Vrednotenje biološkega odziva keratinocitov NCTC-2544 na vlakna steklene in kamene volne

Trivunčević, Daša (avtor), Pišlar, Anja (mentor), Drnovšek, Nataša (komentor)

Pri proizvodnji in delu s stekleno ali kameno volno se delavci pogosto srečujejo z neprijetnimi občutki na koži, kot je srbečica, ki se razvije, ko vlakna pridejo v stik s kožo. Čeprav gre za pogost pojav, mehanizmi, ki vodijo do takšnega kožnega draženja, še niso povsem raziskani. Ker mineralna vlakna ne povzročajo kemične iritacije, se domneva, da je srbenje posledica njihovega mehanskega delovanja. Najverjetneje ga povzročajo drobni delci vlaken, ki ob stiku z zgornjo plastjo kože fizično dražijo povrhnjico. Namen magistrske naloge je bil ugotoviti, ali lahko mineralna vlakna mehansko dražijo keratinocite, celice najbolj zunanje plasti kože, ki prve pridejo v stik z zunanjimi dejavniki. V eksperimentalnem delu magistrske naloge smo ovrednotili vpliv izpostavljenosti različnih vrst mineralnih vlaken človeškim keratinocitom NCTC-2544. Vpliv mineralnih vlaken smo spremljali z različnimi metodami v koncentracijski in časovni odvisnosti. Vpliv vlaken na preživetje celic smo ocenili z metabolnim testom, morebitne poškodbe celične membrane smo določali s testom laktat dehidrogenaze, izražanje proteina akvaporin 3 smo analizirali z imunofluorescenčno detekcijo ter izražanje vnetnih dejavnikov s prenosom western. Spremembe v celični obliki in strukturi smo opazovali s svetlobnim invertnim mikroskopom, medtem ko nam je vrstična elektronska mikroskopija omogočila natančnejši vpogled v spremembe na celični površini. Rezultati so pokazali, da mineralna vlakna vplivajo na metabolno aktivnost keratinocitov, povzročajo poškodbe celične membrane in spremembe v morfologiji celic. Učinki so bili izrazitejši pri višjih koncentracijah in daljši izpostavljenosti, kar se je pokazalo kot zmanjšana vitalnost celic, spremembe oblike, izguba stika s podlago ter povečana izraženost in spremenjena lokalizacija akvaporina 3. Vnetni odziv sicer ni bil vedno zaznan, a opažene spremembe kažejo na vpliv vlaken na celične funkcije. Ti rezultati potrjujejo, da lahko stik z določenimi vrstami mineralnih vlaken vpliva na kožne celice in sproži odzive, ki so značilni za mehansko draženje. Raziskava prispeva k razumevanju vpliva fizikalnih lastnosti vlaken na kožne celice in je lahko v pomoč pri razvoju novih izdelkov.

• Jezik: Slovenski jezik
• Ključne besede: mineralna vlakna, steklena volna, kamena volna, mehansko draženje, keratinociti, poškodba membrane, vnetni odziv
• Vrsta gradiva: Magistrsko delo/naloga
• Tipologija: 2.09 - Magistrsko delo
• Organizacija: FFA - Fakulteta za farmacijo
• Leto izida: 2025
• PID: 20.500.12556/RUL-175187
• COBISS.SI-ID: 254053891
• Datum objave v RUL: 21.10.2025

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Evaluation of biological response of NCTC-2544 keratinocytes to glass and rock wool fibres

Izvleček

During the production and handling of glass or rock wool, workers often experience skin discomfort such as itching, which develops when the fibres come into contact with the skin. Although this is a common occurrence, the mechanisms leading to such skin irritation are not yet fully understood. Since mineral fibres do not cause chemical irritation, it is assumed that the itching results from their mechanical action. Most likely, it is caused by small fibre fragments that physically irritate the outermost layer of the skin. The aim of this Master's thesis was to investigate whether mineral fibres can mechanically irritate keratinocytes, the cells that make up the majority of the outermost layer of the skin and are the first to come into contact with the environment. In the experimental part of the thesis, we evaluated the effects of exposure to different types of mineral fibres on human keratinocytes NCTC-2544. The impact of mineral fibres was monitored using various methods in a concentration and time dependent manner. Cell viability was evaluated using a metabolic assay, while potential cell membrane damage was assessed using the lactate dehydrogenase release assay. Expression of the aquaporin 3 protein was analyzed using immunofluorescence detection, and the expression of inflammatory markers was assessed by western blotting. Changes in cell shape and structure were observed using light inverted microscopy, while scanning electron microscopy provided a more detailed view of changes on the cell surface. The results showed that mineral fibres affect keratinocyte matebolic activity, cause cell membrane damage, and induce changes in cell morphology. The effects were more pronounced at higher concentractions and longer exposure times, manifested as decreased cell viability, altered cell shape, loss of adhesion, and increased expression and altered localization of aquaporin 3. While an inflammatory response was not always detected, the observed changes indicate an effect of the fibres on cellular functions. These findings confirm that contact with certain types of mineral fibres can affect skin cells and trigger responses usually seen with mechanical irritation. The findings offer insight into how fibres interact with skin cells and may help guide the development of more user-friendly materials.

• Ključne besede: mineral fibres, glass wool, stone wool, mechanical irritation, keratinocytes, membrane damage, inflammatory response