DOS 2016

DOS Kongressen 2016 ·

199

Substrate and Surface Guidance of Human

Chondrocytes In Vitro

Natasja Jørgensen, Morten Foss, Nikolaj Gadegaard, Casper Foldager, Martin

Lind, Helle Lysdahl

Ortopædiskforskningslaboratorium,AarhusUniversitetshospital;Interdisciplinary

Nanoscience Center, Aarhus Universitet; School of Engineering, Glasgow

Universitet; Ortopædisk forskningslaboratorium , Aarhus Universitetshospital;

Ortopædisk forskningslaboratorium, Aarhus Universitetshospital; Ortopædisk

forskningslaboratorium , Aarhus Universitetshospital

Background:

The nature of the surface on which chondrocytes are cultured ex

vivo plays an important role for proliferation and differentiation in the field of

cartilage regeneration.

Purpose / Aim of Study:

We aimed to investigate the behaviour of human

chondrocytes on different substrates and surface chemistry.

Materials and Methods:

Human chondrocytes were isolated from cartilage bi-

opsies collected from 3 patients. Chondrocytes were seeded with 2,500 cells/

cm2 on polystyrene (rigid) or polydimethylsiloxane (soft) with surface chem-

istry of oxygen plasma (PL) or fibronectin (FN) and cultured for 1, 4, 7, and 10

days. Proliferation, cell viability, cell size, and gene expression were performed

using methylene blue staining, XTT assay, actin staining, and RT-qPCR, respec-

tively.

Findings / Results:

We found similar proliferative capacity over time for all

substrates and surface chemistry. Cell viability was significantly higher on the

polystyrene compared with PDMS. For surface chemistry, PL and FN, cell vi-

ability was highest in chondrocytes cultured on FN surfaces. The cytoskeleton

of chondrocytes on FN was associated with chondrocyte size > 2000 μm2

compared with PL where chondrocyte sizes were < 1000 μm2. For substrates,

we found significantly higher expression of SOX9 and COL2A1 in chondrocytes

cultured on PDMS compared with polystyrene. For surface chemistry, chondro-

cytes cultured on PL had significantly higher SOX9, COL2A1, ACAN expression

compared with FN.

Conclusions:

Cultivation of human chondrocytes on soft PMDS coated with PL

resulted in chondro- inductive conditions having the lowest cell viability, small-

est cell size, and the highest expression of cartilage specific genes. Constituting

further investigations aiming at elucidating the role of a softer culture substrate

when culturing human chondrocytes ex vivo.

No conflicts of interest reported

150.