Figure 1.
Schematic diagrams of specimens for tensile testing and load-displacement curve.
(A) Schematic diagram of the intervertebral disc and locations of annulus fibrosus (AF) specimens for tensile testing. AF samples (arrow) were dissected from the outer zones of anterior regions, with the longest dimension in the circumferential direction. (B) Schematic diagram of load-displacement curve.
Figure 2.
Representative macroscopic images of AF before and after decellularization.
(A) Triton X-100, (B) SDS, (C) trypsin, (D) control.
Figure 3.
Hematoxylin and eosin (H&E) staining of cross-sections of AF samples.
(A) Triton X-100, (B) SDS, (C) trypsin, (D) control. Collagen fiber fracture (arrows).
Figure 4.
Hoechst 33258 staining of cross-sections of AF samples.
(A) Triton X-100, (B) SDS, (C) trypsin, (D) control. DNA (arrows).
Figure 5.
Toluidine blue staining of cross-sections of AF samples.
(A) Triton X-100, (B) SDS, (C) trypsin, (D) control.
Figure 6.
Safranin O staining of cross-sections of AF samples.
(A) Triton X-100, (B) SDS, (C) trypsin, (D) control.
Figure 7.
Sirius red stain of cross-sections of AF samples.
(A) Triton X-100, (B) SDS, (C) trypsin, (D) control.
Figure 8.
Collagen I immunofluorescent staining of cross-sections of AF samples.
(A) Triton X-100, (B) SDS, (C) trypsin, (D) control.
Figure 9.
Scanning electron micrographs of cross-sections of AF samples.
(A) Triton X-100, (B) SDS, (C) trypsin, (D) control.
Figure 10.
Water (A), collagen (B), and glycosaminoglycan (GAG) content (C) of AF.
Data are mean ± SD. * = p<0.05 compared to control, # = p<0.05 compared to Triton X-100.
Table 1.
The biomechanical properties of annulus fibrosus with decellularization treatments.
Figure 11.
Cytotoxicity of decellularized AF.
MTT assay of proliferation of AF cells cultured with different concentrations of scaffold extracts.
Figure 12.
Recellularization of decellularized AF and Evaluation.
(A) H&E staining of cell-containing constructs. AF cells (arrows). (B) Live/dead staining of cells seeded into decellularized AF. (Green: viable, red: necrotic).