Figure 1.

PDLC activity in peptide-functionalized hydrogels. A) Fold-change ALP activity at 1 week and B) fold-change matrix mineralization at 3 weeks for high single peptide (2 mM) and dual peptide concentrations versus Ctrl. C-D) Regression coefficient estimates for fold-change ALP activity of individual donor PDLCs (C) and pooled values of donor 1–3 PDLCs (D). E-F) Regression coefficient estimates for fold-change mineralization of individual donor PDLCs (E) and pooled values of donor 1–3 PDLCs (F). G-H) Response surfaces for fold-change ALP activity (G) and fold-change matrix mineralization for pooled values of donor 1–3 PDLCs (H). N=3 hydrogels per donor. I) ALP activity and J) matrix mineralization in hydrogels optimized for ALP activity (AO – gray bars) or matrix mineralization (MO – white bars) using PDLCs from pooled donors 4–6. ALP activity was also measured at 1 week on tissue culture polystyrene using the same pooled PDLC population (black bar). K) Representative z-stack images of hydrogel matrix mineralization. Xylenol orange-stained calcium deposits are red, Hoechst 33342-stained PDLC nuclei are blue. Scale bars are 100 μm. L) Total calcium concentration in optimized hydrogels normalized to PDLC DNA content. N=6 hydrogels. A-B) One-way ANOVA with Dunnett post hoc. 1, 2, and 3 indicate p<0.05 for individual donor’s PDLCs versus individual Ctrl, a, b indicates p<0.05 for dual peptide pooled means versus 2 mM RGD (a) and 2 mM GFOGER (b) pooled means, **p<0.01, ***p<0.001 for pooled means versus Ctrl. C-F) ANOVA. I) One-way ANOVA with Tukey post hoc. J, L) unpaired t test. *p<0.05, ***p<0.001, ****p<0.0001 for between group comparisons.