Journal: Bioactive Materials

Article Title: Injection site dictates the immune response to a biodegradable polymer and corresponding collagen regeneration

doi: 10.1016/j.bioactmat.2026.04.004

Figure Lengend Snippet: ID tissue exhibits higher densities of fibroblasts, macrophages, and microvessels compared to SC tissue. Immunofluorescence analysis comparing the expression of Vimentin (a fibroblast marker), CD68 (a macrophage marker), and CD31 (an endothelial cell marker) in ID and SC tissues. For each group, n = 3; data represent mean ± s.d.; ∗P < 0.05 and ∗∗P < 0.01.

Article Snippet: After antigen retrieval and blocking, sections were incubated overnight at 4 °C with primary antibodies targeting vimentin, CD68 (ABclonal, A20803), CD31 (R&D SYSTEMS, AF3628), HMGB1 (Cell Signaling Technology, 3935), HSP70 (ABclonal, A23457), NF-κB p65 (ABclonal, A19653), CD206 (Cell Signaling Technology, 24595), and FAPα (ABclonal, A23789 ).

Techniques: Immunofluorescence, Expressing, Marker

Journal: Bioactive Materials

Article Title: Spatiotemporally engineered microneedle for microenvironment remodeling propels mucosal regeneration after tracheal mucosal injury

doi: 10.1016/j.bioactmat.2026.01.026

Figure Lengend Snippet: Evaluation of mucosal status after tracheal mucosal injury in rabbits . (a) Gross morphology and Masson staining of mucosal defects (n = 3). (b – f) Tracheal samples were collected on Day 10, and RNA sequencing was performed to assess biological differences between Native and Model groups (n = 3). (b) Principal components analysis of samples. (c) Volcano plot of DEGs. (d) KEGG enrichment analysis of DEGs. (e) Chord plot of enriched KEGG terms. (f) Heatmaps of DEGs associated with inflammation and oxidative stress. (g) IF staining of FISH (marker of bacteria, pink), iNOS (marker of M1 macrophages, orange), CD206 (marker of M2 macrophages, green), and immunohistochemical staining of CD31 (marker of endothelial cells) in various samples, blood vessels are denoted by triangles.

Article Snippet: IF staining was performed for iNOS (Abcam, ab283655), CD206 (Proteintech, 18704-1-AP), and CD31 (Abcam, ab28364) to assess inflammatory marker expression, and vascularization, respectively.

Techniques: Staining, RNA Sequencing, Marker, Bacteria, Immunohistochemical staining

Journal: Bioactive Materials

Article Title: Spatiotemporally engineered microneedle for microenvironment remodeling propels mucosal regeneration after tracheal mucosal injury

doi: 10.1016/j.bioactmat.2026.01.026

Figure Lengend Snippet: Gel-AgNA/MgGA MN remodel the microenvironment after TMI by regulating infection, inflammation, oxidative stress, and vascular disruption. (a) FISH (marker of bacteria, red) staining of harvested samples after treated with Gel, Gel-AgNA, Gel-MgGA, and Gel-AgNA/MgGA MN. (b, c) Quantitative analysis of bacterial coverage and fluorescence intensity of FISH (n = 9). (d) Heatmaps of DEGs related to inflammation and oxidative stress. (e) RT-PCR analysis showing the relative expression levels of IL-1, TNF-α, HIF-1α, and IL-6 (n = 3). (f) IF staining of iNOS (marker of M1 macrophages, red) and CD206 (marker of M2 macrophages, green). (g) Quantitative analysis of iNOS and CD206 fluorescence intensity. (h) RT-PCR analysis showing the relative expression levels of Arg1 and CD206. (i) IF staining of CD31 (marker of blood vessels, red). (j, k) RT-PCR analysis showing the relative expression levels of VEGF and HIF-1α (n = 3).

Article Snippet: IF staining was performed for iNOS (Abcam, ab283655), CD206 (Proteintech, 18704-1-AP), and CD31 (Abcam, ab28364) to assess inflammatory marker expression, and vascularization, respectively.

Techniques: Infection, Disruption, Marker, Bacteria, Staining, Fluorescence, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Bioactive Materials

Article Title: Biodegradable Mg 2+ -releasing piezoelectric scaffold for segmental bone defect repair

doi: 10.1016/j.bioactmat.2026.02.017

Figure Lengend Snippet: In vitro evaluation of osteogenic differentiation on Mg 2+ -releasing piezoelectric scaffolds. A) ALP staining of BMSCs cultured with WH Gel and PWH Gel (Scale bar: 1 mm). B) ARS staining of BMSCs cultured with WH Gel and PWH Gel (Scale bar: 1 mm). C-F) RT-qPCR results showing the relative mRNA expression of OPN, RUNX2, OCN, and COL-I in BMSCs cultured with cryogels for 7 days and 14 days. CLSM images showing the expression of (G) OPN, (H) RUNX2, (I) OCN, and (J) COL-I in BMSCs co-cultured with WH Gel and PWH Gel (Scale bar: 50 μm). Data are presented as mean ± S.D. (n = 3 independent replicates). ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; NS, not significant.

Article Snippet: Immunohistochemical staining was carried out for COL-I (Proteintech, 28083-1-AP, USA).

Techniques: In Vitro, Staining, Cell Culture, Quantitative RT-PCR, Expressing

Journal: Bioactive Materials

Article Title: Biodegradable Mg 2+ -releasing piezoelectric scaffold for segmental bone defect repair

doi: 10.1016/j.bioactmat.2026.02.017

Figure Lengend Snippet: In vivo assessments of large segmental bone defect regeneration using Mg 2+ -releasing piezoelectric scaffold. A-B) Schematic showing the surgical procedure for scaffold implantation in rat radial defects (Scale bar = 1 cm). C) Macroscopic images of the defect site at 6- and 12- weeks post-implantation. D) RUS scores for radial repair. E) 3D micro-CT images of the defects at 6- and 12- weeks post-implantation (Scale bar = 3 mm). F-G) Quantitative micro-CT analysis of BV/TV and trabecular number (Tb.N) in cryogel-treated regions at 6- and 12- weeks post-implantation. H) Representative H&E and Masson's trichrome staining images of defect tissues at 6- and 12-weeks post-implantation (Scale bar: 1 mm). I) Immunohistochemical staining for COL-I (Scale bar: 1 mm). J) Representative immunofluorescence staining of CD31 (Scale bar: 1 mm). Data are expressed as mean ± S.D. (n = 3 independent replicates). Statistical significance was determined as ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; NS, not significant.

Article Snippet: Immunohistochemical staining was carried out for COL-I (Proteintech, 28083-1-AP, USA).

Techniques: In Vivo, Micro-CT, Staining, Immunohistochemical staining, Immunofluorescence

Journal: STAR Protocols

Article Title: Protocol for isolating stromal cells from lymphoid tissue for performing scRNA-seq

doi: 10.1016/j.xpro.2026.104501

Figure Lengend Snippet: Cell selection using automated magnetic cell sorting (A) Cells were stained with CD45-biotin and CD31-biotin and sorted using autoMACS® Pro Separator. (B) Number of cells before staining for autoMACS® separation (step 11), and number of cells recovered from positive selection (CD45 + and CD31 + cells) and negative selection (CD45 - and CD31 - cells) in step 23. Each dot represents combined numbers from inguinal, axillary and brachial lymph nodes from 6 mice. Colors indicate biological replicates. (C) Percentages of cells after separation compared to the pre-staining cell count performed in step 11. (D) Purity check of separated cells using flow cytometry and staining for CD45 and CD31. (E) Overlay plots of positive selection (blue) and negative selection (red). (F) Percentage of CD45 + , CD45 - and CD31 + cells recovered (∗∗∗∗ p value < 0.0001, ∗ p value < 0.05). (G) Final viability check of positive and negative selected cells acquired just before performing scRNA-sequencing analysis.

Article Snippet: CD31 antibody, anti-mouse, Biotin (Dilutions in 1:50) , Miltenyi Biotec , Cat# 130-119-562, RRID: AB_2751728.

Techniques: Selection, FACS, Staining, Cell Characterization, Flow Cytometry, Sequencing

Journal: Materials Today Bio

Article Title: Extracellular biogenic nanoscale mitochondria reprogram the wound microenvironment via ROS scavenging independent of cellular uptake

doi: 10.1016/j.mtbio.2026.103023

Figure Lengend Snippet: MSC-mt alleviates oxidative stress and promote tissue regeneration during wound healing (A) In vivo imaging showing the spatial–temporal persistence of fluorescently labeled MSC-mt (mtH) at the wound site at indicated time point, indicating transient but sustained early presence after topical application. (B) Measurement of ATP levels in peri-wound tissues on PWD8 showed enhanced local metabolic activity following mtH treatment. n = 5 ∼ 6 per group. (C) Quantification of malondialdehyde (MDA) levels in peri-wound tissues on PWD8 indicated reduced lipid peroxidation and oxidative stress in both MSC-mt–treated wounds. n = 5 ∼ 6 per group. (D) Laser speckle contrast imaging of blood perfusion at the wound site on PWD8 showed improved microvascular perfusion following mtH treatment. n = 5 per group. (E) Representative immunofluorescence images and quantification of CD31 expression in peri-wound tissues on PWD8, indicating enhanced angiogenesis in mtH–treated wounds. n = 6 per group. (F) Quantitative PCR analysis of angiogenesis-related gene expression in peri-wound tissues on PWD8, indicating transcriptional activation of pro-angiogenic programs following mtH treatment. n = 3 ∼ 5 per group. (G-H) Representative immunohistochemical staining and quantification of Col1a1 in wound tissues on PWD8, showing increased collagen synthesis and matrix remodeling in mtH–treated wounds. n = 6 per group. Scale bar = 100 μm. (I-J) Representative immunofluorescence staining and quantification of Vimentin and TUNEL in wound tissues on PWD8, indicating reduced fibroblast apoptosis following mtH treatment. n = 6 per group. Scale bar = 20 μm. (K-L) Representative immunofluorescence staining and quantification of Vimentin and 8-hydroxyguanosine (8-OHG) in wound tissues on PWD8, indicating attenuated oxidative DNA damage in fibroblasts following mtH treatment. n = 6 per group. Scale bar = 20 μm. Data are presented as mean ± SEM. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.

Article Snippet: Sections were incubated overnight at 4 °C with primary antibodies against CD31 (Servicebio, Cat# GB120005 , 1:200), Vimentin (CST, Cat# 5741, 1:200), and 8-hydroxyguanosine (8-OHG, Rockland, Cat# 200-301-A99, 1:200).

Techniques: In Vivo Imaging, Labeling, Activity Assay, Imaging, Immunofluorescence, Expressing, Real-time Polymerase Chain Reaction, Gene Expression, Activation Assay, Immunohistochemical staining, Staining, TUNEL Assay