Title Utjecaj kemijski funkcionaliziranih jednozidnih ugljikovih nanocjevčica na osobitosti astrocita izloženih in vitro traumatskoj ozljedi Title (english) Influence of chemically functionalized single-walled carbon nanotubes on the characteristics of astrocytes
exposed to in vitro traumatic injury Author Tea Mladenić Mentor Kristina Pilipović (mentor) Committee member Jelena Ban (predsjednik povjerenstva) Committee member Ivana Munitić (član povjerenstva) Granter University of Rijeka Defense date and country 2020-07-31, Croatia Scientific / art field, BIOTECHNICAL SCIENCES Abstract Traumatska ozljeda mozga (engl. traumatic brain injury, TBI) je jedan od vodećih uzroka smrti i invaliditeta širom svijeta, a posebno u osoba mlađih od 45 godina i predstavlja značajan javnozdravstveni problem današnjice. Astrociti, stanice rasprostranjene širom središnjeg živčanog sustava (engl. central nervous system, CNS), značajno su podložne oštećenju nakon TBI. Dosadašnja su istraživanja potvrdila ulogu astrocita u održavanju strukture, funkcije i aktivnosti CNS-a, ali i u potencijalnoj regeneraciji CNS-a nakon ozljede. Do danas učinkoviti terapijski pristup u liječenju TBI nije otkriven, stoga su se istraživači okrenuli ka novom pristupu - tkivnom inženjeringu, točnije primjeni nanomaterijala. Cilj ovog istraživanja bio je istražiti utječu li jednozidne ugljikove nanocjevčice (engl. single walled carbon nanotubes, SWCNTs), kemijski funkcionalizirane s poli-m-aminobenzen sulfonskom kiselinom (engl. poly-m-aminobenzene sulfonic acid, PABS), na preživljavanje i određene karakteristike astrocita u in vitro modelu teške TBI. Istražen je utjecaj PABS-SWCNTs na ekspresiju kiselog fibrilarnog proteina glije (engl. glial fibrillary acidic protein, GFAP) u ozlijeđenim astrocitima. Nije zamijećena značajna razika u intenzitetu GFAP fluorescencije u traumatiziranim stanicama u odnosu na neozlijeđene, a nije uočena ni promjena ekspresije ovog markera u traumatiziranim stanicama tretiranima s PABS-SWCNTs. U drugom je pokusu praćen utjecaj PABS-SWCNTs na preživljenje astrocita nakon traume in vitro te su rezultati istraživanja pokazali da PABS-SWCNTs ne potiču bolje preživljenje, ali nisu ni citotoksične za ozlijeđene astrocite u kulturi. Kako bi se istražilo postoje li promjene u sadržaju sekretoma astrocita nakon ozljede, odnosno primjene istraživanog nanomaterijala, analiziran je sadržaj citokina u mediju za kulturu stanica. Ozljeda stanica uzrokovala je smanjeno otpuštanje Eotaksina-1 i povećanu razinu SDF-1 alfa u mediju, a primjena PABS-SWCNTs nije značajno utjecala na ove promjene. Aplikacija ovog tipa funkcionaliziranih nanocjevčica bila je praćena povećanim otpuštanjem RANTES-a iz ozlijeđenih astrocita u odnosu i na neozlijeđene i ozlijeđene netretirane stanice.
Zaključno, rezultati ovog rada ukazuju da primjena PABS-SWCNTs ne djeluje na preživljenje astrocita podvrgnutih teškoj in vitro TBI unutar prva 24 sata nakon ozljede, no uzrokuje neke promjene u profilu izlučenih citokina koje bi mogle utjecati na funkcije ove vrste stanica u patofiziološkim procesima nakon TBI.
Abstract (english) Traumatic brain injury (TBI) is one of the major causes of death and disability all over the world, especially among people under the age of 45, which is why it is considered a major public health problem of the modern age. Astrocytes are cells located in the central nervous system (CNS), which are significantly susceptible to damage after TBI. Previous researches have confirmed the importance of astrocytes in maintaining the structures, functions and activity of the CNS, but also the potential role in regeneration of the CNS, after TBI. So far, the effective therapeutic approach has not been discovered, which is why researchers have turned to a new approach, which includes tissue engineering, with nanomaterials. The aim of this study was to investigate the effect of single-walled carbon nanotubes (SWCNTs), chemically functionalized with poly-m-aminobenzene sulfonic acids (PABS), to better viability and proliferation of an astrocyte in an in vitro model of severe TBI. The effect of PABS-SWCNT on the expression of glial fibrillary acid (GFAP) was investigated in injured astrocytes. There was no significant difference in GFAP fluorescence intensity in injured cells compared to uninjured ones, also there was no change observed in expression of this market, in traumatized cells compared to ones treated with PABS-SWCNTs. In the second case, experiment monitored the effect of PABS-SWCNT on astrocyte survival after in vitro trauma, and the results of the study showed that PABS-SWCNTs did not lead to better survival, but also did not have cytotoxic effect on injured astrocytes in culture. In order to investigate whether there are changes in the content of astrocyte secretome or in the application of the investigated nanomaterial, cytokine content in the culture cell medium was analyzed. Cell injury caused decrease of Eotaxin-1 release and increase in SDF-1 alpha in the medium, and administration of PABS-SWCNT did not affect these changes. Application of this type of functionalized nanotubes was followed by an increased dismissal of RANTES, from injured astrocytes compared to uninjured and injured untreated cells.
In conclusion, the results of this study indicate that the use of PABS-SWCNTs does not affect the survival of astrocytes subjected to severe in vitro TBI within the first 24 hours after injury, but there was some changes in expression profile of cytokines that could affect on function of this type of cells in pathophysiological process after TBI.
Keywords
Ključne riječi: astrociti
fibrilarni kiseli protein glije
laktatna dehidrogenaza
traumatska ozljeda mozga
Keywords (english)
Key words: astrocytes
carbon nanotubes
glial fibrilary acid protein
lactate dehydrogenase
traumatic brain injury
Language croatian URN:NBN urn:nbn:hr:193:543799 Study programme Title: Biotechnology in medicine Type of resource Text File origin Born digital Access conditions Open access Terms of use Public note Ovo je istraživanje u potpunosti potpomognuto sredstvima Hrvatske zaklade za znanost dodijeljenih uspostavnom istraživačkom projektu UIP-2017-05-9517 "Jednozidne ugljikove nanocjevčice u eksperimentalnoj traumatskoj ozljedi mozga" voditeljice izv. prof. dr. sc. Kristine Pilipović, dr. med. Repository Repository of the University of Rijeka, Faculty of Biotechnology and Drug Development Created on 2020-07-31 08:55:41
