Karakterizacija hidrogelova pripravljenih iz biogenog izvora za primjenu u inženjerstvu tkiva

master's thesis

• Title (english): Characterization of hydrogels made from biogenic source for tissue engineering applications
• Author: Julijan Zrno
• Mentor: Gordana Matijašić (mentor)
• Committee member: Vesna Ocelić Bulatović (predsjednik povjerenstva)
• Committee member: Gordana Matijašić (član povjerenstva)
• Committee member: Jasna Prlić Kardum (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2024-06-07, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering

Abstract

Tkivno inženjerstvo uvelike je istraživano područje kojem znanstvenici posvećuju sve veću pozornost. Ono omogućuje regeneraciju oštećenih tkiva i organa, smanjujući potrebu za transplantacijama i donorima. Decelularizirana izvanstanična matrica (dECM) koristi se u tkivnom inženjerstvu jer pruža prirodnu podlogu koja podržava regeneraciju tkiva, zadržavajući biokemijske i biomehaničke faktore potrebne za rast stanica. Također, smanjuje rizik od imunološkog odbacivanja jer ne sadrži stanične komponente koje bi mogle izazvati imunološku reakciju. Hidrogelovi od decelularizirane izvanstanične matrice kombiniraju biokompatibilnost i prirodnu bioaktivnost dECM-a s mehaničkim svojstvima hidrogela, omogućujući optimalnu podršku za rast i diferencijaciju stanica. Za decelularizaciju jetrenog tkiva korišten je deterdžent natrijev lauret sulfat (SLES). Decelularizirani uzorci dodatno su obrađeni ultrazvukom u trajanju od 1, 2, 4, 8 i 16 sati. Takvim uzorcima kvantificirane su biološke komponente DNK i kolagen. Provedena je i diferencijalna pretražna kalorimetrija kako bi se odredila termička stabilnost proteina. Pripravljeni hidrogelovi od dECM-a karakterizirani su mjerenjem promjene apsorbancije u vremenu kako bi se dobio uvid u kinetiku geliranja te infracrvenom spektroskopijom s Fourierovom transformacijom (FTIR) kako bi se identificirala i potvrdila prisutnost specifičnih biokemijskih komponenti unutar hidrogela. Provedenim ispitivanjima može se zaključiti da je decelularizacija uspješno provedena s obzirom da se kvantifikacijom utvrdio znatno smanjen sadržaj DNK, ali i visok sadržaj kolagena. Najbolje vrijednosti pokazuje uzorak SLES 0.5b 8h, što potvrđuje i FTIR analiza, pa je taj uzorak potencijal za daljnja istraživanja budući da ima i zadovoljavajuću kinetiku geliranja te maksimalnu temperaturu denaturacije.

Abstract (english)

Tissue engineering is a largely researched field to which scientists are paying more and more attention. It enables the regeneration of damaged tissues and organs, reducing the need for transplants and donors. Decellularized extracellular matrix (dECM) is used in tissue engineering because it provides a natural substrate that supports tissue regeneration, retaining biochemical and biomechanical factors necessary for cell growth. Also, it reduces the risk of immune rejection because it does not contain cellular components that could cause an immune reaction. Decellularized extracellular matrix hydrogels combine the biocompatibility and natural bioactivity of dECM with the mechanical properties of hydrogels, enabling optimal support for cell growth and differentiation. The detergent sodium laureth sulfate (SLES) was used to decellularize the liver tissue. Decellularized samples were additionally treated with ultrasound for 1, 2, 4, 8 and 16 hours. The biological components of DNA and collagen were quantified with such samples. Differential scanning calorimetry was also performed to determine the thermal stability of proteins. Prepared dECM hydrogels were characterized by measuring the change in absorbance over time to gain insight into gelation kinetics and by Fourier transform infrared spectroscopy (FTIR) to identify and confirm the presence of specific biochemical components within the hydrogel. Based on the research,
it can be concluded that the decellularization was successful, since the quantification showed a significant decrease in the DNA content, but also a high collagen content. The best values are shown by the sample SLES 0.5b 8h, which is also confirmed by the FTIR analysis, so this sample has the potential to be considered in further research since it has satisfactory gelling kinetics and the maximum denaturation temperature.

• Keywords: tkivno inženjerstvo; decelularizirana izvanstanična matrica (dECM); hidrogelovi; ultrazvuk; SLES
• Keywords (english): tissue engineering; decellularized extracellular matrix (dECM); hydrogels; ultrasound; SLES
• Language: croatian
• URN:NBN: urn:nbn:hr:149:116027
• Study programme: Title: Chemical Engineering - Graduate study; Study programme type: university; Study level: graduate; Academic / professional title: sveučilišni/a magistar/magistra inženjer/inženjerka kemijskog inženjerstva (sveučilišni/a magistar/magistra inženjer/inženjerka kemijskog inženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2024-07-01
• Created on: 2024-06-12 09:52:34