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壳聚糖静电纺丝中结合弹性蛋白以提高机械强度和生物活性的研究。

A Study of Combining Elastin in the Chitosan Electrospinning to Increase the Mechanical Strength and Bioactivity.

作者信息

Su Hengjie, Fujiwara Tomoko, Bumgardner Joel D

机构信息

Department of Biomedical Engineering, UT-UofM Joint Graduate Program in Biomedical Engineering, The University of Memphis, Engineering Technology Bldg #330, Memphis, TN 38152, USA.

Department of Chemistry, The University of Memphis, Smith Hall #409, Memphis, TN 38152, USA.

出版信息

Mar Drugs. 2021 Mar 22;19(3):169. doi: 10.3390/md19030169.

Abstract

While electrospun chitosan membranes modified to retain nanofibrous morphology have shown promise for use in guided bone regeneration applications in in vitro and in vivo studies, their mechanical tear strengths are lower than commercial collagen membranes. Elastin, a natural component of the extracellular matrix, is a protein with extensive elastic property. This work examined the incorporation of elastin into electrospun chitosan membranes to improve their mechanical tear strengths and to further mimic the native extracellular composition for guided bone regeneration (GBR) applications. In this work, hydrolyzed elastin (ES12, Elastin Products Company, USA) was added to a chitosan spinning solution from 0 to 4 wt% of chitosan. The chitosan-elastin (CE) membranes were examined for fiber morphology using SEM, hydrophobicity using water contact angle measurements, the mechanical tear strength under simulated surgical tacking, and compositions using Fourier-transform infrared spectroscopy (FTIR) and post-spinning protein extraction. In vitro experiments were conducted to evaluate the degradation in a lysozyme solution based on the mass loss and growth of fibroblastic cells. Chitosan membranes with elastin showed significantly thicker fiber diameters, lower water contact angles, up to 33% faster degradation rates, and up to seven times higher mechanical strengths than the chitosan membrane. The FTIR spectra showed stronger amide peaks at 1535 cm and 1655 cm in membranes with higher concentrated elastin, indicating the incorporation of elastin into electrospun fibers. The bicinchoninic acid (BCA) assay demonstrated an increase in protein concentration in proportion to the amount of elastin added to the CE membranes. In addition, all the CE membranes showed in vitro biocompatibility with the fibroblasts.

摘要

虽然经过改性以保留纳米纤维形态的电纺壳聚糖膜在体外和体内研究的引导骨再生应用中已显示出应用前景,但其机械撕裂强度低于商业胶原膜。弹性蛋白是细胞外基质的天然成分,是一种具有广泛弹性特性的蛋白质。这项工作研究了将弹性蛋白掺入电纺壳聚糖膜中,以提高其机械撕裂强度,并进一步模拟用于引导骨再生(GBR)应用的天然细胞外成分。在这项工作中,将水解弹性蛋白(ES12,美国弹性蛋白产品公司)添加到壳聚糖纺丝溶液中,添加量为壳聚糖的0至4 wt%。使用扫描电子显微镜(SEM)检查壳聚糖-弹性蛋白(CE)膜的纤维形态,使用水接触角测量法检查其疏水性,在模拟手术固定下测试其机械撕裂强度,并使用傅里叶变换红外光谱(FTIR)和纺丝后蛋白质提取法分析其成分。进行体外实验以基于溶菌酶溶液中的质量损失和成纤维细胞的生长来评估降解情况。与壳聚糖膜相比,含有弹性蛋白的壳聚糖膜显示出明显更粗的纤维直径、更低的水接触角、高达33%的更快降解速率以及高达七倍的更高机械强度。FTIR光谱显示,在弹性蛋白浓度较高的膜中,在1535 cm和1655 cm处的酰胺峰更强,表明弹性蛋白已掺入电纺纤维中。二辛可宁酸(BCA)测定表明,蛋白质浓度的增加与添加到CE膜中的弹性蛋白量成比例。此外,所有CE膜均显示出与成纤维细胞的体外生物相容性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e10/8004263/6604c3857f58/marinedrugs-19-00169-g001.jpg

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