Fig 5. Tethered BP₁₀-T-EGF induces in vitro expansion of hBMSCs cultured on ßTCP scaffolds.

Cells were seeded on control and BP₁₀-T-EGF- treated scaffolds, cultured in expansion medium for 7 days and then subjected to Alamar blue assay to assess cell numbers relative to controls. A 2–2.3 fold increase in total hBMSC number was observed across three different tethered BP₁₀-T-EGF surface densities (N = 3 per condition; one-way ANOVA p-value<0.05 (p-value = 0.02); all pairwise p-values of tethered EGF vs control were <0.05 using Tukey’s multiple comparison test) (A). To parse effects that tethered EGF might have on plating efficiency, MSCs were cultured on ßTCP scaffolds for 12–24 hr and the number of attached cells was then determined using a visual count of nuclei following acid demineralization of agarose-embedded scaffolds. Results from 12-hr and 24-hr cell culture revealed no statistical difference (n.s.; N≥3 per condition; One-way ANOVA p-value>0.05) in the number of hBMSCs as a function of condition, indicating that tethered EGF is not altering the plating efficiency (B).