Edta na2

Chlorpyrifos, a 98% technical product, was provided by Hubei Sharonda Co., Ltd. (Jingzhou, China). Triton X-100 was purchased from Chengdu Haobo Technology Co., Ltd. (Chengdu, China). DTT, PMSF, NADPH, and bovine serum albumin (BSA) were purchased from Beijing Solarbio Science and Technology Co., Ltd. (Beijing, China), as were EDTA-Na₂ and Sodium Dodecyl Sulfate (SDS).

All specimens were decalcified in 10% disodium Ethylenediamine tetraacetic acid (EDTA‐Na2, Solarbio, Beijing, China) at room temperature for ≈30 days. Subsequently, the decalcified samples were longitudinally embedded in paraffin wax and cut into 5 µm sections.
In order to detect phenotype switching of macrophages in vivo, immunofluorescence staining was performed. Tissue sections of 4 and 7 days were incubated with primary antibodies of anti‐CD86 (2 µg mL^−1, NBP2‐25208, Novus) and anti‐CD206 (10 µg mL⁻¹, ab64693, Abcam) at 4 °C overnight followed by incubation with the corresponding fluorescently labeled secondary antibody (CD86: CoraLite594‐conjugated Goat Anti‐Mouse IgG, CD206: CoraLite488‐conjugated Goat Anti‐Rabbit IgG; 1:200; Proteintech) and DAPI. The fluorescent images were obtained with a fluorescence microscope (Olympus), Image J was used to quantify the fluorescence intensity of the stained markers and normalized to DAPI‐stained nuclei counts.

To evaluate effect of these radiopaque agents on the mechanical property of PMMA bone cements, compression tests were carried out using universal mechanical testing machine (E10000, Instron, Hengyi precision instrument Co., Ltd., Shanghai, China) with three repetitions for each group. In order to investigate the effect of adding an amount of porous BaSO₄@PDA/I-CD/Ag microparticles in PMMA bone cements on compressive strength, PMMA bone cements containing 0 wt%, 2.5 wt%, 5 wt%, 7.5 wt% and 10 wt% porous BaSO₄@PDA/I-CD/Ag microparticles were prepared. To compare the effect of different microparticles on compressive strength, PMMA bone cements adding 7.5 wt% commercial BaSO₄, porous BaSO₄, porous BaSO₄@PDA, porous BaSO₄@PDA/Ag, porous BaSO₄@PDA/I-CD and porous BaSO₄@PDA/I-CD/Ag microparticles were then fabricated. The cements were hardened in pillar mold with diameter of 6 mm and height of 12 mm for 1 h and then ejected. The rate for these compression tests were 1 mm min⁻¹. Male sheep lumbar vertebrae (L1-L6), decalcified in 10% 10% EDTA Na₂ (Solarbio, Beijing Solarbio Science & Technology Co., Ltd., Beijing, China) for 20 days, was used to investigate the mechanical property of PMMA bone cement containing 7.5 wt% of porous BaSO₄, and BaSO₄@PDA/I-CD/Ag microparticles. The rate for these compression tests were 5 mm min⁻¹.