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Title:Tkivni inženiring hrustančnega tkiva na biosintetičnem polimernem polyHIPE nosilcu
Authors:ID Naranđa, Jakob (Author)
ID Vogrin, Matjaž (Mentor) More about this mentor... New window
ID Slak Rupnik, Marjan (Comentor)
Files:.pdf DOK_Naranda_Jakob_2017.pdf (24,38 MB)
MD5: 9BD430952820C686F767E6A46F1A2181
 
Language:Slovenian
Work type:Doctoral dissertation
Organization:MF - Faculty of Medicine
Abstract:Tkivni inženiring hrustančnega tkiva še vedno nudi številne možnosti za izboljšavo, navkljub intenzivnim raziskovalnim naporom v zadnjem času. Razvoj umetnih materialov in 3-D celičnih nosilcev ima pomembno vlogo pri regeneraciji hrustančnega tkiva. Zanimiv pristop pri izdelavi celičnih nosilcev predstavlja izgradnja s pomočjo emulzij. Nastali material, imenovan polyHIPE (PHP), je sintetični visoko porozen polimer, ki ga pripravimo s polimerizacijo visokega deleža notranje faze emulzij (HIPEs – high internal phase emulsions). Glavni cilj te doktorske disertacije je raziskati možnosti za tvorbo hrustančnega tkiva znotraj celičnih nosilcev pripravljenih iz PHP materiala. Proizvodnjo PHP nosilcev smo posebej prilagodili tkivnemu inženiringu hrustanca, tako da smo pripravili porozne (85 %) strukture s primarno velikostjo por v območju 50–170 m. Pokazali smo, da je PHP material biokompatibilen s človeškimi sklepnimi hondrociti, kar smo ovrednotili s pomočjo testa za preživetje celic (Live/Dead kit) in histološko analizo. Opazovali smo hondrocite z okroglimi jedri, ki so bili organizirani v večceličnih plasteh na površini PHP nosilca in so rastli približno 300 m v notranjost nosilca. Kopičenje kolagena tipa 2 smo dokazali s pomočjo imunohistokemije, molekularna analiza je pokazala izražanje hrustančno specifičnih genov z ugodnim razmerjem kolagena tipa 2 in tipa 1. Dodatno so bili PHP vzorci biološko razgradljivi, njihove osnovne mehanske lastnosti pa primerljive z nativnim sklepnim hrustancem. Izsledki raziskave dokazujejo, da je zasnovan PHP celični nosilec primeren za nadaljnjo uporabo v tkivnem inženiringu hrustančnega tkiva.
Keywords:humani sklepni hrustanec, tkivni inženiring, biokompatibilni celični nosilec, sintetični polimer, polyHIPE, diferenciacija hondrocitov
Place of publishing:Maribor
Year of publishing:2017
PID:20.500.12556/DKUM-64787 New window
COBISS.SI-ID:6112063 New window
NUK URN:URN:SI:UM:DK:SZV5ID2Y
Publication date in DKUM:17.10.2017
Views:2619
Downloads:303
Metadata:XML DC-XML DC-RDF
Categories:MF
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Secondary language

Language:English
Title:Cartilage tissue engineering on biosynthetic polymer polyHIPE scaffold
Abstract:Cartilage tissue engineering remains a field with possibilities for improvement despite very intense research efforts in recent times. Development of artificial materials and 3-D cellular scaffolds has an important role in facilitation of cartilage regeneration. An interesting approach to scaffold formation presents the so-called emulsion templating technique. The resulting material; termed polyHIPE (PHP), is a synthetic highly porous polymer prepared by the polymerization of high internal phase emulsions (HIPEs). The main objective of this PhD thesis is to investigate the potential for neocartilage formation within cellular scaffolds prepared from PHP material. The fabrication of PHP was specifically tailored to produce a highly porous (85 %) structure with the primary pore size in the range of 50-170 µm for cartilage tissue engineering. The resulting PHP scaffold was proven biocompatible with human articular chondrocytes and viable cells were observed within the materials as evaluated using the Live/Dead assay and histological analysis. Chondrocytes with round nuclei were organized into multicellular layers on the PHP surface and were observed to grow approximately 300 µm into the scaffold interior. The accumulation of collagen type 2 was detected using immunohistochemistry and chondrogenic specific genes were expressed with favorable collagen type 2 to 1 ratio. In addition, PHP samples are biodegradable and their baseline mechanical properties are similar to those of native cartilage. The research results suggested that the designed PHP cellular scaffold is suitable for further use in cartilage tissue engineering.
Keywords:human articular cartilage, tissue engineering, biocompatible cellular scaffold, synthetic polymer, polyHIPE, chondrocyte differentiation


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