Journal: Bioactive Materials

Article Title: AdMSC spheroids encapsulating antioxidant hybrid protein carrier for irradiation-damaged salivary gland repair

doi: 10.1016/j.bioactmat.2026.03.049

Figure Lengend Snippet: Functional characterization of AdMSC spheroids and their integration with GC. (a) Bright-field images and size distribution of spheroids generated in agarose molds targeting a diameter of 300 μm. (b) Representative confocal images of hypoxic core staining in spheroids, showing nuclei (blue) and hypoxic regions (green). (c, d) ELISA quantification of cumulative VEGF and HGF secretion from 2D-cultured AdMSCs and spheroids after 3 days culture. (e) Bright-field and confocal fluorescence images of spheroids alone, spheroids exposed to blue light (B.L), spheroids encapsulating 5% (w/v) gelatin hydrogel, or spheroids encapsulating 4–7% (w/v) GCs and photo-crosslinked under 450 nm blue light. Samples were stained with phalloidin (green, F-actin) and DAPI (blue, nuclei); dotted boxes indicate magnified regions. (f) Quantification of normalized cell proliferation within the GC in each condition over 1, 3, and 5 days incubation (n = 3). (g) Cumulative VEGF secretion from encapsulated spheroids measured at 1, 3, and 5 days (n = 5). (h) Spheroid diameter measurements over the incubation period, indicating the degree of spheroid spreading within each matrix (n = 10). (i) Representative HUVEC tube formation images after treatment with conditioned media from VEGF-free control (-VEGF), VEGF-supplemented media (+VEGF), spheroids alone, spheroid encapsulating 5% (w/v) GC, or spheroid encapsulating 7% (w/v) GC groups. (j–m) Quantitative tube formation parameters, including total tube length, number of meshes, number of junctions, and total mesh area, respectively (n = 4). (n) Representative Annexin V-FITC/propidium iodide flow cytometry plots of H 2 O 2 -treated SG cells in the absence or presence of spheroid encapsulating 5% (w/v) GC or spheroid encapsulating 7% (w/v) GCs. (o) Quantification of live, early apoptotic, and late apoptotic cell populations under each condition (n = 3). (p) Relative M2/M1 polarization of macrophages after LPS stimulation and treatment with conditioned media with spheroids and spheroids encapsulating GCs as determined by ELISA (n = 3). Data are presented as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. ns P > 0.05, ∗ P ≤ 0.05, ∗∗ P ≤ 0.01, and ∗∗∗ P ≤ 0.001.

Article Snippet: VEGF and HGF levels were quantified using human VEGF (ABclonal, RK00023) and human HGF ELISA kits according to the manufacturers’ protocols, and absorbance was read at 450 nm with 540 nm reference using a microplate reader (VersaMax; Molecular Devices, USA).

Techniques: Functional Assay, Generated, Staining, Enzyme-linked Immunosorbent Assay, Cell Culture, Fluorescence, Incubation, Control, Flow Cytometry

Journal: Bioactive Materials

Article Title: AdMSC spheroids encapsulating antioxidant hybrid protein carrier for irradiation-damaged salivary gland repair

doi: 10.1016/j.bioactmat.2026.03.049

Figure Lengend Snippet: Angiogenic effect of spheroids encapsulating GC for IR-damaged salivary glands. (a) Immunofluorescence staining for CD31 (green), an endothelial cell marker, in SG tissue sections to evaluate neovascularization. (b, c) Quantification of CD31-positive areas at 1 and 6 weeks post-treatment. (d) Immunofluorescence staining for VEGF-A (red), an angiogenesis marker, in SG tissue sections. (e, f) Quantification of VEGF-A-positive areas at 1 and 6 weeks post-treatment. (g–n) Relative mRNA expression levels of angiogenesis-related genes, including VEGF, HGF, CD31, and vWF, in mouse SG tissues harvested at 1 and 6 weeks post-treatment, as determined by quantitative PCR. Data are presented as mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. ns P > 0.05, ∗ P ≤ 0.05, ∗∗ P ≤ 0.01, and ∗∗∗ P ≤ 0.001.

Article Snippet: VEGF and HGF levels were quantified using human VEGF (ABclonal, RK00023) and human HGF ELISA kits according to the manufacturers’ protocols, and absorbance was read at 450 nm with 540 nm reference using a microplate reader (VersaMax; Molecular Devices, USA).

Techniques: Immunofluorescence, Staining, Marker, Expressing, Real-time Polymerase Chain Reaction