利用微透析研究皮肤屏障破坏后的早期事件——人离体皮肤模型

K. Baumann, N. P. Knudsen, A. O. Gadsbøll, A. Woetmann, P. Skov
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摘要

磨料损伤后的皮肤屏障修复是由皮肤驻留细胞释放介质促进的,这一过程主要在小鼠或简化的人体系统中进行了研究,之前的研究主要集中在有限数量的生物标志物上。在这里,我们展示了如何在人类离体皮肤模型中研究由皮肤屏障破坏引起的早期事件。通过皮肤微透析从间质液中回收10个相关生物标志物,随后使用多重平台进行样本分析。作为对照,将从微透析取样获得的生物标志物谱与皮肤活检匀浆谱进行比较。我们发现,10种生物标志物中的9种(GM-CSF、CXCL1/GROα、CXCL8/IL-8、CXCL10/IP-10、IL-1α、IL-6、MIF、TNF-α和VEGF)在磨料损伤后显著上调。只有透析液的CCL27/CTACK水平不受皮肤磨损的影响。匀浆中的生物标志物水平与透析液中CCL27/CTACK、CXCL1/GROα、CXCL8/IL-8和IL-6的水平一致。然而,IL-1α对创伤的反应呈相反趋势,MIF的活检水平没有变化。活检匀浆中未检测到GM-CSF、CXCL10/IP-10、TNF-α和VEGF。我们的研究结果表明,人类离体皮肤模型是研究皮肤屏障破坏后早期事件的可靠方法。
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Investigating the Early Events after Skin-Barrier Disruption Using Microdialysis—A Human Ex Vivo Skin Model
Skin-barrier restoration following abrasive trauma is facilitated by mediator release from skin-resident cells, a process that has been investigated primarily in mice or simplified human systems with previous studies focusing on a limited number of biomarkers. Here, we demonstrate how early events caused by skin-barrier disruption can be studied in a human ex vivo skin model. Ten relevant biomarkers were recovered from the interstitial fluid by skin microdialysis with subsequent sample analysis using a multiplex platform. As a control, the biomarker profiles obtained from microdialysis sampling were compared to profiles of skin biopsy homogenates. We found that nine (GM-CSF, CXCL1/GROα, CXCL8/IL-8 CXCL10/IP-10, IL-1α, IL-6, MIF, TNF-α, and VEGF) of the 10 biomarkers were significantly upregulated in response to abrasive trauma. Only dialysate levels of CCL27/CTACK were unaffected by skin abrasion. Biomarker levels in the homogenates corresponded to dialysate levels for CCL27/CTACK, CXCL1/GROα, CXCL8/IL-8, and IL-6. However, IL-1α showed an inverse trend in response to trauma, and biopsy levels of MIF were unchanged. GM-CSF, CXCL10/IP-10, TNF-α, and VEGF were not detected in the biopsy homogenates. Our results suggest that the human ex vivo skin model is a reliable approach to study early events after disruption of the skin barrier.
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