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Application of microscopic methods in the research of atopic dermatitis

Agnieszka Koszewska

Abstract

Atopic dermatitis (AD) is an allergic disease, mainly characterized by dry reddened skin with pruritic lesions. It is very difficult to make an accurate diagnosis of the AD. It consists of several criteria, one of which is a thorough interview with the patient. Moreover, there is also the assessment of the skin lesion (place of occurrence and severity) and also numerous clinical trials. Atopic dermatitis is an incurable disease – the therapy primarily focuses on reliving skin itching and more importantly, ensuring proper skin hydration and preventing inflammation of skin. The treatment depends on the phase and severity of the disease, and is selected individually, although it is crucial to maintain proper skin hydration with the use of emollients. The main aim of this thesis is the application of microscopic methods to thoroughly examine the properties of corneocytes from the epidermis of patients with atopic dermatitis. The tests that were carried out and the obtained results in the future may help us in designing an accurate procedure to assess the differences in both healthy cells and these with eczematous changes. Skin samples were collected by tape-stripping method. The comparative analysis of the obtained results of the tested parameters was conducted using two microscopic methods. Morphological properties of cells were examined using atomic force microscopy (based on topographic images) and phase contrast microscopy (after cell staining with hematoxylin and eosin). The examined morphological parameters were diameter and height of corneocytes. In addition, the mechanical properties of cells were examined with the use of atomic force microscope in force spectroscopy mode. The Young’s modulus value and adhesion force was analyzed. Two layers of cells were collected from each patient from the same place on the skin – the first one for staining and the analysis with inverted microscope, the second one to analyze with AFM. The analysis of the healthy cells collected from several layers was also conducted, with the aim to assess changes in values of examined parameters between layers. The cells diameter values, that were obtain with the use of the optical microscope and topographic images from AFM were compared. Based on the collected results it was found that the value of examined parameter decreases with successive layers of corneocytes. Based on the results, it was observed that regardless of the used method, the diameter value of corneocytes that were taken from places affected with AD was lower, than healthy ones, while their height value was higher. At the same time, the decrease in the volume of corneocytes taken from affected by AD places was observed, compared to corneocytes from healthy skin (6 out of 7 cases). It was also found, that the cytoplasm (from AD cells) binds the dye better than the cytoplasm of cells without eczematous changes. However, it was not possible to make a clear assessment of the dependence in mechanical properties of corneocytes, due to small size of the investigated patient group. In 4 cases the value of Young’s modulus and adhesive force turned out to be higher in AD cells and for others (3 cases) the inverse relation was observed.
Diploma type
Engineer's / Bachelor of Science
Diploma type
Engineer's thesis
Author
Agnieszka Koszewska (FC) Agnieszka Koszewska,, Faculty of Chemistry (FC)
Title in Polish
Wykorzystanie metod mikroskopowych do badania atopowego zapalenia skóry (AZS)
Supervisor
Tomasz Kobiela (FC/CDSB) Tomasz Kobiela,, Chair of Drug and Cosmetics Biotechnology (FC/CDSB)Faculty of Chemistry (FC)
Certifying unit
Faculty of Chemistry (FC)
Affiliation unit
Chair of Drug and Cosmetics Biotechnology (FC/CDSB)
Study subject / specialization
, Biotechnologia
Language
(pl) Polish
Status
Finished
Defense Date
29-01-2019
Issue date (year)
2019
Reviewers
Łukasz Górski (FC/CMB) Łukasz Górski,, Chair of Medical Biotechnology (FC/CMB)Faculty of Chemistry (FC) Tomasz Kobiela (FC/CDSB) Tomasz Kobiela,, Chair of Drug and Cosmetics Biotechnology (FC/CDSB)Faculty of Chemistry (FC)
Keywords in Polish
mikroskop sił atomowych, mikroskop odwróconego pola, atopowe zapalenie skóry, korneocyty, metoda tape-stripping
Keywords in English
atomic force microscopy, inverted microscope, atopic dermatitis, corneocytes, tape-stripping
Abstract in Polish
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Decyzja_o_utajnieniu_Koszewska_Agnieszka_276133.pdf
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Identyfikator pracy APD: 28346

Uniform Resource Identifier
https://repo.pw.edu.pl/info/bachelor/WUTa4095743da884c1eaa7908fb21309995/
URN
urn:pw-repo:WUTa4095743da884c1eaa7908fb21309995

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