Journal: Journal of Nanobiotechnology

Article Title: Nanoparticle-integrated smart hydrogel with H 2 S-triggered antibacterial escalation and sustained oxygen release for periodontal regeneration

doi: 10.1186/s12951-026-04535-7

Figure Lengend Snippet: Osteogenic differentiation of hPDLSCs induced by PDA@Hb/NOCG. ( A ) Representative ALP staining results of hPDLSCs after 7 days of osteogenic differentiation. ( B ) Alizarin Red S staining of hPDLSCs after 21 days of osteogenic induction. ( C ) Quantitative analysis of ALP staining in hPDLSCs. ( D ) Quantification of Alizarin Red S staining in hPDLSCs. ( E-K ) Relative mRNA expression levels of osteogenesis-related genes in hPDLSCs, including ALP, BMP2, COL1A1, OPG, RUNX2, OPN, and OCN, normalized to GAPDH. ( L-O ) Protein expression levels of osteogenesis-related markers in hPDLSCs, including ALP, RUNX2, and β-catenin, and their corresponding quantitative analysis. ( P ) Protein expression levels of osteogenesis-related markers, including ALP, RUNX2, and β-catenin, in hPDLSCs under different treatment conditions with or without DKK1. ( Q ) Corresponding quantitative analysis of ALP, RUNX2, and β-catenin protein expression in each group

Article Snippet: Primary antibodies against ALP (rabbit polyclonal IgG, Cat. No. 18507-1-AP), NF-κB p65 (rabbit polyclonal IgG, Cat. No. 10745-1-AP), and β-catenin (rabbit polyclonal IgG, Cat. No. 17565-1-AP) were purchased from Proteintech (Wuhan, China), while the antibodies against RUNX2 (rabbit polyclonal IgG, Cat. No. HY- P80316 ) and Hb (polyclonal antibody, Cat. No. PAB409Bo01) were obtained from MedChemExpress (MCE, Shanghai, China) and Cloud-Clone Corp. (Wuhan, China), respectively.

Techniques: Staining, Expressing

Journal: Pharmaceutics

Article Title: Dendritic Mesoporous Silica-Modified Decellularized Bone Matrix Scaffold for Sustained Teriparatide Delivery in Bone Defect Repair: Characterization, Drug Release, and In Vitro Biological Performance

doi: 10.3390/pharmaceutics18091067

Figure Lengend Snippet: Osteogenic differentiation-promoting effect of DBM-DMSN@TPTD composite scaffold on BMSCs. ( A ) ALP staining of BMSCs after 7 days of culture. ( B ) Immunofluorescence staining of RUNX2 in BMSCs. ( C ) Immunofluorescence staining of OCN in BMSCs. ( D ) Quantitative analysis of RUNX2 mean fluorescence intensity. ( E ) Quantitative analysis of OCN mean fluorescence intensity. Data are presented as mean ± SD ( n = 3). Comparisons among multiple groups were performed using one-way ANOVA followed by Tukey’s multiple comparisons post hoc test. ns, no significant difference; * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Subsequently, the cells were blocked with 5% bovine serum albumin (BSA, Sigma-Aldrich, St. Louis, MO, USA) at 37 °C for 30 min. Primary antibodies against RUNX2 (ABclonal Technology Co., Ltd., Wuhan, Hubei, China) and OCN (ABclonal Technology Co., Ltd., Wuhan, Hubei, China) were added and incubated overnight at 4 °C in a humidified chamber.

Techniques: Staining, Immunofluorescence, Analysis, Fluorescence

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: Journal of Nanobiotechnology

Article Title: 4D printed hydrogel scaffolds with shape morphing by near-infrared to promote bone regeneration through immune and vascular coupling

doi: 10.1186/s12951-026-04386-2

Figure Lengend Snippet: The biocompatibility assay and osteogenesis assay of AC, AS, PAS, PASN groups. A ) The cell viability assay of AC, AS, PAS, and PASN scaffolds. B ) The osteogenesis-related gene expressions of ALP, COL-1, RUNX2 and OCN at 7 and 14 days. C ) Schematic diagram of BMSCs cocultured with AC, AS, PAS scaffold. D ) Immunofluorescent staining and E ) quantitative analysis of BMSCs co-cultured with different hydrogel scaffolds for 7 days, green for ALP, COL-1, RUNX2, and OCN, red (phalloidin) for F-actin and blue (DAPI) for nuclei. F ) ALP staining of BMSCs for 7 and 14 days. G ) Calcium deposition stained with alizarin red S of BMSCs for 28 days. (*P < 0.05, **P <0.01, and ***P < 0.001; “ns” denoted no significant difference, n = 3, error bars indicate means ± standard deviations)

Article Snippet: Immunofluorescence staining blocking buffer (Beyotime, China), ALP Rabbit Monoclonal Antibody (ABclonal, China), COL-1 Rabbit Monoclonal Antibody (ABclonal, China), OCN Rabbit Monoclonal Antibody (ABclonal, China), RUNX2 Rabbit Monoclonal Antibody (ABclonal, China), and Alexa Fluor 488-labeled Goat Anti-rabbit IgG (Beyotime, China) were used for Immunofluorescence Staining according to the manufacturer’s instructions.

Techniques: Viability Assay, Staining, Cell Culture

Journal: Journal of Nanobiotechnology

Article Title: 4D printed hydrogel scaffolds with shape morphing by near-infrared to promote bone regeneration through immune and vascular coupling

doi: 10.1186/s12951-026-04386-2

Figure Lengend Snippet: Transcriptomics Analysis of BMSCs and verification of the mechanical force sensing pathway in cells. A ) KEGG and B ) GO enrichment bubble diagram of BMSCs [n = 3, adjusted p-value (q value) of <0.05]. C )Topological graph of “focal adhesion signaling pathway mediated by PASN” in the Reactome data. D ) Heatmap analysis of BMSCs, Orange: higher expressions, blue: lower expressions [n = 3, adjusted p-value (q value) of <0.05]. E ) Immunofluorescent staining of BMSCs cultured on different hydrogel scaffolds for 3 days, green for vinculin and G ) YAP, red(phalloidin) for F-actin and blue (DAPI) for nuclei. F ) Quantitative analysis of vinculin area and H ) nucleus/ cytoplasmic ratio of YAP based on fluorescence intensities. I ) Real-time PCR analysis of the mRNA expression of VCL, Lamc, RhoA, ROCK1, ROCK2 for 3 days. J ) Expressions of RhoA, ALP, RUNX2, YAP, Lamin, Vinculin, ROCK1, and COL-1 proteins detected by western blot analysis of BMSCs. K ) Cells sense composite microenvironment by exerting contractile forces with stress fibers through focal adhesions, which activates various proteins, such as focal adhesion kinase (FAK), vinculin, RhoA and ROCK, at the adhesion site. Activation of these proteins leads to adhesion maturation and stress fiber formation and contractility, which in turn transmits forces to the nucleus via the linker of nucleoskeleton and cytoskeleton (LINC) complex, resulting in changes in nuclear envelope tension and nuclear pore opening. This allows the nuclear entry of proteins such as the transcription regulator YAP, leading to a downstream impact on cell phenotype. (*P < 0.05, **P < 0.01, and ***P < 0.001; “ns” denoted no significant difference, n = 3, error bars indicate means ± standard deviations)

Article Snippet: Immunofluorescence staining blocking buffer (Beyotime, China), ALP Rabbit Monoclonal Antibody (ABclonal, China), COL-1 Rabbit Monoclonal Antibody (ABclonal, China), OCN Rabbit Monoclonal Antibody (ABclonal, China), RUNX2 Rabbit Monoclonal Antibody (ABclonal, China), and Alexa Fluor 488-labeled Goat Anti-rabbit IgG (Beyotime, China) were used for Immunofluorescence Staining according to the manufacturer’s instructions.

Techniques: Transcriptomics, Staining, Cell Culture, Fluorescence, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Activation Assay

Journal: Journal of Nanobiotechnology

Article Title: 4D printed hydrogel scaffolds with shape morphing by near-infrared to promote bone regeneration through immune and vascular coupling

doi: 10.1186/s12951-026-04386-2

Figure Lengend Snippet: PASN scaffold induced M2 macrophage polarization which enforces BMSCs osteogenic differentiation in vitro. A ) AC, AS, PAS, PASN scaffold induced M2 macrophage polarization. B ) Flow cytometry exhibiting enhanced expression of the M2 macrophage marker CD206 and decreased expression of the M1 macrophage marker CD86 in the PASN group at 3 days as compared with PAS (n=3). C )Relative mRNA expressions of macrophage polarization-related markers at 3 days (n=3). D ) Schematic diagram of BMSCs cocultured with macrophage-laden AC scaffold, macrophage-laden AS scaffold, macrophage-laden PAS scaffold and macrophage-laden PASN scaffold. E ) ALP staining at 14 days and alizarin red staining at 28 days. F ) Relative mRNA expression of ALP, OCN, RUNX2, and COL-1 in BMSCs at the 7th and 14th day in different groups (n=3). G ) Immunofluorescent staining of BMSCs co-cultured with different hydrogel scaffolds for 7 days, green for ALP, COL-1, RUNX2, and OCN, red(phalloidin) for F-actin and blue (DAPI) for nuclei. (*P < 0.05, **P < 0.01, and ***P < 0.001; “ns” denoted no significant difference, n = 3, error bars indicate means ± standard deviations)

Article Snippet: Immunofluorescence staining blocking buffer (Beyotime, China), ALP Rabbit Monoclonal Antibody (ABclonal, China), COL-1 Rabbit Monoclonal Antibody (ABclonal, China), OCN Rabbit Monoclonal Antibody (ABclonal, China), RUNX2 Rabbit Monoclonal Antibody (ABclonal, China), and Alexa Fluor 488-labeled Goat Anti-rabbit IgG (Beyotime, China) were used for Immunofluorescence Staining according to the manufacturer’s instructions.

Techniques: In Vitro, Flow Cytometry, Expressing, Marker, Staining, Cell Culture

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

Journal: Cureus

Article Title: High Immunohistochemical Expression of Runt-Related Transcription Factor 2 (RUNX2) Is Associated With High Tumor Grade, Muscle Invasion, Lymph Node Metastasis, and Advanced Stage in Urinary Bladder Cancer

doi: 10.7759/cureus.106976

Figure Lengend Snippet: IHC: immunohistochemistry; H&E: hematoxylin and eosin; RUNX2: Runt-related transcription factor 2.

Article Snippet: Immunohistochemistry (IHC) was then performed using the commercially available rabbit polyclonal RUNX2 antibody (Clone: Ab-AF5186#1486, pack size 100 μl, concentration of 1 mg/ml) from Affinity Biosciences, Cincinnati, OH, USA.

Techniques: Immunohistochemistry

Journal: Cureus

Article Title: High Immunohistochemical Expression of Runt-Related Transcription Factor 2 (RUNX2) Is Associated With High Tumor Grade, Muscle Invasion, Lymph Node Metastasis, and Advanced Stage in Urinary Bladder Cancer

doi: 10.7759/cureus.106976

Figure Lengend Snippet: IHC: immunohistochemistry; H&E: hematoxylin and eosin; RUNX2: Runt-related transcription factor 2.

Article Snippet: Immunohistochemistry (IHC) was then performed using the commercially available rabbit polyclonal RUNX2 antibody (Clone: Ab-AF5186#1486, pack size 100 μl, concentration of 1 mg/ml) from Affinity Biosciences, Cincinnati, OH, USA.

Techniques: Immunohistochemistry

Journal: Cureus

Article Title: High Immunohistochemical Expression of Runt-Related Transcription Factor 2 (RUNX2) Is Associated With High Tumor Grade, Muscle Invasion, Lymph Node Metastasis, and Advanced Stage in Urinary Bladder Cancer

doi: 10.7759/cureus.106976

Figure Lengend Snippet: IHC: immunohistochemistry; H&E: hematoxylin and eosin; RUNX2: Runt-related transcription factor 2.

Article Snippet: Immunohistochemistry (IHC) was then performed using the commercially available rabbit polyclonal RUNX2 antibody (Clone: Ab-AF5186#1486, pack size 100 μl, concentration of 1 mg/ml) from Affinity Biosciences, Cincinnati, OH, USA.

Techniques: Immunohistochemistry