Fig 1: Overexpression of WNT16 does not protect against S. aureus-induced arthritis or bone loss. A Kaplan–Meier survival curve showing the percentage of surviving mice over time following S. aureus inoculation (n = 15 per group). Statistical comparison was performed using the log-rank test. Body weight change B and clinical arthritis scores C over the 8-day period post-inoculation. Data are presented as mean ± SEM (n = 13–15 mice per group) and were analyzed using the Mann–Whitney U test. (D-F) µCT analysis of the fifth lumbar vertebra 8 days post-inoculation, showing D bone volume over total volume (BV/TV), E trabecular number (Tb.N), and F trabecular thickness (Tb.Th) in PBS-treated controls and S. aureus-inoculated mice. Statistical analysis was performed using two-way ANOVA. s.a. = S. aureus; ns = non-significant
Fig 2: Overexpression of WNT16 does not protect against PAM2-induced bone loss. A Wnt16 mRNA expression in trabecular-rich vertebral bodies. B Bone volume and C representative 3-dimensional images of calvarial bones analyzed by µCT. D Bone volume over total volume (BV/TV), E trabecular number (Tb. N), and F trabecular thickness (Tb. Th), in femur as measured using µCT. Data were collected from 7-week-old Obl-Wnt16 and wild-type (WT) mice five days after injections of PAM2 or vehicle (control). Values are mean ± SEM (n = 6–13 mice per group). Statistical analyses were performed using a two-way ANOVA. ns = non-significant
Fig 3: TNF-α reduces Wnt16 mRNA expression in osteoblasts, but WNT16 treatment does not prevent periarticular bone loss in antigen-induced arthritis. A Wnt16 mRNA expression in primary mouse calvarial osteoblasts cultured for 24 h in control medium with or without TNF-α (50 ng/ml). Values are presented as mean ± SEM (n = 4). Data were analyzed using an unpaired Student’s t-test. B Schematic representation of the antigen-induced arthritis model. Upper panel: Control group after secondary immunization (left knee = arthritic; right knee = non-arthritic). Lower panel: Liposome-treated group (left knee = arthritic + liposomes with WNT16; right knee = arthritic + empty liposomes). (C-F) Peripheral quantitative computed tomography (pQCT) analysis of bone mineral density (BMD): total BMD in the metaphyseal regions of the distal femur C and proximal tibia D, and trabecular BMD in the same regions of the distal femur E and proximal tibia F. Data were collected from bones adjacent to non-arthritic and antigen-induced arthritic mouse knees treated with PBS, as well as from bones adjacent to arthritic knees treated with liposomes with WNT16 or empty liposomes. Samples were obtained from 9-week-old female wild-type mice. Values are mean ± SEM (n = 5–9 mice per group). Data assessed by paired Student’s t-test. lipo. = liposomes; inj. = injection; w = with; and w/o = without
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