Bovine collagen peptides compounds promote the proliferation and differentiation of MC3T3-E1 pre-osteoblasts

PLoS One. 2014 Jun 13;9(6):e99920. doi: 10.1371/journal.pone.0099920. eCollection 2014.


Objective: Collagen peptides (CP) compounds, as bone health supplements, are known to play a role in the treatment of osteoporosis. However, the molecular mechanisms of this process remain unclear. This study aimed to investigate the effects of bovine CP compounds on the proliferation and differentiation of MC3T3-E1 cells.

Methods: Mouse pre-osteoblast cell line MC3T3-E1 subclone 4 cells were treated with bovine CP compounds. Cell proliferation was analyzed by MTT assays and the cell cycle was evaluated by flow cytometry scanning. Furthermore, MC3T3-E1 cell differentiation was analyzed at the RNA level by real-time PCR and at the protein level by western blot analysis for runt-related transcription factor 2 (Runx2), a colorimetric p-nitrophenyl phosphate assay for alkaline phosphatase (ALP), and ELISA for osteocalcin (OC). Finally, alizarin red staining for mineralization was measured using Image Software Pro Plus 6.0.

Results: Cell proliferation was very efficient after treatment with different concentrations of bovine CP compounds, and the best concentration was 3 mg/mL. Bovine CP compounds significantly increased the percentage of MC3T3-E1 cells in G2/S phase. Runx2 expression, ALP activity, and OC production were significantly increased after treatment with bovine CP compounds for 7 or 14 days. Quantitative analyses with alizarin red staining showed significantly increased mineralization of MC3T3-E1 cells after treatment with bovine CP compounds for 14 or 21 days.

Conclusions: Bovine CP compounds increased osteoblast proliferation, and played positive roles in osteoblast differentiation and mineralized bone matrix formation. Taking all the experiments together, our study indicates a molecular mechanism for the potential treatment of osteoarthritis and osteoporosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Cell Differentiation / drug effects
  • Cell Line
  • Cell Proliferation / drug effects
  • Collagen / pharmacology*
  • Mice
  • Osteoblasts / cytology*
  • Osteoblasts / drug effects*
  • Peptides / pharmacology*


  • Peptides
  • Collagen

Grants and funding

This work was supported by The cooperation project from Technical institute of Physics and Chemistry,CAS (Grant No. LHS- DBSW2013-01).The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.