Hassannejad Zahra, Fendereski Kiarad, Daryabari Seyedeh Sima, Tanourlouee Saman Behboodi, Dehnavi Mehrshad, Kajbafzadeh Abdol-Mohammad
Pediatric Urology and Regenerative Medicine Research Center, Gene, Cell and Tissue Research Institute, Children's Medical Center, Tehran University of Medical Sciences, No. 62, Dr. Gharib's Street, Keshavarz Boulevard, Tehran, 1419733151, Iran.
Cardiovasc Eng Technol. 2024 Dec;15(6):679-690. doi: 10.1007/s13239-024-00744-z. Epub 2024 Aug 12.
Myocardial infarction (MI) is a leading cause of irreversible functional cardiac tissue loss, requiring novel regenerative strategies. This study assessed the potential therapeutic efficacy of recellularized cardiac patches, incorporating fetal myocardial scaffolds with rat fetal cardiomyocytes and acellular human amniotic membrane, in adult Wistar rat models of MI.
Decellularized myocardial tissue was obtained from 14 to 16 week-old human fetuses that had been aborted. Chemical detergents (0.1% EDTA and 0.2% sodium dodecyl sulfate) were used to prepare the fetal extracellular matrix (ECM), which was characterized for bio-scaffold microstructure and biocompatibility via scanning electron microscopy (SEM) and MTT assay, respectively. Neonatal cardiomyocytes were extracted from the ventricles of one-day-old Wistar rats' littermates and characterized through immunostaining against Connexin-43 and α-smooth muscle actin. The isolated cells were seeded onto decellularized tissues and covered with decellularized amniotic membrane. Sixteen healthy adult Wistar rats were systematically allocated to control and MI groups. MI was induced via arterial ligation. Fourteen days post-operation, the MI group was received the engineered patches. Following a two-week post-implantation period, the animals were euthanized, and the hearts were harvested for the graft evaluation.
Histological analysis, DAPI staining, and ultra-structural examination corroborated the successful depletion of cellular elements, while maintaining the integrity of the fetal ECM and architecture. Subsequent histological and immunohistochemichal (IHC) evaluations confirmed effective cardiomyocyte seeding on the scaffolds. The application of these engineered patches in MI models resulted in increased angiogenesis, reduced fibrosis, and restricted scar tissue formation, with the implanted cardiomyocytes remaining viable at graft sites, indicating prospective in vivo cell viability.
This study suggests that multi-layered recellularized cardiac patches are a promising surgical intervention for myocardial infarction, showcasing significant potential by promoting angiogenesis, mitigating fibrosis, and minimizing scar tissue formation in MI models. These features are pivotal for enhancing the therapeutic outcomes in MI patients, focusing on the restoration of the myocardial structure and function post-infarction.
心肌梗死（MI）是导致心脏组织功能不可逆丧失的主要原因，需要新的再生策略。本研究评估了在成年Wistar大鼠心肌梗死模型中，将含有大鼠胎儿心肌细胞和脱细胞人羊膜的再细胞化心脏补片作为潜在治疗方法的疗效。
从14至16周龄人工流产的人类胎儿获取脱细胞心肌组织。使用化学洗涤剂（0.1%乙二胺四乙酸和0.2%十二烷基硫酸钠）制备胎儿细胞外基质（ECM），分别通过扫描电子显微镜（SEM）和MTT试验对其生物支架微观结构和生物相容性进行表征。从1日龄Wistar大鼠同窝仔鼠的心室中提取新生心肌细胞，并通过针对连接蛋白-43和α-平滑肌肌动蛋白的免疫染色进行表征。将分离的细胞接种到脱细胞组织上，并用脱细胞羊膜覆盖。16只健康成年Wistar大鼠被系统地分为对照组和心肌梗死组。通过动脉结扎诱导心肌梗死。术后14天，心肌梗死组接受工程化补片。植入两周后，对动物实施安乐死，并取出心脏进行移植物评估。
组织学分析、4',6-二脒基-2-苯基吲哚（DAPI）染色和超微结构检查证实细胞成分成功去除，同时保持了胎儿ECM和结构的完整性。随后的组织学和免疫组织化学（IHC）评估证实心肌细胞有效接种在支架上。这些工程化补片在心肌梗死模型中的应用导致血管生成增加、纤维化减少和瘢痕组织形成受限，植入的心肌细胞在移植物部位保持存活，表明在体内具有预期的细胞活力。
本研究表明，多层再细胞化心脏补片是一种有前景的心肌梗死手术干预方法，通过促进血管生成、减轻纤维化和最小化心肌梗死模型中的瘢痕组织形成展现出巨大潜力。这些特性对于提高心肌梗死患者的治疗效果至关重要，重点在于恢复梗死后的心肌结构和功能。
