Late-stage Calcific Aortic Valve Disease Within an Aortic Valve-on-a-chip Model
Late-stage Calcific Aortic Valve Disease Within an Aortic Valve-on-a-chip Model
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主动脉瓣芯片模型中的晚期钙化主动脉瓣疾病
DOI：
10.1080/24748706.2021.1900702
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Mahler, Gretchen
中科院分区：
文献类型：
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作者：
Mendoza, Melissa;Chen, Mei-Hsiu;Murray, Bruce;Huang, Peter;Mahler, Gretchen
目的：钙化性主动脉瓣病(CAVD)是一种从主动脉硬化到严重的主动脉狭窄的进展性疾病。目前的治疗方法包括全瓣膜置换术和针对其他心血管疾病量身定制的药物干预。在晚期CAVD中，糖胺多聚糖(GAG)，如硫酸软骨素(CS)，沉积在纤维层。在这里，我们利用3D微流体设备来研究切应力、CS和内皮细胞对主动脉瓣纤维膜模型中钙化的影响，以了解CAVD的进展。方法：利用晶片模具、软光刻和电晕放电制造芯片上的瓣膜装置。设备有一个流动通道和3D基质储存库，只有健康对照或含有1 mg/m L或20 mg/m L CS的胶原蛋白。猪主动脉瓣间质细胞(PAVEC)被包埋在基质中，内皮细胞(PAVEC)种植在基质上。使用蠕动泵在37℃和5%CO2下施加1dyne/cm2和20dyne/cm2的稳定剪应力。结果：PAVEC/PAVEC共培养14天后，随着剪切力和GAG的增加，细胞内钙化程度明显增加。在定量上，ARS随着剪应力的增加而增加，而与GAG无关。在CS存在的情况下，对结节的分析表明，结节的大小随着剪切应力的增加而减小，但它增加了总平均染色面积。与共培养相比，仅含PAVEC的芯片上瓣膜模型显示钙沉积减少。结论：在体外钙化中，存在切应力、PAVEC和增加GAGS浓度时，PAVEC增加。进一步的研究将确定结节的化学成分，并了解早期CAVD的病理生理学。鉴于CAVD没有针对性的治疗方法，创建一个与生理相关的主动脉瓣试验台将为新的治疗方法做出贡献。
Objective: Calcific aortic valve disease (CAVD) is a progressive disease ranging from aortic sclerosis to severe aortic stenosis. Current treatments include total valve replacement and drug interventions tailored to other cardiovascular diseases. In late-stage CAVD, glycosaminoglycans (GAGs), such as chondroitin sulfate (CS), deposit in the fibrosa layer. Here we utilize 3D microfluidic devices to study the effects of shear stress, CS, and endothelial cells on calcification in an aortic valve fibrosa model to understand CAVD progression.Methods: Valve-on-a-chip devices were fabricated utilizing a wafer mold, soft lithography, and corona discharge. Devices have a flow channel and 3D matrix reservoir of collagen-only healthy controls or with 1mg/mL or 20mg/mL CS. Porcine aortic valve interstitial cells (PAVIC) are embedded within and endothelial cells (PAVEC) seeded onto the matrix. Steady shear stress at 1dyne/cm2and 20dyne/cm2was applied using a peristaltic pump at 37°C and 5%CO2. Alizarin Red S (ARS) was used to assess calcific nodule formation after 14-day cultures.Results: PAVIC/PAVEC co-cultures with increasing shear stress and GAGs exhibit increased calcification compared to static controls . Quantitatively, ARS increased as shear stress increased, regardless of GAGs. In the presence of CS, analysis of nodules indicated that nodule size decreased with shear stress, however it increased the total average percent area stained. Valve-on-a-chip models with PAVIC-only show decreased calcium deposition when compared to co-cultures.Conclusions:In vitrocalcification increases in the presence of shear stress, PAVEC, and increased concentrations of GAGs. Further studies will identify the chemical composition of nodules and to understand early-CAVD pathophysiology. Given that CAVD has no targeted therapies, the creation of a physiologically-relevant test-bed of the aortic valve will lead to contributions in new therapeutics.