Molecular Imaging and Quantification of Smooth Muscle Cell and Aortic Tissue Calcification In Vitro and Ex Vivo with a Fluorescent Hydroxyapatite-Specific Probe
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 maj...
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MDPI AG
2022-09-01
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| Series: | Biomedicines |
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| Online Access: | https://www.mdpi.com/2227-9059/10/9/2271 |
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| author | Anna Greco Jaqueline Herrmann Milen Babic Manasa Reddy Gummi Markus van der Giet Markus Tölle Mirjam Schuchardt |
| author_facet | Anna Greco Jaqueline Herrmann Milen Babic Manasa Reddy Gummi Markus van der Giet Markus Tölle Mirjam Schuchardt |
| author_sort | Anna Greco |
| collection | DOAJ |
| description | 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™. 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™ 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. |
| first_indexed | 2024-03-10T00:38:09Z |
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| language | English |
| last_indexed | 2024-03-10T00:38:09Z |
| publishDate | 2022-09-01 |
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| spelling | doaj.art-336a55ff90074082872cf99ff7e70b452023-11-23T15:12:00ZengMDPI AGBiomedicines2227-90592022-09-01109227110.3390/biomedicines10092271Molecular Imaging and Quantification of Smooth Muscle Cell and Aortic Tissue Calcification In Vitro and Ex Vivo with a Fluorescent Hydroxyapatite-Specific ProbeAnna Greco0Jaqueline Herrmann1Milen Babic2Manasa Reddy Gummi3Markus van der Giet4Markus Tölle5Mirjam Schuchardt6Department of Nephrology and Medical Intensive Care, Corporate Member of Freie Universität Berlin and Humboldt Universität zu Berlin, Charité—Universitätsmedizin Berlin, Hindenburgdamm 30, 12203 Berlin, GermanyDepartment of Nephrology and Medical Intensive Care, Corporate Member of Freie Universität Berlin and Humboldt Universität zu Berlin, Charité—Universitätsmedizin Berlin, Hindenburgdamm 30, 12203 Berlin, GermanyDepartment of Nephrology and Medical Intensive Care, Corporate Member of Freie Universität Berlin and Humboldt Universität zu Berlin, Charité—Universitätsmedizin Berlin, Hindenburgdamm 30, 12203 Berlin, GermanyDepartment of Nephrology and Medical Intensive Care, Corporate Member of Freie Universität Berlin and Humboldt Universität zu Berlin, Charité—Universitätsmedizin Berlin, Hindenburgdamm 30, 12203 Berlin, GermanyDepartment of Nephrology and Medical Intensive Care, Corporate Member of Freie Universität Berlin and Humboldt Universität zu Berlin, Charité—Universitätsmedizin Berlin, Hindenburgdamm 30, 12203 Berlin, GermanyDepartment of Nephrology and Medical Intensive Care, Corporate Member of Freie Universität Berlin and Humboldt Universität zu Berlin, Charité—Universitätsmedizin Berlin, Hindenburgdamm 30, 12203 Berlin, GermanyDepartment of Nephrology and Medical Intensive Care, Corporate Member of Freie Universität Berlin and Humboldt Universität zu Berlin, Charité—Universitätsmedizin Berlin, Hindenburgdamm 30, 12203 Berlin, GermanyVessel 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™. 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™ 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.https://www.mdpi.com/2227-9059/10/9/2271fluorescence stainingmineralizationquantificationvascular smooth muscle cell |
| spellingShingle | Anna Greco Jaqueline Herrmann Milen Babic Manasa Reddy Gummi Markus van der Giet Markus Tölle Mirjam Schuchardt Molecular Imaging and Quantification of Smooth Muscle Cell and Aortic Tissue Calcification In Vitro and Ex Vivo with a Fluorescent Hydroxyapatite-Specific Probe Biomedicines fluorescence staining mineralization quantification vascular smooth muscle cell |
| title | Molecular Imaging and Quantification of Smooth Muscle Cell and Aortic Tissue Calcification In Vitro and Ex Vivo with a Fluorescent Hydroxyapatite-Specific Probe |
| title_full | Molecular Imaging and Quantification of Smooth Muscle Cell and Aortic Tissue Calcification In Vitro and Ex Vivo with a Fluorescent Hydroxyapatite-Specific Probe |
| title_fullStr | Molecular Imaging and Quantification of Smooth Muscle Cell and Aortic Tissue Calcification In Vitro and Ex Vivo with a Fluorescent Hydroxyapatite-Specific Probe |
| title_full_unstemmed | Molecular Imaging and Quantification of Smooth Muscle Cell and Aortic Tissue Calcification In Vitro and Ex Vivo with a Fluorescent Hydroxyapatite-Specific Probe |
| title_short | Molecular Imaging and Quantification of Smooth Muscle Cell and Aortic Tissue Calcification In Vitro and Ex Vivo with a Fluorescent Hydroxyapatite-Specific Probe |
| title_sort | molecular imaging and quantification of smooth muscle cell and aortic tissue calcification in vitro and ex vivo with a fluorescent hydroxyapatite specific probe |
| topic | fluorescence staining mineralization quantification vascular smooth muscle cell |
| url | https://www.mdpi.com/2227-9059/10/9/2271 |
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