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细胞外基质作为血管组织工程的先进支架

Extracellular matrices as advanced scaffolds for vascular tissue engineering.

作者信息

Piterina A V, Callanan A, Davis L, Meaney C, Walsh M, McGloughlin T M

机构信息

Centre for Applied Biomedical Engineering Research, Department of Mechanical and Aeronautical Engineering, Materials and Surface Science Institute, University of Limerick, Limerick, Ireland.

出版信息

Biomed Mater Eng. 2009;19(4-5):333-48. doi: 10.3233/BME-2009-0598.

Abstract

An alternative non-vascular extracellular material was obtained by decellularisation of porcine urinary bladder and examined for its potential as scaffold for vascular tissue engineering. Analysis using Scanning Electron Microscopy (SEM), Atomic Force Microscopy (AFM) and Laser Scanning Microscopy (LSCM) showed a porous interconnective microarchitecture, an abundance of well preserved fibers on the abluminal side and a micropatterned flat luminal surface. Uniaxial tensile testing revealed a strength of 1.9+/-0.3 MPa for the rehydrated material in a phosphate buffered saline medium for the ECM-UBM sheet and these results comparable to those of native artery of a middle aged human. Multilamination of the UBM increases the tensile properties in general (9+/-0.45 MPa for 2 layer, 30+/-0.6 MPa for 4 layers construct), with no effect on elongation capacities (38-40%) of the material. Contact-angle measurements indicated that the ECM-UBM surface exhibited a hydrophylic characteristic and better wettability than any vascular synthetic materials. Comparison of the initial adhesion in the multiplayer ECM constructs was evaluated and was found not to be altered by the preparation. The HAECs and HSMC shown an excellent adherence, spread and proliferation on the ECM-UBM material with the preservation of the cell phenotype. The level of MMP-1 and MMP-9 produced by endothelial cells was evaluated in this study and provides some data on the remodelling capacity of the scaffold. Vascular cell seeded-UBM constructs may also provide a suitable and affordable in vitro model for cell-physiology and drug development studies, which may elucidate to the mechanisms of vascular disease formation, and its prevention and treatment.

摘要

通过对猪膀胱进行去细胞处理获得了一种替代性的非血管细胞外材料,并对其作为血管组织工程支架的潜力进行了研究。使用扫描电子显微镜(SEM)、原子力显微镜(AFM)和激光扫描显微镜(LSCM)进行分析,结果显示该材料具有多孔互连的微结构,在腔外侧面有大量保存完好的纤维,以及微图案化的平坦腔面。单轴拉伸试验表明,在磷酸盐缓冲盐水培养基中,用于细胞外基质-猪膀胱黏膜下层(ECM-UBM)片的再水化材料强度为1.9±0.3 MPa,这些结果与中年人的天然动脉相当。一般来说,UBM的多层叠加会提高拉伸性能(两层结构为9±0.45 MPa,四层结构为30±0.6 MPa),而对材料的伸长能力(38-40%)没有影响。接触角测量表明,ECM-UBM表面具有亲水性特征,并且比任何血管合成材料具有更好的润湿性。对多层ECM构建体中的初始粘附进行了比较评估,发现制备过程不会改变其粘附情况。人主动脉内皮细胞(HAECs)和平滑肌细胞(HSMC)在ECM-UBM材料上表现出优异的粘附、铺展和增殖能力,同时保留了细胞表型。本研究评估了内皮细胞产生的基质金属蛋白酶-1(MMP-1)和基质金属蛋白酶-9(MMP-9)水平,并提供了一些关于支架重塑能力的数据。接种血管细胞的UBM构建体还可能为细胞生理学和药物开发研究提供一个合适且经济的体外模型,这可能有助于阐明血管疾病的形成机制及其预防和治疗方法。

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