Tkivni inženiring hrustančnega tkiva na biosintetičnem polimernem polyHIPE nosilcu

doktorska disertacija

Jakob Naranđa (Author), Matjaž Vogrin (Mentor), Ivan Krajnc (Thesis defence commission member), Marjan Rupnik (Co-mentor)

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;

Data

Language:	Slovenian
Year of publishing:	2017
Typology:	2.08 - Doctoral Dissertation
Organization:	UM - University of Maribor
Publisher:	J. Naranđa]
UDC:	612.75-018.3:602.9+606(043.3)
COBISS:	6112063
Views:	1338
Downloads:	158
Average score:	0 (0 votes)
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Other data

Secondary language:	English
Secondary title:	Cartilage tissue engineering on biosynthetic polymer polyHIPE scaffold
Secondary 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.
Secondary keywords:	Cartilage, articular;Sklepni hrustanec;Tissue engineering;Methods;Tkivno inženirstvo;Metode;Biocompatible materials;Biokompatibilni materiali;
URN:	URN:SI:UM:
Type (COBISS):	Dissertation
Thesis comment:	Univ. v Mariboru, Medicinska fak.
Pages:	101 str.
ID:	9236562