Murphy Jack F, Costa Kevin D, Turnbull Irene C
Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, Box 1014, New York City, 10029, NY, USA.
Comput Methods Programs Biomed Update. 2023;3. doi: 10.1016/j.cmpbup.2023.100107. Epub 2023 May 25.
The field of tissue engineering has provided valuable three-dimensional species-specific models of the human myocardium in the form of human Engineered Cardiac Tissues (hECTs) and similar constructs. However, hECT systems are often bottlenecked by a lack of openly available software that can collect data from multiple tissues at a time, even in multi-tissue bioreactors, which limits throughput in phenotypic and therapeutic screening applications.
We developed Rianú, an open-source web application capable of simultaneously tracking multiple hECTs on flexible end-posts. This software is operating system agnostic and deployable on a remote server, accessible via a web browser with no local hardware or software requirements. The software incorporates object-tracking capabilities for multiple objects simultaneously, an algorithm for twitch tracing analysis and contractile force calculation, and a data compilation system for comparative analysis within and amongst groups. Validation tests were performed using in-silico and in-vitro experiments for comparison with established methods and interventions.
Rianú was able to detect the displacement of the flexible end-posts with a sub-pixel sensitivity of 0.555 px/post (minimum increment in post displacement) and a lower limit of 1.665 px/post (minimum post displacement). Compared to our established reference for contractility assessment, Rianú had a high correlation for all parameters analyzed (ranging from to ), demonstrating its high accuracy and reliability.
Rianú provides simultaneous tracking of multiple hECTs, expediting the recording and analysis processes, and simplifies time-based intervention studies. It also allows data collection from different formats and has scale-up capabilities proportional to the number of tissues per field of view. These capabilities will enhance throughput of hECTs and similar assays for in-vitro analysis in disease modeling and drug screening applications.
组织工程领域已通过人类工程心脏组织(hECTs)及类似构建体的形式,提供了有价值的三维物种特异性人类心肌模型。然而,hECT系统常常受到缺乏可公开获取软件的限制,该软件无法一次从多个组织收集数据,即便在多组织生物反应器中也是如此,这限制了表型和治疗筛选应用中的通量。
我们开发了Rianú,这是一款开源网络应用程序,能够在柔性末端柱上同时跟踪多个hECTs。该软件与操作系统无关,可部署在远程服务器上,通过网络浏览器即可访问,无需本地硬件或软件要求。该软件具备同时对多个对象进行跟踪的功能、用于抽搐追踪分析和收缩力计算的算法,以及用于组内和组间比较分析的数据编译系统。使用计算机模拟和体外实验进行验证测试,以便与既定方法和干预措施进行比较。
Rianú能够以0.555像素/柱的亚像素灵敏度(柱位移的最小增量)和1.665像素/柱的下限(柱的最小位移)检测柔性末端柱的位移。与我们既定的收缩性评估参考相比,Rianú对所有分析参数都具有高度相关性(范围从 到 ),证明了其高准确性和可靠性。
Rianú可同时跟踪多个hECTs,加快记录和分析过程,并简化基于时间的干预研究。它还允许从不同格式收集数据,并具有与每个视野中的组织数量成比例的扩大规模能力。这些能力将提高hECTs以及疾病建模和药物筛选应用中体外分析的类似检测的通量。