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In vitro evaluation of osteogenic differentiation capacity . (A–B) Representative photographs of ALP staining on day 14. (C) Quantitative analysis of ALP activity of ADSCs on day 14. (D–E) Representative photographs of ARS staining on day 21. (F–H) RT-qPCR results for the mRNA expression of Collagen-I, <t>Runx2,</t> OPN. Data are presented as the mean ± SEM; n = 3; ∗significant difference between selected groups, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001.
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In vitro evaluation of osteogenic differentiation capacity . (A–B) Representative photographs of ALP staining on day 14. (C) Quantitative analysis of ALP activity of ADSCs on day 14. (D–E) Representative photographs of ARS staining on day 21. (F–H) RT-qPCR results for the mRNA expression of Collagen-I, <t>Runx2,</t> OPN. Data are presented as the mean ± SEM; n = 3; ∗significant difference between selected groups, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001.
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In vitro evaluation of osteogenic differentiation capacity . (A–B) Representative photographs of ALP staining on day 14. (C) Quantitative analysis of ALP activity of ADSCs on day 14. (D–E) Representative photographs of ARS staining on day 21. (F–H) RT-qPCR results for the mRNA expression of Collagen-I, <t>Runx2,</t> OPN. Data are presented as the mean ± SEM; n = 3; ∗significant difference between selected groups, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001.
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In vitro evaluation of osteogenic differentiation capacity . (A–B) Representative photographs of ALP staining on day 14. (C) Quantitative analysis of ALP activity of ADSCs on day 14. (D–E) Representative photographs of ARS staining on day 21. (F–H) RT-qPCR results for the mRNA expression of Collagen-I, <t>Runx2,</t> OPN. Data are presented as the mean ± SEM; n = 3; ∗significant difference between selected groups, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001.
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In vitro evaluation of osteogenic differentiation capacity . (A–B) Representative photographs of ALP staining on day 14. (C) Quantitative analysis of ALP activity of ADSCs on day 14. (D–E) Representative photographs of ARS staining on day 21. (F–H) RT-qPCR results for the mRNA expression of Collagen-I, <t>Runx2,</t> OPN. Data are presented as the mean ± SEM; n = 3; ∗significant difference between selected groups, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001.
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In vitro evaluation of osteogenic differentiation capacity . (A–B) Representative photographs of ALP staining on day 14. (C) Quantitative analysis of ALP activity of ADSCs on day 14. (D–E) Representative photographs of ARS staining on day 21. (F–H) RT-qPCR results for the mRNA expression of Collagen-I, <t>Runx2,</t> OPN. Data are presented as the mean ± SEM; n = 3; ∗significant difference between selected groups, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001.
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In vitro evaluation of osteogenic differentiation capacity . (A–B) Representative photographs of ALP staining on day 14. (C) Quantitative analysis of ALP activity of ADSCs on day 14. (D–E) Representative photographs of ARS staining on day 21. (F–H) RT-qPCR results for the mRNA expression of Collagen-I, Runx2, OPN. Data are presented as the mean ± SEM; n = 3; ∗significant difference between selected groups, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001.

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: In vitro evaluation of osteogenic differentiation capacity . (A–B) Representative photographs of ALP staining on day 14. (C) Quantitative analysis of ALP activity of ADSCs on day 14. (D–E) Representative photographs of ARS staining on day 21. (F–H) RT-qPCR results for the mRNA expression of Collagen-I, Runx2, OPN. 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: Membranes were blocked with 5 % non-fat milk (BD DifcoTM, #232100) in TBST (Beyotime, #ST673) for 1 h at 25 °C and incubated overnight at 4 °C with the following primary antibodies: Osteopontin (OPN) : • Rabbit monoclonal (Proteintech, 22952-1-AP), 1:1000 • Collagen I (COL1A1): Rabbit polyclonal (Proteintech, 14695-1-AP), 1:1000 • RUNX2: Rabbit monoclonal (Proteintech, 20700-1-AP), 1:1000 • NRF2: Rabbit monoclonal (Proteintech, 16396-1-AP), 1:2000 • NQO1: Mouse monoclonal (Proteintech, 67240-1-Ig), 1:2000 • Beta Actin: Mouse monoclonal (Proteintech, 66009-1-Ig), 1:20,000 After three 10-min TBST washes, membranes were incubated with HRP-conjugated goat anti-rabbit/mouse IgG (Beyotime, #A0208/#A0216, 1:5000) for 2 h at 25 °C.

Techniques: In Vitro, Staining, Activity Assay, Quantitative RT-PCR, Expressing

In vitro evaluation of osteogenesis related proteins and genes. (A) The relative protein expression levels of C1, Runx2 and OPN. (B–D) Semi-quantitative analysis of immunoblotting results of C1, Runx2 and OPN. (E) Immunofluorescence staining of OPN. (F) Immunofluorescence staining of OCN. (G) The relative protein expression levels of C1, Runx2 and OPN. (H–J) Semi-quantitative analysis of immunoblotting results of C1, Runx2 and OPN. Data are presented as the mean ± SEM; n = 3; ∗significant difference between selected groups, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001.

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: In vitro evaluation of osteogenesis related proteins and genes. (A) The relative protein expression levels of C1, Runx2 and OPN. (B–D) Semi-quantitative analysis of immunoblotting results of C1, Runx2 and OPN. (E) Immunofluorescence staining of OPN. (F) Immunofluorescence staining of OCN. (G) The relative protein expression levels of C1, Runx2 and OPN. (H–J) Semi-quantitative analysis of immunoblotting results of C1, Runx2 and OPN. 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: Membranes were blocked with 5 % non-fat milk (BD DifcoTM, #232100) in TBST (Beyotime, #ST673) for 1 h at 25 °C and incubated overnight at 4 °C with the following primary antibodies: Osteopontin (OPN) : • Rabbit monoclonal (Proteintech, 22952-1-AP), 1:1000 • Collagen I (COL1A1): Rabbit polyclonal (Proteintech, 14695-1-AP), 1:1000 • RUNX2: Rabbit monoclonal (Proteintech, 20700-1-AP), 1:1000 • NRF2: Rabbit monoclonal (Proteintech, 16396-1-AP), 1:2000 • NQO1: Mouse monoclonal (Proteintech, 67240-1-Ig), 1:2000 • Beta Actin: Mouse monoclonal (Proteintech, 66009-1-Ig), 1:20,000 After three 10-min TBST washes, membranes were incubated with HRP-conjugated goat anti-rabbit/mouse IgG (Beyotime, #A0208/#A0216, 1:5000) for 2 h at 25 °C.

Techniques: In Vitro, Expressing, Western Blot, Immunofluorescence, Staining