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Abstract：
Vascular calcification (VC) is a cardiovascular disease characterized by calcium salt deposition in vascular smooth muscle cells (VSMCs). Standard in vitro models used in VC investigations are based on VSMC monocultures under static conditions. Although these platforms are easy to use, the absence of interactions between different cell types and dynamic conditions makes these models insufficient to study key aspects of vascular pathophysiology. The present study aimed to develop a dynamic endothelial cell-VSMC co-culture that better mimics the in vivo vascular microenvironment. A double-flow bioreactor supported cellular interactions and reproduced the blood flow dynamic. VSMC calcification was stimulated with a DMEM high glucose calcification medium supplemented with 1.9 mM NaH2PO4/Na2HPO4 (1:1) for 7 days. Calcification, cell viability, inflammatory mediators, and molecular markers (SIRT-1, TGFβ1) related to VSMC differentiation were evaluated. Our dynamic model was able to reproduce VSMC calcification and inflammation and evidenced differences in the modulation of effectors involved in the VSMC calcified phenotype compared with standard monocultures, highlighting the importance of the microenvironment in controlling cell behavior. Hence, our platform represents an advanced system to investigate the pathophysiologic mechanisms underlying VC, providing information not available with the standard cell monoculture.
摘要：
血管钙化（VC）是一种以血管平滑肌细胞（VSMC）中钙盐沉积为特征的心血管疾病。VC研究中使用的标准体外模型基于静态条件下的VSMC单一培养。尽管这些平台易于使用，不同细胞类型和动态条件之间缺乏相互作用,使得这些模型不足以研究血管病理生理学的关键方面.本研究旨在开发一种动态的内皮细胞-VSMC共培养物，该培养物可以更好地模拟体内血管微环境。双流生物反应器支持细胞相互作用并再现血流动力学。用补充有1.9mMNaH2PO4/Na2HPO4(1:1)的DMEM高葡萄糖钙化培养基刺激VSMC钙化7天。钙化,细胞活力，炎症介质，并对与VSMC分化相关的分子标志物(SIRT-1、TGFβ1)进行评价。我们的动态模型能够重现VSMC钙化和炎症，并证明与标准单一培养相比，VSMC钙化表型中涉及的效应子的调节差异。强调微环境在控制细胞行为中的重要性。因此，我们的平台代表了一个先进的系统来研究VC的病理生理机制，提供标准细胞单一培养不可用的信息。
