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Title:Preparation of Three Dimensional Structures of Polysaccharide Derivatives for Application in Regenerative Medicine : doctoral disertation
Authors:ID Dobaj-Štiglic, Andreja (Author)
ID Stana-Kleinschek, Karin (Mentor) More about this mentor... New window
ID Maver, Uroš (Mentor) More about this mentor... New window
ID Kargl, Rupert (Comentor)
Files:.pdf DOK_Dobaj_Stiglic_Andreja_2022.pdf (33,63 MB)
MD5: 59CFF2B22ABD9EF57B0DEB5237B5505B
 
Language:English
Work type:Doctoral dissertation
Typology:2.08 - Doctoral Dissertation
Organization:FS - Faculty of Mechanical Engineering
Abstract:Biocompatible polysaccharide scaffolds with controllable pore size, good mechanical properties, and no hazardous chemical crosslinkers are desirable for long-term tissue engineering applications. Despite decades of development of novel scaffolds, there are still many challenges to be solved regarding their production and optimization for specifically engineered tissues. Herein, we have fabricated several three-dimensional (3D) scaffolds using polysaccharide or polysaccharide-protein composite hydrogels or inks for 3D printing, featuring strong shear thinning behavior and adequate printability. The inks, composed of various combinations of chitosan, nanofibrillated cellulose, carboxymethyl cellulose, collagen, and citric acid, were 3D printed, freeze-dried, and dehydrothermally heat-treated to obtain dimensionally and mechanically stable scaffolds. The heat-assisted step induced the formation of covalent amide and ester bonds between the functional groups of chosen polysaccharides and protein collagen. Citric acid was chosen as a non-hazardous and „green” crosslinker to further tailor the mechanical properties and long-term stability of the scaffolds. We have investigated how the complexation conditions, charge ratio, dehydrothermal treatment, and degree of crosslinking influence the scaffolds' chemical, surface, swelling, and degradation properties in the dry and hydrated states. The compressive strength, elastic modulus, dimensional stability and shape recovery of the (crosslinked) scaffolds increased significantly with balanced charge ratio, dehydrothermal treatment, and increased concentrations of citric acid crosslinker and collagen concentrations. The prepared crosslinked scaffolds promoted (clustered) cell adhesion and showed no cytotoxic effects, as determined by cell viability assays and live/dead staining with human bone tissue-derived osteoblasts and human adipose tissue-derived mesenchymal stem cells. The water-based and non-hazardous crosslinking methods presented here can be extended to all polysaccharide- or polysaccharide-protein-based materials to develop cell-friendly scaffolds with tailored properties suitable for various tissue engineering applications.
Keywords:chitosan, carboxymethyl cellulose, nanofibrillated cellulose, citric acid, collagen, freeze drying, 3D printing, dehydrothermal treatment
Place of publishing:Maribor
Place of performance:Maribor
Publisher:[A. Dobaj Štiglic]
Year of publishing:2022
Number of pages:XXIII, 172 str.
PID:20.500.12556/DKUM-81164 New window
UDC:661.728.017:544.773(043.3)
COBISS.SI-ID:125363971 New window
Publication date in DKUM:11.10.2022
Views:900
Downloads:175
Metadata:XML DC-XML DC-RDF
Categories:KTFMB - FS
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Licences

License:CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
Link:http://creativecommons.org/licenses/by-nc-nd/4.0/
Description:The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.
Licensing start date:27.01.2022

Secondary language

Language:Slovenian
Title:Izdelava tridimenzionalnih struktur polisaharidnih derivatov za aplikacijo v regenerativni medicini
Abstract:V zadnjem desetletju je opazen izjemno velik znanstveni napredek v razvoju novih nosilcev za regeneracijo različnih tkiv. Kljub temu je potrebnih še veliko izboljšav, saj trenutni tridimenzionalni (3D) nosilci ne posnemajo dovolj dobro naravnega okolja zunajceličnega matriksa in ne nudijo zadostne mehanske opore, ki bi omogočila dolgotrajno rast in kasnejši razvoj izbranih tkiv. Razvoj novih tkivnih nosilcev tako predstavlja še vedno velik izziv. V tej doktorski disertaciji je predstavljen razvoj novih multifunkcionalnih tkivnih nosilcev iz biokompatibilnih biopolimerov, ki posnemajo strukturo in sestavo zunajceličnega matriksa in so namenjeni za uporabo v hrustančnem in kostnem tkivnem inženirstvu. Tridimenzionalni (3D) nosilci so bili izdelani iz polisaharidnih ali polisaharidno-beljakovinskih kompozitnih hidrogelov ali črnil za 3D tiskanje, ki so se odlikovali z močnim strižnim redčenjem in so bili primerni za 3D tiskanje. Hidrogeli in črnila, pripravljeni iz različnih kombinacij nanofibrilne (NFC) in karboksimetilne (CMC) celuloze, hitozana (CS), kolagena (Coll) in citronske kisline (CA) so bili 3D natisnjeni ali vliti v kalup ter strjevani s kombinacijo sušenja z zamrzovanjem in dehidrotermalne toplotne obdelave (DHT), da smo dobili dimenzijsko in mehansko stabilne nosilce. Termična obdelava nosilcev je povzročila nastanek novih amidnih in esterskih vezi med funkcionalnimi skupinami izbranih polisaharidov in beljakovinskim Coll. CA je služila kot ekološko sprejemljiv zamreževalec, ki je tudi nestrupen za razliko od drugih citotoksičnih zamreževalcev z reaktivnimi kemijskimi skupinami, hkrati pa je tudi izboljšala mehanske lastnosti in dolgotrajno stabilnost nosilcev. Doktorska disertacija podrobneje predstavlja, kako pogoji kompleksiranja, dehidrotermalna obdelava in stopnja zamreženosti vplivajo na poroznost, kemijsko strukturo, nabrekanje in degradacijo suhih in omočenih nosilcev. Tlačna trdnost, modul elastičnosti in dolgotrajna dimenzijska stabilnost nosilcev so se bistveno povečali z uravnoteženim razmerjem naboja, dehidrotermalno obdelavo in z višjimi koncentracijami zamreževalca CA ter Coll. Pripravljeni zamreženi nosilci so tudi izboljšali sposobnost preživetja in proliferacijo mezenhimskih matičnih celic ali osteoblastnih celic, izoliranih iz človeškega kostnega tkiva. V doktorskem delu predstavljene metode fizikalnega in kemijskega zamreženja na vodni osnovi se lahko razširijo na vse materiale na osnovi polisaharidov ali polisaharidov s proteini. S takšnimi pristopi se lahko razvijejo celicam prijazni nosilci s prilagojenimi lastnostmi, primernimi za različne aplikacije v tkivnem inženirstvu.
Keywords:hitozan, karboksimetil celuloza, nanofibrilna celuloza, citronska kislina, kolagen, zamrzovanje s sušenjem, 3D tiskanje


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