The effect of epidermal growth factor (EGF) conjugated with low-molecular-weight protamine (LMWP) on wound healing of the skin
The effect of epidermal growth factor (EGF) conjugated with low-molecular-weight protamine (LMWP) on wound healing of the skin
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DOI：
10.1016/j.biomaterials.2012.07.061
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发表时间：
2012-11-01
期刊：
影响因子：
14
通讯作者：
Park, Jin Woo
中科院分区：
文献类型：
--
作者：
Choi, Jin Kyu;Jang, Ji-Hye;Park, Jin Woo
本研究旨在通过基因重组将高正电荷的鱼精蛋白（LMWP）连接到表皮生长因子（EGF）的N端，构建一种可透皮的重组低分子鱼精蛋白结合EGF（rLMWP-EGF）。我们评价了其生物活性、皮肤渗透性和体内伤口愈合功效。通过连续聚合酶链反应将编码LMWP氨基酸的cDNA克隆到EGF载体上。经表达和纯化后，获得了与LMWP位点特异性结合的重组EGF。体外细胞增殖活性在LMWP缀合后得到很好的保留，并且与rEGF相当。rLMWP-EGF通过三维人造人皮肤结构显示出显著改善的渗透性，并且rLMWP-EGF通过切除的小鼠皮肤的累积渗透比rEGF高约11倍。局部应用rLMWP-EGF显著加速全厚度伤口闭合速率以及糖尿病伤口模型，最可能是由于其增强的皮肤渗透。这些研究结果表明，作为一种新的局部伤口愈合药物的治疗潜力和LMWP的位点特异性共轭肽或蛋白质的基因重组作为一种有用的方法，用于制备高效的生物医学的rLMWP-EGF。(C)2012爱思唯尔有限公司保留所有权利。
This study was designed to develop a skin permeable recombinant low-molecular-weight protamine (LMWP) conjugated epidermal growth factor (EGF) (rLMWP-EGF) by linking a highly positive charged LMWP to the N-terminal of EGF through genetic recombination. We evaluated its biological activity, skin permeability, and wound healing efficacy in vivo. The cDNA for rLMWP-EGF was prepared by serial polymerase chain reaction for encoding amino acids of LMWP to the vector for EGF. After expression and purification, recombinant EGF site-specifically conjugated with LMWP was obtained. The in vitro cell proliferation activity was well preserved after LMWP conjugation and was comparable to that of rEGF. rLMWP-EGF showed markedly improved permeability through the three-dimensional artificial human skin constructs, and the cumulative permeation of rLMWP-EGF across the excised mouse skin was about 11 times higher than that of rEGF. Topically applied rLMWP-EGF significantly accelerated the wound closure rate in full thickness as well as a diabetic wound model most probably due to its enhanced skin permeation. These findings demonstrate the therapeutic potential of rLMWP-EGF as a new topical wound healing drug and the site-specific conjugation of LMWP to peptides or proteins by genetic recombination as a useful method for preparing highly effective biomedicines. (C) 2012 Elsevier Ltd. All rights reserved.