Fig 2: Time-dependent expression of CD18, CD11a, CD11b, CD11c, MyD88 and TIRAP in RAW 264.7 cells following Ti particle treatment using ELISA at (A) 6 h; (B) 24 h; (C) 48 h and (D) 72 h. Data are presented as the mean ± SD; *P<0.05 compared with control. Ti, titanium; TIRAP, TIR-domain-containing adapter protein.

Fig 3: Western blotting and quantitative analysis of integrin levels and inflammation response proteins. (A) Western blot immunoblots results showing CD18, CD11a, CD11b, CD11c and MyD88 TIRAP. (B-D) Quantification of immunoblots using ImageJ software. *P<0.05. Ti, titanium; TIRAP, TIR-domain-containing adapter protein.

Fig 4: MPs induce inflammation by recognizing TLR2/4 in BMDMs. (A) Representative Western blot images of BMDMs treated with MPs or LPS (positive control) for 24 h. TLR2, TLR4, TIRAP, MyD88, TRAF6, P-JNK, and P-c-Jun protein expression levels, analyzed by Western blotting. β-actin expression was used as a loading CTL. (B) Relative mRNA expression levels of Il-1β, Il-6, and Tnf-α, measured in the presence of MP. Gapdh was used for normalization. (C) Representative flow cytometry plots presenting BMDM M1 and M2 polarization upon MP treatment. The M1 and M2 macrophage populations were quantified as CD86+CD206– and CD86–CD206+, respectively. (D) Representative immunofluorescence images of BMDMs treated with or without MPs for 24 h, stained with MPO (green), CitH3 (red), and DAPI (blue). Scale bars, 10 μm. Fluorescence intensity-based MPO and CitH3 quantification in BMDMs. a.u., arbitrary unit. The data represent at least three independent experiments and are expressed as the mean ± SEM, using one-way ANOVA. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05.