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Micropatterning and characterization of electrospun poly(ε-caprolactone)/gelatin nanofiber tissue scaffolds by femtosecond laser ablation for tissue engineering applications

机译:飞秒激光消融电纺聚(ε-己内酯)/明胶纳米纤维组织支架的微图案和表征,用于组织工程应用

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Experimental investigations aimed at assessing the effectiveness of femtosecond (FS) laser ablation for creating microscale features on electrospun poly(ε-caprolactone) (PCL)/gelatin nanofiber tissue scaffold capable of controlling cell distribution are described. Statistical comparisons of the fiber diameter and surface porosity on laser-machined and as-spun surface were made and results showed that laser ablation did not change the fiber surface morphology. The minimum feature size that could be created on electrospun nanofiber surfaces by direct-write ablation was measured over a range of laser pulse energies. The minimum feature size that could be created was limited only by the pore size of the scaffold surface. The chemical states of PCL/gelatin nanofiber surfaces were measured before and after FS laser machining by attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy and X-ray photoelectron spectroscopy (XPS) and showed that laser machining produced no changes in the chemistry of the surface. In vitro, mouse embryonic stem cells (mES cells) were cultured on as-spun surfaces and in laser-machined microwells. Cell densities were found to be statistically indistinguishable after 1 and 2 days of growth. Additionally, confocal microscope imaging confirmed that spreading of mES cells cultured within laser-machined microwells was constrained by the cavity walls, the expected and desired function of these cavities. The geometric constraint caused statistically significant smaller density of cells in microwells after 3 days of growth. It was concluded that FS laser ablation is an effective process for microscale structuring of these electrospun nanofiber tissue scaffold surfaces.
机译:描述了旨在评估飞秒(FS)激光烧蚀在能够控制细胞分布的电纺聚(ε-己内酯)(PCL)/明胶纳米纤维组织支架上形成微尺度特征的有效性的实验研究。对激光加工和初纺表面上的纤维直径和表面孔隙率进行统计比较,结果表明激光烧蚀不会改变纤维的表面形态。在一定范围的激光脉冲能量范围内,测量了通过直接写入烧蚀可在电纺纳米纤维表面上产生的最小特征尺寸。可以产生的最小特征尺寸仅受支架表面的孔径限制。在FS激光加工之前和之后,通过衰减全反射傅里叶变换红外(ATR-FTIR)光谱和X射线光电子能谱(XPS)测量了PCL /明胶纳米纤维表面的化学状态,结果表明激光加工不会改变化学性质表面。在体外,将小鼠胚胎干细胞(mES细胞)培养在初生表面和激光加工的微孔中。发现在生长1天和2天后,细胞密度在统计学上是无法区分的。此外,共聚焦显微镜成像证实,在激光加工的微孔中培养的mES细胞的扩散受腔壁,这些腔的预期功能和期望功能的约束。生长3天后,几何约束在微孔中引起统计学上显着较小的细胞密度。结论是,FS激光烧蚀是这些电纺纳米纤维组织支架表面的微观结构的有效过程。

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