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将人源肝 ECM 与具有拓扑特征的静电纺丝支架结合用于构建肝微环境。

Combining human liver ECM with topographically featured electrospun scaffolds for engineering hepatic microenvironment.

机构信息

Institute for Bioengineering, School of Engineering, University of Edinburgh, Edinburgh, UK.

Foundation of Liver Research, The Roger Williams Institute of Liver Study, London, UK.

出版信息

Sci Rep. 2024 Oct 5;14(1):23192. doi: 10.1038/s41598-024-73827-5.

Abstract

Liver disease cases are rapidly expanding worldwide, and transplantation remains the only effective cure for end-stage disease. There is an increasing demand for developing potential drug treatments, and regenerative therapies using in-vitro culture platforms. Human decellularized extracellular matrix (dECM) is an appealing alternative to conventional animal tissues as it contains human-specific proteins and can serve as scaffolding materials. Herein we exploit this with human donor tissue from discarded liver which was not suitable for transplant using a synergistic approach to combining biological and topographical cues in electrospun materials as an in-vitro culture platform. To realise this, we developed a methodology for incorporating human liver dECM into electrospun polycaprolactone (PCL) fibres with surface nanotopographies (230-580 nm). The hybrid scaffolds were fabricated using varying concentrations of dECM; their morphology, mechanical properties, hydrophilicity and stability were analysed. The scaffolds were validated using HepG2 and primary mouse hepatocytes, with subsequent results indicating that the modified scaffolds-maintained cell growth and influenced cell attachment, proliferation and hepatic-related gene expression. This work demonstrates a novel approach to harvesting the potential from decellularized human tissues in the form of innovative in-vitro culture platforms for liver.

摘要

肝脏疾病在全球范围内迅速蔓延,而移植仍然是治疗终末期疾病的唯一有效方法。人们对开发潜在药物治疗方法和使用体外培养平台的再生疗法的需求不断增加。与人源去细胞细胞外基质(dECM)相比,传统的动物组织更具吸引力,因为它含有人类特异性蛋白,并可用作支架材料。在这里,我们利用来自废弃肝脏的人类供体组织,这些组织不适合移植,采用协同方法将生物和地形线索结合到电纺材料中,作为体外培养平台。为了实现这一点,我们开发了一种将人源肝 dECM 纳入具有表面纳米形貌(230-580nm)的电纺聚己内酯(PCL)纤维中的方法。使用不同浓度的 dECM 来制备混合支架;分析了它们的形态、机械性能、亲水性和稳定性。使用 HepG2 和原代小鼠肝细胞对支架进行了验证,随后的结果表明,修饰后的支架保持了细胞的生长,并影响了细胞的附着、增殖和与肝脏相关的基因表达。这项工作展示了一种从去细胞化的人类组织中获取潜在价值的新方法,即将其转化为用于肝脏的创新性体外培养平台。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebcf/11455933/bc3b21ca6284/41598_2024_73827_Fig1_HTML.jpg

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