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血小板衍生伤口愈合因子中的免疫球蛋白 G。

Immunoglobulin G in Platelet-Derived Wound Healing Factors.

机构信息

Department of Physiology and Biochemistry, Faculty of Medicine and Surgery, Centre of Molecular Medicine and Biobanking, University of Malta and Division of Pathology, Mater Dei Hospital, Malta MSD2080.

Department of Surgery, Faculty of Medicine and Surgery, University of Malta Medical School and Mater Dei Hospital, Malta MSD2080.

出版信息

Biomed Res Int. 2021 Jan 28;2021:4762657. doi: 10.1155/2021/4762657. eCollection 2021.

Abstract

We intended to reformulate an existing platelet-derived wound healing formula to target each phase of the healing wound with the appropriate phase-specific molecules. A decreased perfusion of the skin, often associated with conditions such as thalassemia, sickle cell disease, diabetes mellitus, and chronic vascular disease, is the most common etiology of cutaneous ulcers and chronic wounds. We had previously shown that a PDWHF topically applied to a chronic nonhealing ulcer of a -thalassemia homozygote stimulated and accelerated closure of the wound. The PDWHF was prepared from a pooled platelet concentrate of a matching blood group, consisting of a combination of platelet -granule-derived factors. Processing of the apheresis-pooled platelets yielded various amounts of proteins (3.36 g/mL ± 0.25 (SD) ( = 10)) by the better lysis buffer method. Immunoglobulin G was found to be the most abundant -granule-secreted protein. Equally broad quantities of the IgG (10.76 ± 12.66% (SD) ( = 10)) and IgG/albumin ratios (0.6 ± 0.4 (SD) ( = 10)) were quantified. We have developed a method using a reformulated lysis buffer followed by size exclusion chromatography and affinity chromatography to extract, identify, quantify, and purify IgG from activated platelets. IgG purification was confirmed by Western blot and flow cytometry. It was thought unlikely that the platelet IgG could be accounted for by adsorption of plasma protein, though the variable quantities could account for diversity in wound healing rates. The IgG could protect the wound even from subclinical infections and functionally advance healing. It may be useful in the management of skin ulcers in the early phase of wound healing.

摘要

我们旨在重新配方现有的血小板衍生伤口愈合配方,以便用适当的特定于相的分子靶向愈合伤口的每个阶段。皮肤灌注减少,通常与地中海贫血、镰状细胞病、糖尿病和慢性血管疾病等情况相关,是皮肤溃疡和慢性伤口最常见的病因。我们之前曾表明,局部应用于 - 地中海贫血纯合子慢性未愈合溃疡的 PDWHF 可刺激并加速伤口闭合。PDWHF 是从匹配血型的血小板浓缩物中制备的,包含血小板 - 颗粒衍生因子的组合。通过更好的裂解缓冲液方法处理血小板分离浓缩物可产生不同量的蛋白质(3.36 g/mL ± 0.25(SD)(= 10))。免疫球蛋白 G 被发现是最丰富的 - 颗粒分泌蛋白。同样广泛的 IgG 量(10.76 ± 12.66%(SD)(= 10))和 IgG/白蛋白比(0.6 ± 0.4(SD)(= 10))被定量。我们已经开发了一种使用重新配方的裂解缓冲液,然后进行大小排阻色谱和亲和色谱的方法,从激活的血小板中提取、鉴定、定量和纯化 IgG。通过 Western blot 和流式细胞术证实了 IgG 的纯化。虽然血小板 IgG 的量可能会导致伤口愈合率的差异,但不太可能是由于血浆蛋白的吸附造成的。这种 IgG 甚至可以保护伤口免受亚临床感染,并在功能上促进愈合。它可能对伤口愈合早期的皮肤溃疡管理有用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6b99/7861922/7f664e9c910d/BMRI2021-4762657.001.jpg

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