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采用山羊尸体肺组织制备支架及其在皮肤组织工程中的应用。

Fabrication and characterization of scaffold from cadaver goat-lung tissue for skin tissue engineering applications.

机构信息

Department of Polymer and Process Engineering, Indian Institute of Technology, Roorkee, India.

出版信息

Mater Sci Eng C Mater Biol Appl. 2013 Oct;33(7):4032-8. doi: 10.1016/j.msec.2013.05.045. Epub 2013 May 29.

Abstract

The present study aims to fabricate scaffold from cadaver goat-lung tissue and evaluate it for skin tissue engineering applications. Decellularized goat-lung scaffold was fabricated by removing cells from cadaver goat-lung tissue enzymatically, to have cell-free 3D-architecture of natural extracellular matrix. DNA quantification assay and Hematoxylin and eosin staining confirmed the absence of cellular material in the decellularized lung-tissue. SEM analysis of decellularized scaffold shows the intrinsic porous structure of lung tissue with well-preserved pore-to-pore interconnectivity. FTIR analysis confirmed non-denaturation and well maintainance of collagenous protein structure of decellularized scaffold. MTT assay, SEM analysis and H&E staining of human skin-derived Mesenchymal Stem cell, seeded over the decellularized scaffold, confirms stem cell attachment, viability, biocompatibility and proliferation over the decellularized scaffold. Expression of Keratin18 gene, along with CD105, CD73 and CD44, by human skin-derived Mesenchymal Stem cells over decellularized scaffold signifies that the cells are viable, proliferating and migrating, and have maintained their critical cellular functions in the presence of scaffold. Thus, overall study proves the applicability of the goat-lung tissue derived decellularized scaffold for skin tissue engineering applications.

摘要

本研究旨在利用山羊肺组织制备支架,并评估其在皮肤组织工程中的应用。通过酶消化从山羊尸体肺组织中去除细胞,从而制备脱细胞山羊肺支架,以获得具有天然细胞外基质 3D 结构的无细胞支架。DNA 定量分析和苏木精-伊红染色证实脱细胞肺组织中不存在细胞物质。脱细胞支架的 SEM 分析显示肺组织具有固有多孔结构,且孔与孔之间的连通性得到很好的保留。FTIR 分析证实脱细胞支架中非变性和胶原蛋白结构的良好保持。在脱细胞支架上接种人皮肤来源间充质干细胞的 MTT 检测、SEM 分析和 H&E 染色证实了干细胞在脱细胞支架上的附着、活力、生物相容性和增殖。脱细胞支架上的人皮肤来源间充质干细胞表达角蛋白 18 基因,以及 CD105、CD73 和 CD44,表明细胞在支架存在的情况下是存活、增殖和迁移的,并保持其关键的细胞功能。因此,总体研究证明了山羊肺组织来源的脱细胞支架在皮肤组织工程中的应用可行性。

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