Journal: Bioactive Materials

Article Title: MSC-mimicking nanovesicle embedded bio-adhesive hydrogel for dual immunomodulation and osteogenesis to promote maxillofacial bone regeneration

doi: 10.1016/j.bioactmat.2026.02.032

Figure Lengend Snippet: In vivo evaluation of bone regeneration after hydrogel implantation in the mandibular bone defect mouse model. a) Schematic illustration of the mandibular bone defect mouse model. b) Micro-CT 3D reconstruction images of the mandibular bone samples at 4 and 8 weeks post-surgery. Scale bar = 1 mm. c) Semi-quantitative analysis of BV/TV, bone surface, Tb.N and Tb.sp (n = 6) in mouse mandibular bone defects implanted with different hydrogels at 8 weeks post-surgery. d) H&E staining and Masson trichrome staining of tissue sections of mandibular defects at 8 weeks post-surgery. Scale bar = 100 μm. e, f) Immunofluorescent staining images and corresponding semi-quantitative analysis of the expression levels of RUNX2 and OCN in mandibular bone defect areas at 4 and 8 weeks post-surgery (n = 3). Scale bar = 50 μm. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( a was created with bioRender. com).

Article Snippet: After blocking with 5% skimmed milk (diluted with TBST), the membranes were probed with the primary antibodies for CD90 (1:1000, Cat. ab92574, Abcam, UK), CD146 (1:1000, Cat. ab75769, Abcam, London, UK), CD105 (1:500, Cat. sc18838, Santa, USA), CXCR4 (1:250, Cat. 35-8800, Invitrogen, CA, USA), Na + /K + ATPase (1:1000, Cat. sc28800, Santa, USA), BMP2 (1:1000, Cat. ab284387, Abcam, UK), RUNX2 (1:1000, Cat. 12556, Cell Signaling Technology, USA) and β -actin (1:10000, Cat. T0022, Affinity, China) overnight at 4 °C, followed by bathing with horseradish peroxidase (HRP) conjugated secondary antibodies (1:5000, Cat. S0002 or Cat. S0001, Affinity, China).

Techniques: In Vivo, Micro-CT, Staining, Expressing

Journal: Bioactive Materials

Article Title: MSC-mimicking nanovesicle embedded bio-adhesive hydrogel for dual immunomodulation and osteogenesis to promote maxillofacial bone regeneration

doi: 10.1016/j.bioactmat.2026.02.032

Figure Lengend Snippet: The PEG-pp@nMSC@MT hydrogel effectively promotes BMMSCs' osteogenesis in vitro . a) Schematic illustration of co-culture BMMSCs with hydrogels. b) ALP staining of BMMSCs co-cultured with different scaffolds after 7 days. Scale bar = 500 μm. c) Semi-quantitative analysis of ALP staining (n = 3). d) ARS staining of BMMSC co-cultured with different scaffolds after 21 days. Scale bar = 500 μm. e) Semi-quantitative analysis of ARS staining (n = 3). f) mRNA expression of osteogenic genes (BMP2, OCN, and RUNX2) of BMMSCs treated with different hydrogels in the MMP condition after 5 and 10 days (n = 3). g) Western blot analysis of osteogenic protein (BMP2 and RUNX2) of BMMSCs treated with different hydrogels in the MMP condition after 7 and 14 days. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( a was created with bioRender. com).

Article Snippet: After blocking with 5% skimmed milk (diluted with TBST), the membranes were probed with the primary antibodies for CD90 (1:1000, Cat. ab92574, Abcam, UK), CD146 (1:1000, Cat. ab75769, Abcam, London, UK), CD105 (1:500, Cat. sc18838, Santa, USA), CXCR4 (1:250, Cat. 35-8800, Invitrogen, CA, USA), Na + /K + ATPase (1:1000, Cat. sc28800, Santa, USA), BMP2 (1:1000, Cat. ab284387, Abcam, UK), RUNX2 (1:1000, Cat. 12556, Cell Signaling Technology, USA) and β -actin (1:10000, Cat. T0022, Affinity, China) overnight at 4 °C, followed by bathing with horseradish peroxidase (HRP) conjugated secondary antibodies (1:5000, Cat. S0002 or Cat. S0001, Affinity, China).

Techniques: In Vitro, Co-Culture Assay, Staining, Cell Culture, Expressing, Western Blot

Journal: Bioactive Materials

Article Title: A multimodal ROS logic-gated therapeutic platform disrupts the vicious cycle of senescence to promote aged bone defect repair

doi: 10.1016/j.bioactmat.2026.02.002

Figure Lengend Snippet: Therapy platform enhances osteogenic potential of senescent BMSCs in vitro . (A) Representative ALP staining images at day 7 (scale bars: top-10 mm, and bot-200 μm). (B) Quantitative analysis of ALP activity ( n = 3). (C) Representative ARS staining images at day 21 (scale bars: top-5 mm and bot-200 μm). (D) Quantification of mineralized matrix formation via ARS staining ( n = 3). (E-H) qRT-PCR analysis of osteogenic markers ( Alp , Osx , Runx2 , and Opn ) in BMSCs ( n = 3). (I) Representative immunofluorescence images of RUNX2 staining (scale bars: 50 μm). (J) Quantitative analysis of RUNX2 fluorescence intensity ( n = 3). (K) Representative immunofluorescence images of OPN staining (scale bars: 50 μm). (L) Quantitative analysis of OPN fluorescence intensity ( n = 3). Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Protein expression of RUNX2 (Proteintech, 20700-1-AP; 1:500) and OPN (Abcam, ab63856; 1:500) were detected by immunofluorescence staining on day 14, and fluorescence intensities were quantified using ImageJ software (Media Cybernetics).

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

Journal: Bioactive Materials

Article Title: A multimodal ROS logic-gated therapeutic platform disrupts the vicious cycle of senescence to promote aged bone defect repair

doi: 10.1016/j.bioactmat.2026.02.002

Figure Lengend Snippet: In vivo evaluation of the therapeutic platform for aged bone defect repair. (A) Representative 3D-reconstructed micro-CT images of calvarial bone defects in different groups (scale bars: 2 mm). (B-C) Quantification of bone volume fraction (BV/TV) and bone mineral density (BMD) based on micro-CT analysis ( n = 5). (D-E) Representative histological images of H&E staining (D) and Masson's trichrome staining (E). Low-magnification images (scale bars: 500 μm) show the overall defect region, while high-magnification images (scale bars: 100 μm) highlight the new bone (blue box) and defect-host bone interface (red box). Green arrows indicate defect boundaries; FT: fiber tissue; NB: new formed bone; HB: host bone. (F-G) Immunofluorescence images and quantification of RUNX2 expression in defect areas (scale bars: 50 μm, n = 3). (H-I) Immunofluorescence images and quantification of OCN expression (scale bars: 50 μm, n = 3). (J) Schematic illustration of the programmed ROS bidirectional modulation strategy for infection control and repair of aged bone defects, depicting acousto-optic dynamic ROS generation for antimicrobial effects followed by antioxidant modulation to rejuvenate senescent BMSCs and promote aged bone regeneration. Data are expressed as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Protein expression of RUNX2 (Proteintech, 20700-1-AP; 1:500) and OPN (Abcam, ab63856; 1:500) were detected by immunofluorescence staining on day 14, and fluorescence intensities were quantified using ImageJ software (Media Cybernetics).

Techniques: In Vivo, Micro-CT, Staining, Immunofluorescence, Expressing, Infection, Control

Journal: Bioactive Materials

Article Title: MSC-mimicking nanovesicle embedded bio-adhesive hydrogel for dual immunomodulation and osteogenesis to promote maxillofacial bone regeneration

doi: 10.1016/j.bioactmat.2026.02.032

Figure Lengend Snippet: In vivo evaluation of bone regeneration after hydrogel implantation in the mandibular bone defect mouse model. a) Schematic illustration of the mandibular bone defect mouse model. b) Micro-CT 3D reconstruction images of the mandibular bone samples at 4 and 8 weeks post-surgery. Scale bar = 1 mm. c) Semi-quantitative analysis of BV/TV, bone surface, Tb.N and Tb.sp (n = 6) in mouse mandibular bone defects implanted with different hydrogels at 8 weeks post-surgery. d) H&E staining and Masson trichrome staining of tissue sections of mandibular defects at 8 weeks post-surgery. Scale bar = 100 μm. e, f) Immunofluorescent staining images and corresponding semi-quantitative analysis of the expression levels of RUNX2 and OCN in mandibular bone defect areas at 4 and 8 weeks post-surgery (n = 3). Scale bar = 50 μm. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( a was created with bioRender. com).

Article Snippet: Briefly, tissue sections underwent antigen retrieval (37 °C, 30 min), blocked by 5% bovine serum albumin (BSA, RT, 1 h), then sequentially incubated (4 °C) with lineage-specific probes: macrophage marker F4/80 (1:200, Cat. sc-52664, Santa Cruz Biotechnology) and CD68 (1:250, Cat. 14-0681-81, ThermoFisher, USA), CD206 (1:500, Cat. 24595T, Cell Signaling Technology, USA) and Arg-1 (1:250, Cat. 82975, Proteintech, China) for a M2 marker, iNOS (1:500, Cat. ab178945, Abcam, UK) and CD86 (1:300, Cat. DF6332, Affinity, China) for a M1 marker, RUNX2 (1:150, Cat. sc390351, Santa Cruz Biotechnology, USA) and OCN (1:150, Cat. sc390877, Santa Cruz Biotechnology, USA) for osteogenesis markers, VEGF (1:50, Cat. sc57496, Santa Cruz Biotechnology, USA) and CD31 (1:50, Cat. sc20071, Santa Cruz Biotechnology, USA) for angiogenesis markers, and CD146 (1:200, Cat. Ab75769, Abcam, UK) for stem cell surface marker, overnight at 4 °C.

Techniques: In Vivo, Micro-CT, Staining, Expressing

Journal: Bioactive Materials

Article Title: MSC-mimicking nanovesicle embedded bio-adhesive hydrogel for dual immunomodulation and osteogenesis to promote maxillofacial bone regeneration

doi: 10.1016/j.bioactmat.2026.02.032

Figure Lengend Snippet: The PEG-pp@nMSC@MT hydrogel effectively promotes BMMSCs' osteogenesis in vitro . a) Schematic illustration of co-culture BMMSCs with hydrogels. b) ALP staining of BMMSCs co-cultured with different scaffolds after 7 days. Scale bar = 500 μm. c) Semi-quantitative analysis of ALP staining (n = 3). d) ARS staining of BMMSC co-cultured with different scaffolds after 21 days. Scale bar = 500 μm. e) Semi-quantitative analysis of ARS staining (n = 3). f) mRNA expression of osteogenic genes (BMP2, OCN, and RUNX2) of BMMSCs treated with different hydrogels in the MMP condition after 5 and 10 days (n = 3). g) Western blot analysis of osteogenic protein (BMP2 and RUNX2) of BMMSCs treated with different hydrogels in the MMP condition after 7 and 14 days. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( a was created with bioRender. com).

Article Snippet: Briefly, tissue sections underwent antigen retrieval (37 °C, 30 min), blocked by 5% bovine serum albumin (BSA, RT, 1 h), then sequentially incubated (4 °C) with lineage-specific probes: macrophage marker F4/80 (1:200, Cat. sc-52664, Santa Cruz Biotechnology) and CD68 (1:250, Cat. 14-0681-81, ThermoFisher, USA), CD206 (1:500, Cat. 24595T, Cell Signaling Technology, USA) and Arg-1 (1:250, Cat. 82975, Proteintech, China) for a M2 marker, iNOS (1:500, Cat. ab178945, Abcam, UK) and CD86 (1:300, Cat. DF6332, Affinity, China) for a M1 marker, RUNX2 (1:150, Cat. sc390351, Santa Cruz Biotechnology, USA) and OCN (1:150, Cat. sc390877, Santa Cruz Biotechnology, USA) for osteogenesis markers, VEGF (1:50, Cat. sc57496, Santa Cruz Biotechnology, USA) and CD31 (1:50, Cat. sc20071, Santa Cruz Biotechnology, USA) for angiogenesis markers, and CD146 (1:200, Cat. Ab75769, Abcam, UK) for stem cell surface marker, overnight at 4 °C.

Techniques: In Vitro, Co-Culture Assay, Staining, Cell Culture, Expressing, Western Blot

Journal: JBMR Plus

Article Title: Follistatin-like protein 1 (FSTL1) modulates bone remodeling and attenuates bone loss in a mouse model of postmenopausal osteoporosis

doi: 10.1093/jbmrpl/ziag037

Figure Lengend Snippet: Fstl1 enhances the osteogenic differentiation potential of MC3T3-E1 cells. (A) Representative alkaline-phosphatase (ALP) staining at 7 d. (B) Quantitative ALP activity. (C) Representative Alizarin Red S (ARS) staining at 14 d. (D) Quantification of ARS absorbance (OD 562 ). (E-H) Relative mRNA expression of Runx2, Sp7, Ocn, and Opn at 14 d determined by RT-qPCR. (I) Western-blot bands for RUNX2 and SP7. (J and K) Densitometric ratios of RUNX2/GAPDH and SP7/GAPDH. Sample sizes: con ( n = 3), OE-Fstl1 ( n = 3), and sh-Fstl1 ( n = 3). Data are presented as mean ± SD; inter-group comparisons were performed using 1-way ANOVA. * p < .05, ** p < .01, *** p < .001, **** p < .0001; ns, not significant.

Article Snippet: Antibodies against RUNX2, SP7, NFATC1, and CTSK were purchased from Abcam, while the anti-GAPDH antibody and anti-FSTL1 antibody were obtained from Proteintech.

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