Title Toksičnost 3-piridilporfirina u in vivo modelu Drosophila melanogaster
Title (english) Toxicity of 3-pyridylporphyrins in an in vivo model of Drosophila melanogaster
Author Sara Čabrijan
Mentor Nela Malatesti (mentor)
Mentor Ana Filošević (komentor)
Committee member Ivana Ratkaj (predsjednik povjerenstva)
Committee member Rozi Andretić Waldowski (član povjerenstva)
Committee member Nela Malatesti (član povjerenstva)
Committee member Ana Filošević (član povjerenstva)
Granter University of Rijeka (Faculty of Biotechnology and Drug Development) Rijeka
Defense date and country 2020-09-08, Croatia
Scientific / art field, discipline and subdiscipline BIOTECHNICAL SCIENCES Biotechnology
Abstract Porfirini su skupina organskih spojeva, koja se posljednjih godina sve više koristi kao fotosenzibilizator (PS) u fotodinamičkoj terapiji (PDT). PDT se bazira na mehanizmu fototoksične ekscitacije PS-a uz pomoć svjetlosti prikladne valne duljine, čime nastaju reaktivne kisikove vrste odgovorne za citotoksičnost. U pretkliničkim istraživanjima korištenjem in vitro i in vivo pristupa potrebno je dokazati fototoksičnost PS-a samo u prisustvu ekscitacijske valne duljine, kao i netoksičnost u odsutstvu svjetla. Ovaj diplomski rad nastavak je istraživanja (foto)toksičnosti novosintetiziranih N-oksidiranih i N-metiliranih (3-piridil)porfirina, provedenih in vitro na humanim stanicama karcinoma. Cilj rada je razviti protokol ispitivanja stabilnosti i toksičnosti N-oksidiranih i N-metiliranih (3-piridil)porfirina u in vivo modelu D. melanogaster, koristeći TMPyP3. Spektroskopskim metodama potvrđena je kemijska stabilnost TMPyP3 u metanolu, destiliranoj vodi, PBS-u te otopinama proteina i fizioloških kationa. Utvrđeno je kako TMPyP3 ostaje kemijski stabilan samo u hranjivom mediju za D. melanogaster bez dodatka kvasca. Koncentracija apsorbiranog spoja iznosi oko 1nM, a veća je u prethodno izgladnjelim jedinkama hranjenim 180 min s 40 μM TMPyP3 u usporedbi s jedinkama koje nisu izgladnjele, a hranjene su 24 sata. TMPyP3 nije se pokazao akutno toksičnim u mraku za jedinke D. melanogaster. Pokazano je kako PDT LED izvorom crvene svjetlosti pri intenzitetu 40 mW/cm2 nije letalna, ali je potencijalno neurotoksična za D. melanogaster hranjene 40 μM TMPyP3 180 min neovisno o trajanju osvjetljavanja. Protokol ispitan ovim radom pokazao se efikasnim za detekciju fototoksičnosti PDT terapije 3-piridilporfirinima u in vivo modelu D. melanogaster. Daljnjim razvojem protokola, D. melanogaster mogla bi postati važan model u pretkliničkim studijama 3-piridilporfirina, omogućujući brzo i ekonomično testiranje toksičnosti velikog broja ovih spojeva, nakon čega će samo najbolji kandidati pristupiti klasičnim studijama na modelima sisavaca.
Abstract (english) Porphyrins are a group of organic compounds, which have been increasingly used as photosensitizers (PS) in photodynamic therapy (PDT) in recent years. PDT is based on the mechanism of phototoxic excitation of PS with the help of light of appropriate wavelength, thus producing reactive oxygen species responsible for cytotoxicity. In preclinical studies using the in vitro and in vivo approaches, it is necessary to demonstrate the phototoxicity of PS only in the presence of excitation wavelength, as well as non-toxicity in the absence of light. This thesis is a continuation of the (photo)toxicity study of newly synthesized N-oxidized and N-methylated (3-pyridyl)porphyrins, conducted in vitro on human cancer cells. The aim of this study was to develop a protocol for testing the stability and toxicity of N-oxidized and N-methylated (3-pyridyl) porphyrins in an in vivo model of D. melanogaster, using TMPyP3. Spectroscopic methods confirmed the chemical stability of TMPyP3 in methanol, distilled water, PBS and solutions of proteins and physiological cations. TMPyP3 was found to remain chemically stable only in nutrient medium for D. melanogaster without yeast addition. The concentration of the absorbed compound is about 1 nM, and is higher in pre-starved flies fed for 180 minutes with 40 μM TMPyP3 compared to non-starved flies fed for 24 hours. TMPyP3 showed no acute dark toxicity for D. melanogaster. It has been shown that the PDT therapy using LED red light source at an intensity of 40 mW/cm2 is not lethal, but is potentially neurotoxic to D. melanogaster fed with 40 μM TMPyP3 for 180 minutes regardless of the duration of illumination. The protocol examined in this study proved effective for detecting the phototoxicity of PDT therapy with 3-pyridylporphyrins in an in vivo model of D. melanogaster. With further development of the protocol, D. melanogaster could become an important model in preclinical studies of 3-pyridylporphyrins, enabling rapid and cost-effective toxicity testing of a large number of these compounds, after which only the best candidates will access classical studies in mammalian models.
Keywords
porfirini
stabilnost
fototoksičnost
fotodinamička terapija
Drosophila melanogaster
Keywords (english)
porphyrins
stability
phototoxicity
photodynamic therapy
Drosophila melanogaster
Language croatian
URN:NBN urn:nbn:hr:193:019548
Study programme Title: Biotechnology in medicine Study programme type: university Study level: graduate Academic / professional title: magistar/magistra biotehnologije u medicini (magistar/magistra biotehnologije u medicini)
Type of resource Text
File origin Born digital
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Terms of use
Created on 2020-10-08 11:38:45