Journal: Bioactive Materials

Article Title: Composite hydrogel-microsphere delivery system promotes early nerve-mediated bone regeneration and late-stage mechanotransduction-driven bone remodeling via sequential release of NGF and Yoda1

doi: 10.1016/j.bioactmat.2025.10.040

Figure Lengend Snippet: Effects on osteogenesis and angiogenesis of the NGF-G/P@Yoda1-stimulated neural cells. (A) Schematic diagram illustrating co-culture models of different composite hydrogels with RSC96 and PC-12, along with the collection of co-culture supernatant as the condition medium for BMSCs and HUVECs cultivation. (B–D) ALP staining at day 7 to assess early osteogenic capacity and Alizarin Red S (ARS) staining at day 21 to assess late-stage calcified nodule formation of BMSCs and semi-quantitative analyses. (E–H) Immunofluorescence staining at day 7 to evaluate expression of osteogenic marker proteins COL1A and OPN and semi-quantitative analysis of mean flourscence indensity (MFI). (I, J) Western blot to evaluate the expression of downstream proteins and osteogenic markers mediated by CGRP and semi-quantitative analysis. (K, L) Wound healing assay and healing ratio quantitative analysis to asses migration capacity of HUVECs. (M, N) Transwell assay and relative quantitative analysis of HUVECs' migration. (O–Q) Tube formation assay and quantification of number of nodes and number of tubes to assess angiogenic capacity of HUVECs. The quantitative data (n ≥ 3) in the figures represent as the averages ± SD, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: Primary antibodies included IL-1β (GB11113, Servicebio, Wuhan, China), CD31 (ab222783, Abcam, UK), EMCN ( GB112648 , Servicebio, Wuhan, China), OPN (22952-1-AP, Proteintech, Wuhan, China), OCN (A6205, ABclonal, Wuhan, China), TUBB3, CGRP, p-YAP1, β-catenin, COL1A, and RUNX2.

Techniques: Co-Culture Assay, Staining, Immunofluorescence, Expressing, Marker, Western Blot, Wound Healing Assay, Migration, Transwell Assay, Tube Formation Assay

Journal: Bioactive Materials

Article Title: Composite hydrogel-microsphere delivery system promotes early nerve-mediated bone regeneration and late-stage mechanotransduction-driven bone remodeling via sequential release of NGF and Yoda1

doi: 10.1016/j.bioactmat.2025.10.040

Figure Lengend Snippet: Effect of NGF-G/P@Yoda1 on promoting bone regeneration and bone remodeling in femur def ect model. (A–D) Representative immunohistochemical staining images at week 8 for RUNX2 and OPN and semi-quantitative analysis of positive areas to evaluate the osteogenic capacity of composite hydrogels. (E, F) Representative immunofluorescence (IF) staining images at week 8 of OCN and semi-quantitative analysis. (G–J) Representative IF staining images at week 8 of YAP1and β-catenin and semi-quantitative analyses. (K, L) Representative TRAP staining images at week 8 and semi-quantitative analysis of positive areas to assess bone resorption and remodeling. The quantitative data (n ≥ 3) in the figures represent as the averages ± SD, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: Primary antibodies included IL-1β (GB11113, Servicebio, Wuhan, China), CD31 (ab222783, Abcam, UK), EMCN ( GB112648 , Servicebio, Wuhan, China), OPN (22952-1-AP, Proteintech, Wuhan, China), OCN (A6205, ABclonal, Wuhan, China), TUBB3, CGRP, p-YAP1, β-catenin, COL1A, and RUNX2.

Techniques: Immunohistochemical staining, Staining, Immunofluorescence