Fig. 7

Deletion mutant 2 (Δ2-RUNX1-iMSCs) showed enhanced bone formation and repair of cranial defects in vivo: a Study design of calvarial defect in 8-week-old NOD-SCID mice. Wild type (BD-1-4)-iMSCs, homozygous deletion mutant Δ1-iMSCs (#9) and Δ2-iMSCs (#46) were implanted in defect site at 3 different time during surgery (day 0), and week 4 and week 8; b Surgical procedure involved in creation of 2.3 mm sub-critical-sized defects in the parietal bones of 8-week-old mice and delivery of iMSCs encapsulated with 4% PEG-MAL hydrogels; c Quantification of regenerated bone volume within the defect site at 12-week post surgery and μCT reconstructions of skull and defect size are shown; d Quantification of differences in regenerated bone volume (%BV/TV) within the defect and compared them between wildtype and Δ1-iMSCs (#9) and Δ2-iMSCs (#46) by μCT analysis. μCT reconstructions of defects in each experimental group were shown. Data are presented as mean ± SD with P-values reported. P ≤ 0.01 indicate values are statistically different in Δ-RUNX2-iMSCs as compared to their wild type control iMSCs. n = 10 mice per group as shown in Figure D. Data were analyzed by paired t-Test using Graph-pad Prism (V9.0); e Representative sections of the calvarial defect area on mice skulls from all three experimental groups were stained with hematoxylin and eosin (H&E) staining. Dashed boxed areas in the upper panel are shown at a higher magnification (10X) in the bottom panel for each sample; f Immunofluorescence staining of implanted iMSCs for detection of human vimentin to distinguish implanted human cells from host mice cells. Nuclear DNA was labeled with DAPI. Scale bars, 170 μm