Human intervertebral disc-derived cells are recruited by human serum and form nucleus pulposus-like tissue upon stimulation with TGF-beta3 or hyaluronan in vitro.
The aims of this work were to test whether human intervertebral disc-derived nucleus pulposus cells (hNP-cells) are attracted by human serum and to analyze if matrix generation from hNP-cells is promoted under the influence of transforming growth factor-beta3 (TGF-beta3) or hyaluronan (HA) in vitro. Using the multi-well chemotaxis assay to determine cell migration under the influence of different concentrations of human serum, it was demonstrated that dedifferentiated hNP-cells are able to migrate towards a serum fraction gradient in a concentration-dependent manner. Re-differentiation capacity of hNP-cells in 3D micro-masses under the influence of TGF-beta3 or hyaluronan was also tested. Gene expression analysis of types I, II, III and IX collagen, as well as aggrecan, COMP and LINK of hNP-cells in 3D-micro-mass cell-culture revealed a strong increase of these markers in TGF-beta3 treated cells. Furthermore, histochemical and immuno-histochemical staining after 28d showed proteoglycan and type II collagen-rich matrix for both, the TGF-beta3 and the hyaluronan treated cells. These findings show that TGF-beta3 or hyaluronan are able to induce the differentiation and that human serum stimulates the migration of hNP-cells in vitro. Therefore, hyaluronan and serum are suited for cell-free biomaterials as cell migration and differentiation inducing factors intended for biological treatment strategies of the intervertebral disc.
Haberstroh
K
K
Charité-University Medical Center Berlin, Department of Rheumatology, Berlin, Germany.
Enz
A
A
Zenclussen
M L
ML
Hegewald
A A
AA
Neumann
K
K
Abbushi
A
A
Thomé
C
C
Sittinger
M
M
Endres
M
M
Kaps
C
C
D054337:Cell Dedifferentiation / Q000187:drug effects / Q000502:physiology; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis; D006801:Humans; D006820:Hyaluronic Acid / Q000494:pharmacology*; D007403:Intervertebral Disc / Q000166:cytology* / Q000378:metabolism*; D044967:Serum; D053782:Transforming Growth Factor beta3 / Q000494:pharmacology*; D055675:Viscosupplements / Q000494:pharmacology*
D016326:Extracellular Matrix Proteins; D053782:Transforming Growth Factor beta3; D055675:Viscosupplements; D006820:Hyaluronic Acid