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从人皮肤中回收富含核心蛋白聚糖的组分提取物(D):一种加速方案。

Recovery of the Decorin-Enriched Fraction, Extract (D), From Human Skin: An Accelerated Protocol.

作者信息

Wheatley Denys N, Graham Emma, McMaster R Shannon, Muir Ian F K, Holmes John D, Davies Michaela

出版信息

J Biomed Biotechnol. 2004;2004(4):211-218. doi: 10.1155/S1110724304308089.

Abstract

The original extraction procedure of Engel and Catchpole [1] has often been used to recover decorin-enriched material from the skin. This material has a strong inhibitory effect on fibroblast proliferation, and clearly suppresses it in skin except after the first 5-6 days of wounding when new scaffold material is required. The aim of our present study has been to find and evaluate the product of a faster recovery method, and to check its consistency as a more reliable means of regularly obtaining sufficient material for topical application in wounds that might become hypertrophic. Modifications of the original Toole and Lowther [2] extraction procedure have been carefully evaluated in an attempt to cut preparation time without compromising biological activity of the inhibitory extract. We have devised a faster recovery procedure without compromising biological activity, even if initial recovery has been somewhat reduced. The latter problem could be offset by repeated cycles of the final extraction step. The main inhibitory activity is shown to be within the decorin-enriched "extract D," as the core protein and DSPG II. Adjustment of the extract towards neutrality after dialysis against water keeps most of the extracted protein in solution and yielded a decorin-enriched preparation that had a specific activity equivalent to that of the old method. It also yielded a fraction that was readily lyophilised to give a small amount of material that could be stored indefinitely without loss of activity and readily redissolved in aqueous solution. A reliable and relatively quick method is presented for the production, from human skin, of a decorin-enriched preparation that has strong fibroblast inhibitory action. The value of the procedure is that it is inexpensive and can produce the quantities that might be used topically in reducing hypertrophic scarring of wounds.

摘要

恩格尔和卡奇波尔[1]最初的提取方法常被用于从皮肤中回收富含核心蛋白聚糖的物质。这种物质对成纤维细胞增殖具有强烈的抑制作用,在皮肤中能明显抑制其增殖,但在伤口愈合的最初5 - 6天除外,此时需要新的支架材料。我们当前研究的目的是找到并评估一种更快回收方法的产物,并检查其一致性,作为一种更可靠的手段,以便定期获得足够的材料用于可能形成肥厚性瘢痕的伤口的局部应用。我们仔细评估了对图尔和洛瑟[2]原始提取方法进行的修改,试图在不损害抑制性提取物生物活性的前提下缩短制备时间。我们设计了一种更快的回收方法,即使初始回收率有所降低,也不会损害生物活性。后一个问题可以通过最终提取步骤的重复循环来弥补。主要的抑制活性显示在富含核心蛋白聚糖的“提取物D”中,即核心蛋白和双糖链蛋白聚糖II。用水透析后将提取物调至中性,可使大部分提取的蛋白质保持在溶液中,得到一种富含核心蛋白聚糖的制剂,其比活性与旧方法相当。它还产生了一种易于冻干的组分,得到少量可无限期储存而不失活性且能迅速重新溶解于水溶液中的材料。本文介绍了一种可靠且相对快速的方法,用于从人皮肤中制备具有强烈成纤维细胞抑制作用的富含核心蛋白聚糖的制剂。该方法的价值在于成本低廉,能够生产出可用于局部减少伤口肥厚性瘢痕的数量。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/99a4/555769/5bdae86f7c7b/30808.fig.001.jpg

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