University of Groningen
Increasing the versatility of an ex vivo model in nanosafety studies and fibrosis
Bartucci, Roberta
DOI:
10.33612/diss.119127385
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Publication date: 2020
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Bartucci, R. (2020). Increasing the versatility of an ex vivo model in nanosafety studies and fibrosis. University of Groningen. https://doi.org/10.33612/diss.119127385
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SAMENVATTING
Met nano-technologie wordt het ontwikkelen, gebruiken, bestuderen en controleren van materialen bedoeld die een dimensie in de orde van grootte van nanometers (10-9 m) hebben. Deze technologie en materi-alen hebben veel toepassingen gevonden, en het gebruik neemt alleen maar toe. Nano-materialen hebben bijgedragen aan de vooruitgang van onder andere elektronische apparatuur maar ook op allerlei andere vlakken in onze samenleving, zoals voedingsmiddelen, medicijnen, kleding, cosmetica, verzorgingsproducten etc. Nanotechnologie voor medicijnen heeft met name een grote impact op de mensheid, genaamd nano-medicijnen.
Nano-materialen gebruikt voor medicinale doeleinden zorgen niet alleen dat medicijnen beter op de bedoelde plek in het lichaam komen, maar ook voor weefsel engineering, beeldvormende technieken en diagnostiek doeleinden. Vergeleken met conventionele medicijnen hebben nano-medicijnen vaak een grotere efficiëntie.
Waarom zijn nano-materialen eigenlijk zo speciaal?
Materialen krijgen unieke eigenschappen wanneer ze op nano-schaal gemaakt worden, deze eigenschappen zijn anders wanneer de mate-rialen op een grotere schaal gemaakt worden. Dankzij hun grootte kunnen ze zich gemakkelijk verspreiden in het menselijk lichaam, en cellen binnen dringen. Omdat nano-medicijnen groter zijn dan con-ventionele medicijnen, kunnen ze niet via diffusie over de membraan. Nano-deeltjes moeten daarom actief worden opgenomen worden door de cellen, hiervoor maken ze gebruik van de zogenoemde endocytose. Ondanks dat nano-medicijnen veel voordelen hebben moeten we an-dere nadelige factoren niet vergeten. Namelijk dat nano-medicijnen ook zouden kunnen ophopen in andere organen of weefsels dan het eigenlijke doelorgaan. Dit zou kunnen leiden tot toxiciteit in andere or-ganen. Dezelfde factoren die nano-medicijnen zo veelbelovend maken, kunnen er voor zorgen dat nano-materialen ook potentieel gevaarlijk zijn voor de gezondheid. Bovendien komen we niet alleen in aanraking met nano-deeltjes door nano-medicijnen, maar ook door blootstellin-gen aan nano-technologie, zoals producten die nano-deeltjes bevatten. Nano-materialen zijn overal en mensen kunnen er op verschillende manieren aan blootgesteld worden zoals via water, lucht, voeding enzo-voort, maar ook van binnen uit als er bijvoorbeeld implantaten gebruikt
zijn, zoals bijvoorbeeld nano-medicijnen, of nano-materiaal gebruikt bij bijvoorbeeld beeldvormende diagnostiek.
Daarom is het van groot belang dat we gaan begrijpen hoe cellen om-gaan met nano-materialen. Wat de impact op cellulair niveau en het hele organisme is, dit is essentiële informatie om nieuwe en veilige nano-materialen te ontwikkelen.
Nano-veiligheid heet het wetenschappelijke vakgebied dat zich hiermee bezighoudt. Studies wordt bemoeilijkt door alle verschillende soorten van nano-materielen die er zijn, zo maken de precieze grootte, de vorm, de zuiverheid, de stabiliteit van nano-deeltjes dat ze zich verschillend kunnen gedragen. De manier van omgaan met nano-materialen kan tot verschillende resultaten in veiligheids-studies leiden. Ook op ver-schillende cellen, of verver-schillende weefsel kan het effect anders zijn. De primaire effecten van nano-materialen komen tot uiting in de eer-ste organen die eraan blootgeeer-steld worden, bijvoorbeeld de longen bij inademen. Secundaire effecten worden gevonden in de organen waar de nano-materialen zich ophopen, bijvoorbeeld de lever of de nieren. Waar nano-materialen ook voor bedoeld zijn, het goed in kaart brengen van de veiligheid ervan is ontzettend belangrijk. Dit is het belangrijkste doel van nano-veiligheid.
Een van de grote obstakels in deze studie is het vinden van een be-trouwbaar, kosteneffectief, niet tijdrovend in vitro model voor de vei-ligheids-testen zodat dierproeven kunnen worden voorkomen. In vivo, dierproeven, bestuderen voornamelijk de ophoping op orgaan-niveau. Uitkomsten van in vivo experimenten in proefdieren zijn echter niet altijd direct te koppelen aan hoe de nano-materialen bij de mensen werken. Bij in vitro testen wordt meestal maar één soort cel getest, daardoor gaat de complexiteit van een orgaan verloren. Er zijn on-geveer 200 verschillende celtypes in het menselijk lichaam, dus het is ook heel moeilijk om een in vitro analyse methode te vinden die al deze verschillen kan bevatten. Het is dus belangrijk verschillende technieken te combineren.
Voor dit doel zijn er nieuwe in vitro systemen ontwikkeld zoals 2D en 3D cel culturen, co-culturen, organoids en organen op een chip. Dit met altijd in het achterhoofd om minder dierproeven te hoeven uitvoeren.
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Ex vivo modellen zouden ook een goed alternatief kunnen zijn.
Pre-cision-Cut Tissue Slices (PCTS) zijn een 3D ex vivo model. Nadat een orgaan uitgenomen is worden kleine pluggen gemaakt (5mm diameter), deze worden vervolgens in een speciaal apparaat (Krumdieck slicer) tot dunne plakjes van ongeveer 200 μm gesneden. Tissue slices kunnen van verschillende organismen, inclusief mensen, en verschillende organen gemaakt worden. Als PCTS van menselijke afkomst is, dan is dat chirurgisch restmateriaal.
PCTS is een model wat veel gebruikt wordt voor medicijnmetabolisme en toxicologie studies. Bovendien wordt het gebruikt bij fibrose studies, zoals bijvoorbeeld de studie van een specifiek eiwit tijdens de ontwik-keling van fibrose, zoals vanin-1 in dit proefschrift (Hoofdstuk 6). Recentelijk worden PCTS ook gebruikt voor nano-toxicologie studies. Ze zijn een beter alternatief dan celcultures, omdat ze een betere in-vivo achtige omgeving hebben. Het is een echt stukje weefsel, hebben een hoge reproduceerbaarheid, en zijn gemakkelijk te hanteren in het lab. In dit proefschrift worden PCTS als ex vivo model voor nano- veiligheid studies gepresenteerd (Deel I) en als een model om de rol van vanin-1, een eiwit betrokken bij oxidatieve stress en ontstekingen, in verschillenden stadia van fibrose te bestuderen (Deel II).
Deel I
In hoofdstuk 2 worden rat, muis, en menselijke PCTS van de lever bestudeerd na behandeling met nano-deeltjes. Er is gekozen voor de lever omdat bekend is daar veel nano-deeljes kunnen ophopen. Ge-carboxyleerde en amino-gemodificeerde polystyrene nano-deeltjes worden gebruikt omdat het al bekend is dat die een effect op cellen hebben. Na 72 uur behandeling van de PCTS met nano-deeltjes is de levensvatbaarheid gemeten, en dode cellen geteld. Amino-gemodifi-ceerde polystyrene nano-deeltjes laten een toxiciteit zien in de slices die overeenkomt met in vitro studies, daarnaast werd ook apoptose aangetoond. Ook hebben we gekeken hoe nano-deeltjes over de tijd opgenomen worden. Dit hebben we bekeken met fluorescentie micro-scopie en voor het eerst is de nano-deeltjes opname in cellen gekwan-tificeerd na weefsel digestie en gebruikmakend van flow cytometry. Met name macrofagen (Kupffer cellen) nemen de nano-deeltjes op,
wat overeenkomt met resultaten van gepubliceerde in vivo studies. De resultaten van de rat, muis en de mens komen overeen.
In hoofdstuk 3 gaan we een stap verder, namelijk wordt het gedrag van verschillende nano-deeltjes in PCTS van de lever maar ook van de longen en de nieren bestudeerd. Dat zijn andere belangrijke orga-nen waar nano-deeltjes kunorga-nen ophopen. Toxicologie, morfologie, opname en distributie zijn bekeken in deze organen. Amino-gemo-dificeerde polystyrene nano-deeltjes waren wederom toxisch in de PCTS van verschillende weefsels en de gecarboxyleerde polystyrene en de silica nano-deeltjes niet. Voor de stukjes longen is er ook weer gebruik gemaakt van flow cytometry om opname in de afzonderlijke cellen te bestuderen, en mRNA expressie van bekende ontstekings-ei-witten, en macrophage polarisatie zijn gemeten. Nano-deeltjes ver-zamelen zich wederom in de macrofagen, alleen sneller dan in de lever, er is echter geen verhogende concentratie van markers voor ontsteking gemeten.
Daarna is bekeken of het ex vivo model, PCTS, ook gebruikt kan worden om lange termijn blootstelling van nano-deeltjes te bestuderen. Chroni-sche blootstelling kan leiden tot veiligheids-zorgen. Hoofdstuk 4 gaat over muis PCTS van de lever die worden blootgesteld aan verschillende nano-deeltjes in verschillende condities en na blootstelling worden fibrose en ontsteking markers bestudeerd. Verschillende markers voor fibrose werden gemeten tijdens het langdurig kweken van PCTS in aan-wezigheid van verschillende nano-deeltjes. Er is met name aandacht besteed aan het optimaliseren van de condities van nano-deeltjes om blootstelling aan de PCTS te bestuderen.
Onder andere hebben we de effecten op de lever bekeken na bloot-stelling aan nano-deeltjes in de aanwezigheid of afwezigheid van een overmaat van “vrije” eiwitten in de oplossing. Ook hebben we het effect van een eiwitomhulsel (corona) om de nano-deeltjes bestudeerd. Ook is het effect van incubatietijd van de nano-deeltjes op de slices bestudeerd. Zonder eiwitten leiden amino-gemodificeerde polystyrene nano-deel-tjes als ze elke 24 uur ververst worden tot tekenen van ontsteking, maar niet van fibrose. Maar als er een corona om de nano-deeltjes was waren deze tekenen van ontsteking niet zichtbaar. Silica nano-deeltjes, gecar-boxyleerde polystyrene nano-deeltjes, en titanium nano-deeltjes lieten geen tekenen van ontsteking of fibrose zien.
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Onze resultaten laten zien dat het belangrijk is onder welke condities je PCTS blootstelt aan nano-deeltjes en dat rijp? worden van nano-deeltjes in de oplossing zeer sterk de reacties van het weefsel kan beïnvloeden. Samenvattend, in het eerste gedeelte zijn PCTS gebruikt als een complex
ex-vivo model om de opname en impact en de potentiele effecten van
nano-deeltjes te bestuderen. Hiervoor zijn methodes geoptimaliseerd en de resultaten laten zien dat PCTS een uitstekend ex vivo model zijn om nano-deeltjes distributie te bestuderen.
Deel II
In het tweede gedeelte van dit proefschrift zijn PCTS gebruikt om de expressie van het eiwit vanin 1 in verschillende organen te bestuderen in in vivo fibrose modellen.
Vanin 1 is een ectoenzyme GPI verankerd eiwit dat tot expressie komt in lever, nieren, darmen en longen. Gedurende de laatste jaren is va-nin 1 bestudeerd met betrekking tot oxidatieve stress, ontsteking, en metabole ziekten. In zieke organen kan vanin 1 zowel een beschermend als verergerend effect hebben. Dat hangt af van de aard en het stadium van de ziekte en welk orgaan. In hoofdstuk 5 is de rol van vanin 1 in de lever, de nieren, de darmen en de longen voor zowel fysiologische als psychopathologische condities bediscussieerd.
In hoofdstuk 6 hebben we de verschillende mate van expressie van vanin 1 gemeten op zowel mRNA niveau als eiwit niveau in verschil-lende organen. PCTS zijn gebruikt als model omdat ze spontaan fibrose ontwikkelen na 48 uur kweken. Verschillen in vanin 1 op zowel mRNA als eiwit niveau werden gevonden in de verschillende organen. De conclusie is dat PCTS geschikt zijn voor het bestuderen van de vei-ligheid van nano-deeltjes, en PCTS ook gebruikt kunnen worden om de rol van een eiwit, vanin1, bij fibrose beter te kunnen begrijpen.
ACKNOWLEDGMENTS
The Italian factor has been always present…when the review about Vanin 1 has been completely accepted I was in Rogliano, the same goes
for the first accepted experimental paper in SMALL and now, when I am writing these acknowledgments at the end of my PhD journey.
These 4 years have been amazing, with ups and downs (gastritis in-cluded), and they went so fast as I could never have expected. It has been intense, but with the right people around and a bit of optimism, everything landed with a great end.
First of all, I want to thank my supervisors Prof. A. Salvati and Prof. P. Olinga, and my co-promoter Dr. Y. L. Boersma, who guided me through these four years. Dear Anna, I still remember our first inter-view via skype…your enthusiasm and curiosity to truly get to know me as potential PhD candidate for your group, broke the screen. “If I talk too fast and too much please let me know” and I thought “ok, we may have already something in common :)” and at the same time I thought “secondo me è una tosta! Mi piace!”. I came then to visit in July and you showed me around and introduced me to some potential colleagues. Everything felt spontaneous and on the right place for me. After visiting the city and let the following weekend passed, I had clear
in my mind that I wanted to start the PhD with you and the other PI’s. You played a fundamental role for my scientific and personal growth! I felt we found our click after the end of the 1st year. We shared, scien-tifically speaking and non, delicate, happy, down and funny moments, which I will always remember. I am truly grateful for the fact that you have been always present and reachable (doesn’t matter if it was August or Christmas or if you were in maternity leave), keen to solve problems, give advices, chase me when necessary and cheer me up with nice words or gestures! There are many things that I would like to mention and to say thanks for, but for now I have to keep it short and I really hope we do keep in touch, for scientific purposes and non…I would love to see Sophia e Matteo growing up :). Dear Peter, hartelijk bedankt voor alles! You have been for me always a reference point both for scientific discussions, suggestions, ideas, help for data interpretation, for lab work (we even sliced human tissue together… and this says already everything about you!) and for informal chatting. I felt always comfortable to talk with you (sometimes I even forgot I was talking with my supervisor), listened and always supported. Your optimism, your advices to breath in and breath out and, last but not least, to take holidays, gave me the right comfort and confidence to
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proceed this journey with pleasure! Really…thanks! You always showed interest in what I was thinking and which was my point of view. I really hope we stay in touch also after the end of my PhD period...anyway, I will be around for sure! Dear Ykelien L. (for all the times you have corrected your name in my abstracts, presentation, papers and so on…I know now how important the initial of the second name is for you :)) thanks to have been part of the supervisors team and to have brought Vanin 1, a lot of English grammar and Western Blotting…in my life and in this project. After publishing the review on Vanin 1, we even got an email of compliments from a professor of University of South Carolina for our work. Thanks for organizing since the first months of the PhD, dinners at your place, and for the nice chats we used to have on the 9th floor. I wish you a lot of success for your future!
A big thanks also to Prof. G. Groothuis, who was head of the PTT department when I started. It was such a pleasure to have scientific discussions with you, but also share anecdotes during breaks!
I want to thank Prof. K. Poelstra, actual head of the PTT department. Although I have not been directly your student, you have been always
prone to help, discuss scientific topics, give nice suggestions and com-fort me when you were passing by the office and finding me still behind the screen the day before summer holidays. Thanks for your support! A special thanks to the reading committee Prof. R. Gosens, Prof. R. A. Bank and Prof. A. Duschl to have read and approved my thesis.
To my guardians in this special day, Daphne and Nadia, my paranim-phs. Dear Daphnina, I am really happy you have accepted the task to be my paranimph today as friend and as representative of the RoDaOliDa :). We started almost together and we went through this together, from the first till the last day also in the same office. We got to know each other day by day and we became friends, on top of being colleagues in the same research group. I was so happy to share with you beautiful moments as THE one with Jordi, the day at the Zoo, lekker BBQ or simply a drink after shopping. You have been always honest with me, opened to listen, give advices and thoughtful! After nice chats you always double-check whether you said something too much, it is so sweet of you and it says everything about your pure soul! Good luck for these last months…it is going to be all fine, you will graduate and you will do great as always! I will be here to support you! My dear Nadia, friend since ever and ever! We met for the first time when we were both 6 years old, unbelievable! We did elementary school, middle school and high school together… each time I think about it and now we are here, still together in the 2020. Even more here in Groningen to support me in this special occasion. I
will never forget this moment! You are the friend that everyone would like to have…sweet, always present (also in silence, but always there), trustful, supportive, never intended to hurt you, smart, auto ironic, fun…one big pillar of our Rogliano crew, that you greatly represent! Thanks to be my friend. I am entirely sure we will be in each other life for long and this makes me feel really comforted! Grazie nostro NN!
My next thanks go to Prof. B. N. Melgert: dear Barbro, thanks to have been part of my PhD journey, for the helpful scientific discussion during the Monday meetings, for each time I sought help from you, for the pleasant chats during lunch break, for receiving appreciated emails when I was suddenly in Italy for health problems or when I could not get back my deposit of the rented house! All things that I remember and I appreciated then, and now! I take this occasion to say thanks to all your research group for the nice time spent together and specially Anienke and Fransien. Anienke, thanks for your help when I needed to figure the flow cytometry protocol out, for your being direct but never rude towards me, and for the great chats we have had, about work, life and future. I wish you a lot of success for these last months, believe in yourself and you will do a great job! Fransien, you are smart and mod-est, helpful and truly caring for colleagues around you. I won’t forget when that day with Anienke, you brought me chocolate to cheer me up during the last weeks of my contract, where you saw me immersed in the thesis..maybe a little bit too much. I did not need to say anything, you both just understood that it was a bit heavier than other moments. Small gesture that made my day. Thanks!
Thanks to Dr. L. Beljaars and Dr. I. De Graaf…Dear Leonie and Inge thanks to have “chased” me during work discussions, which spurred me to do better and better. You had always good points, good questions and constructive criticism. I appreciate a lot your being friendly and always open to help!
A great help was given by Dr. Christoffer Åberg, therefore to him it goes a huge thanks. Really appreciated your professionalism, punc-tuality and precision.
But…what is a PhD without a special secretary?! Dear Gillian, if you were not there, someone had to invent you! You are not just the secre-tary of the department…I officially declare you as a supportive member of the working team, friend team, e fun time, and I could continue! :) You have been always present with your optimism, jokes and with your heart. You did me a good turn!
…and what is a lab without technicians? They play maybe the most im-portant role in a PhD student life and for this I want to say a big thanks to
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Catharina, Dorenda, Eduard, and Marina! Catharina…precise, hard worker (you do so many things in one day), honest, direct, helpful… thanks to be always present and to be able to count on you! thanks to make me practice my Dutch…zo aardig van jou! :) Dorenda…thanks for all the time you have planned animal experiments for me, to check on me if I was doing well and for your help in the slice lab! Eduard…hard worker, always and I say always keen to help, never rude (although you try your best with your jokes…but I know you are basically just a sweet person) and fun! Marina…echt gezellig! Nice to work with you, peaceful and patient…also with my dutch! Ilse, I had the pleasure to be with you in the group only for short, but I immediately recognized your empathic personality and your being eager to learn! I am sure you will do great. Good luck!
Sometimes I think I have been so lucky to have had such wonderful colleagues…for me this factor has been for sure as important as my PhD project. Without them it would have not been the same.
A special thanks to Aldy, with your suggestions to follow #PhDlife, you made my days…thanks for the LOL together!; Harita, I know what is to be shared PhD…keep up like this!; Keni, we started this journey the same day, so…yes, we have something special together! Good luck for these last months…almost there!; Natasha, when you started your brought a nice wind of change! Good luck for your new endeavors!; Valentina, the first colleague I met even before to come to Groningen…
thanks to have introduced me to the city and to people when I was new here. A lot of success in Canada…I hope there the food is more for your taste…:p kidding!) and Hector. Hectorino, gracias por ser como eres. It was great to have you in the office in the last months…you are fun and chill, really enjoy to talk with you and thanks to “scold” me and Daphne when we were lost in deep conversations…”these two”! You are such a hard worker and nerdy smart…you deserve all the best! Good luck for your future!
Being a shared PhD means in principle that you are part of different groups, in my special case three. Therefore, I want to thank everyone of Peter’s group and CBP department, who has already finished and who started recently, and all the colleagues I worked with at the Slice lab. Thanks for the time spent together, your help and for the produc-tive scientific discussions in each weekly meeting. However, there are some special persons that I want to acknowledge: Gerian, Emma, Mitchel and Magda. Gerian, thanks to be friendly and gentle in the working environment, to be the one always prone to help colleagues and make sure things in the lab are organized. Thanks for all the time
you have sacrificed mice for me, namely at least once per week. It is not your priority and I felt so bad when people were not understanding that,given your presence for granted. Emma, thanks for your help to process human tissue and for your quick response when I was seeking help or samples from you. Mitchel, I really enjoy(ed) our conversations… they start from science, they take a huge turn (around hobbies, life and love) and they go back to science! Love it! Thanks for all the time you have inflated lungs for me! Magda, we unbelievably shared so many things in these 4 years! Happy and fun moments, deep downs and troubles, and last but not least our weekly Zumba hour! You can be proud of me…I am still attending each week, even alone! I wish you all the best because this is what you deserve!
When you have colleagues, who become also friends…then I think nothing can go wrong! Laura & Olivia, you have been such important pieces of my puzzle! Laurita, I remember when we were together in our first big office and I already thought we may have had so many things in common…and indeed! How much we did together? How many times we had “our” talks in which we confronted each other and comforted each other? How many times we cried and then laughed, and then cried, and laughed!? We supported each other in everything…work related staff, personal issues or accomplishments! You are a such enthusiastic person about your job and life, you like to give advices and to get advices…most importantly you care for people around you! I feel like I know you since longer time (maybe for our common background?! ;) )! It was super fun to share the office with you, with great LOL and at the same time with seriousness. Our common working path is about to be divided, but I truly hope that our path of friendship will stay the same! Oly, you are the sweetest person on this earth! I think always I have been lucky to have met you. Without knowing it, you have taught me a lot…that we can be fragile and strong at the same time! You are kind and gentle also in the No day, deeply honest and full of love to give without expecting anything in return. You appreciate so much when someone does something nice to you and for you… it is clear on your face! I am so happy you are going to stay longer in The Netherlands, because I am sure we will spend much more time together and we will celebrate many more birthdays together as last year! Stay as you are, and believe in yourself, you are strong…I see that!
Viktoriia and Natalia, former colleagues and friends. Thanks for the help you gave me in the lab during the first years of my PhD! Although our working relationship ended, I am really happy to see that we are
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still in contact with each other, that we have fun outside and enjoy together nice Moments! I am sure we will keep like this!
I want at this point to thank Dr. M. Houweling, who was my super-visor during the short research stay at the Utrecht University during my master. Dear Martin, you do not know but you played a fundamental role in my decision to start the PhD. I admired your way to be scientist… always enthusiastic, curious, attentive, patient, happy to do your job well and to teach me step by step all the techniques I used for that project with the help of Martijn. I want to take this occasion to say thank you for how you welcomed me in your research group in Utrecht, teaching me a lot but also making me enjoy my first dutch summer “today there is sun Roberta, go read the article in the Wilhelminapark!”. I also remember the first Saturday I was in Utrecht, before to start in the lab, you came with your wife to pick me up by bike and you both showed me the way to arrive at the university the following Monday. So nice of you!
Four years in Groningen are also made by memories outside the lab and working environment. I cannot really say that I did not enjoy my time here. The music played a big role…therefore, I decided to mention for each chapter artists who I have seen live in The Netherlands during concerts and festivals in these last years. The music of these artists has had always the power to clear my mind, reset my brain, and remind me that life must be enjoyed also during the PhD! Thanks to Aska, who converged this music in kaleidoscopic fluorescence imaging of this thesis, in the cover.
All the people I have met have contributed to make this journey un-forgettable, also a special Dutch teacher as Chris, understanding when that week I had maybe too much in my head and lowering the amount of dutch grammar rules for that day and instead talking about music, concerts, books and food! Obviously still in dutch. New and old friends have been constantly present in this part of my life, too. My dutch non-dutch semi non-dutch friends Jana, Simon, Lara, Alberto and Aska have been and some still are a fundamental chapter of my life in Groningen. We enjoy(ed) great moments together, our biertje on Thursday evening, political conversations, international dinners, board games evenings, parties, etc. and above all quality time together! Jana, my super inter-national friend…you know 1000 languages, you are always updated to any news in the world, you are the most versatile and eclettic person I know, and always ready to have fun together. I admire you for your being always thirst of knowledge. Thanks for these years together!
I want to say thanks to Kristina, Silvan, Mark and Alwine, Aike’s friends who made me feel always welcome in the group and part of it
since almost 6 years! I enjoy your company and I am always happy to spend time together!
I met new people, I have new friends, but my friends from Rogliano have been and are present always in my life. Alessia, Angelo, Anton-ella, Corrado, Fernando, Giuseppe, Katia, Laura, Luigi, Manuela, Marilena, Marta, Martina, Matteo, Serena, Simona and Tom-maso....voi siete non la mia seconda famiglia, ma parte della mia fa-miglia! Con alcuni di voi ci conosciamo da sempre, vero Martinuzza (30 anni e non sentirli amica mia...so come avresti voluto essere qui, e lo sei!)? Con alcuni di voi abbiamo fatto addirittura l’asilo insieme, che mi dici Tommy? Persino nella giunta comunale dei giovani insieme, o no Matte? Abbiamo fatto di tutto...dalle vacanze estive al momento studio in biblioteca..confermi,Ale (la mia dottoressa!)? So quanto avreste vo-luto essere a Groningen per la fine di questo mio percorso, e non è stato possibile, ma so che sarete qui con il pensiero quel giorno! “Una vera amicizia non è vedersi tutti i giorni. Con un vero amico puoi stare anche mesi senza sentirti, ma sai perfettamente che lui ci sarà comunque e nel momento in cui vi rivedrete sarà sempre tutto uguale”. Grazie! Vi voglio strabene!
Een speciale dank aan Kristianne en Ninne, mijn Nederlandse fa-milie. Ik voelde mij altijd welkom bij jullie, sinds de eerste keer toen we 6 jaar geleden elkaar ontmoetten. Soms is het moeilijk om ver weg te zijn van je eigen familie, maar het is minder moeilijk met jullie hier. Ik geniet van onze weekenden en vakantie samen, avondeten, Sinter-klaas pakketjes avond en wandelen! Ik vind we een heel speciale relatie hebben. Dankjulliewel voor alles!
Come sarei potuta arrivare fin qui senza di voi, mamma, papà e Armando? Voi che mi avete sempre appoggiata in tutto e per tutto,
che mi avete dato gli strumenti, le possibilità e l’apertura mentale di capire cosa avrei voluto fare e dove...tutti voi ognuno a modo proprio. Vi siete sempre fidati di me, mi avete creduta e stimolata in maniera
sottile e mai pesante, senza intromettervi troppo nelle mie scelte. Mi avete insegnato il valore della Famiglia, Amore, Amicizia, Onestà in-tellettuale e d’animo, della Libertà senza mai intaccare quella del mio vicino, del Rispetto per gli altri e per il Lavoro. Ci siete sempre stati nei momenti di sconforto e nei momenti felici...e so quanto la distanza sia pesante anche per voi, ma non me lo fate mai capire o pesare. Ogni volta che ci incontriamo sono felice 1000 e triste 2000, perchè penso già che il distacco mi peserà per almeno le prime 48h..non importa da quanto tempo siamo a distanza ormai! Vi sarò riconoscente per tutto quello che avete fatto, fate e farete....e spero che oggi un poco poco io vi abbia
A
ckno
wledgments
reso orgogliosi! Un’ultima cosa...Army, il mio “piccolo” fratellino, ti voglio un bene infinito! In bocca al lupo per questa nuova avventura... un PhD student finisce e un altro inizia!
Now it is time to close this chapter, and I cannot really do that without mention you, il mio Aikino! We met almost 6 years ago in the lab in Utrecht, you were already PhD student (but no, he was not my super-visor) and it took quite some time to start to talk to each other. But since then, we never left each other, not even when I went back to Italy to finish the other project started in UNICAL for almost one year, not even when you went to Houston for part of your PhD, not even when we could see each other in The Netherlands only in the weekend…still we did see each other each weekend for 3 years! Now, since 1 year and half we are finally living in the same city, and house, after you found a cool PostDoc position in Zernike! You have been truly supportive in these 4 years, understanding when I needed some comfort words, comfort food or when instead was better just to get some distractions. You look at me and you understand me...I do not need to add anything! We try to keep scientific discussions out of the table in our house, but yea, not always easy with nerds ;). I am glad to have you in my life…as partner, friend, family, colleague and companion of funny moments! I enjoy really a lot our holidays, cycling, hiking and cooking together… just fun! Thanks to make me feel the best sensations ever...to be my half and know that I can always count on you!
ABOUT THE AUTHOR
Roberta was born in Rogliano, Cosenza (Italy) on 19-08-1989. She attended her secondary education in Cosenza at the Liceo Classico Bernardino Telesio where she finished with 100 out of 100 as final mark. She then pursued the master degree in Chemistry and pharma-ceutical technology at the University of Calabria (Italy), with the final mark of 106 out of 110. As part of her master, Roberta did a short internship of 3 months at the Utrecht University, a longer internship at the University of Calabria and a 6-month internship in a pharmacy. Afterwards, she passed the national Italian exam to become officially
pharmacist. However, Roberta decided to continue her academic ca-reer as PhD student at the University of Groningen, at the Groningen Research Institute of Pharmacy (GRIP). She started in October 2015 and termed with the promotion in March 2020. During these years, Roberta presented her project in the three departments, where she was part of, Pharmacy days and at national and international conferences, collecting different prizes. She also participated with a video about Nanosafety to the Accomplissh impact award competition; to stimulate societal impact from scientific research (https://www.youtube.com/ watch?v=DPtGCmYC984&t=5s). Roberta will continue her career as scientist in an international company that provides bioanalytical sup-port from Pre-clinical to Phase IV studies.
Confer
ence att
endance & pr
esentations
AWARDS AND HONORS
(2018) Prize for oral talk at the Pharmacy day organized by the Gronin-gen Research Institute of Pharmacy (GRIP), GroninGronin-gen (NL);
(2018) Poster prize at the annual Dutch meeting of Toxicology (NVT), Hilversum (NL);
(2018) “Precision-Cut Liver Slices as a promising ex-vivo model for nanosafety studies” awarded with Travel grant at the NANOTOX 2018, Neuss (DE).
CONFERENCE ATTENDANCE & PRESENTATIONS
(2016) FIGON-DMD, Dutch Medicine Days, Poster presentation (2016) NMSA – Nanomaterials safety assessments conference, Malaga (SPAIN) – Poster Presentation
(2017) NVT 2017 – Dutch annual meeting of Toxicology – Oral presentation
(2018) CLINAM 2018 (Basel, SWITZERLAND) – 11th European and Global Summit for Clinical Nanomedicine, Targeted Delivery and Pre-cision Medicine – Poster presentation
(2018) NVT 2018 – Dutch annual meeting of Toxicology – Poster presentation
(2018) NANOTOX conference 2018 in Neuss (GERMANY) – Oral presentation
LICENSES, CERTIFICATIONS AND COURSES
ATTENDANCE DURING THE PHD
Qualification for the profession of pharmacist in Italy; Laboratory Animal Science (Art.9 Netherlands); Cellular Imaging light and basic courses; Presentation skills course;
Advanced drug delivery and drug targeting course; Scientific integrity and writing courses;
Dutch language course (up to B1).
TEACHING
(2017) FaTeM students supervision;
(2016–2018) Supervision of two master students, from the lab work to the correction of the final report;
