期刊
BIOMEDICINES
卷 10, 期 9, 页码 -出版社
MDPI
DOI: 10.3390/biomedicines10092271
关键词
fluorescence staining; mineralization; quantification; vascular smooth muscle cell
资金
- Bundesministerium fur Bildung und Forschung [FK031L0231]
- Sonnenfeld Stiftung
This study aims to establish a rapid and efficient working protocol for specific detection of hydroxyapatite (HAP) in cells and tissue, using a fluorescence dye, OsteoSense (TM). This method combines high specificity and improved sensitivity, making it valuable for researching the pathogenesis of vascular calcification and testing therapeutic inhibitors.
Vessel calcification is characterized by the precipitation of hydroxyapatite (HAP) in the vasculature. Currently, no causal therapy exists to reduce or prevent vessel calcification. Studying the underlying pathways within vascular smooth muscle cells and testing pharmacological intervention is a major challenge in the vascular research field. This study aims to establish a rapid and efficient working protocol for specific HAP detection in cells and tissue using the synthetic bisphosphonate fluorescence dye OsteoSense (TM). This protocol facilitates especially early quantification of the fluorescence signal and permits co-staining with other markers of interest, enabling smaller experimental set-ups with lesser primary cells consumption and fast workflows. The fluorescence-based detection of vascular calcification with OsteoSense (TM) combines a high specificity with improved sensitivity. Therefore, this methodology can improve research of the pathogenesis of vascular calcification, especially for testing the therapeutic benefit of inhibitors in the case of in vitro and ex vivo settings.
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