The effect of novel nitrogen-rich plasma polymer coatings on the phenotypic profile of notochordal cells.
BACKGROUND
The loss of the notochordal cells from the nucleus pulposus is associated with ageing and disc degeneration. However, understanding the mechanisms responsible for the loss of these cells has been hampered in part due to the difficulty of culturing and maintaining their phenotype. Furthermore, little is known about the influence of the substratum on the molecular markers of notochordal cells.


METHODS
Notochordal cells were isolated from lumbar spine of non-chondrodystrophoid dogs and cultured on N-rich plasma polymer layers, so-called "PPE:N" (N-doped plasma-polymerised ethylene, containing up to 36% [N]) surfaces, for 3, 7 or 14 days. Gene expression of vimentin (VIM), pleiotrophin (PTN), matrix Gla protein (MGP), cartilage oligomeric matrix protein (COMP), keratin 18 (KRT 18), aggrecan (AGG), collagen type 1 (COL1A2), collagen type 2 (COL2A1) was analyzed through semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR).


RESULTS
Notochordal cells were maintained in culture on PPE:N for up to 14 days with no loss in cell viability. Except for VIM, gene expression varied depending on the culture periods and [N] concentration of the substratum. Generally, PPE:N surfaces altered gene expression significantly when cells were cultured for 3 or 7 days.


CONCLUSIONS
The present study has shown that notochordal cells from dogs can attach to and grow on PPE:N surfaces. Analysis of the expression of different genes in these cells cultured on different N-functionalized surfaces indicates that cellular behaviour is gene-specific and time-dependent. Further studies are required to better understand the roles of specific surface functionalities on receptor sites, and their effects on cellular phenotypes.
Mwale
Fackson
F
Division of Orthopaedic Surgery, McGill University, and Lady Davis Institute for Medical Research, 3755, Chemin de la Cote Ste-Catherine, Montreal, QC H3T 1E2, Canada. fmwale@ldi.jgh.mcgill.ca
Wang
Hong Tian
HT
Petit
Alain
A
Girard-Lauriault
Pierre-Luc
PL
Hunter
Christopher J
CJ
Ouellet
Jean A
JA
Wertheimer
Michael R
MR
Antoniou
John
J
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D001698:Biomedical Engineering / Q000379:methods; D004285:Dogs; D005030:Ethylenes / Q000737:chemistry; D005786:Gene Expression Regulation; D006801:Humans; D009584:Nitrogen / Q000097:blood / Q000737:chemistry* / Q000378:metabolism; D009672:Notochord / Q000166:cytology* / Q000378:metabolism; D010641:Phenotype; D010949:Plasma / Q000378:metabolism; D011108:Polymers / Q000737:chemistry*