Within the projetc group we had transfered the methodological expertise from senior researchers to younger collegues; from protocols of synthesis and functionalisation to protocols for visualization and quantification of NPs uptake. PI has established already strong collaboration with several Slovenian groups with complementary expertise: Nanotesla Institute, Laboratory for biotechnology - KI, Laboratory for electrochemistry of materials- KI, Institute of cell biology and Laboratory for molecular neurobiology from Faculty of Medicine UL Within the project the two PhD thesis (one awarded with Krka's prize, one proposed for the Krka's prize and Institut Jozef Stfean gold medal), one masters and two BsC thesis were finished, in addition there were five poster presentations of Undergraduate students. The young researched Jasna Lojk owas awarded Krka award for 2015 for her PhD thesis (mentor Mojca Pavlin, MR mentor VB Bregar) with the doctoral dissertation titled Analysis of interactions between cells and magnetic nanoparticles. Mentorships: PhD: LOJK, Jasna. Analysis of interactions between cells and magnetic nanoparticles : doctoral dissertation. Ljubljana: [J. Lojk], 2015.. [COBISS.SI-ID 11039572] STROJAN, Klemen. Protoemska karakterizacija nanodelcev za ovrednotenje njihove cititoksičnosti in imunotoksičnosti ter razvoj novih vrst nanodelcev : doktorska disertacija. Ljubljana: [K. Strojan], 2018 [COBISS.SI-ID 11948372] Masters degree: STROJAN, Klemen. Analiza vpliva površinske plasti magnetnih nanodelcev na preživetje celic in vitro : magistrsko delo = Effect of magnetic nanoparticles surface layer on in vitro cell viability : M. Sc. thesis, (Biotehniška fakulteta, Študij biotehnologije, Magistrsko delo magistrskega študija - 2. stopnja Biotehnologija, 19). Ljubljana: [K. Strojan], 2014. XI, 49 f., ilustr. [COBISS.SI-ID 8023929] BSc degrees: PUŠ, Urška. Dolgoročna biodistribucija različnih nanodelcev in vivo : diplomski seminar = Long term biodistribution of different nanoparticles in vivo : B. Sc. thesis, (Biotehniška fakulteta, Študij biotehnologije, Diplomski seminar univerzitetni študij - 1. stopnja, 240). Ljubljana: [U. Puš], 2015. VI[COBISS.SI-ID 8252025] DEKLEVA, Dominik. Vpliv srebrovih nanodelcev na sesalske celice v kulturi : diplomsko delo na univerzitetnem študiju Biokemije. Ljubljana: [D. Dekleva], 2016. 33 str., ilustr. [COBISS.SI-ID 1537116611] Student research project published on conferences: 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]
E.01 National awards
COBISS.SI-ID: 11039572The invention relates to the method for in vitro loading of biological eukaryotic cells with polymer coated magnetic nanoparticles that enables high loading of nanoparticles into biological cells where the cells retain high viability and where the loaded nanoparticles exhibit multimodal function involving optical and transmission electron imaging, and allow external manipulation with magnetic field. The method describes protocol for in vitro loading with nanoparticles having magnetic core and have coating with at least one or several polymer layers. The protocol and developed nanoparticles enable high loading of NPs inside the cell with small effect on cellular viability and can be thus used for cell labeling and tracking in different biomedical and biotechnological applications like tissue regeneration or tracking of stem cells.
F.32 International patent
COBISS.SI-ID: 10825556European commission identified nanoparticle risk assessment as a potential problem The thorough testing of nanotoxicity are 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. The knowledge obtained with the proposed project can be used to develop safer nanoparticles, nanocarriers and other nanomaterials. This knowledge can be applied in different biotechnological and pharmaceutical companies as well as industry. 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] Together with a working group we have established a Nanoportal webpage within Ministry of Health, that presents main informations and EU guidelines regarding potential nanotoxicity of nanoparticles. It present crucial information about nanomaterials that are of interest for nanotechnology industry and academia. Mojca Pavlin was also invited by the Ministry of Health, Office for chemicals to present her lecture at the Chemical safety meeting, ahe also presented her talk regarding nanotoxicity on the main Slovenian popular science portal Znanost na cesti with the title: “The promises of nanotechnology and safty of nanoparticles”. Znanost na cesti: dr. Mojca Pavlin, Skupina za nano in biotehnološke aplikacije, Fakulteta za elektrotehniko UL moderator: Luka Hvalc, Val 202 14. junij 2017, Kavarna Union PAVLIN, Mojca. Obeti nanotehnologij in varnost nanodelcev : prispevek predavanje na dogodku Znanost na cesti, 14. jun. 2017, Ljubljana. http://www.znanostnacesti.si/obeti-nanotehnologij.aspx. [COBISS.SI-ID 11782484] Furthermore, it is important to share the knowledge also to wider scientific public therefore we published in in Slovenian journal Medicinski razgledi 2013;52: 371-85 (target group are clinicians and medical students) »Endocytic pathways: their role in the development and treatment of diseases«,a review paper (authors: Jasna Lojk, Mojca Pavlin, Mateja Erdani Kreft), where we have gathered so far known facts about different types of endocytosis and endocytic pathways, we also discuss their role in the development of diseases and new treatments.
F.18 Transfer of new know-how to direct users (seminars, fora, conferences)
COBISS.SI-ID: 28137511We 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. In field of Nanotoxicity and Neurotocicity Mojca Pavlin is connected within the COST project BIONECA as a vice-chair of WG3.
B.03 Paper at an international scientific conference
COBISS.SI-ID: 11335252In year 2015 we have established new laboratory within our department of Faculty of electrical engineering. Our facilities consist of i) fully equipped laboratory for synthesis of nanoparticles and ii) fully equipped cell culture laboratory. Methods available in our laboratories: •Synthesis of magnetic and gold nanoparticles, Synthesis of liposomes •Culturing of numerous mammalian cell lines and cell models, •Viability and proliferation assays (MTS, Propidium iodide, trypan blue exclusion), •Reactive oxygen species measurements, ••Western blot analysis •Microscopy •Endotoxin analysis
D.02 Establishment of a research centre, laboratory, study course, association