Greco Anna, Herrmann Jaqueline, Babic Milen, Gummi Manasa Reddy, van der Giet Markus, Tölle Markus, Schuchardt Mirjam
Department of Nephrology and Medical Intensive Care, Corporate Member of Freie Universität Berlin and Humboldt Universität zu Berlin, Charité-Universitätsmedizin Berlin, Hindenburgdamm 30, 12203 Berlin, Germany.
Biomedicines. 2022 Sep 14;10(9):2271. doi: 10.3390/biomedicines10092271.
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.
血管钙化的特征是血管中羟基磷灰石（HAP）的沉淀。目前，尚无减少或预防血管钙化的因果疗法。研究血管平滑肌细胞内的潜在途径并测试药物干预是血管研究领域的一项重大挑战。本研究旨在建立一种快速有效的工作方案，用于使用合成双膦酸盐荧光染料OsteoSense™在细胞和组织中进行特异性HAP检测。该方案特别有助于早期荧光信号定量，并允许与其他感兴趣的标记物进行共染色，从而实现更小的实验设置，减少原代细胞消耗并实现快速工作流程。使用OsteoSense™基于荧光的血管钙化检测结合了高特异性和更高的灵敏度。因此，该方法可以改善血管钙化发病机制的研究，特别是在体外和离体情况下测试抑制剂的治疗效果。
