Table 1.
Sequence of primers.
Table 2.
Histological grading scale by Masuda et al [31].
Figure 1.
Sox9-transduced BMSCs in monolayer.
(A) P3 BMSCs exhibited a uniform fibroblast-like shape, and fluorescence microscopy showed Sox9 transduction efficiency at days 3, 7 and 14. (B) Sox9 expression was evaluated by western blot at days 3, 7, 10 and 14 after transduction. (C) Real-time PCR for the expressions of Sox9, Aggrecan, Col II, Col I and Col X was performed at day 21 after transduction using GAPDH as a housekeeping gene. All results represent the mean±SD of triplicate samples. *P<0.05, #P<0.01. Scale bar = 200 μm.
Figure 2.
Sox9-transduced BMSCs in C/Gp gels.
(A) Scanning electron microscopy observation of the C/Gp gels. (B) The C/Gp was liquid at room temperature and gelled at 37°C. (C) Live/dead assay of the BMSCs encapsulated in C/Gp gels after 3 days of culture. (D) The MTT assay was used to assess the BMSC proliferation in C/Gp gels over 14 days. (E) The gene expression analysis of the BMSCs in C/Gp gels after 21 days of culture. #P<0.01. Scale bar = 25 μm.
Figure 3.
(A) T2-weighted images of the treated discs (outlined by the red box) and adjacent control discs at 6 and 12 weeks post-transplantation. (B) The mean T2 of NP at 6 and 12 weeks post-transplantation. *P<0.05, #P<0.01 vs. the Sox9 group. Coronal and sagittal CT reconstruction images for the GFP (C) and Sox9 (D) groups.
Figure 4.
Macroscopic views of the representative discs at 12 weeks post-transplantation:
: (A) normal control group; (B) degeneration group; (C) C/Gp group; (D) GFP group and (E) Sox9 group.
Figure 5.
Haematoxylin and eosin staining of sections from the 5 groups at 6 (A–J) and 12 weeks (K–T) post-transplantation.
Scale bar = 500 μm.
Figure 6.
Safranin O staining of sections from the 5 groups at 6 (A–J) and 12 weeks (K–T) post-transplantation.
Scale bar = 200 μm.
Figure 7.
Type II collagen immunohistological staining and histological scores.
Representative sections from the 5 groups at 6 (A–E) and 12 weeks (F–J) post-transplantation. (K) Quantitative analysis of the type II collagen staining using mean density. (L) Histological scores of discs from the 5 groups at 6 and 12 weeks post-treatment. *P<0.05, #P<0.01 vs. the Sox9 group. Scale bar = 200 μm.
Figure 8.
Double staining of the transplanted BMSCs using antibodies against GFP (green) and Sox9 (red) at 12 weeks post-transplantation.
DAPI was used to stain the cell nucleus (blue), and the colours are labelled in the image. Scale bar = 200 μm.