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. 2025 Oct 7;13:1607469. doi: 10.3389/fbioe.2025.1607469

FIGURE 6.

The figure is composed of fluorescence microscopy images and corresponding bar graphs showing mean pixel intensity for COL2A1 and ACAN in 2D and 3D-like cultures. Panels A and B display 2D cultures, with nuclei stained blue, cytoskeleton red, and COL2A1 or ACAN green. Panels C and D present bar graphs quantifying COL2A1 or ACAN intensity in 2D samples. Panels E and F show 3D-like cultures with the same staining. Panels G and H provide bar graphs of COL2A1 and ACAN intensity in 3D samples. Differences between HHA/BC and HHA/BC+PRP samples are visible across all panels.

IF staining and quantification of COL2A1 and ACAN in 2D and 3D-like in vitro culture. (A) IF images of COL2A1 in 2D culture; (B) IF images of ACAN in 2D culture; (C) quantification of COL2A1 mean pixel fluorescence intensity in 2D culture; (D) quantification of ACAN mean pixel fluorescence intensity in 2D culture; (E) IF images of COL2A1 in 3D-like culture; (F) IF images of ACAN in 3D-like culture; (G) quantification of COL2A1 mean pixel fluorescence intensity in 3D-like culture; (H) quantification of ACAN mean pixel fluorescence intensity in 3D-like culture. Representative micrographs: FITC green antibody was used for collagen and aggrecan detection, while nuclei were stained in blue and cytoskeleton in red. Scale bar 10 μm, objective magnification ×40. Data are presented as mean ± SD. Statistical analysis was performed using a t-test: *p < 0.05 vs. pCTR; #p < 0.05 for HHA/BC+PRP vs. HHA/BC.