Journal: Materials Today Bio

Article Title: Ultrasound-activated piezoelectric Silk-PVDF hydrogel reprograms the osteoimmune microenvironment via NRF2 signaling for accelerated bone regeneration

doi: 10.1016/j.mtbio.2026.102779

Figure Lengend Snippet: Piezoelectric hydrogel activates NRF2 to attenuate ROS and macrophages polarization for osteogenesis . (A–B) RT-qPCR results for the mRNA expression of pro-inflammatory differentiation of macrophages. (C–D) RT-qPCR results for the mRNA expression of anti-inflammatory differentiation of macrophages. (E) The relative protein expression levels of INOS, CD206. (F–G) Semi-quantitative analysis of immunoblotting results of INOS, CD206. (H) ROS staining of ADSCs. (I) Mean intensity of ROS staining. (J) The relative protein expression levels of NRF2, NQO1, GPX4. (K–M) Semi-quantitative analysis of immunoblotting results of NRF2, NQO1, GPX4. Data are presented as the mean ± SEM; n = 3; ∗significant difference between selected groups, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001.

Article Snippet: Cells were fixed with 4 % paraformaldehyde (Servicebio, #G1101) for 15 min at 25 °C, permeabilized with 0.3 % Triton X-100 in PBS for 15 min, and blocked with 5 % BSA (Sigma, #A7906) containing 10 % normal goat serum (Servicebio, #G5009) for 1 h. Primary antibodies were diluted in antibody diluent (Servicebio, #G1212) and incubated overnight at 4 °C: • Osteopontin (OPN): Rabbit monoclonal (Proteintech, #22952-1-AP), 1:500 • Osteocalcin (OCN): Rabbit polyclonal (Proteintech, #20277-1-AP), 1:500 • iNOS: Mouse anti-iNOS (Proteintech, #22226-1-AP), 1:500 • CD206: Rabbit anti-CD206 (Proteintech, #18704-1-AP), 1:500 After three PBS washes, species-matched secondary antibodies were applied for 1 h at 25 °C in the dark: • Alexa Fluor 488: Goat anti-rabbit IgG (Servicebio, #GB25303), 1:500 • Alexa Fluor 488: Goat anti-mouse IgG (Servicebio, #GB25301), 1:500 • Cy3: Goat anti-rabbit IgG (Servicebio, # GB21303), 1:500 Nuclei were counterstained with DAPI (Servicebio, #G1407).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Staining