Development of an Efficient Extraction Methodology to Analyse Potential Inflammatory Biomarkers from Sebum.

IF 2.8 4区 医学 Q2 DERMATOLOGY Skin Pharmacology and Physiology Pub Date : 2023-01-01 DOI:10.1159/000528653
Hemalatha Jayabal, Dan L Bader, Peter Worsley
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引用次数: 3

Abstract

Introduction: Proteins, such as cytokines and chemokines, are present in varying concentrations in a range of biofluids, with an important signalling role in maintaining homeostasis. Commercial tapes have been employed to non-invasively collect these potential biomarkers in sebum from the skin surface to examine their concentrations in conditions including acne, atopic dermatitis, and pressure ulcers. However, the identification of robust biomarker candidates is limited by the low abundance of specific proteins extracted by current methodologies. Therefore, this study was designed to develop an optimized extraction method for potential inflammatory biomarkers in sebum collected with Sebutapes.

Methods: Commercial tapes (Sebutapes) coated with synthetic sebum were used to systematically evaluate the effects of chemical and mechanical stimuli on extraction efficiency. Varying concentrations of high- and low-abundance biomarkers (IL-1α, IL-6, IL-8, INF-γ, TNF-α, and IL-1RA) were used to spike the synthetic sebum samples. Methodological variables included different surfactants, mechanical stimuli, and buffer volume. Extraction efficiency was estimated using immunoassay kits from the extracted buffer.

Results: The results revealed that the use of a surfactant, i.e., β-dodecyl maltoside, in addition to the mechanical stimuli, namely, sonication and centrifugation, resulted in an increased recovery of cytokines, ranging from 80% for high-abundant cytokines, such as IL-1α and IL-1RA, and up to 50% for low-abundance cytokines, including TNF-α, IL-6 and IL-8. Compared to previous methods, the new extraction protocol resulted in between a 1.5-2.0-fold increase in extraction efficiency.

Conclusion: The study revealed that there was a high degree of variability in the extraction efficiency of different cytokines. However, improved efficiency was achieved across all cytokines with selective surfactants and mechanical stimuli. The optimised protocol will provide means to detect low levels of potential biomarkers from skin surface, enabling the evaluation of local changes in pro- and anti-inflammatory cytokines present in different skin conditions.

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开发一种有效的提取方法来分析皮脂中潜在的炎症生物标志物。
蛋白质,如细胞因子和趋化因子,以不同的浓度存在于一系列生物体液中,在维持体内平衡中起着重要的信号作用。商业胶带已被用于非侵入性地收集皮肤表面皮脂中的这些潜在生物标志物,以检查其在痤疮、特应性皮炎和压疮等情况下的浓度。然而,现有方法提取的特异性蛋白质丰度较低,限制了生物标志物候选物的鉴定。因此,本研究旨在开发一种优化的提取方法,从油脂贴片收集的皮脂中提取潜在的炎症生物标志物。方法:采用人造皮脂包被商用胶带(油脂胶带),系统评价化学和机械刺激对提取效率的影响。使用不同浓度的高丰度和低丰度生物标志物(IL-1α, IL-6, IL-8, INF-γ, TNF-α和IL-1RA)来刺突合成皮脂样品。方法变量包括不同的表面活性剂、机械刺激和缓冲体积。用免疫测定试剂盒从提取的缓冲液中估计提取效率。结果:结果显示,使用表面活性剂,即β-十二烷基麦芽糖苷,加上机械刺激,即超声和离心,导致细胞因子的回收率增加,从80%的高丰度细胞因子,如IL-1α和IL-1RA,到高达50%的低丰度细胞因子,包括TNF-α, IL-6和IL-8。与以前的方法相比,新的提取方案的提取效率提高了1.5-2.0倍。结论:不同细胞因子的提取效率存在较大差异。然而,在选择性表面活性剂和机械刺激下,所有细胞因子的效率都得到了提高。优化后的方案将提供检测皮肤表面低水平潜在生物标志物的方法,从而能够评估不同皮肤状况下存在的促炎性和抗炎性细胞因子的局部变化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Skin Pharmacology and Physiology
Skin Pharmacology and Physiology 医学-皮肤病学
CiteScore
5.20
自引率
7.40%
发文量
23
审稿时长
>12 weeks
期刊介绍: In the past decade research into skin pharmacology has rapidly developed with new and promising drugs and therapeutic concepts being introduced regularly. Recently, the use of nanoparticles for drug delivery in dermatology and cosmetology has become a topic of intensive research, yielding remarkable and in part surprising results. Another topic of current research is the use of tissue tolerable plasma in wound treatment. Stimulating not only wound healing processes but also the penetration of topically applied substances into the skin, this novel technique is expected to deliver very interesting results.
期刊最新文献
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