Department of Tissue Engineering and Regenerative Medicine, Faculty of Medicine, National University of Malaysia, Kuala Lumpur 56000, Malaysia.
Graduate School of Medicine, KPJ Healthcare University, Nilai 71800, Negeri Sembilan, Malaysia.
Int J Mol Sci. 2024 Oct 30;25(21):11697. doi: 10.3390/ijms252111697.
Revascularization procedures such as percutaneous coronary intervention (PCI) and coronary artery bypass grafting (CABG) are crucial to restore blood flow to the heart and are used in the treatment of myocardial infarction (MI). However, these techniques are known to cause myocardial reperfusion injury in the ischemic heart. The present study aims to mimic ischemia-reperfusion injury in vitro on primary human cardiomyocytes (HCMs) and use the established injury model to study the rescue mechanism of skeletal muscle cell (SkM)-seeded electrospun fiber-coated human amniotic membrane scaffold (EF-HAM) on injured cardiomyocytes through paracrine secretion. An in vitro ischemia-reperfusion injury model was established by exposing the HCM to 5 h of hypoxia, followed by a 6 h reoxygenation period. Six different conditioned media (CM) including three derived from SkM-seeded EF-HAMs were introduced to the injured cells to investigate the cardioprotective effect of the CM. Cell survival analysis, caspase-3 and XIAP expression profiling, mitochondrial membrane potential analysis, and measurement of reactive oxygen species (ROS) were conducted to evaluate the outcomes of the study. The results revealed a significant increase in the viability of HCM exposed to H/R injury by 77.2% ( < 0.01), 111.8% ( < 0.001), 68.7% ( < 0.05), and 69.5% ( < 0.05) when supplemented with HAM CM, EF-HAM 3 min CM, EF-HAM 5 min CM, and EF-HAM 7 min CM, respectively. Furthermore, CM derived from SkM-seeded EF-HAM scaffolds positively impacted hypoxia-/reoxygenation-induced changes in caspase-3 expression, mitochondrial membrane potential, and reactive oxygen species generation, but not in XIAP expression. These findings suggest that EF-HAM composite scaffolds can exert antiapoptotic and cardioregenerative effects on primary human cardiomyocytes through the paracrine mechanism.
血运重建术，如经皮冠状动脉介入治疗（PCI）和冠状动脉旁路移植术（CABG），对于恢复心脏血流至关重要，用于心肌梗死（MI）的治疗。然而，这些技术已知会导致缺血性心脏中的心肌再灌注损伤。本研究旨在体外模拟原发性人心肌细胞（HCM）的缺血再灌注损伤，并使用建立的损伤模型，通过旁分泌研究骨骼肌细胞（SkM）-种子电纺纤维包被人羊膜支架（EF-HAM）对损伤心肌细胞的挽救机制。通过将 HCM 暴露于 5 小时缺氧，然后进行 6 小时复氧，建立体外缺血再灌注损伤模型。将六种不同的条件培养基（CM）引入受损细胞，包括三种源自 SkM-种子 EF-HAM 的 CM，以研究 CM 的心脏保护作用。进行细胞存活率分析、caspase-3 和 XIAP 表达谱分析、线粒体膜电位分析和活性氧（ROS）测量，以评估研究结果。结果显示，暴露于 H/R 损伤的 HCM 的存活率显著增加，分别为 77.2%（ < 0.01）、111.8%（ < 0.001）、68.7%（ < 0.05）和 69.5%（ < 0.05），当补充 HAM CM、EF-HAM 3 分钟 CM、EF-HAM 5 分钟 CM 和 EF-HAM 7 分钟 CM 时。此外，CM 源自 SkM-种子 EF-HAM 支架，可对缺氧/复氧诱导的 caspase-3 表达、线粒体膜电位和活性氧生成的变化产生积极影响，但对 XIAP 表达没有影响。这些发现表明，EF-HAM 复合支架可通过旁分泌机制对原代人心肌细胞发挥抗凋亡和心脏再生作用。
