Fig. 5: Endogenous stem cells recruitment by HCLS.
a1 Schematic depiction of in vitro BMSCs recruitment by incubating various scaffolds in a total rabbit cranial bone marrow for 48 h. a2 CD44 immunofluorescent staining imaged by CLSM. a3 Schematic depiction of transwell assay in studying BMSCs migration to various scaffolds. a4 Quantification of migrated BMSCs by dissolving crystal violet and spectrophotometrically measured at 573 nm, the optical density (OD) resulted was normalized by control. BMSCs alone was used as control (**p = 0.0028, ***p = 0.0001, ***p = 0.0002, ***p = 6.7070 × 10−5). n = 3 biologically independent replicates. b SEM images of endogenous cells on the surface of various scaffolds. c CLSM images (rhodamine-phalloidin/DAPI staining) of endogenous cells on the surface of various scaffolds. d CLSM images of CD44/F-actin/DAPI staining of various scaffolds. e1 Confocal quantitative images (CD44/DAPI staining) of ESCs in scaffolds. e2 High-magnification images (CD44/DAPI staining) of ESCs in scaffolds. e3 Quantitative analysis of the total number of cells in scaffolds (*p = 0.0470, **p = 0.0023). e4 Quantitative analysis of the number of ESCs in scaffolds (***p = 0.0008, ***p = 0.0003). n = 3 biologically independent replicates. f1–3 BMP-2 and Runx2 immunofluorescence staining at one week of different treatments and semiquantitative analysis (f1: ***p = 0.0009, ***p = 5.5298 × 10−5; f2: **p = 0.0078, ***p = 5.8847 × 10−5). g1 CLSM images of CD31 immunofluorescence staining. g2 High-magnification images (CD31/DAPI staining) in scaffolds. g3 Quantitative analysis of the distribution of blood vessels inside various scaffolds (**p = 0.0044, **p = 0.0023). n = 3 cells examined three independent experiments. h H&E staining of different scaffolds after one week (V: new vessels. N: new bone tissue. S: scaffolds). (Two-sided comparison, error bars represent standard deviation, *p < 0.05, **p < 0.01, and ***p < 0.001).
