Effect of nitrogen-rich cell culture surfaces on type X collagen expression by bovine growth plate chondrocytes

Biomed Eng Online. 2011 Jan 18:10:4. doi: 10.1186/1475-925X-10-4.

Abstract

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.

Conclusion: 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aggrecans / metabolism
  • Animals
  • Cattle
  • Cell Culture Techniques
  • Cell Division
  • Cells, Cultured
  • Chondrocytes / cytology
  • Chondrocytes / metabolism*
  • Collagen Type II / metabolism
  • Collagen Type X / genetics
  • Collagen Type X / metabolism*
  • Culture Media, Conditioned
  • Cyclin B2 / metabolism
  • Down-Regulation
  • Growth Plate / cytology*
  • Growth Plate / metabolism
  • Hypertrophy / metabolism
  • Matrix Metalloproteinase 13 / metabolism
  • Nitrogen / metabolism*
  • RNA, Messenger
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Aggrecans
  • Collagen Type II
  • Collagen Type X
  • Culture Media, Conditioned
  • Cyclin B2
  • RNA, Messenger
  • Matrix Metalloproteinase 13
  • Nitrogen