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用于半月板组织工程的由脱细胞半月板细胞外基质和聚己内酯组成的电纺纳米纤维的制备与表征

Fabrication and characterization of electrospun nanofibers composed of decellularized meniscus extracellular matrix and polycaprolactone for meniscus tissue engineering.

作者信息

Gao Shuang, Guo Weimin, Chen Mingxue, Yuan Zhiguo, Wang Mingjie, Zhang Yu, Liu Shuyun, Xi Tingfei, Guo Quanyi

机构信息

Center for Biomedical Material and Tissue Engineering, Academy for Advanced Interdisciplinary Studies, Peking University, Yi He Yuan Road No. 5, HaiDian District, Beijing 100871, China.

出版信息

J Mater Chem B. 2017 Mar 28;5(12):2273-2285. doi: 10.1039/c6tb03299k. Epub 2017 Mar 8.

Abstract

Many kinds of scaffolds have been produced in meniscus tissue engineering, but few have matched the mechanical properties of native meniscus, making it impossible for them to sustain large stress at initial implantation. In this study, we used a differential centrifugation method to obtain decellularized meniscus extracellular matrix (DMECM) and combined the DMECM with polycaprolactone (PCL) via electrospinning to fabricate random and aligned microfibers. The FTIR results and biochemical assays demonstrated the successful mixing of these two elements, and the addition of DMECM improved the hydrophilicity of the microfibers. The blending of DMECM also enhanced the tensile modulus of the microfibers, and aligned fibers had tensile moduli ranging from 132.27 to 331.40 MPa, which match that of human meniscus. In addition, we defined yield stress as the lose-efficacy point. The results showed that DMECM/PCL fibers had higher yield stresses than the pure PCL fibers, and the aligned fibers had higher yield stress values than the randomly oriented fibers. Nanoindentation results showed that adding DMECM had no significant impact on modulus and hardness with the exception of fibers containing 80% DMECM, which exhibited an obvious increase in modulus. In vitro assay demonstrated that the DMECM/PCL fibers had no hemolysis or cytotoxicity. Meniscus cells could attach and proliferate on the fibers, and the fiber orientation had a direct influence on cell arrangement. RT-PCR results showed that meniscus cells had higher gene expressions of aggrecan, collagen I, collagen II and Sox 9 when seeded on fibers with higher DMECM contents.

摘要

在半月板组织工程中已制备出多种支架,但很少有支架能与天然半月板的力学性能相匹配,这使得它们在初始植入时无法承受较大应力。在本研究中,我们采用差速离心法获得脱细胞半月板细胞外基质(DMECM),并通过静电纺丝将DMECM与聚己内酯(PCL)结合,制备出随机排列和定向排列的微纤维。傅里叶变换红外光谱(FTIR)结果和生化分析表明这两种成分成功混合,并且添加DMECM提高了微纤维的亲水性。DMECM的混合还提高了微纤维的拉伸模量,定向排列的纤维拉伸模量在132.27至331.40MPa之间,与人类半月板的拉伸模量相匹配。此外,我们将屈服应力定义为失效点。结果表明,DMECM/PCL纤维的屈服应力高于纯PCL纤维,并且定向排列的纤维比随机取向的纤维具有更高的屈服应力值。纳米压痕结果表明,除了含有80% DMECM的纤维模量明显增加外,添加DMECM对模量和硬度没有显著影响。体外实验表明,DMECM/PCL纤维没有溶血或细胞毒性。半月板细胞可以在纤维上附着并增殖,并且纤维取向对细胞排列有直接影响。逆转录-聚合酶链反应(RT-PCR)结果表明,当接种在DMECM含量较高的纤维上时,半月板细胞中聚集蛋白聚糖、I型胶原、II型胶原和Sox 9的基因表达较高。

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