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脱细胞支架联合负载纳米颗粒在体外心脏瓣膜组织工程中的应用。

Application of decellularized scaffold combined with loaded nanoparticles for heart valve tissue engineering in vitro.

作者信息

Deng Cheng, Dong Nianguo, Shi Jiawei, Chen Si, Xu Lei, Shi Feng, Hu Xingjian, Zhang Xianzheng

机构信息

Department of Cardiovascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.

Key Laboratory of Biomedical Polymers of Ministry of Education and Department of Chemistry, Wuhan University, Wuhan, 430072, China.

出版信息

J Huazhong Univ Sci Technolog Med Sci. 2011 Feb;31(1):88-93. doi: 10.1007/s11596-011-0156-2. Epub 2011 Feb 19.

Abstract

The purpose of this study was to fabricate decelluarized valve scaffold modified with polyethylene glycol nanoparticles loaded with transforming growth factor-β1 (TGF-β1), by which to improve the extracellular matrix microenvironment for heart valve tissue engineering in vitro. Polyethylene glycol nanoparticles were obtained by an emulsion-crosslinking method, and their morphology was observed under a scanning electron microscope. Decelluarized valve scaffolds, prepared by using trypsinase and TritonX-100, were modified with nanoparticles by carbodiimide, and then TGF-β1 was loaded into them by adsorption. The TGF-β1 delivery of the fabricated scaffold was measured by asing enzyme-linked immunosorbent assay. Whether unseeded or reseeded with myofibroblast from rats, the morphologic, biochemical and biomechanical characteristics of hybrid scaffolds were tested and compared with decelluarized scaffolds under the same conditions. The enzyme-linked immunosorbent assay revealed a typical delivery of nanoparticles. The morphologic observations and biological data analysis indicated that fabricated scaffolds possessed advantageous biocompatibility and biomechanical property beyond decelluarized scaffolds. Altogether this study proved that it was feasible to fabricate the hybrid scaffold and effective to improve extracellular matrix microenvironment, which is beneficial for an application in heart valve tissue engineering.

摘要

本研究的目的是制备负载转化生长因子-β1(TGF-β1)的聚乙二醇纳米颗粒修饰的去细胞瓣膜支架,以此改善体外心脏瓣膜组织工程的细胞外基质微环境。通过乳液交联法制备聚乙二醇纳米颗粒,并在扫描电子显微镜下观察其形态。使用胰蛋白酶和曲拉通X-100制备去细胞瓣膜支架,通过碳二亚胺用纳米颗粒对其进行修饰,然后通过吸附将TGF-β1负载到其中。通过酶联免疫吸附测定法测量制备支架的TGF-β1释放情况。无论未接种还是接种大鼠成肌纤维细胞,均测试混合支架的形态、生化和生物力学特性,并在相同条件下与去细胞支架进行比较。酶联免疫吸附测定显示纳米颗粒有典型的释放。形态学观察和生物学数据分析表明,制备的支架具有优于去细胞支架的生物相容性和生物力学性能。总之,本研究证明制备混合支架是可行的,并且有效改善细胞外基质微环境,这有利于在心脏瓣膜组织工程中的应用。

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