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. 2022 Jul 7;13:3919. doi: 10.1038/s41467-022-31646-0

Fig. 2. Potential inhibitory role of AnxA1 in inflammatory osteolysis induced by implant debris.

Fig. 2

a Neutrophil count in Ly6G antibody- and control antibody-treated mice. Detection of neutrophils using IHC in granulomatous tissue formed on calvarial bone after the implantation of UHMWPE debris and treatment by antibodies. Treatment with a Ly6G antibody reduced the number of Elastase positive cells. Results represent the mean of relative expression values ± SEM of 3 mice. Significant difference between the two groups was determined by two-tailed Student’s t test. Scale bars are 100 µm. b AnxA1 expression in calvarial bone tissues of neutrophil-depleted mice analyzed by qRT-PCR. Results represent the mean of relative expression values ± SEM of 4 mice. c–f Neutrophil depletion exaggerates the osteolytic activity triggered by UHMWPE and VE-UHMWPE polyethylene debris in a murine model. c Quantification of the lytic area in calvarial bone tissues analyzed by micro-CT. Results represent the mean ± SEM of 6 mice. d, e Quantification of TRAP-stained areas and inflammatory infiltrate in calvarial bone sections. Significant difference between the two groups was determined by two-tailed Student’s t test and for multiple groups by one-way ANOVA, followed by Tukey’s multiple-comparison procedure. f Representative images for micro-CT and histological observations of bone sections stained by TRAP and H&E. Scale bar 100 µm. g–k Adoptive transfer of bone marrow neutrophils (BMN) suppress the osteolytic activity triggered by UHMWPE debris. BMN were isolated from donor mice and then injected into calvarial bone after the implantation of UHMWPE debris. Transferred cells were non-stimulated (BMN) or stimulated with TNF-α (aBMN). g Quantification of the lytic area in calvarial bone tissues analyzed by micro-CT. h, i Quantification of TRAP-stained areas and inflammatory infiltrate in calvarial bone sections. Results represent the means of values ± SEM of 6 mice. Significant difference among groups was determined by one-way ANOVA, followed by Tukey’s multiple-comparison procedure. k Representative images for micro-CT and histological observations of bone sections stained by TRAP and H&E. Scale bar 100 µm. *p < 0.05, **p < 0.001, ***p < 0.0001, ****p < 0.00001. ns indicates no significant difference. Source data are provided as a Source Data file.