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.

1.
FDA-NIH Biomarker Working Group. BEST (Biomarkers, EndpointS, and other tools) resource. In: GroupNBW F-NBW, editor. Silver spring (MD): Food and Drug Administration (US); 2016.
2.
Califf RM. Biomarker definitions and their applications. Exp Biol Med. 2018;243(3):213–21.
3.
Niemeyer-van der Kolk T, van der Wall HEC, Balmforth C, Van Doorn MBA, Rissmann R. A systematic literature review of the human skin microbiome as biomarker for dermatological drug development. Br J Clin Pharmacol. 2018 2018/10/01;84(10):2178–93.
4.
EPUAP; NPIA; PPPPIA. Prevention and treatment of pressure ulcers/injuries: clinical practice guideline. The international guideline. In: Haesler E, editor. European pressure ulcer advisory Panel, national pressure injury advisory Panel and Pan pacific pressure injury alliance; 2019.
5.
Kottner J, Balzer K. Do pressure ulcer risk assessment scales improve clinical practice? J Multidiscip Healthc. 2010;3:103–11.
6.
Moore ZE, Patton D. Risk assessment tools for the prevention of pressure ulcers. Cochrane Database Syst Rev. 2019;1(1):CD006471–CD71.
7.
Larsen CM, Faulenbach M, Vaag A, Vølund A, Ehses JA, Seifert B, et al. Interleukin-1–Receptor antagonist in type 2 diabetes mellitus. N Engl J Med. 2007;356(15):1517–26.
8.
Alnek K, Kisand K, Heilman K, Peet A, Varik K, Uibo R. Increased blood levels of growth factors, proinflammatory cytokines, and Th17 cytokines in patients with newly diagnosed type 1 diabetes. PLoS One. 2015;10(12):e0142976.
9.
de Mooij CEM, Netea MG, van der Velden WJFM, Blijlevens NMA. Targeting the interleukin-1 pathway in patients with hematological disorders. Blood. 2017 Jun 15;129(24):3155–64.
10.
Mantovani A, Barajon I, Garlanda C. IL-1 and IL-1 regulatory pathways in cancer progression and therapy. Immunol Rev. 2018 Jan;281(1):57–61.
11.
Soetens JFJ, Worsley PR, Bader DL, Oomens CWJ. Investigating the influence of intermittent and continuous mechanical loading on skin through non-invasive sampling of IL-1α. J Tissue Viability. 2019 Feb;28(1):1–6.
12.
Soetens JFJ, Worsley PR, Herniman JM, Langley GJ, Bader DL, Oomens CWJ. The expression of anaerobic metabolites in sweat and sebum from human skin subjected to intermittent and continuous mechanical loading. J Tissue Viability. 2019;28(4):186–93.
13.
Rashmi R, Rao KS, Basavaraj KH. A comprehensive review of biomarkers in psoriasis. Clin Exp Dermatol. 2009;34(6):658–63.
14.
Minematsu T, Nakagami G, Sari Y, Akase T, Sugama J, Nagase T, et al. Candidate biomarkers for deep tissue damage from molecular biological and biochemical aspects. J Tissue Viability. 2010 May;19(2):77–83.
15.
de Wert LA, Bader DL, Oomens CW, Schoonhoven L, Poeze M, Bouvy ND. A new method to evaluate the effects of shear on the skin. Wound Repair Regen. 2015 Nov–Dec;23(6):885–90.
16.
Hemmes B, de Wert LA, Brink PRG, Oomens CWJ, Bader DL, Poeze M. Cytokine IL1α and lactate as markers for tissue damage in spineboard immobilisation. A prospective, randomised open-label crossover trial. J Mech Behav Biomed Mater. 2017 Nov;75:82–8.
17.
Kimura N, Nakagami G, Minematsu T, Sanada H. Non-invasive detection of local tissue responses to predict pressure ulcer development in mouse models. J Tissue Viability. 2020;29(1):51–7.
18.
Andersson AM, Sølberg J, Koch A, Skov L, Jakasa I, Kezic S, et al. Assessment of biomarkers in pediatric atopic dermatitis by tape strips and skin biopsies. Allergy. 2022;77(5):1499–509.
19.
Peeters EA, Oomens CW, Bouten CV, Bader DL, Baaijens FP. Mechanical and failure properties of single attached cells under compression. J Biomech. 2005;38(8):1685–93.
20.
Bronneberg D, Spiekstra SW, Cornelissen LH, Oomens CW, Gibbs S, Baaijens FP, et al. Cytokine and chemokine release upon prolonged mechanical loading of the epidermis. Exp Dermatol. 2007 Jul;16(7):567–73.
21.
Worsley PR, Prudden G, Gower G, Bader DL. Investigating the effects of strap tension during non-invasive ventilation mask application: a combined biomechanical and biomarker approach. Med Devices (Auckl). 2016;9:409–17.
22.
Koudounas S, Bader DL, Voegeli D. Investigating the release of inflammatory cytokines in a human model of incontinence-associated dermatitis. J Tissue Viability. 2021;30(3):427–33.
23.
Bronneberg D. Biochemical markers for early detection of superficial pressure ulcers. In: Soft tissue engineering and mechanobiology: Technische Universiteit Eindhoven; 2007.
24.
van der Lans MTC. Biochemical marker for early and objective detection of grade 1 pressure ulcers. Biomed Eng. 2007.
25.
Perkins MA, Osterhues MA, Farage MA, Robinson MK. A noninvasive method to assess skin irritation and compromised skin conditions using simple tape adsorption of molecular markers of inflammation. Skin Res Technol. 2001;7(4):227–37.
26.
Lee JM, Carson R, Arce C, Mahajan M, Lobst S. Development of a minimally invasive epidermal abrasion device for clinical skin sampling and its applications in molecular biology. Int J Cosmet Sci. 2009;31(1):27–39.
27.
Minematsu T, Horii M, Oe M, Sugama J, Mugita Y, Huang L, et al. Skin blotting: a noninvasive technique for evaluating physiological skin status. Adv Skin Wound Care. 2014 Jun;27(6):272–9.
28.
Piérard GE, Piérard-Franchimont C, Paquet P, Hermanns-Lê T, Radermacher J, Delvenne P. Cyanoacrylate skin surface stripping and the 3S-Biokit advent in tropical dermatology: a look from Liège. ScientificWorldJournal. 2014;2014:462634.
29.
Clausen ML, Slotved HC, Krogfelt KA, Agner T. Tape stripping technique for stratum corneum protein analysis. Sci Rep. 2016;6(1):19918.
30.
Falcone D, Spee P, Salk K, Peppelman M, van de Kerkhof PCM, van Erp PEJ. Measurement of skin surface biomakers by Transdermal Analyses Patch following different in vivo models of irritation: a pilot study. Skin Res Technol. 2017;23(3):336–45.
31.
Perkins MA, Cardin CW, Osterhues MA, Robinson MK. A non-invasive tape absorption method for recovery of inflammatory mediators to differentiate normal from compromised scalp conditions. Skin Res Technol. 2002 Aug;8(3):187–93.
32.
Bostan LE, Worsley PR, Abbas S, Bader DL. The influence of incontinence pads moisture at the loaded skin interface. J Tissue Viability. 2019 Aug;28(3):125–32.
33.
Renert-Yuval Y, Thyssen JP, Bissonnette R, Bieber T, Kabashima K, Hijnen D, et al. Biomarkers in atopic dermatitis-a review on behalf of the International eczema council. J Allergy Clin Immunol. 2021;147(4):1174–90.e1.
34.
Qiu Z, Zhu Z, Liu X, Chen B, Yin H, Gu C, et al. A dysregulated sebum-microbial metabolite-IL-33 axis initiates skin inflammation in atopic dermatitis. J Exp Med. 2022 Oct 3;219(10):e20212397.
35.
Bader D, Oomens C. The potential of biomarkers in the early detection of pressure ulcers. In: Romanelli M, Clark M, Gefen A, Ciprandi G, editors. Science and practice of pressure ulcer management. London: Springer; 2018. p. 1–15.
36.
Nakai A, Minematsu T, Tamai N, Sugama J, Urai T, Sanada H. Prediction of healing in category I pressure ulcers by skin blotting with plasminogen activator inhibitor 1, interleukin-1α, vascular endothelial growth factor C, and heat shock protein 90α: a pilot study. J Tissue Viability. 2019;28(2):87–93.
37.
Seddon AM, Curnow P, Booth PJ. Membrane proteins, lipids and detergents: not just a soap opera. Biochim Biophys Acta. 2004;1666(1–2):105–17.
38.
Andersen KK, Oliveira CL, Larsen KL, Poulsen FM, Callisen TH, Westh P, et al. The role of decorated SDS micelles in sub-CMC protein denaturation and association. J Mol Biol. 2009;391(1):207–26.
39.
Arachea BT, Sun Z, Potente N, Malik R, Isailovic D, Viola RE. Detergent selection for enhanced extraction of membrane proteins. Protein Expr Purif. 2012 Nov;86(1):12–20.
40.
Lu GW, Valiveti S, Spence J, Zhuang C, Robosky L, Wade K, et al. Comparison of artificial sebum with human and hamster sebum samples. Int J Pharm. 2009 Feb 9;367(1–2):37–43.
41.
Palazzo G, Lopez F, Mallardi A. Effect of detergent concentration on the thermal stability of a membrane protein: the case study of bacterial reaction center solubilized by N,N-dimethyldodecylamine-N-oxide. Biochim Biophys Acta. 2010 Jan;1804(1):137–46.
42.
de Jager W, Bourcier K, Rijkers GT, Prakken BJ, Seyfert-Margolis V. Prerequisites for cytokine measurements in clinical trials with multiplex immunoassays. BMC Immunol. 2009;10:52.
43.
Simpson S, Kaislasuo J, Guller S, Pal L. Thermal stability of cytokines: a review. Cytokine. 2020 Jan;125:154829.
44.
Krishnan S, Raibekas AA. Multistep aggregation pathway of human interleukin-1 receptor antagonist: kinetic, structural, and morphological characterization. Biophys J. 2009;96(1):199–208.
45.
Yang Z, Wang C, Zhou Q, An J, Hildebrandt E, Aleksandrov LA, et al. Membrane protein stability can be compromised by detergent interactions with the extramembranous soluble domains. Protein Sci. 2014;23(6):769–89.
46.
Goldenzweig A, Fleishman SJ. Principles of protein stability and their application in computational design. Annu Rev Biochem. 2018;87:105–29.
47.
Gray RJ. Impaired lymphatic clearance and pro-inflammatory cytokines in mechanically loaded tissue - implications in pressure ulcer aetiology. Southampton: Faculty of Health SciencesUniversity of Southampton; 2017.
48.
Koudounas S, Abbas S, Voegeli D. The effect of absorbent pad design on skin wetness, skin/pad microclimate, and skin barrier function: a quasi-experimental open cohort study. J Wound Ostomy Continence Nurs. 2020;47(5):497–506.
49.
Wei JCJ, Haridass IN, Crichton ML, Mohammed YH, Meliga SC, Sanchez WY, et al. Space- and time-resolved investigation on diffusion kinetics of human skin following macromolecule delivery by microneedle arrays. Sci Rep. 2018;8(1):17759.
50.
Jankovskaja S, Engblom J, Rezeli M, Marko-Varga G, Ruzgas T, Björklund S. Non-invasive skin sampling of tryptophan/kynurenine ratio in vitro towards a skin cancer biomarker. Sci Rep. 2021 Jan 12;11(1):678.
51.
Schaap MJ, Bruins FM, He X, Orro K, Peppelman M, van Erp PEJ, et al. Skin surface protein detection by transdermal analysis patches in pediatric psoriasis. Skin Pharmacol Physiol. 2021;34(5):271–80.
52.
Shi C, Shao W, Xiong Y, Tian C. A gas chromatographic method for quantification of detergents frequently used in membrane protein structural studies. Anal Biochem. 2008 Dec 15;383(2):326–8.
53.
Sheu HM, Chao SC, Wong TW, Yu-Yun Lee J, Tsai JC. Human skin surface lipid film: an ultrastructural study and interaction with corneocytes and intercellular lipid lamellae of the stratum corneum. Br J Dermatol. 1999;140(3):385–91.
54.
Ludovici M, Kozul N, Materazzi S, Risoluti R, Picardo M, Camera E. Influence of the sebaceous gland density on the stratum corneum lipidome. Sci Rep. 2018;8(1):11500.
55.
Ji W, Zhu J, Wu W, Wang N, Wang J, Wu J, et al. Wearable sweat biosensors refresh personalized health/medical diagnostics. Research (Wash D C). 2021;2021:9757126.
56.
Abiakam N, Jayabal H, Abbas S, Bader D, Worsley P. The effects of moistened incontinence pads on loaded skin with reference to biophysical and biochemical parameters. J Tissue Viability. 2022. Submitted for publication.
57.
Jayabal H, Abiakam NS, Bader D, Worsley P. An optimised method to analyse inflammatory markers from sebum and its role in detecting skin damage. Prague: EPUAP Annual Meeting; 2022.
You do not currently have access to this content.