DOS Kongressen 2014 ·
133
Human chondrocytes cultivated on modified
polystyrene conserve their chondrogenic phenotype
in vitro
Natasja Leth Jørgensen, Dang Le, Casper Foldager, Martin Lind, Helle Lysdahl
Orthopaedic Research Laboratory, Aarhus University Hospital; Interdisciplinary
Nanoscience Center, iNANO, Aarhus University; Sports Trauma Clinic, Aarhus
University Hospital
Background:
Autologous articular chondrocytes tend to dedifferentiate under
prolonged expansion culture ex vivo. Porous scaffolds have been widely used to
guide cells and grow new tissue.
Purpose / Aim of Study:
In this study, we investigated the influence of pre-
cipitant induced porosity augmentation (PIPA) modified polystyrene surface on
human chondrocytes (HCs) cultured in vitro. We hypothesized, that culturing
HCs on 2D PIPA modified surfaces would conserve their phenotype.
Materials and Methods:
The modification of polystyrene was made by the
technique precipitant induced porosity augmentation (PIPA). Polystyrene was
immersed with 1,4 dioxane and ozone treated prior to cell seeding. HCs were
enzymatically isolated from cartilage biopsies collected from the inter-condylar
groove in the distal femur. Isolated chondrocytes were expanded to passage
1 (P1) in DMEM/F12 supplemented with 10% FCS, 5 ng/mL bFGF, 1 ng/mL
TGFβ3, and 1:100 penicillin-streptomycin (P/S). Chondrocytes in P1 were then
seeded on PIPA modified polystyrene surfaces or on traditional monolayer (on a
plan polystyrene surface) with 10,000 cells/cm2 in DMEM/F12 supplemented
with 10% FCS and 1:100 P/S. HCs were cultivated until P4 and samples were
collected for quantitative RT-PCR at each passages P1-P4. Toluidine blue and
collagen II stains were performed on P2 HCs cultivated in pellets, 1×106 cells/
mL, for 28 days.
Findings / Results:
The PIPA surface promoted chondrogenic differentiation
of HCs compared with traditional monolayer culture evident by higher gene ex-
pression of COL II, and the differentiation indices COLII/COLI and AGG/COLI.
HCs expanded on PIPA surfaces prior to pellet formation revealed a better chon-
drogenicity by more synthesis of proteoglycans and collagen II.
Conclusions:
Cultivation of HCs on the PIPA modified surface seems to con-
serve the chondrogenic phenotype.
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