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用于骨组织工程的电纺取向明胶-羟基磷灰石纤维支架

Electrospun oriented gelatin-hydroxyapatite fiber scaffolds for bone tissue engineering.

作者信息

Salifu Ali A, Lekakou Constantina, Labeed Fatima H

机构信息

Advanced Materials Group, University of Surrey, Guildford, Surrey, GU2 7XH, United Kingdom.

Centre of Biomedical Engineering, University of Surrey, Guildford, Surrey, GU2 7XH, United Kingdom.

出版信息

J Biomed Mater Res A. 2017 Jul;105(7):1911-1926. doi: 10.1002/jbm.a.36058. Epub 2017 Apr 3.

Abstract

Tissue engineering of human fetal osteoblast cells was investigated on gelatin-hydroxyapatite (HA), crosslinked, electrospun oriented fiber scaffolds at the different HA concentrations of 0, 10, 20, and 25 wt % in the dry fibers and different fiber diameter, pore size and porosity of scaffolds. Rheological tests and proton nuclear magnetic resonance spectroscopy were conducted for all solutions used for electrospinning. It was found that 25 wt % HA-gelatin scaffolds electrospun at 20 kV led to the greatest cell attachment, cell proliferation and extracellular matrix (ECM) production while fiber orientation improved the mechanical properties, where crosslinked electrospun 25 wt % HA-gelatin fiber scaffolds yielded a Young's modulus in the range of 0.5-0.9 GPa and a tensile strength in the range of 4-10 MPa in the fiber direction for an applied voltage of 20-30 kV, respectively, in the electrospinning of scaffolds. Biological characterization of cell seeded scaffolds yielded the rate of cell growth and ECM (collagen and calcium) production by the cells as a function of time; it included cell seeding efficiency tests, alamar blue cell proliferation assay, alkaline phosphate (ALP) assay, collagen assay, calcium colorimetric assay, fluorescence microscopy for live and dead cells, and scanning electron microscopy for cell culture from 1 to 18 days. After 18 days, cells seeded and grown on the 25 wt % HA-gelatin scaffold, electrospun at 20 kV, reached production of collagen at 370 μg/L and calcium production at 0.8 mM. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1911-1926, 2017.

摘要

研究了人胎儿成骨细胞在明胶-羟基磷灰石(HA)交联电纺取向纤维支架上的组织工程情况,该支架在干纤维中的HA浓度分别为0、10、20和25 wt%,且具有不同的纤维直径、孔径和孔隙率。对所有用于电纺的溶液进行了流变学测试和质子核磁共振光谱分析。结果发现,在20 kV电压下电纺的25 wt% HA-明胶支架导致最大的细胞附着、细胞增殖和细胞外基质(ECM)产生,而纤维取向改善了力学性能,其中在支架电纺过程中,交联电纺的25 wt% HA-明胶纤维支架在20 - 30 kV的施加电压下,在纤维方向上的杨氏模量范围为0.5 - 0.9 GPa,拉伸强度范围为4 - 10 MPa。接种细胞的支架的生物学特性给出了细胞生长速率和细胞产生的ECM(胶原蛋白和钙)随时间的函数关系;它包括细胞接种效率测试、alamar蓝细胞增殖测定、碱性磷酸酶(ALP)测定、胶原蛋白测定、钙比色测定、活细胞和死细胞的荧光显微镜检查以及1至18天细胞培养的扫描电子显微镜检查。18天后,接种在20 kV下电纺的25 wt% HA-明胶支架上生长的细胞,胶原蛋白产量达到370 μg/L,钙产量达到0.8 mM。© 2016威利期刊公司。《生物医学材料研究杂志》A部分:105A:1911 - 1926,2017年。

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