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Comparison of suction blistering and tape stripping for analysis of epidermal genes, proteins and lipids

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Abstract

Analysis of epidermal genes, proteins and lipids is important in the research and diagnosis of skin diseases. Although punch biopsy is the first-choice technique for the skin sampling, it is unnecessarily invasive for obtaining a sample just for the epidermal analysis. Here we compare two less invasive methods, suction blistering (SB) and tape stripping (TS), for the analysis of selected epidermal genes (quantitative real-time reverse transcription PCR, qRT-PCR), proteins (western blotting, WB), and lipids in ten healthy volunteers. TS provided significantly less material than SB and no viable epidermal layers could be obtained according to the reflectance confocal microscopy. Consistently, only the SC protein filaggrin and housekeeping GAPDH together with FLG and RPL13A mRNA were detected by TS. In the SB samples, WB and qRT-PCR could easily detect all the selected proteins (claudin-1, occludin, filaggrin, laminin and GAPDH) and genes (CLDN1, OCLN, FLG, LAMA3 and RPL13A), respectively. A single SB sample further provided enough of material for immunohistochemistry and lipid analyses, which was not feasible with the TS samples. Immunohistochemistry of the SB samples showed intact epidermal structure and a characteristic expression of claudin-1. Infrared spectroscopy showed well-ordered lipids with both orthorhombic and hexagonal packing and high-performance thin layer chromatography confirmed all lipid classes (including ceramide subclasses) in correct proportions. Taken together, SB represents a reliable sampling technique that can be utilized for multipurpose epidermal analyses in various studies.

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Abbreviations

HPTLC:

High performance thin layer chromatography

IHC:

Immunohistochemistry

IR:

Infrared

qRT-PCR:

Quantitative real-time reverse transcription PCR

PL:

Phospholipids

RCM:

Reflectance confocal microscopy

SB:

Suction blistering

SC:

Stratum corneum

SG:

Stratum granulosum

TS:

Tape stripping

WB:

Western blotting

References

  1. Benfeldt E, Serup J, Menné T (1999) Microdialysis vs. suction blister technique for in vivo sampling of pharmacokinetics in the human dermis. Acta Derm Venereol 79:338–342

    Article  CAS  PubMed  Google Scholar 

  2. Bleck O, Abeck D, Ring J et al (1999) Two ceramide subfractions detectable in Cer(AS) position by HPTLC in skin surface lipids of non-lesional skin of atopic eczema. J Invest Dermatol 113:894–900

    Article  CAS  PubMed  Google Scholar 

  3. Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37:911–917

    Article  CAS  PubMed  Google Scholar 

  4. Boncheva M, Damien F, Normand V (2008) Molecular organization of the lipid matrix in intact Stratum corneum using ATR-FTIR spectroscopy. Biochim Biophys Acta 1778:1344–1355

    Article  CAS  PubMed  Google Scholar 

  5. Broccardo CJ, Mahaffey SB, Strand M et al (2009) Peeling off the layers: skin taping and a novel proteomics approach to study atopic dermatitis. J Allergy Clin Immunol 124:1113–1115

    Article  PubMed  Google Scholar 

  6. Brönneke S, Brückner B, Söhle J et al (2015) Genome-wide expression analysis of wounded skin reveals novel genes involved in angiogenesis. Angiogenesis 18:361–371

    Article  PubMed  Google Scholar 

  7. Camera E, Ludovici M, Galante M et al (2010) Comprehensive analysis of the major lipid classes in sebum by rapid resolution high-performance liquid chromatography and electrospray mass spectrometry. J Lipid Res 51:3377–3388

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Chao YC, Nylander-French LA (2004) Determination of keratin protein in a tape-stripped skin sample from jet fuel exposed skin. Ann Occup Hyg 48:65–73

    CAS  PubMed  Google Scholar 

  9. Chapman SJ, Walsh A, Jackson SM, Friedmann PS (1991) Lipids, proteins and corneocyte adhesion. Arch Dermatol Res 283:167–173

    Article  CAS  PubMed  Google Scholar 

  10. Clausen ML, Slotved HC, Krogfelt KA, Agner T (2016) Tape stripping technique for stratum corneum protein analysis. Sci Rep 6:19918

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Clausen ML, Jungersted JM, Andersen PS et al (2013) Human β-defensin-2 as a marker for disease severity and skin barrier properties in atopic dermatitis. Br J Dermatol 169:587–593

    Article  CAS  PubMed  Google Scholar 

  12. Egawa G, Doi H, Miyachi Y, Kabashima K (2013) Skin tape stripping and cheek swab method for a detection of filaggrin. J Dermatol Sci 69:263–265

    Article  CAS  PubMed  Google Scholar 

  13. Gupta S, Shroff S, Gupta S (1999) Modified technique of suction blistering for epidermal grafting in vitiligo. Int J Dermatol 38:306–309

    Article  CAS  PubMed  Google Scholar 

  14. Jacobi U, Weigmann HJ, Ulrich J et al (2005) Estimation of the relative stratum corneum amount removed by tape stripping. Skin Res Technol 11:91–96

    Article  CAS  PubMed  Google Scholar 

  15. Jungersted JM, Bomholt J, Bajraktari N et al (2013) In vivo studies of aquaporins 3 and 10 in human stratum corneum. Arch Dermatol Res 305:699–704

    Article  CAS  PubMed  Google Scholar 

  16. Kendall AC, Nicolaou A (2013) Bioactive lipid mediators in skin inflammation and immunity. Prog Lipid Res 52:141–164

    Article  CAS  PubMed  Google Scholar 

  17. Kendall AC, Pilkington SM, Massey KA et al (2015) Distribution of bioactive lipid mediators in human skin. J Invest Dermatol 135:1510–1520

    Article  CAS  PubMed  Google Scholar 

  18. Kováčik A, Opálka L, Šilarová M et al (2016) Synthesis of 6-hydroxyceramide using ruthenium-catalyzed hydrosilylation–protodesilylation. Unexpected formation of a long periodicity lamellar phase in skin lipid membranes. RSC Adv 6:73343–73350

    Article  Google Scholar 

  19. Lademann J, Ilgevicius A, Zurbau O et al (2006) Penetration studies of topically applied substances: optical determination of the amount of stratum corneum removed by tape stripping. J Biomed Opt 11:54026

    Article  CAS  Google Scholar 

  20. Leitch CS, Natafji E, Yu C et al (2016) Filaggrin-null mutations are associated with increased maturation markers on Langerhans cells. J Allergy Clin Immunol 138:482–490

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Leung DY (2013) New insights into atopic dermatitis: role of skin barrier and immune dysregulation. Allergol Int 62:151–161

    Article  CAS  PubMed  Google Scholar 

  22. Macdonald N, Cumberbatch M, Singh M et al (2006) Proteomic analysis of suction blister fluid isolated from human skin. Clin Exp Dermatol 31:445–448

    Article  CAS  PubMed  Google Scholar 

  23. Mazereeuw-Hautier J, Gres S, Fanguin M et al (2005) Production of lysophosphatidic acid in blister fluid: involvement of a lysophospholipase D activity. J Invest Dermatol 125:421–427

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Nachat R, Méchin MC, Takahara H et al (2005) Peptidylarginine deiminase isoforms 1–3 are expressed in the epidermis and involved in the deimination of K1 and filaggrin. J Invest Dermatol 124:384–393

    Article  CAS  PubMed  Google Scholar 

  25. Nischal U, Nischal K, Khopkar U (2008) Techniques of skin biopsy and practical considerations. J Cutan Aesthet Surg 1:107–111

    Article  PubMed  PubMed Central  Google Scholar 

  26. Oikarinen A, Savolainen ER, Tryggvason K et al (1982) Basement membrane components and galactosylhydroxylysyl glucosyltransferase in suction blisters of human skin. Br J Dermatol 106:257–266

    Article  CAS  PubMed  Google Scholar 

  27. Opálka L, Kováčik A, Sochorová M et al (2015) Scalable synthesis of human ultralong chain ceramides. Org Lett 17:5456–5459

    Article  PubMed  Google Scholar 

  28. Panoutsopoulou IG, Wendelschafer-Crabb G, Hodges JS, Kennedy WR (2009) Skin blister and skin biopsy to quantify epidermal nerves: a comparative study. Neurology 72:1205–1210

    Article  PubMed  Google Scholar 

  29. Panoutsopoulou IG, Luciano CA, Wendelschafer-Crabb G et al (2015) Epidermal innervation in healthy children and adolescents. Muscle Nerve 51:378–384

    Article  PubMed  PubMed Central  Google Scholar 

  30. Pinkus H (1952) Examination of the epidermis by the strip method II. Biometric data on regeneration of the human epidermis. J Invest Dermatol 19:431–447

    Article  CAS  PubMed  Google Scholar 

  31. Proksch E, Brandner JM, Jensen JM (2008) The skin: an indispensable barrier. Exp Dermatol 17:1063–1072

    Article  PubMed  Google Scholar 

  32. Puig S, Carrera C, Salerni G, Rocha-Portela J (2012) Epidermis, dermis and epidermal appendages. In: Hofmann-Wellenhof R, Pellacani G, Malvehy J, Soyer HP (eds) Reflectance confocal microscopy for skin diseases. Springer, New York, pp 23–31

    Chapter  Google Scholar 

  33. Saksela O, Alitalo K, Kiistala U, Vaheri A (1981) Basal lamina components in experimentally induced skin blisters. J Invest Dermatol 77:283–286

    Article  CAS  PubMed  Google Scholar 

  34. Sandby-Møller J, Poulsen T, Wulf HC (2003) Epidermal thickness at different body sites: relationship to age, gender, pigmentation, blood content, skin type and smoking habits. Acta Derm Venereol 83:410–413

    Article  PubMed  Google Scholar 

  35. Schreiner V, Gooris GS, Pfeiffer S et al (2000) Barrier characteristics of different human skin types investigated with X-ray diffraction, lipid analysis, and electron microscopy imaging. J Invest Dermatol 114:654–660

    Article  CAS  PubMed  Google Scholar 

  36. Serup J, Jemec GBE, Grove GL (eds) (2006) Handbook of non-invasive methods and the skin, 2nd edn. CRC Press, Boca Raton

    Google Scholar 

  37. Tagami H (2008) Location-related differences in structure and function of the stratum corneum with special emphasis on those of the facial skin. Int J Cosmet Sci 30:413–434

    Article  CAS  PubMed  Google Scholar 

  38. Thiele JJ, Hsieh SN, Briviba K, Sies H (1999) Protein oxidation in human stratum corneum: susceptibility of keratins to oxidation in vitro and presence of a keratin oxidation gradient in vivo. J Invest Dermatol 113:335–339

    Article  CAS  PubMed  Google Scholar 

  39. van Smeden J, Hoppel L, van der Heijden R et al (2011) LC/MS analysis of stratum corneum lipids: ceramide profiling and discovery. J Lipid Res 52:1211–1221

    Article  PubMed  PubMed Central  Google Scholar 

  40. Varila E, Rantala I, Oikarinen A et al (1995) The effect of topical oestradiol on skin collagen of postmenopausal women. Br J Obstet Gynaecol 102:985–989

    Article  CAS  PubMed  Google Scholar 

  41. Vávrová K, Henkes D, Strüver K et al (2014) Filaggrin deficiency leads to impaired lipid profile and altered acidification pathways in a 3D skin construct. J Invest Dermatol 134:746–753

    Article  PubMed  Google Scholar 

  42. Wallmeyer L, Lehnen D, Eger N et al (2015) Stimulation of PPARα normalizes the skin lipid ratio and improves the skin barrier of normal and filaggrin deficient reconstructed skin. J Dermatol Sci 80:102–110

    Article  CAS  PubMed  Google Scholar 

  43. Weigmann HJ, Lademann J, Meffert H et al (1999) Determination of the horny layer profile by tape stripping in combination with optical spectroscopy in the visible range as a prerequisite to quantify percutaneous absorption. Skin Pharmacol Appl Skin Physiol 12:34–45

    Article  CAS  PubMed  Google Scholar 

  44. Wong R, Tran V, Morhenn V et al (2004) Use of RT-PCR and DNA microarrays to characterize RNA recovered by non-invasive tape harvesting of normal and inflamed skin. J Invest Dermatol 123:159–167

    Article  CAS  PubMed  Google Scholar 

  45. Wong R, Tran V, Talwalker S, Benson NR (2006) Analysis of RNA recovery and gene expression in the epidermis using non-invasive tape stripping. J Dermatol Sci 44:81–92

    Article  CAS  PubMed  Google Scholar 

  46. Yuki T, Hachiya A, Kusaka A et al (2011) Characterization of tight junctions and their disruption by UVB in human epidermis and cultured keratinocytes. J Invest Dermatol 131:744–752

    Article  CAS  PubMed  Google Scholar 

  47. Zoschke C, Ulrich M, Sochorová M et al (2016) The barrier function of organotypic non-melanoma skin cancer models. J Control Release 233:10–18

    Article  CAS  PubMed  Google Scholar 

  48. Zuber TJ (2002) Punch biopsy of the skin. Am Fam Phys 65:1155–1158 (1161–1162, 1164)

    Google Scholar 

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Acknowledgements

We would like to thank Soňa Moravčíková for the technical assistance. This work was supported by Grant no. SGS_2017_004, Czech Science Foundation (16-25687J) and Charles University (SVV260401).

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Authors and Affiliations

  1. Department of Biological and Biochemical Sciences, Faculty of Chemical-Technology, University of Pardubice, Pardubice, Czech Republic

    Marek Svoboda

  2. Department of Research and Development, Contipro a.s., Dolní Dobrouč, Czech Republic

    Marek Svoboda, Marta Hlobilová, Markéta Marešová & Iva Dolečková

  3. Skin Barrier Research Group, Faculty of Pharmacy, Charles University, Hradec Králové, Czech Republic

    Michaela Sochorová, Andrej Kováčik & Kateřina Vávrová

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  1. Marek Svoboda
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Correspondence to Marek Svoboda.

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Svoboda, M., Hlobilová, M., Marešová, M. et al. Comparison of suction blistering and tape stripping for analysis of epidermal genes, proteins and lipids. Arch Dermatol Res 309, 757–765 (2017). https://doi.org/10.1007/s00403-017-1776-6

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  • DOI: https://doi.org/10.1007/s00403-017-1776-6

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