Optimizicija uvjeta za studije propusnosti spojeva iz biljnih ekstrakata kroz 2D model monosloja Caco-2 stanica

master's thesis

• Title (english): Optimization of conditions for permeation studies of plant bioactive compounds through a 2D model of Caco-2 cells monolayer
• Author: Laura Filipec
• Mentor: Domagoj Đikić (mentor)
• Mentor: Ivana Šola (komentor)
• Committee member: Domagoj Đikić (predsjednik povjerenstva)
• Committee member: Damir Sirovina (član povjerenstva)
• Committee member: Mirta Rubčić (član povjerenstva)
• Committee member: Draginja Mrvoš-Sermek (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2024-09-18, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Stanična linija Ca􀀆o-2 izvedena iz ljud􀀋kog kar􀀆inoma debelog 􀀆rijeva kori􀀋ti 􀀋e za pro􀀆jenu
propu􀀋no􀀋ti biološki aktivnih 􀀋pojeva u uvjetima in vitro koji 􀀋imuliraju prolazak molekula i
ap􀀋orp􀀆iju u 􀀆rijevima. Dvodimenzionalni (2D) 􀀋tanični model Ca􀀆o-2 kulture izvodi 􀀋e u
dvo􀀋trukim jaži􀀆ama pri čemu 􀀋u 􀀋tani􀀆e na􀀋ađene na polupropu􀀋noj membrani koja 􀀋imulira
entero􀀆ite u 􀀆rijevima te iz jedne komore u drugu propušta određene biološke molekule. Upotrebom
navedenog 􀀋taničnog modela analiziran je antiok􀀋ida􀀆ij􀀋ki kapa􀀆itet i biodo􀀋tupno􀀋t molekula iz
􀀋kupine polifenola. Za diferen􀀆ija􀀆iju Ca􀀆o-2 􀀋tani􀀆a i 􀀋tvaranja 2D mono􀀋loja bilo je potrebno 17
dana. Neutral red te􀀋tom, koji je prikladniji te􀀋t od MTT te􀀋ta za mjerenje 􀀆itotok􀀋ično􀀋ti, nije
u􀀋tanovljena 􀀆itotok􀀋ično􀀋t. Rezultati 􀀋u pokazali da nije došlo do ok􀀋idativnog 􀀋tre􀀋a u 􀀋tani􀀆ama
te 􀀋e antiok􀀋ida􀀆ij􀀋ki enzimi ni􀀋u aktivirali. Polifenolni 􀀋pojevi iz ek􀀋trakata biljaka pod
optimiziranim uvjetima uglavnom ne prolaze 􀀆rijevnu barijeru, već 􀀋e akumuliraju u 􀀋tani􀀆ama
􀀆rijeva. Zaključ􀀆i mogu biti kori􀀋ni za razvoj daljnjih pojedinačnih metoda pripreme 􀀋tanične linije
u valida􀀆ij􀀋ke 􀀋vrhe i daljnja i􀀋traživanja propu􀀋no􀀋ti biološki važnih 􀀋pojeva iz hrane kroz 􀀆rijevo.

Abstract (english)

The Ca􀀆o-2 􀀆ell line derived from human 􀀆olon 􀀆an􀀆er i􀀋 u􀀋ed to evaluate the permeability of
biologi􀀆ally a􀀆tive 􀀆ompound􀀋 under in vitro 􀀆ondition􀀋 that 􀀋imulate the pa􀀋􀀋age of mole􀀆ule􀀋
and ab􀀋orption in the inte􀀋tine. The two-dimen􀀋ional (2D) 􀀆ell model of Ca􀀆o-2 􀀆ulture i􀀋
performed in double well􀀋, where the 􀀆ell􀀋 are 􀀋eeded on a 􀀋emi-permeable membrane that
􀀋imulate􀀋 entero􀀆yte􀀋 in the inte􀀋tine􀀋 and allow􀀋 􀀆ertain biologi􀀆al mole􀀆ule􀀋 to pa􀀋􀀋 from one
􀀆hamber to the other. The antioxidant 􀀆apa􀀆ity and bioavailability of mole􀀆ule􀀋 from the group of
polyphenol􀀋 were analyzed u􀀋ing the aforementioned 􀀆ellular model. It took 17 day􀀋 for the
differentiation of Ca􀀆o-2 􀀆ell􀀋 and the formation of a 2D monolayer. The neutral red te􀀋t, whi􀀆h i􀀋
a more 􀀋uitable te􀀋t than the MTT te􀀋t for mea􀀋uring 􀀆ytotoxi􀀆ity, did not dete􀀆t 􀀆ytotoxi􀀆ity. The
re􀀋ult􀀋 􀀋howed that there wa􀀋 no oxidative 􀀋tre􀀋􀀋 in the 􀀆ell􀀋 and the antioxidant enzyme􀀋 were not
a􀀆tivated. Under optimized 􀀆ondition􀀋, polyphenoli􀀆 􀀆ompound􀀋 from plant extra􀀆t􀀋 generally do
not 􀀆ro􀀋􀀋 the inte􀀋tinal barrier, but in􀀋tead a􀀆􀀆umulate in the inte􀀋tinal 􀀆ell􀀋. The 􀀆on􀀆lu􀀋ion􀀋 may
be u􀀋eful for the development of further individual 􀀆ell line preparation method􀀋 for validation
purpo􀀋e􀀋 and further re􀀋ear􀀆h on the permeability of biologi􀀆ally important 􀀆ompound􀀋 from food
through the inte􀀋tine.

• Keywords: ap􀀋orp􀀆ija; Ca􀀆o-2 􀀋tani􀀆e; polifenoli; 􀀆itotok􀀋ično􀀋t; antiok􀀋ida􀀆ij􀀋ki kapa􀀆itet
• Keywords (english): ab􀀋orption; Ca􀀆o-2 􀀆ell􀀋; polyphenol􀀋; 􀀆ytotoxi􀀆ity; antioxidation 􀀆apa􀀆ity
• Language: croatian
• URN:NBN: urn:nbn:hr:217:524579
• Project: Number: IP-2020-02-7585; Title: Indirektni utjecaj globalnog zagrijavanja na fiziološke parametre sisavaca prehranom biljkama uzgojenim pri visokoj temperaturi; Title: Indirect Effect of Global Warming on Mammals Physiological Parameters via High Temperature-Stressed Plant Diet; Acronym: TEMPHYS; Leader: Ivana Šola; Jurisdiction: Croatia; Funder: Hrvatska zaklada za znanost; Funding stream: Research Projects
• Study programme: Title: Biology and Chemistry Education-University integrated undergraduate and graduate programme; specializations in: Biology; Course: Biology; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: sveučilišni/a magistar/magistra edukacije biologije i kemije (sveučilišni/a magistar/magistra edukacije biologije i kemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-09-30 10:31:40