Fig 2: miR-29b-3p regulated cartilaginous degeneration-related molecules, including MMP-1, MMP-13, COL II and COL X, and direct targeted at 3' UTR of rat GRN (rGRN) mRNA. A-D. Concentrations of secreted MMP-1, MMP-13, COL II and COL X in the media of rat primary chondrocytes or SW-1353 human chondrosarcoma cells after transfection with the miR-29b-3p mimic or the inhibitor measured by ELISA assay. E, F. mRNA expression of COL2A1 and COL10A1 of rat primary chondrocytes or SW-1353 human chondrosarcoma cells after transfection with the miR-29b-3p mimic or the inhibitor measured by qRT–PCR assay. G. Schematic representation of miR-29b-3p's predicted binding site in the 3'UTR of rGRN mRNAs. H, I. The relative luciferase activity after the cotransfection of miR-29b-3p mimic and 3' UTR of GRN mRNA into HEK293 and SW1353 cells respectively. *P < 0.05, **P < 0.01, compared with NC (negative control). NS: not significant.

Fig 3: KAT6B promotes chondrocyte proliferation. a qPCR was performed to measure the expression of KAT6B in primary chondrocytes from CS tissues after transfection. b Western blot was performed to measure the expression of KAT6B in primary chondrocytes from CS tissues after transfection. c EdU staining was performed to test cell proliferation in primary chondrocytes from CS tissues after transfection. DAPI, blue; EdU, red. Bar, 100 µm. d Western blotting was performed to measure the expression of RUNX2, COL10A1, ß-catenin, c-Myc, and VEGF in primary chondrocytes in primary chondrocytes from CS tissues after transfection. CS, congenital scoliosis. *p < 0.05

Fig 4: Gene expression levels of chondrogenic and hypertrophic markers of ADSCs and ACs cultured in 3D scaffolds for 4 weeks.ADSCs cultured with RAD16-I, RAD/CS and RAD/Decorin scaffolds in chondrogenic medium were analyzed by qRT-PCR for collagen type I (COL1, A), collagen type II (COL2, B), SOX9 (C), aggrecan (ACAN, D), collagen type X (COL10, E) and RUNX2 (F). ACs cultured with RAD16-I, RAD/CS and RAD/Decorin scaffolds in expansion (exp) and chondrogenic (ch) medium were analyzed by qRT-PCR for COL1 (G), COL2 (H), SOX9 (I), ACAN (J), COL10 (K) and RUNX2 (L). Ct values relative to ribosomal protein L22 (RPL22) were obtained and reported as the fold increase (??Ct) relative to 2D cultures (Significant differences are indicated as * for p<0.05, ** for p<0.01, and *** for p<0.001, One-way ANOVA, N = 2 n = 3).

Fig 5: Gene expression of chondrogenic and hypertrophic markers for PCL, PCL/RAD and RAD samples under expansion or chondrogenic media for 30 days. (a) Collagen type I (COL1A1), (b) collagen type II (COL2A1), (c) collagen type X (COL10A1), (d) RUNX2, (e) SOX9 and (f) aggrecan (ACAN) were determined through qRT-PCR. Ct values relative human beta actin (ACTB) were obtained and reported as fold increase (??Ct) relative to 2D cultures (Statistical differences are indicated as: *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001, two-way ANOVA). n = 3.