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Micromechanical force promotes aortic valvular calcification
The Journal of Thoracic and Cardiovascular Surgery ( IF 6 ) Pub Date : 2021-08-14 , DOI: 10.1016/j.jtcvs.2021.08.014
Yefan Jiang 1 , Jinjie Chen 2 , Fuxiang Wei 3 , Yixuan Wang 2 , Si Chen 2 , Geng Li 2 , Nianguo Dong 2
Affiliation  

Objective

Calcified aortic valvular disease is known as an inflammation-related process related to force. The purpose of this study was to determine whether micromechanical force could induce valve calcification of porcine valvular interstitial cells and to examine the role of integrin αvβ3 in valvular calcification by using a novel method: magnetic twisting cytometry.

Methods

Porcine valvular interstitial cells were cultured in vitro, and micromechanical force was applied to porcine valvular interstitial cells using magnetic twisting cytometry. Changes in calcification-related factors osteopontin and RUNX2 were detected. By using the calcification medium, the optimal magnetic twisting cytometry parameters for inducing valvular interstitial cell calcification were determined, and a magnetic twisting cytometry calcification promotion model was established. The role of αvβ3 in calcification was studied by using αvβ3 antagonists to block the function of αvβ3.

Results

Reverse transcription polymerase chain reaction assays showed that the expression of osteopontin was enhanced 30 minutes after 25G-1Hz 5 minutes of stimulation. Western blotting assays showed that the expression of osteopontin and RUNX2 was upregulated 24 hours after 25G-1Hz 5 minutes of stimulation. The optimal magnetic twisting cytometry parameter for inducing porcine valvular interstitial cell calcification was 25G-2Hz for 10 minutes. The expression of osteopontin and RUNX2 decreased significantly after the addition of αvβ3 antagonist. Clinically, patients with bicuspid aortic valves had high expression of RUNX2 and β3 in the aortic valve, and β3 significantly correlated with RUNX2.

Conclusions

By using magnetic twisting cytometry, we established a porcine valvular interstitial cell calcification model by micromechanical force stimulation and obtained the optimal parameters. Integrin αvβ3 plays a key role in the aortic valve calcification process.



中文翻译:

微机械力促进主动脉瓣膜钙化

客观的

钙化主动脉瓣膜病被称为与力有关的炎症相关过程。本研究的目的是确定微机械力是否可以诱导猪瓣膜间质细胞的瓣膜钙化,并通过使用一种新方法:磁扭转细胞术来检查整合素 αvβ3 在瓣膜钙化中的作用。

方法

体外培养猪瓣膜间质细胞,利用磁力扭转细胞仪对猪瓣膜间质细胞施加微机械力。检测钙化相关因子骨桥蛋白和RUNX2的变化。利用钙化介质确定了诱导瓣膜间质细胞钙化的最佳磁力扭曲流式细胞术参数,建立了磁力扭曲细胞术钙化促进模型。通过使用αvβ3拮抗剂阻断αvβ3的功能来研究αvβ3在钙化中的作用。

结果

逆转录聚合酶链反应检测表明,在 25G-1Hz 刺激 5 分钟后,骨桥蛋白的表达增强了 30 分钟。Western blotting 检测显示,在 25G-1Hz 刺激 5 分钟后 24 小时,骨桥蛋白和 RUNX2 的表达上调。用于诱导猪瓣膜间质细胞钙化的最佳磁力扭转细胞仪参数为 25G-2Hz 10 分钟。添加αvβ3拮抗剂后骨桥蛋白和RUNX2的表达显着降低。临床上,二尖瓣主动脉瓣患者主动脉瓣RUNX2和β3高表达, β3与RUNX2显着相关。

结论

我们采用磁力扭转细胞仪,通过微机械力刺激建立了猪瓣膜间质细胞钙化模型,并获得了最佳参数。整合素αvβ3在主动脉瓣钙化过程中起关键作用。

更新日期:2021-08-14
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