Razvoj novega in vitro modela trebušne slinavke z uporabo tehnike 3D tiska

doktorska disertacija

Marko Milojević (Avtor), Uroš Maver (Mentor), Andraž Stožer (Komentor)

Povzetek

Celični nosilci iz polisaharidov posnemajo strukturo in sestavo ekstracelularnega matriksa, v katerem celice trebušne slinavke proliferirajo. Kljub vsesplošni uporabi polisaharidnih hidrogelov v tkivnem inženirstvu večina metod izdelave ni uspešna pri izdelavi stabilnih hidrogelnih nosilcev, ki bi omogočili dolgotrajno rast in ohranitev fenotipa specifičnih celic. Ključnega pomena za uspešno uporabo posameznega materiala v tkivnem inženirstvu je tudi uravnavanje lastnosti nosilcev, izdelanih iz njega (nabrekanje, degradacija, reološke lastnosti, mehanske lastnosti, poroznost, hidrofilnost, mikro- in nano-površinske lastnosti), tako da te posnemajo izvorno tkivo. V doktorski nalogi smo razvili nove polisaharidne formulacije, ki so primerne za 3D tisk visoke ločljivosti, sočasno pa ohranjajo dolgoročno strukturno stabilnost v pogojih celičnih kultur, kar je ključno za njihovo dolgotrajno uporabo. Z vgradnjo nikelj-bakrovih nanodelcev smo dokazali, da lahko celičnim nosilcem spreminjamo osnovne materialne lastnosti, tako pa postavili temelje za izgradnjo nosilcev, ki natančneje posnemajo lastnosti izvornega tkiva. Sočasno smo razvili hibridno tehniko 3D tiska, ki omogoča hkraten tisk termoplastičnih polimerov s hidrogeli in izgradnjo mehansko stabilnejših nosilcev. Izdelali smo tudi novo šobo in protokol core/shell tiska, ki zagotavljata izdelavo nosilcev s pretočnimi filamenti, ki posnemajo osnovno funkcionalnost žilja in vitro. Po uspešni izolaciji celic trebušne slinavke, smo nazadnje izdelali novo biočrnilo z vključenimi celicami in situ, ki ga lahko natisnemo v želene geometrijske oblike. S funkcionalnimi celičnimi testi smo dokazali, da so vse razvite formulacije biokompatibilne, tiskani nosilci pa podpirajo dolgoročno rast in viabilnost izoliranih celic trebušne slinavke ter tako omogočajo izvedbo dolgotrajnejših eksperimentov.

Ključne besede

3D tisk;celični nosilci;polisaharidni hidrogeli;celice trebušne slinavke;model in vitro;Patofiziologija;Disertacije;Tkivno inženirstvo;In vitro modeli;Tkivni modeli;Bolezenski modeli;Bolezni prebavil;Trebušna slinavka;Model trebušne slinavke;3D tiskanje;Polisaharidne molekule;

Podatki

Jezik:	Slovenski jezik
Leto izida:	2021
Tipologija:	2.08 - Doktorska disertacija
Organizacija:	UM MF - Medicinska fakulteta
Založnik:	M. Milojević]
UDK:	612.34+004.356.2(043.3)
COBISS:	85011715
Št. ogledov:	307
Št. prenosov:	61
Ocena:	0 (0 glasov)
Metapodatki:

Ostali podatki

Sekundarni jezik:	Angleški jezik
Sekundarni naslov:	Development of a novel in vitro model of the pancreas using 3D printing techniques
Sekundarni povzetek:	Polysaccharide-based scaffolds replicate the structure and composition of the extracellular matrix in which pancreatic cells readily proliferate. Despite the widespread use of polysaccharide hydrogels in tissue engineering, most manufacturing techniques have failed to produce stable hydrogel scaffolds, which are key to long-term growth and phenotype preservation of specific cells. Moreover, controlling the scaffold properties made from specific materials (swelling, degradation, rheological properties, mechanical properties, porosity, hydrophilicity, micro- and nano-topographical properties) is also crucial for its effective mimicking of respective native tissues. In this dissertation, we developed novel polysaccharide formulations suitable for high-resolution 3D printing while exhibiting long-term structural stability under cell culture conditions. By incorporating nickel-copper nanoparticles, we have shown that we can alter the fundamental properties of the scaffolds, which provides the basis for constructing scaffolds that better mimic the properties of the targeted native tissue. In parallel, we have developed a hybrid 3D printing method that enables the simultaneous printing of thermoplastic polymers with hydrogels and the construction of more mechanically stable scaffolds, enabling further growth control of the incorporated cells. We have also developed a novel nozzle and core/shell printing protocol that enables the fabrication of scaffolds with hollow filaments that mimic the basic functionality of the vasculature in vitro, which is necessary to provide long-term perfusion of artificial tissues. Finally, following the successful isolation of pancreatic cells, we have developed a novel bioink with in situ incorporated cells, which can be used to print 3D structures with desired geometries. Using functional cell-based assays, we have demonstrated that all developed formulations are biocompatible and that the printed scaffolds support the long-term growth and viability of the isolated pancreatic cells and as such enable their use as part of long-term experiments.
Sekundarne ključne besede:	3D printing;scaffolds;polysaccharide hydrogels;pancreatic cells;in vitro model;Trebušna slinavka;
Vrsta dela (COBISS):	Doktorska disertacija
Komentar na gradivo:	Univ. v Mariboru, Medicinska fak.
Strani:	X, 148 str.
ID:	13283441