MC3T3 cells were cultured for 10 days in osteogenic medium until mineralization started, at which time 300μM pVTK peptide was added (indicated by arrow on timeline (A)), and cultured for 4 additional days. (B) Calcium levels (quantified from demineralized matrices) and (C) Enpp1 enzyme activity were significantly reduced with peptide treatment (D) TNAP enzyme activity was significantly higher at day 14 with peptide treatment (* p < 0.05 Osteo vs. Osteo + Peptide at a specific day, t-test, ** p < 0.001 Osteo vs. Osteo + Peptide at a specific day, t-test, ^ p < 0.05 compared to day 10 within Osteo or Osteo + Peptide treatment group, ^^ p < 0.001 compared to day 10 within Osteo or Osteo + Peptide treatment group, % p < 0.05 compared to day 12 within Osteo or Osteo + Peptide treatment group, 1 way ANOVA or 1 way ANOVA on Ranks, Student-Newman-Keuls post hoc test)