Utjecaj detergenata na djelotvornost decelularizacije tkiva

undergraduate thesis

• Title (english): The impact of detergents on the tissue decellularization efficacy
• Author: Dinka Omerčić
• Mentor: Gordana Matijašić (mentor)
• Committee member: Gordana Matijašić (predsjednik povjerenstva)
• Committee member: Jasna Prlić Kardum (član povjerenstva)
• Committee member: Danijela Ašperger (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2024-09-09, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering

Abstract

Sve veća potreba za nadomjestkom određenog tkiva ili organa, uz manjak donora, za sobom povlači novu sferu interesa kroz tkivno inženjerstvo i razvoj tkiva in vitro. Takav se poduhvat bazira na nacjepljivanju stanica na nosač izvanstanične matrice od biomaterijala izražene biokompaktibilnosti. Pri tome se stvara trodimenzionalno izgrađeno kompleksno tkivo, koje se potom transplantira. Kako bi se izvanstanična matrica izolirala, postala nosačem za stanice te zadržala svoju strukturu, potrebno je proći kroz proces decelularizacije. U tom je procesu tkivo podvrgnuto fizikalnim, kemijskim i enzimatskim postupcima. U ovome je radu provedena decelularizacija svinjske jetre putem kemijske metode, gdje su upotrebljavani detergenti: natrijev lauril sulfat (SDS), natrijev lauret sulfat (SLES), Triton-X 100 i Tween 20.
Nakon provedenih decelularizacija, postupak se nastavio kvantifikacijom bioloških komponenata kojima je utvrđena koncentracija prisutne deoksiribonukleinske kiseline (DNK), kolagena i glikozaminoglikana (GAG). Dodatno je analizirana struktura tkiva svjetlosnim mikroskopom i stereomikroskopom. Rezultati istraživanja pokazali su varijabilnu učinkovitost detergenata poput SDS-a, SLES-a, Triton-X 100 i Tween 20 u procesu decelularizacije svinjske jetre. Kvantifikacija sadržaja DNK, kolagena i glikozaminoglikana (GAG-a) u decelulariziranim uzorcima tkiva otkrila je različitu sposobnost detergenata u očuvanju ovih ključnih komponenata. Mikrografije tkiva dodatno su ilustrirale oštećenja i puknuća u strukturi izvanstanične matrice, s nekim detergentima koji su uzrokovali veća oštećenja od drugih.
Ovi rezultati naglašavaju potrebu za pažljivim odabirom detergenata i optimizacijom procesa decelularizacije kako bi se minimizirala oštećenja strukture tkiva i osigurala njihova biokompatibilnost za buduću primjenu.

Abstract (english)

Increasing need for the replacement of a specific tissue or organ, with a smaller number of donors, entail a new sphere of interest through tissue engineering and tissue development in vitro. Such an undertaking is based on the grafting of a cell on an extracellular matrix carrier made of biomaterials with pronounced biocompatibility. In doing so, a three-dimensional complex tissue is created, which is then transplanted. In order for the extracellular matrix to be isolated, become a carrier for cells and retain its structure, it is necessary to go through the decellularization process. In this process, the tissue is subjected to physical, chemical and enzymatic procedures. In this work, decellularization of porcine liver was carried out by a chemical method using 1% solutions of detergents sodium lauryl sulfate (SDS), sodium laureth sulfate (SLES), Triton-X 100 and Tween 20. After the decellularization, the obtained samples were tested using quantification methods, in order to measure the presence of DNA, collagen and GAG for the detection of residual biological material. In addition to various methods of quantification, the tissue structure was analyzed using a light microscope and a stereomicroscope. The research results showed variable effectiveness of detergents such as SDS, SLES, Triton-X 100, and Tween 20 in the decellularization process of porcine liver. Quantification of DNA, collagen, and glycosaminoglycan (GAG) content in decellularized tissue samples revealed the different abilities of detergents in preserving these key components. Tissue micrographs further illustrated damages and ruptures in the extracellular matrix structure, with some detergents causing greater damage than others. These findings emphasize the need for careful selection of detergents and optimization of the decellularization process to minimize tissue structure damage and ensure their biocompatibility for future applications.

• Keywords: tkivno inženjerstvo; izvanstanična matrica; decelularizacija; detergent
• Keywords (english): tissue engineering; extracellular matrix; decellularization; detergent
• Language: croatian
• URN:NBN: urn:nbn:hr:149:001686
• 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: 2024-11-12 09:22:12