심장 흥분-수축 연결의 지속적인 모니터링을 위한 조직-센서 플랫폼의 바이오하이브리드 3차원 프린팅 및 중개 연구로의 응용

Abstract

Drug-induced cardiotoxicity is a major concern during drug development, as it can lead to adverse effects on the heart such as including arrhythmias and heart failure. Although there are various methods for preclinical cardiotoxicity tests, they often lack physiological relevance and may not accurately predict the cardiotoxic potential of a compound. In recent years, the field of stem cell biology has made significant strides in the development of in vitro model for drug testing. Specifically, the use ofhuman induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CMs) has emerged as a promising alternative to traditional methods of toxicity screening. In addition, 3D engineered hearttissue (EHT) has also shown promise as an in vitro model for investigating cardiac muscle functionsand pharmacological effects. EHT exhibits physiological auxotonic contractions and electrical cues,making it an ideal model for studying cardiotoxicity. With the growing demand for research on 3D cardiac models, various approaches have been suggested; however, there is currently no adequate platform available for the continuous monitoring of acute and chronic pharmacological effects in vitro. Specifically, simultaneous electrical recordings of contractile force and electrical signals in dynamically beating 3D cardiac tissues had not been satisfactorily developed before. To address this, I haveintroduced a biohybrid 3D printing method for fabricating a tissue–sensor platform that enables thesimultaneous recordings of contractile force and electrical signals of EHT. The tissue–sensor platform is composed of an EHT along with two distinct sensors: bipillar-grafted strain gauge (BPSG) sensorand rib-shaped multielectrode (RME). The BPSG sensor is three-dimensionally printed with a structure of two pillars as grafts onto a strain gauge-embedded substrate. The bipillar design promotesEHTcontractility and guides its self-assembly, facilitating the strain gauge-embedded substrate to bend in response to the applied force of EHT contraction. The RME, also printed in three dimensions,hasa stacked structure consisting of multiple passivated electrodes arranged in two layers. This configuration enables real-time 3D mapping of the electrical signals of the EHT. Furthermore, the incorporation of a wireless multi-channel device can provide continuous monitoring of EHT contractile forceand electrical signals, simultaneously. This ability allows for the continuous monitoring of cardiac excitation-contraction coupling (ECC), a complexprocess that regulates cardiac muscle contraction in response to electrical signals, making the platform a valuable tool for investigating the molecular mechanisms underlying cardiac muscle physiology and pathology. The major objective of this dissertation isbiohybrid 3D printing of a tissue–sensor platform for continuously monitoring of thecardiac ECCand its application for translational research. The specific objectives include: (1) developing a BPSG sensor to monitor EHT contractile force; (2) developingREs to monitor EHT electricalsignals; (3)integrating both the BPSG sensor and REs to monitor both physiological parameters; and(4) utilizingthe integrated tissue–sensor platform for translational research. Overall, the tissue–sensorplatform represents a significant advancement in the field of in vitro cardiac modeling anddrug screening. With further development and optimization, the platform has the potential to become awidely used tool for studying cardiac muscle functions and developing new therapeutic approaches forheart disease.