Qualifying X-ray and Stimulated Raman Spectromicroscopy for Mapping Cutaneous Drug Penetration
Qualifying X-ray and Stimulated Raman Spectromicroscopy for Mapping Cutaneous Drug Penetration
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DOI：
10.1021/acs.analchem.9b00519
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发表时间：
2019-06-04
影响因子：
7.4
通讯作者：
Schaefer-Korting, Monika
中科院分区：
文献类型：
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作者：
Wanjiku, Barbara;Yamamoto, Kenji;Schaefer-Korting, Monika
局部给药的研究除了依赖离体的人类和动物皮肤外，还依赖于重建的人类皮肤（RHS），每种皮肤都具有特定的生理特征。在这里，我们比较了地塞米松从乙醇羟乙基纤维素凝胶到离体人体皮肤、小鼠皮肤和 RHS 的渗透情况。为了全面了解皮肤形态和渗透增强机制，结合了扫描透射 X 射线显微镜 (STXM)、液相色谱串联质谱 (LC-MS/MS) 和受激拉曼光谱显微镜 (SRS)。 STXM 提供高空间分辨率和无标记药物检测，因此对组织损伤敏感。尽管样品制备和数据分析存在差异，但通过 STXM 和 LC-MS/MS 检测和定量的 RHS 中地塞米松的含量非常相似，并且在暴露的前 100 分钟内有所增加。 SRS 揭示了凝胶和角质层之间的相互作用，或者更具体地说，它的蛋白质和脂质结构。与两种类型的离体皮肤类似，RHS 角质层内较高的蛋白质与脂质比率表明乙醇暴露 30 分钟后脂质量减少。延长乙醇暴露导致离体基质中的脂质持续减少，而 RHS 中的蛋白质完整性似乎受到损害，从而导致蛋白质信号下降。总之，LC-MS/MS 证明了 STXM 对于无标记药物检测的预测能力。将 STXM 与 SRS 相结合，精确剖析了乙醇的渗透增强作用。关于局部给药的进一步研究应考虑这些补充技术的潜力。
Research on topical drug delivery relies on reconstructed human skin (RHS) in addition to ex vivo human and animal skin, each with specific physiological features. Here, we compared the penetration of dexamethasone from an ethanolic hydroxyethyl cellulose gel into ex vivo human skin, murine skin, and RHS. For comprehensive insights into skin morphology and penetration enhancing mechanisms, scanning transmission X-ray microscopy (STXM), liquid chromatography tandem mass spectrometry (LC-MS/MS), and stimulated Raman spectromicroscopy (SRS) were combined. STXM offers high spatial resolution with label-free drug detection and is therefore sensitive to tissue damage. Despite differences in sample preparation and data analysis, the amounts of dexamethasone in RHS, detected and quantified by STXM and LC-MS/MS, were very similar and increased during the first 100 min of exposure. SRS revealed interactions between the gel and the stratum corneum or, more specifically, its protein and lipid structures. Similar to both types of ex vivo skin, higher protein-to-lipid ratios within the stratum corneum of RHS indicated reduced lipid amounts after 30 min of ethanol exposure. Extended ethanol exposure led to a continued reduction of lipids in the ex vivo matrixes, while protein integrity appeared to be compromised in RHS, which led to declining protein signals. In conclusion, LC-MS/MS proved the predictive capability of STXM for label-free drug detection. Combining STXM with SRS precisely dissected the penetration enhancing effects of ethanol. Further studies on topical drug delivery should consider the potential of these complementary techniques.