Ispitivanje različitih uvjeta decelularizacije tkiva u pripravi hidrogelova izvanstanične matrice

undergraduate thesis

• Title (english): Influence of different decellularization parameters in the production of hydrogels from extracellular matrix
• Author: Valentina Erceg
• Mentor: Gordana Matijašić (mentor)
• Committee member: Gordana Matijašić (predsjednik povjerenstva)
• Committee member: Jasna Prlić Kardum (član povjerenstva)
• Committee member: Vesna Ocelić Bulatović (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2024-04-29, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering

Abstract

Inženjerstvo tkiva je grana inženjerstva koja u novije vrijeme poprima sve veći značaj jer pruža alternativu transplantaciji organa iz ljudskih donora. Zbog rastuće potrebe za transplantacijom organa i dugim listama čekanja te sve starijom populacijom očekuje se da će potreba za transplantacijom organa u budućnosti rasti. Inženjerstvo tkiva bi također moglo riješiti pitanje etičnosti ispitivanja lijekova na životinjama jer bi nosači nastali ovim tehnologijama trebali bolje oponašati ljudsko tkivo. U tu svrhu razvijeni su nosači napravljeni od različitih biokompatibilnih materijala koji posjeduju različita fizička i kemijska svojstva.
Od tih biomaterijala, poseban se značaj pridaje decelulariziranoj izvanstaničnoj matrici jer ona ima manju mogućnost izazivanja imunološke reakcije te joj trodimenzionalna arhitektura početnih tkiva ostaje bolje očuvana nakon decelularizacije. U ovom radu ispitano je šest različitih uvjeta decelularizacije svinjske jetre kako bi se utvrdio njihov utjecaj na strukturu i sastav biomaterijala. Korištene su 1 %-tne i 0,5 %-tne otopine natrijeva lauret sulfata (SLES), kao i 0,5 %-tna otopina detergenta SLES-a u kombinaciji obrade ultrazvukom u vremenskom periodu od 1, 2, 4 i 8 h. Kako bi se procijenila očuvanost strukture dobivene izvanstanične matrice provedena je karakterizacija bioloških komponenata i analiza strukture tkiva svjetlosnim mikroskopom te stereomikroskopom. Dobiveni rezultati pokazuju da je decelularizacija uspješno provedena u svim uzorcima jer se udio DNK značajno smanjio.
Vrlo dobri uvjeti decelularizacije postižu se s 0,5 %-tnom otopinom SLES-a i 0,5 %-tnom otopinom SLES-a u ultrazvučnoj kupelji u vremenskom trajanju od 1 h. Prilikom decelularizacije u ovim uvjetima postignut je najmanji udio DNK, a struktura izvanstanične matrice je ostala očuvana. Povećanje koncentracije SLES-a imalo je blago djelovanje na strukturu tkiva te je značajan udio bioloških komponenata ostao očuvan. Nadalje, duljom ultrazvučnom obradom uzoraka nastala su značajna oštećenja strukture izvanstanične matrice. Iz dobivenih rezultata također je vidljivo da je u svim uzorcima tijekom decelularizacije udio glikozaminoglikana pao ispod granice detekcije.

Abstract (english)

Tissue engineering is a branch of engineering that has recently become increasingly important because it offers an alternative to organ transplantation from human donors.
Due to the growing demand for organ transplants and the long waiting lists, as well as the aging population, the need for organ transplants is expected to increase in the future. Tissue engineering could also solve the ethical problem of drug testing on animals, as the scaffolds produced by these technologies should better mimic human tissue. To this end, scaffolds made of various biocompatible materials with different physical and chemical properties have been developed. Among these biomaterials, decellularized extracellular matrix is of particular importance because it has a lower potential to induce immune responses and its three-dimensional architecture is better preserved after decellularization. In this study, six different conditions for decellularization of porcine liver were investigated to determine their effects on the structure and composition of the biomaterial. 1 and 0.5 wt% solutions of sodium laureth sulfate (SLES) and a 0.5 wt% solution of SLES detergent in combination with sonication for 1, 2, 4, and 8 hours were used. To evaluate the preservation of the structure of the obtained extracellular matrix, the biological components were characterized, and the tissue structure was analyzed using a light microscope and a stereomicroscope. The obtained results show that decellularization was successfully performed in all samples, as the DNA content decreased significantly. Very good decellularization conditions were obtained with a 0.5% SLES solution and a 0.5% SLES solution with 1-hour sonication. Under these conditions, the lowest DNA content was obtained, and the structure of the extracellular matrix was preserved. Increasing the SLES concentration had only a minor effect on tissue structure, as a significant portion of the biological components remained intact. Moreover, prolonged ultrasonic treatment of the samples resulted in significant damage to the extracellular matrix structure. It is also evident from the results obtained that the content of glycosaminoglycans in all samples fell below the detection limit during decellularization.

• Keywords: inženjerstvo tkiva; nosač; decelularizirana izvanstanična matrica; biomaterijal; SLES
• Keywords (english): tissue engineering; scaffold; decellularized extracellular matrix; biomaterial; SLES
• Language: croatian
• URN:NBN: urn:nbn:hr:149:273733
• Study programme: Title: Chemical Engineering - Undergraduate study; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka kemijskog inženjerstva (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka kemijskog inženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-05-08 10:19:48