Vpliv različnih nanodelcev na preživetje, stresni odziv in diferenciacijo celic Caco-2

Ahačič, Špela (author), Pavlin, Mojca (mentor)

V našem delu smo analizirali vpliv 48-urne izpostavitve nanodelcev (ND) food grade (FG) nano TiO2, nano SiO2 in nano srebra, v zmernih (2 in 10 µg/mL) in višjih koncentracijah (50 in 100 µg/mL) na celično linijo Caco-2. Z metodo diferencialnega barvanja po Hoechstu in s propidijevim jodidom (PI) smo prikazali koncentracijsko odvisen vpliv izbranih ND na preživetje celic. S pomočjo fluorometričnih metod smo analizirali nastanek reaktivnih kisikovih spojin (ROS) v tretiranih nediferenciranih celicah Caco-2 z reagentom CM-H2DCFDA ter analizirali aktivnost alkalne fosfataze (ALP) v diferenciranih celicah Caco-2 preko zaznavanja signala od MUP (metilumbeliferil fosfat) odcepljenih fosfatnih skupin. Delež ROS je bil povečan v primeru celic tretiranih s FG TiO2 in SiO2 pri koncentracijah > 50 µg/mL, vendar nikjer ni prišlo do statistično značilnih razlik v primerjavi s kontrolo. Rezultati ne nakazujejo na vpliv izbranih ND na diferenciacijo celic, kaže pa se od koncentracije odvisen vpliv preučevanih nanodelcev na aktivnost membranske ALP, ki se sintetizira v diferenciranih celicah Caco-2. Prikazali smo vpliv izbranih ND na preživetje celic in vpliv višjih koncentracij izbranih ND na nastanek ROS v tretiranih nediferenciranih celicah Caco-2. Za določitev vpliva izbranih ND na aktivnost ALP v diferenciranih celicah Caco-2 bi morali poskus izvesti v večih bioloških ponovitvah.

• Language: Slovenian
• Keywords: Biotehnologija, nanodelci, Caco-2, nanoTiO2, nanoSiO2, nanoAg
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Place of publishing: Ljubljana
• Publisher: [Š. Ahačič]
• Year: 2022
• PID: 20.500.12556/RUL-135029
• UDC: 601.2:620.3:606:61(043.2)
• COBISS.SI-ID: 97741571
• Publication date in RUL: 18.02.2022

Secondary language

• Language: English
• Title: Effect of different nanoparticles on viability, stress response and differentitation of cell line Caco-2

Abstract

In our thesis we have analysed the effect of acute treatment of Caco-2 with three nanoparticles (NP) – food grade (FG) nano TiO2, nano SiO2 and nano silver NP with moderate (2 and 10 µg/mL) and high (50 in 100 µg/mL) concentrations. With colouring by Hoechst and propidium iodide (PI) we observed concentration dependent effect of selected NPs on cell viability. With spectrophotometric methods we have analysed the quantity of reactive oxygen species (ROS) in treated undifferentiated cells Caco-2 with reagent CM-H2DCFDA and also analysed the activity of alkaline phosphatase (ALP) in treated differentiated cells by detecting from methylumbelliferyl phosphate derived phosphate groups. Albeit ROS was elevated in with high concentrations FG TiO2 and SiO2 (> 50 µg/mL) treated undifferentitated cells, no statistically significant differences were observed. Although results do not indicate concentration dependent effect of selected NPs on cell differentiation, there could be an effect on alkaline phosphatase activity in treated differentiated cells Caco-2. We have shown an effect of selected NPs on cell viability and effect of high concentrations FG TiO2 and SiO2 NPs on ROS generation in treated undifferentiated cells Caco-2. The effect of selected NPs on ALP activity in treated differentiated cells Caco-2 could be established with additional biological repetitions.

• Keywords: Biotechnology, nanoparticles, Caco-2, nanoTiO2, nanoSiO2, nanoAg