Osteopontin Antibody - #AF0227

Figure 4. Biocompatibility and osteogenic differentiation ability of HN25. G) Immunofluorescence staining of ALP and OPN after culturing 14 d on different samples. Scale bar = 100 μm. H) Volcano plot showing differently expressed genes in MSCs from HN25 under US irradiation compared to the control.

Figure 5. In vitro biocompatibilities and osteogenic activities of the substrates. (A) CCK-8 assay of hBMSCs on the indicated surfaces after 1, 3 and 7 days of culture. * denotes p < 0.05 and ** denotes p < 0.01 compared to Ti-NTs, # denotes p < 0.05 compared to the corresponding control group without peptides. Error bars denote the standard deviations over quadruplicate measurements with separately implants. (B) Confocal fluorescence microscopy images of hBMSCs stained with vinculin, F-actin and DAPI after being cultured for 24 h. Scale bar, 50 μm. (C-F) qRT-PCR assay of osteogenic gene expression of (C) ALP, (D) RUNX-2, (E) COL1 and (F) OPN of hBMSCs after 7 and 14 days of culture. * denotes p < 0.05, ** denotes p < 0.01 and *** denotes p < 0.001 compared to Ti-NTs-P-A; # denotes p < 0.05, & denotes p < 0.01 and $ denotes p < 0.001 compared to the corresponding control group without peptides. All error bars denote the standard deviations over quadruplicate measurements with separately implants. (G) Immunofluorescence staining of hBMSCs cultured on Ti-NTs-P-A for 7 days (ALP and RUNX-2) and 14 days (OPN). The images were obtained by confocal fluorescence microscopy. Scale bar, 50 μm. (H) Western blotting of hBMSCs cultured on the substrates for 7 and 14 days. At each time point, left lane was Ti-NTs, middle lane was Ti-NTs-A and right lane was Ti-NTs-P-A. * denotes p < 0.05, ** denotes p < 0.01 and *** denotes p < 0.001 compared to Ti-NTs-P-A. Error bars denote the standard deviations over triplicate measurements with separately Western blotting results.

Fig. 6 VSIG4 promoted SPP1 expression through enhancing the histone H3 lysine 18 lactylation and promoting the transcription activity of STAT3. (A) Co-culture of mATC with BMDMs isolated from VSIG4-KO and WT mice to examine the lactate concentration. (B) Isolation of TAMs from mATC-derived tumors in VSIG4-KO (n = 3) and WT mice (n = 3) on day 32 to examine the lactate concentration. Data are presented as mean ± S.E.M. (C-D) Co-culture of mATC with BMDMs isolated from VSIG4-KO and WT mice to examine the expressions of histone H3 lysine 18 lactylation (H3K18la). (E) Genome browser track analysis using dataset GSE192358 showed the H3K18la lactylation levels in the binding region of Spp1 with or without the lactate treatment, where the left area of the dotted line represented the binding region of H3K18la on the Spp1 promotor. The mRNA (F) and protein (G-H) level of Spp1 in BMDM with or without the lactate treatment (n = 3). (I-J) The expression of Stat1, pStat1, Stat3, pStat3, and pStat6 in BMDMs after VSIG4 knockout (n = 3). (K) The JASPAR database predicted the presence of STAT3 binding sites in the SPP1 promoter region. The mRNA (L) and protein (M-N) level of Spp1 in BMDMs after STAT3 inhibitor Stattic treatment (n = 3). (O) The transcriptional activity of STAT3 on SPP1 was determined by dual luciferase reporter gene assay (n = 3). Data are presented as mean ± S.D. *P 

Figure 9 Enhanced angiogenesis through restored mechanobiological and piezoelectric microenvironment in vivo. A) CAM assay showing vascular networks formed around different implants. B) Semiquantitative analysis of vascular parameters from the CAM assay. C) Macroscopic analysis of implant size, shape, and morphology. D) Surgical site showing implant placement in rat femur. E) Methylene blue/basic fuchsin-stained sections at 4 weeks postimplantation (new bone=red, fibrous tissue=blue, implants=black). F) Quantitative analysis of BIC% at 4 weeks. G) Immunohistochemical staining of Col1 in peri-implant tissue at 4 weeks. H) Immunohistochemical staining of OPN in peri-implant tissue at 4 weeks. I) Immunohistochemical staining of CD31 in peri-implant tissue at 4 weeks. J) Immunohistochemical staining of VEGF in peri-implant tissue at 4 weeks. Target proteins appear deep brown. ANOVA followed by Tukey's post hoc test was performed for statistical analysis (error bar: ±SD; n = 3; *p 

Fig. 5 The impact of GaPP@FSB on the differentiation of BMSCs, Chondrocytes and TSPCs. Confocal laser scanning microscopy (CLSM) images showing fluorescence-labelled (A) OPN in the BMSCs (scale bar = 200 μm), and the Q-PCR data (B) revealed the expression of osteogenic differentiation-related genes in BMSCs cultured on the GaPP@FSB scaffold. Additionally, the CLSM images also showed the fluorescence-labelled (C) Tnmd in TSPCs (bar 100 μm), and Q-PCR data (D) revealed the expression of tenogenic differentiation-related genes in TSPCs cultured on the GaPP@FSB scaffold. The investigation approach for the influence of scaffolds on chondrocytes is uniform. (E) CLSM reveals the level of ACAN, a chondrocyte matrix-related proteoglycan, and (F) the Q-PCR results demonstrate the expression of genes related to chondrogenic differentiation. Data are presented as mean values ± SD, *p 

Fig. 7 OHADN fiber@Yoda1 hydrogel promoted alveolar bone regeneration in vivo. (a) Immunohistochemical staining and quantification of average optical density for RUNX2 and OPN of the alveolar bone defect. (b) Immunohistochemical staining and quantification of average optical density for PIEZO1, ITGα5 and TNS1 of the alveolar bone defect. (c) Immunohistochemical co-staining of PIEZO1 (green) and ITGα5 (red). Yellow indicates their spatial co-localization. Right panel: Quantitative curve of the fluorescence assay analyzed by Image J software. (**P