Malmö University Publications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Non-invasive monitoring of low molecular weight biomarkers relevant to skin inflammation and cancer
Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
2021 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Development of skin inflammation and cancer in viable epidermis and dermis involve slow molecular weight (LMW) metabolites. We hypothesize that these LMW compounds can be collected on the surface of the skin and used for non-invasive diagnostics of skin disorders. Keeping in mind that substantial transdermal penetrationis achieved only for molecules of < 500 Da, we focused on topical monitoring of LMW biomarkers. In this thesis we investigated non-invasive, topical methods for monitoring LMW biomarkers by relevant in vitro and in vivo experiments. The LMW biomarkers were:

- reactive oxygen species (ROS), specifically, hydrogen peroxide, H2O2

- amino acids and their derivatives, i.e., tryptophan (Trp), kynurenine (Kyn; a Trpderivative), phenylalanine (Phe), and tyrosine (Tyr; a Phe derivative).

Initially, we have carried out in vitro experiments using dermatomed porcine skin and cell cultures. We characterized permeability of the biomarkers through skin and assessed methods of their monitoring. By using Prussian white particles, deposited on porcine skin, we demonstrated that hydrophilic biomarkers, such as H2O2, permeate the skin mainly through hair follicle pathways (Paper I). In paper II, we have showed that the enzymes transforming Trp to the inflammation and cancer biomarker Kyn, are expressed in the basal layer of epidermis. The magnitude of changes of the Trp/Kyn ratio in the cell culture model was assessed. In paper III, we have characterized Trp and Kyn permeability through skin in vitro, concluding that their permeabilities through stratum corneum are comparable. By in vivo experiments outlined in Paper IV, we have demonstrated the feasibility of topical, non-invasive sampling of Trp and Kyn, in relation to other amino acids. Kyn detection was compromised by its low abundance on the skin. In paper V, we performed a proof-of-concept study in vivo and confirmed that non-invasive sampling of Trp and amino acids of similar abundance, such as Phe and Tyr, is more robust. We concluded that Phe/Trp ratio might be equally good biomarker of skin disorders as a predicted Trp/Kyn ratio. Summarizing, the results of this thesis provide basic knowledge for deeper clinical studies of non-invasive, topical sampling of hydrophilic LMW biomarkers of skin inflammation and cancer.

Place, publisher, year, edition, pages
Malmö: Malmö universitet, 2021. , p. 66
Series
Malmö University Health and Society Dissertations, ISSN 1653-5383 ; 2021:9
Keywords [en]
Tryptophan, Kynurenine, Phenylalanine, Tyrosine, Low molecular Weight Biomarkers, Non-Invasive, Skin Surface Sampling, Skin Permeability, Hydrogels, Prussian White
National Category
Health Sciences
Research subject
Health and society
Identifiers
URN: urn:nbn:se:mau:diva-46859DOI: 10.24834/isbn.9789178772230ISBN: 978-91-7877-222-3 (print)ISBN: 978-91-7877-223-0 (electronic)OAI: oai:DiVA.org:mau-46859DiVA, id: diva2:1611579
Public defence
2021-11-29, Live stream and on location at HS assembly hall, Jan Waldenströms gata 25, ö, 09:00
Opponent
Supervisors
Note

Paper III published in dissertation as manuscript with title Non-invasive, Topical Sampling of Potential Skin Cancer Biomarkers,Kynurenine and Tryptophan: Study on Healthy Volunteers

Available from: 2021-11-15 Created: 2021-11-15 Last updated: 2022-11-09Bibliographically approved
List of papers
1. Visualisation of H2O2 penetration through skin indicates importance to develop pathway-specific epidermal sensing
Open this publication in new window or tab >>Visualisation of H2O2 penetration through skin indicates importance to develop pathway-specific epidermal sensing
Show others...
2020 (English)In: Microchimica Acta, ISSN 0026-3672, E-ISSN 1436-5073, Vol. 187, no 12, article id 656Article in journal (Refereed) Published
Abstract [en]

Elevated amounts of reactive oxygen species (ROS) including hydrogen peroxide (H2O2) are observed in the epidermis in different skin disorders. Thus, epidermal sensing of H2O2 should be useful to monitor the progression of skin pathologies. We have evaluated epidermal sensing of H2O2 in vitro, by visualising H2O2 permeation through the skin. Skin membranes were mounted in Franz cells, and a suspension of Prussian white microparticles was deposited on the stratum corneum face of the skin. Upon H2O2 permeation, Prussian white was oxidised to Prussian blue, resulting in a pattern of blue dots. Comparison of skin surface images with the dot patterns revealed that about 74% of the blue dots were associated with hair shafts. The degree of the Prussian white to Prussian blue conversion strongly correlated with the reciprocal resistance of the skin membranes. Together, the results demonstrate that hair follicles are the major pathways of H2O2 transdermal penetration. The study recommends that the development of H2O2 monitoring on skin should aim for pathway-specific epidermal sensing, allowing micrometre resolution to detect and quantify this ROS biomarker at hair follicles. Graphical abstract

Place, publisher, year, edition, pages
Springer, 2020
Keywords
Epidermal sensing, Hydrogen peroxide, Prussian blue, Hair follicles, Skin penetration
National Category
Dermatology and Venereal Diseases
Identifiers
urn:nbn:se:mau:diva-37075 (URN)10.1007/s00604-020-04633-9 (DOI)000589701400001 ()33188446 (PubMedID)2-s2.0-85095947121 (Scopus ID)
Available from: 2020-12-03 Created: 2020-12-03 Last updated: 2024-06-17Bibliographically approved
2. Effect of IFN-γ on the kynurenine/tryptophan ratio in monolayer-cultured keratinocytes and a 3D reconstructed human epidermis model
Open this publication in new window or tab >>Effect of IFN-γ on the kynurenine/tryptophan ratio in monolayer-cultured keratinocytes and a 3D reconstructed human epidermis model
Show others...
2020 (English)In: Journal of dermatological science (Amsterdam), ISSN 0923-1811, E-ISSN 1873-569X, Vol. 99, no 3, p. 177-184, article id S0923-1811(20)30234-6Article in journal (Refereed) Published
Abstract [en]

BACKGROUND: Interferon-gamma (IFN-γ) represents a potent inducer for keratinocyte inflammatory and immune activation in vitro. Since tryptophan (trp) conversion to kynurenine (kyn) is involved in inflammation, the topical kyn/trp ratio may serve as a biomarker of skin inflammation. However, the trp metabolism in keratinocytes exposed to IFN-γ is not yet fully understood.

OBJECTIVE: The aim of this study was to establish a human epidermis model in order to quantify cytokine and kyn/trp secretion from IFN-γ stimulated cells and tissues. Moreover, to compare the cell response of 2D-cultured keratinocytes and the 3D epidermis model.

METHODS: Polycarbonate filters were used on which primary keratinocytes could attach and stratify in order to form the typical layers of reconstructed human epidermis (RHE). After IFN-γ treatment, secretion of kyn/trp was measured by high performance liquid chromatography. Gene and protein expression of indoleamine 2,3-dioxygenase 1 (IDO) was analyzed with real-time PCR and immunohistochemistry. The secretion of cytokines was quantified with ELISA.

RESULTS: Trp catabolism to kyn was significantly increased (P < 0.01) in the 2D culture in response to IFN-γ treatment. Before kyn secretion, IDO was strongly upregulated (P < 0.001). IFN-γ treatment also increased the secretion of IL-6 and IL-8 from the keratinocytes. In the RHE, IDO was upregulated by IFN-γ, and kyn secretion could be detected. Interestingly, IDO expression was only present in the basal cells of the RHE.

CONCLUSION: Our results suggest that IFN-γ acts as an inducer of trp degradation preferentially in undifferentiated keratinocytes, indicated by the IDO expression in the basal layer of the RHE.

Place, publisher, year, edition, pages
Japanese Society for Investigative Dermatology, 2020
Keywords
IDO, Kynurenine, Pro-inflammatory cytokines, Reconstructed human epidermis, Tryptophan
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:mau:diva-18014 (URN)10.1016/j.jdermsci.2020.07.005 (DOI)000582365800005 ()32782183 (PubMedID)2-s2.0-85089257839 (Scopus ID)
Available from: 2020-08-18 Created: 2020-08-18 Last updated: 2024-02-05Bibliographically approved
3. Non-invasive skin sampling of tryptophan/kynurenine ratio in vitro towards a skin cancer biomarker
Open this publication in new window or tab >>Non-invasive skin sampling of tryptophan/kynurenine ratio in vitro towards a skin cancer biomarker
Show others...
2021 (English)In: Scientific Reports, E-ISSN 2045-2322, Vol. 11, no 1, article id 678Article in journal (Refereed) Published
Abstract [en]

The tryptophan to kynurenine ratio (Trp/Kyn) has been proposed as a cancer biomarker. Non-invasive topical sampling of Trp/Kyn can therefore serve as a promising concept for skin cancer diagnostics. By performing in vitro pig skin permeability studies, we conclude that non-invasive topical sampling of Trp and Kyn is feasible. We explore the influence of different experimental conditions, which are relevant for the clinical in vivo setting, such as pH variations, sampling time, and microbial degradation of Trp and Kyn. The permeabilities of Trp and Kyn are overall similar. However, the permeated Trp/Kyn ratio is generally higher than unity due to endogenous Trp, which should be taken into account to obtain a non-biased Trp/Kyn ratio accurately reflecting systemic concentrations. Additionally, prolonged sampling time is associated with bacterial Trp and Kyn degradation and should be considered in a clinical setting. Finally, the experimental results are supported by the four permeation pathways model, predicting that the hydrophilic Trp and Kyn molecules mainly permeate through lipid defects (i.e., the porous pathway). However, the hydrophobic indole ring of Trp is suggested to result in a small but noticeable relative increase of Trp diffusion via pathways across the SC lipid lamellae, while the shunt pathway is proposed to slightly favor permeation of Kyn relative to Trp.

Place, publisher, year, edition, pages
Nature Publishing Group, 2021
National Category
Cancer and Oncology
Identifiers
urn:nbn:se:mau:diva-39748 (URN)10.1038/s41598-020-79903-w (DOI)000621920400026 ()33436784 (PubMedID)2-s2.0-85099386099 (Scopus ID)
Available from: 2021-01-25 Created: 2021-01-25 Last updated: 2024-02-05Bibliographically approved
4. Non-Invasive, Topical Sampling of Potential, Low-Molecular Weight, Skin Cancer Biomarkers: A Study on Healthy Volunteers.
Open this publication in new window or tab >>Non-Invasive, Topical Sampling of Potential, Low-Molecular Weight, Skin Cancer Biomarkers: A Study on Healthy Volunteers.
Show others...
2022 (English)In: Analytical Chemistry, ISSN 0003-2700, E-ISSN 1520-6882, Vol. 94, no 15, p. 5856-5865Article in journal (Refereed) Published
Abstract [en]

Monitoring of low-molecular weight cancer biomarkers, such as tryptophan (Trp) and its derivative kynurenine (Kyn), might be advantageous to non-invasive skin cancer detection. Thus, we assessed several approaches of topical sampling of Trp and Kyn, in relation to phenylalanine (Phe) and tyrosine (Tyr), on the volar forearm of six healthy volunteers. The sampling was performed with three hydrogels (made of agarose or/and chitosan), hydrated starch films, cotton swabs, and tape stripping. The biomarkers were successfully sampled by all approaches, but the amount of collected Kyn was low, 20 ± 10 pmol/cm2. Kyn quantification was below LOQ, and thus, it was detected only in 20% of topical samples. To mitigate variability problems of absolute amounts of sampled amino acids, Tyr/Trp, Phe/Trp, and Phe/Tyr ratios were assessed, proving reduced inter-individual variation from 79 to 45% and intra-individual variation from 42 to 21%. Strong positive correlation was found between Phe and Trp, pointing to the Phe/Trp ratio (being in the 1.0–2.0 range, at 95% confidence) being least dependent on sampling materials, approaches, and sweating. This study leads to conclusion that due to the difficulty in quantifying less abundant Kyn, and thus the Trp/Kyn ratio, the Phe/Trp ratio might be a possible, alternative biomarker for detecting skin cancers.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2022
National Category
Cancer and Oncology
Identifiers
urn:nbn:se:mau:diva-51301 (URN)10.1021/acs.analchem.1c05470 (DOI)000792814500018 ()35394278 (PubMedID)2-s2.0-85128387453 (Scopus ID)
Available from: 2022-05-04 Created: 2022-05-04 Last updated: 2023-11-29Bibliographically approved
5. Hydrogels and Cubic Liquid Crystals for Non-Invasive Sampling of Low-Molecular-Weight Biomarkers-An Explorative In Vivo Study
Open this publication in new window or tab >>Hydrogels and Cubic Liquid Crystals for Non-Invasive Sampling of Low-Molecular-Weight Biomarkers-An Explorative In Vivo Study
Show others...
2022 (English)In: Pharmaceutics, E-ISSN 1999-4923, Vol. 14, no 2, article id 313Article in journal (Refereed) Published
Abstract [en]

The molecular composition of human skin is altered due to diseases, which can be utilized for non-invasive sampling of biomarkers and disease diagnostics. For this to succeed, it is crucial to identify a sampling formulation with high extraction efficiency and reproducibility. Highly hydrated skin is expected to be optimal for increased diffusion of low-molecular-weight biomarkers, enabling efficient extraction as well as enhanced reproducibility as full hydration represents a well-defined endpoint. Here, the aim was to explore water-based formulations with high water activities, ensuring satisfactory skin hydration, for non-invasive sampling of four analytes that may serve as potential biomarkers, namely tryptophan, tyrosine, phenylalanine, and kynurenine. The included formulations consisted of two hydrogels (chitosan and agarose) and two different liquid crystalline cubic phases based on the polar lipid glycerol monooleate, which were all topically applied for 2 h on 35 healthy subjects in vivo. The skin status of all sampling sites was assessed by electrical impedance spectroscopy and transepidermal water loss, enabling explorative correlations between biophysical properties and analyte abundancies. Taken together, all formulations resulted in the successful and reproducible collection of the investigated biomarkers. Still, the cubic phases had an extraction capacity that was approximately two times higher compared to the hydrogels.

Place, publisher, year, edition, pages
MDPI, 2022
Keywords
low-molecular-weight biomarker, tryptophan, kynurenine, tyrosine, phenylalanine, tryptophan-to-kynurenine ratio, skin barrier integrity, stratum corneum, natural moisturizing factor, electrical impedance spectroscopy, transepidermal water loss
National Category
Cancer and Oncology
Identifiers
urn:nbn:se:mau:diva-50952 (URN)10.3390/pharmaceutics14020313 (DOI)000765124800001 ()35214046 (PubMedID)2-s2.0-85124460011 (Scopus ID)
Available from: 2022-04-06 Created: 2022-04-06 Last updated: 2024-07-04Bibliographically approved

Open Access in DiVA

errata(340 kB)33 downloads
File information
File name ERRATA01.pdfFile size 340 kBChecksum SHA-512
e2e8a89a54f879912b7e804ed89751adc228f044d87ab7a86cb215d795c80e6e9908b0e7750acf8a03353fd3038b22c2d094ab102a6978c5c6f5c9c3f013f1aa
Type errataMimetype application/pdf
fulltext(19418 kB)89 downloads
File information
File name FULLTEXT03.pdfFile size 19418 kBChecksum SHA-512
dcac1db6c29f49979f8880b2f9d561fa135fd1f0a8b63d61bf2f380e626a323614ffc074274ae2c39fd06174bde02010f7b275784f157ef45c14bf34096ecd93
Type fulltextMimetype application/pdf

Other links

Publisher's full text

Authority records

Jankovskaja, Skaidre

Search in DiVA

By author/editor
Jankovskaja, Skaidre
By organisation
Department of Biomedical Science (BMV)Biofilms Research Center for Biointerfaces
Health Sciences

Search outside of DiVA

GoogleGoogle Scholar
Total: 218 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

doi
isbn
urn-nbn

Altmetric score

doi
isbn
urn-nbn
Total: 1489 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf