Journal: Bioactive Materials

Article Title: An intelligent nanoliposome alleviates disc degeneration and discogenic pain by inhibiting neurovascular ingrowth via a “Soil-conditioning, seed-modulating, and weeds-suppressing” strategy

doi: 10.1016/j.bioactmat.2025.11.048

Figure Lengend Snippet: NM-LP TK /RSV-MnCDs alleviates discogenic pain in IVDD. (A) Experimental timeline schematic. (B–C) Open-field behavioral analysis showing locomotor activity parameters (active time, central mean speed, total distance, and mean speed) across groups (n = 5). (D–E) PWL and PWT test showing alleviation of thermal and mechanical allodynia by NM-LP TK /RSV-MnCDs in LSI mice (n = 5). (F–G) IHC staining and quantification analyses of DRG sections showing expression of sensory neuropeptides CGRP and SP (n = 5). Scale bar, 100 μm. (H–I) IF co-labeling of PGP9.5 and CGRP/SP in DRG, and corresponding quantification analyses (n = 5). Scale bar, 100 μm. Data are presented as mean ± SD. For comparisons among multiple groups at a single time point, one-way ANOVA followed was employed to determine statistical significance. For datasets involving multiple groups evaluated over time, including PWL and PWT assessments, a two-way repeated measures ANOVA was utilized. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; ns = not significant.

Article Snippet: For immunofluorescence (IF) staining, antigen retrieval was conducted using citrate buffer (0.1 M, pH 6.0), followed by blocking in 10 % normal goat serum (Solarbio, Cat. No. SL038) at room temperature for 1 h. Sections were incubated overnight at 4 °C with primary antibodies targeting ACAN (Cat. No. DF7561, Affinity), COL2A1 (Cat. No. AF0135, Affinity), MMP3 (Cat. No. AF0217, Affinity), MMP13 (Cat. No. 18165-1-AP, Proteintech), CD31 (Cat. No. AF0077, Affinity), p-LATS1 (Cat. No. AF7170, Affinity), p-YAP (Cat. No. AF3328, Affinity), Tuj1 (Cat. No. 66375, Proteintech), CGRP (Cat. No. DF7386, Affinity), SP (Cat. No. DF7522, Affinity), and PGP9.5 (Cat. No. AF5490, Affinity) followed by incubation with appropriate fluorescent secondary antibodies for 2 h at room temperature and counterstaining with DAPI (Beyotime, Cat. No. C1006).

Techniques: Activity Assay, Immunohistochemistry, Expressing, Labeling

Journal: Cell reports

Article Title: CGRP signaling links tumor-associated pain to immune evasion in oral squamous cell carcinoma

doi: 10.1016/j.celrep.2026.116994

Figure Lengend Snippet: (A and B) Representative images of OSCC tumor serial sections stained with either anti-Tubb3 (top, green outline), anti-TRPV1 (middle, red outline), or anti-CGRP (bottom, blue outline) from a patient with (A) and without (B) CGRPα + nerve innervation. Positive stains are indicated by red (TRPV1) or blue (CGRP) arrows. Tumor margins are indicated by a gray dashed line. Image magnification: 6× (left) and 40× (right). Scale bar: 200 μm. (C) Quantification of the percentage of total TRPV1 and CGRP-IR nerve area relative to total Tubb3-IR nerve area across tumor tissue sections from 23 patients with HNSCC. Density is reported as a stacked bar graph. (D) Quantification of the percentage of total CD8 T cell density per square millimeter. (E) Representative images of OSCC tumor serial sections with either a large nerve bundle and low anti-CD8 or small nerve presence and high anti-CD8 immunoreactivity. Scale bar: 150 μm. (F and G) Simple linear regressions were run between patient-reported pain, percentage of total CGRP-IR nerve area relative to total Tubb3-IR, and CD8 + T cell density relative to tumor area. Pain was measured by the FACT-HN additional question 12, “I have pain in my mouth, throat or neck” (FACT-HN10). The response to this question is rated on a scale of 0 (not at all) to 4 (very much). Spearman correlation r coefficients are listed on each graph; p < 0.05. Patient demographics are located in .

Article Snippet: Rabbit Anti-CGRP , Cell Signaling , 14959 S; RRID: AB_2798662.

Techniques: Staining

Journal: Cell reports

Article Title: CGRP signaling links tumor-associated pain to immune evasion in oral squamous cell carcinoma

doi: 10.1016/j.celrep.2026.116994

Figure Lengend Snippet: (A) Representative images of CGRP-IR in 300 μm optically cleared sagittal tongue sections from naive (left), MOC1 (middle), and MOC2 (right) tumor-bearing mice (10× magnification stitch, full focus z stack). Scale bar: 0.5 mm. (B) Representative images of co-staining with anti-CGRP and the pan-neuronal marker PGP9.5 in a 20 μm sagittal tongue section to demonstrate CGRP antibody specificity. Scale bar: 50 μm. (C) Representative images of neurite outgrowth in dissociated TG neurons in response to 24 h incubation in cell culture medium, MOC2 CM, or MOC2 medium + 1 μM anti-NGF monoclonal antibody (αNGF). A TG from one mouse was used for each treatment group, and the experiment was replicated five times ( n = 3 males, 2 females). For analysis, dissociated neurons were stained with anti-Tubb3 to visualize neurites and anti-NeuN to visualize cell bodies. Scale bar: 50 μm. (D–F) Integrative density of Tubb3 + neurites relative to the number of cell bodies in each field in response to medium and CM with or without αNGF; 12 areas of each chamber were imaged under 20× magnification, and analysis was completed within cell culture type (i.e., PEK, MOC1, and MOC2). Data are represented as mean ± SEM. One-way ANOVA; * p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.001.

Article Snippet: Rabbit Anti-CGRP , Cell Signaling , 14959 S; RRID: AB_2798662.

Techniques: Staining, Marker, Incubation, Cell Culture

Journal: Cell reports

Article Title: CGRP signaling links tumor-associated pain to immune evasion in oral squamous cell carcinoma

doi: 10.1016/j.celrep.2026.116994

Figure Lengend Snippet: (A) Representative image of a coronal tongue section from non-tumor bearing mice 1 week following treatment with vehicle (top) or RTX (bottom), stained with anti-CGRP to demonstrate loss of CGRP-expressing fibers following RTX treatment. Scale bar: 100 μm. (B) Schematic of the trigeminal CGRP release assay. (C) CGRP protein quantification in TG isolated from mice 1 week after RTX injection into the tongue ( n = 5 F/group). Data are represented as mean ± SEM. Independent t test, ** p < 0.01. (D) Representative ATF3 staining of in TRPV1-IR neurons in the trigeminal mandibular branch in TG sections from vehicle- and RTX-treated mice. Scale bar: 50 μm. (E) Percentage of ATF3 + TRPV1 + neurons relative to total TRPV1 + neurons in the V3 region from across 9 sections/mouse of mice treated with vehicle or RTX ( n = 3 vehicle, 4 RTX). Tissue was collected 7 days after treatment. There was no difference in the number of TRPV1 neurons between groups. Data are represented as mean ± SEM. Independent t test, *** p < 0.005. (F) RTX treatment did not affect body weight compared to vehicle-treated mice. Change in body weight was calculated as (post-treatment day 7) – (weight on first day of treatment). Independent t test. (G) Representative H&E-stained 5 μm tongue sections from vehicle- and RTX-treated mice 21 days after treatment. The tumor boundary is indicated by a black dashed line. Scale bar: 1 mm. (H) MOC1 tumor volume between groups over time ( n = 5 F/group). Data are represented as mean ± SEM. two-way ANOVA, * p < 0.05. (I and J) Example dot plots and quantification of CD3 + T cell subtype, CD19 + B cells, and NK1.1 + natural killer (NK) cells between groups ( n = 3 F/group). Within-subtype t test comparison, **** p < 0.0001.

Article Snippet: Rabbit Anti-CGRP , Cell Signaling , 14959 S; RRID: AB_2798662.

Techniques: Staining, Expressing, Release Assay, Isolation, Injection, Comparison