共聚焦拉曼光谱在皮肤屏障功能评估和经皮吸收研究中的应用

doi:10.3969/j.issn.2097-2806.2025.04.013

摘要/Abstract

摘要：

在皮肤研究领域，共聚焦拉曼光谱是一项新兴的分析技术。它可以用于皮肤生理学分析，获得有关皮肤分子组成和结构的数据，并且该技术还具有高空间分辨率、非侵入性以及无需提前对目标物质进行标记（如使用荧光染料标记）的优点。基于以上优点，这项技术具有成为实验室标准化分析技术的巨大潜力。但共聚焦拉曼光谱技术目前仍处在一个不断发展和完善的阶段，文章介绍了共聚焦拉曼光谱的原理，重点强调了该技术的优点，并概述了共聚焦拉曼光谱技术在皮肤研究，特别是皮肤屏障功能和活性物质经皮吸收研究中的应用。同时分析了共聚焦拉曼光谱的局限性，并展望了共聚焦拉曼光谱的未来发展趋势。

关键词: 经皮吸收, 共聚焦拉曼光谱, 皮肤屏障, 局限性

Abstract:

Confocal Raman spectroscopy （CRS） is a new analytical technology in the field of percutaneous absorption, which enables physiology analysis to obtain information on the molecular composition and structure of the skin. Due to the advantages of high spatial resolution, non-invasive and non-pretreatment of sample compared to conventional methods, CRS possesses great potential to become a standardized laboratory analytical technique. Currently, the development and application of CRS is still in a stage of improvement. This review focuses on elucidating the principles and the advantages of this technology, and summarizes the application of CRS technique in skin research, including the skin barrier function assessment and permeation research. Furthermore, the limitations of CRS, such as the attenuation of signal and the interference of skin autofluorescence are analyzed, and the challenges and opportunities in CRS technology are also discussed.

Key words: percutaneous absorption, confocal Raman spectroscopy, skin barrier, limitation

中图分类号:

TQ658

杨武成, 谢宇, 魏剑, 范瑞芳, 谭建华, 席绍峰. 共聚焦拉曼光谱在皮肤屏障功能评估和经皮吸收研究中的应用[J]. 日用化学工业（中英文）, 2025, 55(4): 508-515.

Wucheng Yang, Yu Xie, Jian Wei, Ruifang Fan, Jianhua Tan, Shaofeng Xi. Application of confocal Raman spectroscopy in the evaluation of skin barrier function and permeability[J]. China Surfactant Detergent & Cosmetics, 2025, 55(4): 508-515.

图/表 5

图1

图2

图3

图4

表1

参考文献 56

[1]	Rajkumar J, Chandan N, Lio P, et al. The skin barrier and moisturization: function, disruption, and mechanisms of repair[J]. Skin Pharmacology and Physiology, 2023, 36(4): 174-185. doi: 10.1159/000534136 pmid: 37717558
[2]	Egawa M. Raman microscopy for skin evaluation[J]. The Analyst, 2021, 146(4): 1142-1150.
[3]	Bouwstra J A, Nădăban A, Bras W, et al. The skin barrier: an extraordinary interface with an exceptional lipid organization[J]. Progress in Lipid Research, 2023, 92: 101252
[4]	Gorzelanny C, Mess C, Schneider S W, et al. Skin barriers in dermal drug delivery: which barriers have to be overcome and how can we measure them?[J]. Pharmaceutics, 2020, 12(7): 684.
[5]	Mohammed D, Yang Q, Guy R H, et al. Comparison of gravimetric and spectroscopic approaches to quantify stratum corneum removed by tape-stripping[J]. European Journal of Pharmaceutics and Biopharmaceutics, 2012, 82(1): 171-174. doi: 10.1016/j.ejpb.2012.05.018 pmid: 22713518
[6]	Ali S M, Bonnier F, Lambkin H, et al. A comparison of Raman, FT IR and ATR-FT IR micro spectroscopy for imaging human skin tissue sections[J]. Analytical Methods, 2013, 5(9): 2281-2291.
[7]	Perticaroli S, Yeomans D J, Wireko F C, et al. Translating chemometric analysis into physiological insights from in vivo confocal Raman spectroscopy of the human stratum corneum[J]. Biochimica Biophysica Acta, 2019, 1861(2): 403-409.
[8]	Brzozowski K, Matuszyk E, Pieczara A, et al. Stimulated Raman scattering microscopy in chemistry and life science-development, innovation, perspectives[J]. Biotechnology Advances, 2022, 60: 108003.
[9]	Serebrennikova K V, Berlina A N, Sotnikov D V, et al. Raman scattering-based biosensing: new prospects and opportunities[J]. Biosensors, 2021, 11(12): 512.
[10]	Tancrède-Bohin E, Baldeweck T, Brizion S, et al. In vivo multiphoton imaging for non-invasive time course assessment of retinoids effects on human skin[J]. Skin Research and Technology, 2020, 26(6): 794-803.
[11]	Caspers P J, Lucassen G W, Puppels G J. Combined in vivo confocal Raman spectroscopy and confocal microscopy of human skin[J]. Biophysical Journal, 2003, 85(1): 572-580. pmid: 12829511
[12]	Sharma A, Sharma S, Zarrow A, et al. Raman spectroscopy: incorporating the chemical dimension into dermatological diagnosis[J]. Indian Journal of Dermatology, 2016, 61(1): 1-8. doi: 10.4103/0019-5154.173978 pmid: 26955087
[13]	Franzen L, Windbergs M. Applications of Raman spectroscopy in skin research—From skin physiology and diagnosis up to risk assessment and dermal drug delivery[J]. Advanced Drug Delivery Reviews, 2015, 89: 91-104. doi: 10.1016/j.addr.2015.04.002 pmid: 25868454
[14]	Nakagawa N, Matsumoto M, Sakai S. In vivo measurement of the water content in the dermis by confocal Raman spectroscopy[J]. Skin Research and Technology, 2010, 16(2): 137-141. doi: 10.1111/j.1600-0846.2009.00410.x pmid: 20456092
[15]	Kourbaj G, Bielfeldt S, Seise M, et al. Measurement of dermal water content by confocal Raman spectroscopy to investigate intrinsic aging and photoaging of human skin in vivo[J]. Skin Research and Technology, 2020, 27(3): 404-413.
[16]	Rigal A, Michael-Jubeli R, Nkengne A, et al. Raman confocal microscopy and biophysics multiparametric characterization of the skin barrier evolution with age[J]. Journal of Biophotonics, 2021, 14(9).
[17]	Caspers P J, Lucassen G W, Carter E A, et al. In vivo confocal Raman microspectroscopy of the skin: noninvasive determination of molecular concentration profiles[J]. The Journal of Investigative Dermatology, 2001, 116(3): 434-442.
[18]	Caspers P J, Lucassen G W, Bruining H A, et al. Automated depth-scanning confocal Raman microspectrometer for rapid in vivo determination of water concentration profiles in human skin[J]. Journal of Raman Spectroscopy, 2000, 31: 813-818.
[19]	Wu J, Polefka T G. Confocal Raman microspectroscopy of stratum corneum: a pre-clinical validation study[J]. International Journal of Cosmetic Science, 2008, 30(1): 47-56. doi: 10.1111/j.1468-2494.2008.00428.x pmid: 18377630
[20]	Egawa M, Kajikawa T. Changes in the depth profile of water in the stratum corneum treated with water[J]. Skin Research and Technology, 2009, 15(2): 242-249. doi: 10.1111/j.1600-0846.2009.00362.x pmid: 19622134
[21]	Egawa M, Tagami H. Comparison of the depth profiles of water and water-binding substances in the stratum corneum determined in vivo by Raman spectroscopy between the cheek and volar forearm skin: effects of age, seasonal changes and artificial forced hydration[J]. British Journal of Dermatology, 2007, 158(2): 251-260.
[22]	Rawlings A V, Harding C R. Moisturization and skin barrier function[J]. Dermatologic Therapy, 2004, 17: 43-48.
[23]	Van Mierlo M M F, Caspers P J, Jansen M S, et al. Natural moisturizing factor as a biomarker for filaggrin mutation status in a multi-ethnic paediatric atopic dermatitis cohort[J]. Clinical and Experimental Allergy, 2021, 51(11): 1510.
[24]	Zhang L, Cambron T, Niu Y, et al. Mcr approach revealing protein, water, and lipid depth profile in atopic dermatitis patients’ stratum corneum via in vivo confocal Raman spectroscopy[J]. Analytical Chemistry, 2019, 91(4): 2784-2790.
[25]	Rinnov M R, Halling A S, Gerner T, et al. Skin biomarkers predict development of atopic dermatitis in infancy[J]. Allergy, 2022, 78(3): 791-802. doi: 10.1111/all.15518 pmid: 36112082
[26]	Zolotas M, Schleusener J, Lademann J, et al. Atopic dermatitis: molecular alterations between lesional and non-lesional skin determined noninvasively by in vivo confocal Raman microspectroscopy[J]. International Journal of Molecular Sciences, 2023, 24(19): 14636.
[27]	Koppes S A, Kemperman P, Van Tilburg I, et al. Determination of natural moisturizing factors in the skin: Raman microspectroscopy versus HPLC[J]. Biomarkers, 2017, 22(6): 502-507. doi: 10.1080/1354750X.2016.1256428 pmid: 27805415
[28]	Caspers P J, Lucassen G W, Wolthuis R, et al. In vivo Raman spectroscopy of human skin: determination of the composition of natural moisturizing factor[J]. Biomedical Applications of Raman Spectroscopy, 1999, 3608: 99-102.
[29]	Richters R J H, Falcone D, Uzunbajakava N E, et al. Sensitive skin: assessment of the skin barrier using confocal Raman microspectroscopy[J]. Skin Pharmacology and Physiology, 2017, 30(1): 1-12. doi: 10.1159/000452152 pmid: 28122376
[30]	Stamatas G N, Roux P F, Boireau-Adamezyk E, et al. Skin maturation from birth to 10 years of age: Structure, function, composition and microbiome[J]. Experimental Dermatology, 2023, 32(9): 1420-1429.
[31]	Jacobi U, Kaiser M, Toll R, et al. Porcine ear skin: an in vitro model for human skin[J]. Skin Research and Technology, 2006, 13(1): 19-24.
[32]	Ashtikar M, Matthäus C, Schmitt M, et al. Non-invasive depth profile imaging of the stratum corneum using confocal Raman microscopy: First insights into the method[J]. European Journal of Pharmaceutical Sciences, 2013, 50(5): 601-608. doi: 10.1016/j.ejps.2013.05.030 pmid: 23764946
[33]	Hoppel M, Baurecht D, Holper E, et al. Validation of the combined ATR-FT IR/tape stripping technique for monitoring the distribution of surfactants in the stratum corneum[J]. International Journal of Pharmaceutics, 2014, 472(1/2): 88-93.
[34]	Egawa M, Hirao T, Takahash M. In vivo estimation of stratum corneum thickness from water concentration profiles obtained with Raman spectroscopy[J]. Acta Dermato-Venereologica, 2007, 87(1): 4-8. pmid: 17225007
[35]	Böhling A, Bielfeldt S, Himmelmann A, et al. Comparison of the stratum corneum thickness measured in vivo with confocal Raman spectroscopy and confocal reflectance microscopy[J]. Skin Research and Technology, 2013, 20(1): 50-57.
[36]	Tfayli A, Piot O, Pitre F, et al. Follow-up of drug permeation through excised human skin with confocal Raman microspectroscopy[J]. European Biophysics Journal, 2007, 36(8): 1049-1058.
[37]	Kourbaj G, Gaiser A, Bielfeldt S, et al. Assessment of penetration and permeation of caffeine by confocal Raman spectroscopy in vivo and ex vivo by tape stripping[J]. International Journal of Cosmetic Science, 2022, 45(1): 14-28. doi: 10.1111/ics.12820 pmid: 36350131
[38]	Krombholz R, Liu Y, Lunter D J. In-line and off-line monitoring of skin penetration profiles using confocal Raman spectroscopy[J]. Pharmaceutics, 2021, 13(1): 67.
[39]	Mélot M, Pudney P D A, Williamson A-M, et al. Studying the effectiveness of penetration enhancers to deliver retinol through the stratum cornum by in vivo confocal Raman spectroscopy[J]. Journal of Controlled Release, 2009, 138(1): 32-39. doi: 10.1016/j.jconrel.2009.04.023 pmid: 19401210
[40]	Mateus R, Moore D J, Hadgraft J, et al. Percutaneous absorption of salicylic acid-in vitro and in vivo studies[J]. International Journal of Pharmaceutics, 2014, 475(1/2): 471-474.
[41]	Iliopoulos F, Tang C F, Li Z, et al. Confocal Raman spectroscopy for assessing bioequivalence of topical formulations[J]. Pharmaceutics, 2023, 15(4): 1075.
[42]	Mohammed D, Matts P J, Hadgraft J, et al. In vitro-in vivo correlation in skin permeation[J]. Pharmaceutical Research, 2013, 31(2): 394-400.
[43]	Kichou H, Munnier E, Dancik Y, et al. Estimating the analytical performance of Raman spectroscopy for quantification of active ingredients in human stratum corneum[J]. Molecules, 2022, 27(9): 2843.
[44]	Caspers P J, Williams A C, Carter E A, et al. Monitoring the penetration enhancer dimethyl sulfoxide in human stratum corneum in vivo by confocal Raman spectroscopy[J]. Pharmaceutical Research, 2002, 19(10): 1577-1580. pmid: 12425479
[45]	Caspers P J, Nico C, Bakker Schut T C, et al. Method to quantify the in vivo skin penetration of topically applied materials based on confocal Raman spectroscopy[J]. Translational Biophotonics, 2019, 1(1/2).
[46]	He Y, Wu W, Li J, et al. In vivo Raman spectroscopy study on the stimulation mechanism of surfactant[J]. Skin Research and Technology, 2020, 26(6): 898-904.
[47]	Iliopoulos F, Caspers P J, Puppels G J, et al. Franz cell diffusion testing and quantitative confocal Raman spectroscopy: in vitro-in vivo correlation[J]. Pharmaceutics, 2020, 12(9): 887.
[48]	Singer S, Karrer S, Berneburg M. Modern sun protection[J]. Current Opinion in Pharmacology, 2019, 46: 24-28. doi: S1471-4892(18)30124-3 pmid: 30731327
[49]	Supe S, Takudage P. Methods for evaluating penetration of drug into the skin: a review[J]. Skin Res Technol, 2020, 27(3): 299-308.
[50]	Wang M, Tan J, Qi Z, et al. A combined study of skin penetration by confocal Raman spectroscopy and human metabolism: a case of benzophenone-3 in sunscreen[J]. Environmental Pollution, 2023, 340: 122368.
[51]	Tippavajhala V K, De Oliveira Mendes T, Martin A A. In vivo human skin penetration study of sunscreens by confocal Raman spectroscopy[J]. AAPS PharmSciTech, 2017, 19(2): 753-760.
[52]	Benson H a E. Assessment and clinical implications of absorption of sunscreens across skin[J]. American Journal of Clinical Dermatology, 2000, 1(4): 217-224. pmid: 11702366
[53]	Dai T, Pikkula B M, Wang L V, et al. Comparison of human skin opto-thermal response to near-infrared and visible laser irradiations: a theoretical investigation[J]. Physics in Medicine and Biology, 2004, 49(21): 4861-4877. pmid: 15584524
[54]	Ramírez-Elías M G, Alda J, González F J. Noise and artifact characterization of in vivo Raman spectroscopy skin measurements[J]. Applied Spectroscopy, 2012, 66(6): 650-655. doi: 10.1366/11-06495 pmid: 22732535
[55]	Schleusener J, Carrer V, Patzelt A, et al. Confocal Raman imaging of skin sections containing hair follicles using classical least squares regression and multivariate curve resolution-alternating least squares[J]. Quantum Electronics, 2019, 49(1): 6-12. doi: 10.1070/QEL16901
[56]	Wang H, Zhao J, Lee A M D, et al. Improving skin Raman spectral quality by fluorescence photobleaching[J]. Photodiagnosis and Photodynamic Therapy, 2012, 9(4): 299-302. doi: 10.1016/j.pdpdt.2012.02.001 pmid: 23200009

部位	人数	均值/μm	标准差
脸颊	15	16.8	2.84
上臂	15	21.8	3.63
前臂	14	22.6	4.33
手背	13	29.3	6.84
手掌	15	173.0	36.96