Journal: Cell Reports Medicine

Article Title: A biomimetic hybrid membrane vesicle nanoplatform attenuates tendinopathy through neuroinflammation modulation and tendon regeneration

doi: 10.1016/j.xcrm.2026.102937

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Article Snippet: Rabbit Polyclonal anti-CGRP , ABclonal , Cat# A5542; RRID: AB_2766323.

Techniques: Recombinant, Lysis, Hydroxyproline Assay, Enzyme-linked Immunosorbent Assay, Isolation, SYBR Green Assay, Bicinchoninic Acid Protein Assay, Staining, Software

Journal: Stem Cell Research & Therapy

Article Title: Activation of the G-protein coupled estrogen receptor 1 (GPER1) reduces transient receptor potential vanilloid 1 (TRPV1) activity and human iPSC-derived nociceptive neuron firing

doi: 10.1186/s13287-026-05174-3

Figure Lengend Snippet: Generation of iPSC line BO-VC1 -derived nociceptive neurons. Differentiation of ( A ) human iPSC line BO-VC1 into nociceptive neurons was done by dual SMAD inhibition, suppressing TGF-ß and BMP4 signaling, in knockout serum replacement (KSR) medium (day 0–4). This was followed by an overlapping inhibition of glycogen synthase kinase-3ß (GSK-3ß), vascular endothelial growth factor (VEGF), and Notch signaling (day 2–12), with KSR medium incremented with N2/B27 medium every second day by 25% from day 4 on. After reseeding of the ( B ) immature sensory neurons, final maturation was performed in N2/B27 medium infused with the neurotrophic factors glial cell line-derived neurotrophic factor (GDNF), nerve growth factor (NGF), and brain-derived neurotrophic factor (BDNF) (day 13 - approx. 60) according to the protocols of Chambers and Schoepf leading to the differentiation of ( C ) intermediate and finally ( D ) mature nociceptors (Scale bar: 500 μm). Mature nociceptive neurons express ( E ) the neuron specific microtubule element βIII-tubulin (βIIITUB) and vGLUT2, the sensory neuron markers ( F ) insulin gene enhancer protein 1 (ISL1) as well as the brain-specific homeobox/POU domain protein 3 A (BRN3A). Additionally, iPSC-derived differentiated cells express the nociceptive markers ( G ) transient receptor potential cation channel subfamily V member 1(TRPV1), H voltage-gated sodium ion channel 1.9 (Na V 1.9) and I TWIK-related spinal cord potassium channel (TRESK). Moreover, co-expression of ( J ) the calcitonin gene-related peptide (CGRP), the nociceptive marker high affinity nerve growth factor receptor (TRKA) and the estrogen receptor G-protein coupled estrogen receptor 1 (GPER1) was confirmed (Scale bars: 50 μm) ( K ) Quantification of the percentage of positive cells for the respective markers was 95.96 ± 2,495% for CGRP, 89.68 ± 6,137% for GPER1, 91.67 ± 5.270% for Na V 1.9, 95.83 ± 4.167% for TRKA, 80.38 ± 7.143% for TRPV1 and 98.55 ± 0.2899% for TRESK. L Realtime-PCR analysis in order to quantify nociceptive neuron marker and GPER1 gene expression of HDFa-derived iPSCs (line BO-VC1) and corresponding nociceptive neurons relative to HDFa cell samples mean value. Means ± SEM (standard error of the mean) from three independent passages were statistically analyzed by one-way ANOVA with Turkey’s multiple comparison test (L). n = 3 (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001)

Article Snippet: Nevertheless, incubation with the first antibodies CGRP (1:500; # 414 004, Synaptic Systems GmbH, Göttingen, Germany), TRKA (1:500; # MA515509 , Thermo Fisher Scientific, Darmstadt, Germany), GPER1 (1:500; # ab39742, Abcam, ), BRN3A (1:100, # ab245230, Abcam, Cambridge, UK), ISL1 (1:100, #H00003670-M05, Abnova, Taipeh, Taiwan), TRPV1 (1:1000, # SAB5700857, Sigma-Aldrich, Merck, Darmstadt, Germany), vGlut2 (1:400, # MAB5504, Sigma-Aldrich, Merck, Darmstadt, Germany), ßIIITUB (1:300, # PA1-41331, Thermo Fisher Scientific, Darmstadt, Germany or 1:250, #302 304, Synaptic Systems GmbH, Göttingen, Germany) and Na V 1.9 (1:100, # ab65160, Abcam, Cambridge, UK) was done at 4 °C overnight.

Techniques: Derivative Assay, Inhibition, Knock-Out, Expressing, Marker, Gene Expression, Comparison

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