Effect of nitrogen-rich cell culture surfaces on type X collagen expression by bovine growth plate chondrocytes.
BACKGROUND
Recent evidence indicates that osteoarthritis (OA) may be a systemic disease since mesenchymal stem cells (MSCs) from OA patients express type X collagen, a marker of late stage chondrocyte hypertrophy (associated with endochondral ossification). We recently showed that the expression of type X collagen was suppressed when MSCs from OA patients were cultured on nitrogen (N)-rich plasma polymer layers, which we call "PPE:N" (N-doped plasma-polymerized ethylene, containing up to 36 atomic percentage (at.% ) of N.


METHODS
In the present study, we examined the expression of type X collagen in fetal bovine growth plate chondrocytes (containing hypertrophic chondrocytes) cultured on PPE:N. We also studied the effect of PPE:N on the expression of matrix molecules such as type II collagen and aggrecan, as well as on proteases (matrix metalloproteinase-13 (MMP-13) and molecules implicated in cell division (cyclin B2). Two other culture surfaces, "hydrophilic" polystyrene (PS, regular culture dishes) and nitrogen-containing cation polystyrene (Primaria®), were also investigated for comparison.


RESULTS
Results showed that type X collagen mRNA levels were suppressed when cultured for 4 days on PPE:N, suggesting that type X collagen is regulated similarly in hypertrophic chondrocytes and in human MSCs from OA patients. However, the levels of type X collagen mRNA almost returned to control value after 20 days in culture on these surfaces. Culture on the various surfaces had no significant effects on type II collagen, aggrecan, MMP-13, and cyclin B2 mRNA levels.


CONCLUSIONS
Hypertrophy is diminished by culturing growth plate chondrocytes on nitrogen-rich surfaces, a mechanism that is beneficial for MSC chondrogenesis. Furthermore, one major advantage of such "intelligent surfaces" over recombinant growth factors for tissue engineering and cartilage repair is potentially large cost-saving.
Lady Davis Institute for Medical Research, SMBD-Jewish General Hospital, Montreal, QC, Canada.
Alain
A
Petit
Caroline N
CN
Demers
Pierre-Luc
PL
Girard-Lauriault
Dorothy
D
Stachura
Michael R
MR
Wertheimer
John
J
Antoniou
Fackson
F
Mwale
D053674:Aggrecans / Q000378:metabolism; D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques; D002455:Cell Division; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D024043:Collagen Type II / Q000378:metabolism; D024121:Collagen Type X / Q000235:genetics / Q000378:metabolism*; D017077:Culture Media, Conditioned; D056765:Cyclin B2 / Q000378:metabolism; D015536:Down-Regulation; D006132:Growth Plate / Q000166:cytology* / Q000378:metabolism; D006984:Hypertrophy / Q000378:metabolism; D053509:Matrix Metalloproteinase 13 / Q000378:metabolism; D009584:Nitrogen / Q000378:metabolism*; D012333:RNA, Messenger; D020133:Reverse Transcriptase Polymerase Chain Reaction
D053674:Aggrecans; D024043:Collagen Type II; D024121:Collagen Type X; D017077:Culture Media, Conditioned; D056765:Cyclin B2; D012333:RNA, Messenger; D053509:Matrix Metalloproteinase 13; D009584:Nitrogen
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