Endokrino disruptivno djelovanje smjesa nanočestica plastike i odabranih lijekova in vitro

doctoral thesis

• Title (english): Endocrine disrupting effects of mixtures containing plastic nanoparticles and selected pharmaceuticals in vitro
• Author: Lucija Božičević
• Mentor: Ivana Vinković Vrček (mentor)
• Mentor: Valerije Vrček (komentor)
• Committee member: Dubravka Vitali Čepo (predsjednik povjerenstva)
• Committee member: Jozsef Petrik (član povjerenstva)
• Committee member: Dajana Kučić Grgić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of organic chemistry); Zagreb
• Defense date and country: 2024-07-18, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy
• Universal decimal classification (UDC): 615 - Pharmacology. Therapeutics. Toxicology

Abstract

Pretjerano korištenje i loše gospodarenje plastikom, doveli su do onečišćenja okoliša nanočesticama plastike (engl. plastic nanoparticles, PNP). Iako broj istraživanja o štetnim učincima PNP raste, nedostaju pouzdane informacije o učincima PNP na ljudsko zdravlje, posebice interakcijama s endokrinim sustavom. Osim PNP, okoliš je onečišćen i brojnim lijekovima. Paracetamol, ibuprofen, karbamazepin i fluoksetin svrstavaju su među lijekove koji su u okolišu kvantificirani u znatnim količinama. PNP mogu adsorbirati lijekove te činiti složene smjese čije djelovanje još nije istraženo. Unatoč tome, prisutnost okolišnih onečišćivala u složenim smjesama zanemaruje se te se primarno ispituje učinak pojedinačnih tvari. Cilj istraživanja bio je utvrditi endokrino disruptivne učinke složenih smjesa PNP i lijekova. Istraživanje se temelji na ispitivanju učinaka na aktivnost estrogenskog receptora (ER) i proces steroidogeneze. Složene smjese pripremljene su miješanjem lijekova i/ili nanočestica polistirena, polipropilena i polietilena. PNP su karakterizirane obzirom na oblik, veličinu, raspodjelu veličina i ζ potencijal. Učinci pojedinih tvari i njihovih smjesa na preživljenje stanica ispitani su pomoću MTS testa i protočnom citometrijom. Unos PNP u stanice određen je metodom protočne citometrije. Učinci na ER aktivaciju ispitani su luciferaznim testom prema OECD smjernici 455, a utjecaj na proces steroidogeneze qPCR-om i LC-MS/MS metodom prema OECD smjernici 456. Dobiveni rezultati pokazali su da se učinci pojedinačnih tvari značajno razlikuju u odnosu na učinke složenih smjesa. Smjese PNP i lijekova pokazale su znatno jače toksične i endokrino disruptivne učinke. Binarne i tercijarne smjese lijekova pokazuju složeni odnos doze i biološkog odgovora koji ovisi o sastavu smjese.

Abstract (english)

Excessive use and inadequate management of plastic have led to environmental pollution with plastic nanoparticles (PNP). Although the number of studies on the adverse effects of PNP is increasing, there is a lack of reliable information on the effects of PNP on human health, especially their interactions with the endocrine system. In addition to PNP, the environment is also polluted with numerous drugs. Paracetamol, ibuprofen, carbamazepine and fluoxetine are among the drugs that are quantified in the environment in significant quantities. PNP can adsorb drugs and form complex mixtures whose action has not been investigated. Despite this, the presence of environmental pollutants in complex mixtures is neglected and the primary focus is on the effect of individual substances. The aim of the research was to determine endocrine disrupting effects of complex mixtures of PNP and drugs. Research is based on the examination of the effects on the estrogen receptor (ER) acitivity and the process of steroidogenesis. Complex mixtures were prepared by mixing drugs and/or nanoparticles of polystyrene, polypropylene and polyethylene. PNPs are characterized by means of shape, size, size distribution and ζ potential. The effects of individual substances and their mixtures on cell survival were tested using the MTS assay and flow cytometry. PNP uptake into cells was determined by flow cytometry. The effects on ER activation were tested with the luciferase assay and OECD Guideline No. 455, and the influence on the steroidogenesis with qPCR and the LC-MS/MS method and OECD Guideline No. 456. The obtained results showed that the effects of individual substances differ significantly compared to the effects of complex mixtures. Mixtures of PNP and drugs showed significantly stronger toxic and endocrine disrupting effects. Binary and ternary mixtures of drugs show a complex dose-biological response relationship that depends on the composition of the mixture.

• Keywords: nanočestice plastike; lijekovi; složene smjese; endokrini disruptori; estrogenski receptor; steroidogeneza
• Keywords (english): plastic nanoparticles; pharmaceuticals; mixtures; endocrine disruptors; estrogen receptor; steroidogenesis
• Language: croatian
• URN:NBN: urn:nbn:hr:163:818326
• Promotion: 2024
• Study programme: Title: Pharmceutical-biochemical sciences - doctoral study - university study; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor/doktorica znanosti u području biomedicine i zdravstva (doktor/doktorica znanosti u području biomedicine i zdravstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-09-20 11:29:49