Dr. Mojca Pavlin was invited by the Ministry of Health, Office for chemicals to present her lecture at the Chemical safety meeting and she also presented her talk regarding nanotoxicity on the main Slovenian popular science portal “Znanost na cesti” (Science on the Street) with the title: “The promises of nanotechnology and safety of nanoparticles”. European commission identified nanoparticle risk assessment as a potential problem so thorough testing of nanotoxicity is essential for implementation of any new nanoparticle based application and could serve as industry guidelines. Better understanding of mechanisms together with testing will improve the evaluation of new nanomaterials industrial applications in the design stage and greatly improve the safety of industrial nanomaterials before they reach the market. This knowledge can be applied in different biotechnological and pharmaceutical companies as well as industry. Together with a working group we have established a Nanoportal webpage within Ministry of Health that presents information and EU guidelines regarding potential nanotoxicity of nanoparticles. REMŠKAR, Maja, LAVRENČIČ ŠTANGAR, Urška, DROBNE, Damjana, PAVLIN, Mojca. Portal za nanomateriale : Nanoportal RS, Ministrstvo za zdravje, Urad RS za kemikalije. http://www.uk.gov.si/. [COBISS.SI-ID 28137511] It presents crucial information about nanomaterials that are of interest for nanotechnology industry and academia. The research group was also patircipated as a organizer for specific workshop for children’s summer camp (12-16 years of age) of innovative technologies, organized yearly by the Faculty of Electrical Engineering (University of Ljubljana). We also published several papers in Slovenian journals “Medicinski razgledi” (Medical Outlooks) and “Zdravniški Vestnik” (Slovenian Medical Journal), peer-reviewed journals aimed at publishing research by Slovenian physicians and generally researchers working on topics related to medicine. Dr. Saša Novak is editor of the portal "Science on the road" . Novak, Saša (urednik 2013-). Ljubljana: SATENA, 2013-. ISSN 2385-9997. http://www.znanostnacesti.si/. [COBISS.SI-ID 3421519]
B.04 Guest lecture
COBISS.SI-ID: 11782484The project also enables transfer of the state-of-the-art knowledge into educational system through undergraduate and graduate student courses in the field of biomedical engineering and nanobiotechnology (graduate programs Biology, PhD program Biosciences, UL). Group leaders have been mentors of several graduate and doctoral students, and young researchers in different fields. Involvement of graduate students, young researchers and postdoctoral researchers enabled them hands-on approach in high-tech nanobiotechnological and biomolecular methods. Within the project 1 doctoral thesis(Klemen Strojan) 2 Masters thesis (Ajda Demšar, Jan Gregorec) and several undergraduate students research work was performed, their work was presented with several conference presentations. STROJAN, Klemen. Proteomska karakterizacija nanodelcev za ovrednotenje njihove citotoksičnosti in imunotoksičnosti ter razvoj novih vrst nanodelcev : doktorska disertacija. Ljubljana: [K. Strojan], 2018. X, 167 str. mentor (M.Pavlin), [COBISS.SI-ID 11948372] OBLAK, Blaž, STROJAN, Klemen, SVETE, Jurij, PAVLIN, Mojca. Interakcija med magnetnimi nanodelci oplaščenimi s polietileniminom in označenimi z rodaminom B ter celicami CHO = Interaction between polyethyleneimine coated rhodamine B labelled magnetic nanoparticles and CHO cells. V: PREMATE, Ester (ur.), et al. Zbornik konference, Biosfera - konferenca študentov bioloških znanosti, Ljubljana, 23. 9. 2016. Ljubljana: Društvo študentov biologije. 2016, str. 5-6, ilustr. http://biosfera.si/files/2016/09/Biosfera_zbornik.pdf?e9270b KLADNIK, Jerneja, STROJAN, Klemen, PAVLIN, Mojca. Vpliv prostega polietilenimina na transfekcijo celic CHO z magnetnimi nanodelci = Effect of free polyethyleneimine on the transfection of CHO cells with magnetic nanoparticles. V: PREMATE, Ester (ur.), et al. Zbornik konference, Biosfera - konferenca študentov bioloških znanosti, Ljubljana, 23. 9. 2016. Ljubljana: Društvo študentov biologije. 2016, str. 3-4, ilustr. http://biosfera.si/files/2016/09/Biosfera_zbornik.pdf?e9270b. [COBISS.SI-ID 11536212] GRANDA, Špela, LOJK, Jasna, PAVLIN, Mojca. Vpliv nanodelcev titanovega dioksida na preživetje celic THP-1 = Effect of titanium dioxide nanoparticles on survival of THP-1 cells. V: PREMATE, Ester (ur.), et al. Zbornik konference, Biosfera - konferenca študentov bioloških znanosti, Ljubljana, 23. 9. 2016. Ljubljana: Društvo študentov biologije. 2016, str. 33-34, ilustr. http://biosfera.si/files/2016/09/Biosfera_zbornik.pdf?e9270b. [COBISS.SI-ID 11536468] GREGOREC, Jan. Vpliv ouabaina na preživetje in rast človeških celic raka dojke MDA-MB-231 in MCF-7 v celični kulturi in na modelu tumorskih sferoidov in vitro = Effects of ouabain on survival and proliferation of human breast cancer cell lines MDA-MB-231 and MCF-7 in cell culture and spheroid models in vitro : magistrski študijski program Industrijska farmacija. Ljubljana: [J. Gregorec], 2018. VI, 57 str., graf. prikazi, ilustr. http://www.ffa.uni-lj.si/docs/default-source/knjiznica-doc/magistrske/2018/gregorec_jan_mag_nal_2018.pdf?sfvrsn=2. [COBISS.SI-ID 4612465]
D.10 Educational activities
COBISS.SI-ID: 11536468We present new in vitro models for assessing long-term nanotoxicity. Nanoparticles (NPs) have to be appropriately characterized which is also one of the main reasons for the difficult implementation of NPs into biomedicine. Most in vitro nanotoxicology studies analyze only short-term cytotoxicity due to lack of in vitro models that enable long-term exposure, We used biomimetic in vitro urothelium (porcine) model and L6 rat myoblasts cells with differentiation capability to assess viability after short-term (24h-72h) and long-term (10 days, 3 months) acute and continuous exposure to i) NPs which we are developing for biomedical applications: polyacrylic acid (PAA) and polyethileneimine (PEI) coated magnetic NPs) and ii) SiO2, TiO2 as commercial types of NPs.. Overall we observed minimal toxicity on urothelium model except for magnetic PEI NPs confirming that urothelium represents one of the strongest tissue barriers. Cell viability did not drop significantly in case of 31-day of sub chronic exposure of urothelium cells to NPs. As expected the rat myoblasts L6 cells were much more sensitive to NDs than porcine urothelial cells. Interestingly, elevated levels of mRNA for nuclear transcriptional factor Nrf2 involved in regulation of expression of antioxidant proteins indicating induced stress response for both TiO2 and PAA NPs after continuous exposure. The effect of some NPs on differentiation level of L6 cells was additionally confirmed by measuring mRNA levels of myogenin (MyoG) and myosin I (MyhI), characteristic biomarkers of differentiated rat myoblasts L6 cells. While for PAA NPs no effect was observed, TiO2 NPs clearly decreased MyoG and MyhI. However, it is important to stress that 50 µg/ml is still very high concentration that is difficulty to be achieved in vivo, therefore further experiments are needed to asses potential stress repose at lower concentrations. Altogether we show that presented in vitro models of differentiating myoblast and urothelial cells can contribute to the field of nanotoxicology and testing of newly designed NPs. The analysis of cytokines induction is in progress.
B.03 Paper at an international scientific conference
COBISS.SI-ID: 11335252During the project we have established several advance protocols that enabled us to performe s-t-o-a reasearch: • Protocols for protein corona formation and analysis using MS • Protocols for isolation of PBMC cells from blood, for isolation of monocytes, protocols for flow cytometry analsis of PBMC cells • Protocols of formation of predefined corona for mechanistical analysis • Protocols for western blot for analysis of efefcts of NPs on cell signalling
F.01 Acquisition of new practical knowledge, information and skills
COBISS.SI-ID: 11736404We have presented the results of nanotoxciology and immunogenicy on several international and national conferences with posters and oral talks. PAVLIN, Mojca, LOJK, Jasna, SKOČAJ, Matej, STROJAN, Klemen, PIRKMAJER, Sergej, MIŠ, Katarina, VERANIČ, Peter, MARŠ, Tomaž. Primary human myoblasts as an in vitro model for gene electrotransfer and nanotoxicity studies. V: KACIN, Alan (ur.), PIRKMAJER, Sergej (ur.), PODBREGAR, Matej (ur.). Skeletal muscle research - from cell to human 2017 : book of abstracts : symposium and workshop, University of Ljubljana, Faculty of Medicine, Slovenia, 28th - 30th May 2017. Ljubljana: Faculty of Medicine: Faculty of Health Sciences. 2017, str. 58-59. [COBISS.SI-ID 33277913] kategorija: SU (S) točke: 0.25, št. avtorjev: 8 . LOJK, Jasna, STROJAN, Klemen, MIŠ, Katarina, BREGAR, Vladimir Boštjan, HAFNER BRATKOVIČ, Iva, BIZJAK, Maruša, PIRKMAJER, Sergej, PAVLIN, Mojca. Stress and immune response of primary human myoblast to two types of polymer coated magnetic nanoparticles. V: LÖFFLER, Beat (ur.). Witnessing the solutions and tackling the hurdles : conference proceedings, CLINAM 10/2017, European & Global Summit for Clinical Nanomedicine and Targeted Medicine, Basel, May 7-10, 2017, (Conference proceedings (Clinam), ISSN 2296-9667). Basel: European Foundation for Clinical Nanomedicine. 2017, str. 184-185, ilustr. [COBISS.SI-ID 11736148] kategorija: SU (S) točke: 0.25, št. avtorjev: 8 PAVLIN, Mojca, SKOČAJ, Matej, BIZJAK, Maruša, STROJAN, Klemen, LOJK, Jasna, ERDANI-KREFT, Mateja, VERANIČ, Peter, PIRKMAJER, Sergej, MIŠ, Katarina, HAFNER BRATKOVIČ, Iva, BREGAR, Vladimir Boštjan. Differentiating human myoblasts and urothelial cells as a model for study long-term nanotoxicity in vitro. V: LÖFFLER, Beat (ur.). Witnessing the solutions and tackling the hurdles : conference proceedings, CLINAM 10/2017, European & Global Summit for Clinical Nanomedicine and Targeted Medicine, Basel, May 7-10, 2017, (Conference proceedings (Clinam), ISSN 2296-9667). Basel: European Foundation for Clinical Nanomedicine. 2017, str. 198-199, ilustr. [COBISS.SI-ID 11736660] kategorija: SU (S) točke: 0.19, št. avtorjev: 11 STROJAN, Klemen, LEONARDI, Adrijana, BREGAR, Vladimir Boštjan, KRIŽAJ, Igor, SVETE, Jurij, LOJK, Jasna, PAVLIN, Mojca. Dispersion of nanoparticles in different media importantly determines the composition of their protein corona. V: LÖFFLER, Beat (ur.). Witnessing the solutions and tackling the hurdles : conference proceedings, CLINAM 10/2017, European & Global Summit for Clinical Nanomedicine and Targeted Medicine, Basel, May 7-10, 2017, (Conference proceedings (Clinam), ISSN 2296-9667). Basel: European Foundation for Clinical Nanomedicine. 2017, str. 209-210, ilustr. [COBISS.SI-ID 11736404] kategorija: SU (S) točke: 0.29, št. avtorjev: 7 PAVLIN, Mojca, SKOČAJ, Matej, LOJK, Jasna, STROJAN, Klemen, ERDANI-KREFT, Mateja, VERANIČ, Peter, PIRKMAJER, Sergej, MIŠ, Katarina, BREGAR, Vladimir Boštjan. Evaluation of potential short- and long-term toxicity of commercial and biomedical nanoparticles. V: BioNanoMed 2016 : congress documentation, 7th International Congress Nanotechnology in Medicine & Biology, 6-8 April 2016, Krems, Austria. [S. l.: s. n.]. 2016, str. [77], ilustr. [COBISS.SI-ID 11335252] kategorija: SU (S) točke: 0.22, št. avtorjev: 9 LOJK, Jasna, STROJAN, Klemen, MIŠ, Katarina, BREGAR, Vladimir Boštjan, HAFNER BRATKOVIČ, Iva, BIZJAK, Maruša, JERALA, Roman, PIRKMAJER, Sergej, PAVLIN, Mojca. Stress and immune responses to polymer coated nanoparticles. V: BioNanoMed 2016 : congress documentation, 7th International Congress Nanotechnology in Medicine & Biology, 6-8 April 2016, Krems, Austria. [S. l.: s. n.]. 2016, str. [98]. [COBISS.SI-ID 11335764]
F.18 Transfer of new know-how to direct users (seminars, fora, conferences)
COBISS.SI-ID: 33277913