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用于半月板组织工程的脱细胞半月板细胞外基质和明胶/壳聚糖支架的研发。

Development of decellularized meniscus extracellular matrix and gelatin/chitosan scaffolds for meniscus tissue engineering.

作者信息

Yu Zhang, Lili Jiang, Tiezheng Zheng, Li Sha, Jianzhuang Wang, Haichao Dong, Kedong Song, Tianqing Liu

机构信息

State Key Laboratory of Fine Chemicals, Dalian R&D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian, China.

Department of Physical Education, Dalian University of Technology, Dalian, China.

出版信息

Biomed Mater Eng. 2019;30(2):125-132. doi: 10.3233/BME-191038.

Abstract

BACKGROUND

Meniscus tissue engineering has provided a great potential treatment for meniscal injuries. However, few scaffolds in meniscus tissue engineering have matched the mechanical properties of native meniscus.

OBJECTIVE

In this study, we developed a composite scaffold using decellularized meniscus extracellular matrix (DMECM) and gelatin/chitosan (G/C) to explore a preferable ratio to enhance the elastic modulus and cytotoxicity properties of scaffolds.

METHODS

The microstructure, porosity, cytotoxicity, and strength of the composite scaffolds were evaluated. The micro-architectures of the samples were evaluated using scanning electron microscope (SEM). Fourier Transform Infrared analysis (FTIR) was used to confirm the chemical structure with different type composite scaffolds. The compressive elastic modulus of all the scaffolds were measured by the universal tensile testing machine DNS300. Calcein-AM (fluorescent green) and propidium iodide (fluorescent red) were used to stain live cells and dead cells. Morphology and spatial distribution of cells within scaffolds were observed by confocal laser scanning microscopy FV 1000.

RESULTS

SEM showed that the composite scaffolds had suitable porous structure. CCK-8 and live/dead staining demonstrated that the composite scaffolds had no cytotoxicity and could promote bone marrow mesenchymal stem cells (BMSCs) proliferation. The FTIR results demonstrated the successful mixing of these two elements, and the addition of DMECM improved the elastic modulus and cytotoxicity of G/C composite scaffolds.

CONCLUSIONS

This study developed a composite scaffold using DMECM and G/C, and demonstrated that it might be suitable for meniscal tissue engineering application.

摘要

背景

半月板组织工程为半月板损伤提供了极具潜力的治疗方法。然而,半月板组织工程中很少有支架能与天然半月板的力学性能相匹配。

目的

在本研究中,我们开发了一种使用脱细胞半月板细胞外基质(DMECM)和明胶/壳聚糖(G/C)的复合支架,以探索能提高支架弹性模量和细胞毒性特性的最佳比例。

方法

对复合支架的微观结构、孔隙率、细胞毒性和强度进行评估。使用扫描电子显微镜(SEM)评估样品的微观结构。采用傅里叶变换红外分析(FTIR)来确认不同类型复合支架的化学结构。所有支架的压缩弹性模量通过万能拉伸试验机DNS300进行测量。使用钙黄绿素 - AM(绿色荧光)和碘化丙啶(红色荧光)对活细胞和死细胞进行染色。通过共聚焦激光扫描显微镜FV 1000观察支架内细胞的形态和空间分布。

结果

SEM显示复合支架具有合适的多孔结构。CCK - 8和活/死染色表明复合支架无细胞毒性,且能促进骨髓间充质干细胞(BMSCs)增殖。FTIR结果表明这两种成分成功混合,并且添加DMECM提高了G/C复合支架的弹性模量和细胞毒性。

结论

本研究开发了一种使用DMECM和G/C的复合支架,并证明其可能适用于半月板组织工程应用。

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