Institute of Bioengineering, School of Engineering, The University of Edinburgh, Edinburgh EH10 SHF, United Kingdom.
ACS Appl Bio Mater. 2023 Jun 19;6(6):2158-2171. doi: 10.1021/acsabm.3c00032. Epub 2023 Jun 7.
Donor liver shortage is a crucial global public health problem as whole-organ transplantation is the only definitive cure for liver disease. Liver tissue engineering aims to reproduce or restore function through in vitro tissue constructs, which may lead to alternative treatments for active and chronic liver disease. The formulation of a multifunctional scaffold that has the potential to mimic the complex extracellular matrix (ECM) and their influence on cellular behavior, are essential for culturing cells on a construct. The separate employment of topographic or biological cues on a scaffold has both shown influences on hepatocyte survival and growth. In this study, we investigate both of these synergistic effects and developed a new procedure to directly blend whole-organ vascular perfusion-decellularized rat liver ECM (dECM) into electrospun fibers with tailored surface nanotopography. Water contact angle, tensile test, and degradation studies were conducted to analyze scaffold hydrophilicity, mechanical properties, and stability. The results show that our novel hybrid scaffolds have enhanced hydrophilicity, and the nanotopography retained its original form after hydrolytic degradation for 14 days. Human hepatocytes (HepG2) were seeded to analyze the scaffold biocompatibility. Cell viability and DNA quantification imply steady cell proliferation over the culture period, with the highest albumin secretion observed on the hybrid scaffold. Scanning electron microscopy shows that cell morphology was distinctly different on hybrid scaffolds compared to control groups, where HepG2 began to form a monolayer toward the end of the culture period; meanwhile, typical hepatic markers and ECM genes were also influenced, such as an increasing trend of albumin appearing on the hybrid scaffolds. Taken together, our findings provide a reproducible approach and utilization of animal tissue-derived ECM and emphasize the synergism of topographical stimuli and biochemical cues on electrospun scaffolds in liver tissue engineering.
供体肝脏短缺是一个全球性的公共卫生难题,因为全器官移植是治疗肝脏疾病的唯一有效方法。肝脏组织工程旨在通过体外组织构建来复制或恢复功能,这可能为活跃和慢性肝脏疾病提供替代治疗方法。构建多功能支架,具有潜在的能力来模拟复杂的细胞外基质 (ECM) 及其对细胞行为的影响,对于在构建物上培养细胞是必不可少的。在支架上单独使用形貌或生物线索都对肝细胞的存活和生长有影响。在这项研究中,我们研究了这两种协同效应,并开发了一种新的方法,即将整个器官血管灌注去细胞化大鼠肝脏 ECM(dECM)直接混入具有定制表面纳米形貌的电纺纤维中。通过水接触角、拉伸试验和降解研究来分析支架的亲水性、机械性能和稳定性。结果表明,我们的新型混合支架具有增强的亲水性,并且纳米形貌在水解降解 14 天后仍保留其原始形式。接种人肝癌细胞 (HepG2) 来分析支架的生物相容性。细胞活力和 DNA 定量表明,细胞在培养期间稳定增殖,在混合支架上观察到最高的白蛋白分泌。扫描电子显微镜显示,与对照组相比,混合支架上的细胞形态明显不同,在培养期末,HepG2 开始形成单层;同时,还影响了典型的肝标志物和 ECM 基因,例如在混合支架上出现白蛋白的趋势增加。总之,我们的研究结果提供了一种可重复的方法,利用了动物组织衍生的 ECM,并强调了形貌刺激和生化线索在肝脏组织工程中对电纺支架的协同作用。