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海藻酸盐敷料作为伤口愈合潜在蛋白质递送系统的开发与功能表征

Development and functional characterization of alginate dressing as potential protein delivery system for wound healing.

作者信息

Momoh Frederick U, Boateng Joshua S, Richardson Simon C W, Chowdhry Babur Z, Mitchell John C

机构信息

Department of Pharmaceutical Chemical and Environmental Sciences, Faculty of Engineering and Science, University of Greenwich at Medway, Central Avenue, Chatham Maritime, ME4 4TB Kent, UK.

Department of Pharmaceutical Chemical and Environmental Sciences, Faculty of Engineering and Science, University of Greenwich at Medway, Central Avenue, Chatham Maritime, ME4 4TB Kent, UK.

出版信息

Int J Biol Macromol. 2015 Nov;81:137-50. doi: 10.1016/j.ijbiomac.2015.07.037. Epub 2015 Jul 26.

Abstract

This study aimed to develop and characterize stable films as potential protein delivery dressings to wounds. Films were prepared from aqueous gels of sodium alginate (SA) and glycerol (GLY) (SA:GLY 1:0, 1:1, 1:2, 2:3, 2:1, 4:3). Purified recombinant glutathione-s-transferase (GST), green fluorescent protein (GFP) and GST fused in frame to GFP (GST-GFP) (model proteins) were characterized (SDS PAGE, Western blotting, immune-detection, and high sensitivity differential scanning calorimetry) and loaded (3.3, 6.6 and 30.2mg/g of film) into SA:GLY 1:2 film. These were characterized using texture analysis, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), scanning electron microscopy, swelling, adhesion, dissolution and circular dichroism (CD). The protein loaded dressings were uniform, with a good balance between flexibility and toughness. The films showed ideal moisture content required for protein conformation (TGA), interactions between proteins and film components (DSC), indicating stability which was confirmed by CD. Swelling and adhesion showed that formulations containing 6.6mg/g of protein possessed ideal characteristics and used for in vitro dissolution studies. Protein release was rapid initially and sustained over 72h and data fitted to various kinetic equations showed release followed zero-order and Fickian diffusion. The results demonstrate the potential of SA dressings for delivering therapeutic proteins to wounds.

摘要

本研究旨在开发并表征稳定的薄膜,作为潜在的伤口蛋白质递送敷料。薄膜由海藻酸钠(SA)和甘油(GLY)的水凝胶制备而成(SA:GLY比例为1:0、1:1、1:2、2:3、2:1、4:3)。对纯化的重组谷胱甘肽 - S - 转移酶(GST)、绿色荧光蛋白(GFP)以及与GFP读码框融合的GST(GST - GFP)(模型蛋白)进行表征(SDS - PAGE、蛋白质印迹法、免疫检测和高灵敏度差示扫描量热法),并将其载入SA:GLY 1:2的薄膜中(载入量为3.3、6.6和30.2mg/g薄膜)。使用质地分析、差示扫描量热法(DSC)、热重分析(TGA)、扫描电子显微镜、溶胀、粘附、溶解和圆二色性(CD)对这些薄膜进行表征。载入蛋白质的敷料均匀,在柔韧性和韧性之间取得了良好平衡。薄膜显示出蛋白质构象所需的理想水分含量(TGA)、蛋白质与薄膜成分之间的相互作用(DSC),CD证实了其稳定性。溶胀和粘附表明,含有6.6mg/g蛋白质的制剂具有理想特性,并用于体外溶解研究。蛋白质释放最初迅速,并持续72小时,拟合各种动力学方程的数据表明释放遵循零级和菲克扩散。结果证明了SA敷料向伤口递送治疗性蛋白质地潜力。

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