstitial Cells: A New Model to Study Aortic Valve Calcification
Published on: November 20, 2017
Hyperglycemia-simulating environment attenuated experimentally induced calcification in cultured human aortic valve
Arsenii Zabirnyk1,2, Daria Evensen1,2, John-Peder Escobar Kvitting3,4
1Department of Molecular Medicine, Division of Physiology, Institute of Basic Medical Science, University of Oslo, Oslo, Norway.
Abstract:
Objectives: The role of diabetes mellitus as a risk factor for the development of calcific aortic valve disease has not been fully clarified. Aortic valve interstitial cells (VICs) have been suggested to be crucial for calcification of the valve. Induced calcification in cultured VICs is a good in vitro model for aortic valve calcification. The purpose of this study was to investigate whether increased glucose levels increase experimentally induced calcification in cultured human VICs. Design: VICs were isolated from explanted calcified aortic valves after valve replacement. Osteogenic medium induced calcification of cultured VICs at different glucose levels (5, 15, and 25 mM). Calcium deposits were visualized using Alizarin Red staining and measured spectrophotometrically. Results: The higher the glucose concentration, the lower the level of calcification. High glucose (25 mM) reduced calcification by 52% compared with calcification at a physiological (5 mM) glucose concentration (correlation and regression analysis: r = -0.55, p = .025 with increased concentration of glucose). Conclusions: In vitro hyperglycemia-like conditions attenuated calcification in VICs. High glucose levels may trigger a series of events that secondarily stimulate calcification of VICs in vivo.
Insights
Diabetes mellitus and calcific aortic valve disease are linked. High glucose levels paradoxically reduced calcification in human valve cells in vitro, suggesting complex in vivo mechanisms.
Area of Science:
- Cardiovascular Biology
- Metabolic Disease Research
- Cellular Pathophysiology
Background:
- The role of diabetes mellitus in calcific aortic valve disease (CAVD) remains unclear.
- Aortic valve interstitial cells (VICs) are implicated in CAVD pathogenesis.
- In vitro models of VIC calcification are essential for studying CAVD.
Purpose of the Study:
- To investigate the effect of elevated glucose concentrations on experimentally induced calcification in cultured human VICs.
- To determine if hyperglycemia influences the calcification process in VICs.
Main Methods:
- Human VICs were isolated from explanted calcified aortic valves.
- VICs were cultured in osteogenic medium with varying glucose concentrations (5, 15, and 25 mM).
- Alizarin Red staining and spectrophotometry were used to quantify calcium deposits.
Main Results:
- Increased glucose concentration inversely correlated with VIC calcification.
- High glucose (25 mM) significantly reduced calcification by 52% compared to physiological levels (5 mM).
- Statistical analysis showed a significant negative correlation between glucose concentration and calcification (r = -0.55, p = 0.025).
Conclusions:
- Hyperglycemia-like conditions in vitro attenuated VIC calcification.
- These findings suggest that high glucose may indirectly promote VIC calcification in vivo through complex mechanisms.
