Journal: Biomaterials Research

Article Title: Injectable Poloxamer and Hyaluronic Acid Hydrogel for Sustained Co-Delivery of Dexamethasone and Lidocaine Ameliorates Neuropathic Pain

doi: 10.34133/bmr.0373

Figure Lengend Snippet: TRPV1-mediated nociceptive sensitization in the DRG and spinal cord dorsal horn (SC) following CCI and treatment. (A) Schematic illustration of the proposed mechanism: Injury-induced TRPV1 ion channel activation triggers calcium influx and downstream CGRP release, which activates adenylate cyclase/PKA signaling to amplify neuropathic pain sensitization. (B) Representative immunofluorescence images of the DRG stained for TRPV1 (red), NeuN (green), and DAPI (blue). The Injury group shows marked up-regulation of TRPV1 in sensory neurons. Dex/Lid@PLX/HA treatment substantially reduces TRPV1 expression, restoring it to near-Naive levels. Scale bars, 200 μm (overview) and 50 μm (inset). (C) Immunofluorescence staining for TRPV1 (red) and NeuN (green) in the spinal dorsal horn. The dashed line indicates the dorsal horn boundary. Dex/Lid@PLX/HA significantly suppresses injury-induced central TRPV1 up-regulation. Scale bar, 200 μm. (D to F) Quantitative analysis of the relative TRPV1 + area in the DRG (top) and spinal cord (middle), and the SGC/neuron ratio. Dex/Lid@PLX/HA shows marked suppression of TRPV1 overexpression relative to the injury group. Data are presented as mean ± SEM. **** P < 0.0001, ** P < 0.01, * P < 0.05, ns: not significant.

Article Snippet: The sections were then incubated overnight at 4 °C with the following primary antibodies: TRPV1 (Alomone Labs, catalog number ACC-030-GP), Iba-1 (Abcam, catalog number ab5076), NeuN (Abcam, catalog number ab104224), CD68 (Abcam, catalog number ab31630), CD163 (Abcam, catalog number ab182422), CGRP (Abcam, catalog number ab47027), GFAP (Millipore, catalog number MAB360), and NF200 (Abcam, catalog number ab8135).

Techniques: Activation Assay, Immunofluorescence, Staining, Expressing, Over Expression

Journal: bioRxiv

Article Title: A novel role for TRPV1 in macrophage giant cell formation

doi: 10.64898/2026.05.11.724406

Figure Lengend Snippet: (A) Immunoblot analysis shows TRPV1, TRPA1, and actin levels in WT BMDMs 48 h following treatment with or without IL-4 plus GMCSF (25 ng/ml). (B-C) Densitometric quantification of immunoblot data from (A) (n = 3 biological replicates; one-way ANOVA, **p < 0.01, ***p < 0.001). (D) Representative immunofluorescence images of WT BMDMs stained for TRPV1 (red) using anti-TRPV1 IgG (original magnification, 60 x; scale bar, 2 μm; n = 10 cells per condition). Statistical analysis by Student’s t-test, ***p < 0.001.

Article Snippet: C57BL/6 and TRPV1 KO mice were purchased from The Jackson Laboratory (ME, USA).

Techniques: Western Blot, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: A novel role for TRPV1 in macrophage giant cell formation

doi: 10.64898/2026.05.11.724406

Figure Lengend Snippet: (A) Representative Giemsa-stained images of multinucleated FBGCs in WT BMDMs left untreated or stimulated with IL-4 plus GMCSF (25 ng/ml, 96 h), in the presence or absence of the TRPV1 antagonist AMG. (B-D) Quantitative analysis of FBGC formation from (A): (B) number of FBGCs per high-power field, (C) percentage of fused BMDMs, and (D) average FBGC size. Data represent n = 3 biological replicates with 5 images per group; scale bar, 100 μm; Student’s t-test, ***p < 0.001, ****p < 0.0001. (E) Representative immunofluorescence images of WT BMDMs transfected with scramble or TRPV1-targeting siRNA, stained for TRPV1 (red) using anti-TRPV1 IgG (original magnification, 60x; scale bar, 2 μm). (F) Quantification of TRPV1 fluorescence intensity (n = 10 cells per condition; Student’s t-test, ***p < 0.001). (G) Immunoblot showing TRPV1 expression in WT BMDMs 48 h after transfection with scramble or TRPV1 siRNA. (H-J) Quantification of FBGC formation following TRPV1 knockdown: (H) number of FBGCs per high-power field, (I) percentage of fused BMDMs, and (J) average FBGC size. Data represent n = 3 biological replicates with 5 images per group; Student’s t-test, ***p < 0.001.

Article Snippet: C57BL/6 and TRPV1 KO mice were purchased from The Jackson Laboratory (ME, USA).

Techniques: Staining, Immunofluorescence, Transfection, Fluorescence, Western Blot, Expressing, Knockdown

Journal: bioRxiv

Article Title: A novel role for TRPV1 in macrophage giant cell formation

doi: 10.64898/2026.05.11.724406

Figure Lengend Snippet: (A) Representative images of FBGCs formed by WT BMDMs cultured on collagen-coated (10 μg/ml) PA hydrogels of defined stiffness (1 kPa and 50 kPa), left untreated or stimulated with IL-4 plus GMCSF (25 ng/ml, 96 h), in the presence or absence of the TRPV1 antagonist AMG. (B-C) Quantification of FBGC formation from (A): (B) number of FBGCs per high-power field and (C) percentage of fused BMDMs. (D) Representative images of FBGCs in WT and TRPV1 KO BMDMs cultured on Permanox slides under untreated or IL-4 plus GMCSF-stimulated conditions (25 ng/ml, 96 h). (E-F) Quantification of FBGC formation from (D): (E) number of FBGCs per high-power field and (F) percentage of fused BMDMs. (G) Representative images of FBGCs in WT and TRPV1 KO BMDMs cultured on collagen-coated (10 μg/ml) 50 kPa PA hydrogels, with or without IL-4 plus GMCSF stimulation (25 ng/ml, 96 h). (H-I) Quantification of FBGC formation from (G): (H) number of FBGCs per high-power field and (I) percentage of fused BMDMs. Data represent n = 3 biological replicates with 5 images per group. Scale bar, 20 μm; statistical analysis by one-way ANOVA, ***p < 0.001, ****p < 0.0001.

Article Snippet: C57BL/6 and TRPV1 KO mice were purchased from The Jackson Laboratory (ME, USA).

Techniques: Cell Culture

Journal: bioRxiv

Article Title: A novel role for TRPV1 in macrophage giant cell formation

doi: 10.64898/2026.05.11.724406

Figure Lengend Snippet: (A) Spinning-disk confocal images of WT and TRPV1 KO BMDMs showing Ca 2+ influx (red) under untreated (UT) conditions or following IL-4 plus GMCSF stimulation (25 ng/ml) in response to the TRPV1 agonist capsaicin; scale bar, 50 μm. (B) Quantification of fluorescence intensity from (A) (n = 4 fields per condition). (C) FlexStation 3 measurements of Ca 2+ influx in TRPV1 KO BMDMs stimulated with the TRPV4-specific agonist GSK1016790A (GSK101) under untreated (buffer control) or IL-4 plus GMCSF-treated conditions (25 ng/ml). (D) Quantification of Ca 2+ responses from (C). Experiments were performed three times with quadruplicate measurements. RFU, relative fluorescence units. Statistical significance was determined by Student’s t -test (**p < 0.01, ***p < 0.001).

Article Snippet: C57BL/6 and TRPV1 KO mice were purchased from The Jackson Laboratory (ME, USA).

Techniques: Fluorescence, Control

Journal: bioRxiv

Article Title: A novel role for TRPV1 in macrophage giant cell formation

doi: 10.64898/2026.05.11.724406

Figure Lengend Snippet: (A) Immunoblot analysis shows TRPV1, TRPA1, and actin levels in WT BMDMs 48 h following treatment with or without IL-4 plus GMCSF (25 ng/ml). (B-C) Densitometric quantification of immunoblot data from (A) (n = 3 biological replicates; one-way ANOVA, **p < 0.01, ***p < 0.001). (D) Representative immunofluorescence images of WT BMDMs stained for TRPV1 (red) using anti-TRPV1 IgG (original magnification, 60 x; scale bar, 2 μm; n = 10 cells per condition). Statistical analysis by Student’s t-test, ***p < 0.001.

Article Snippet: Primary antibodies comprised anti-actin (cat# 4970S; Cell Signaling Technology, Danvers, MA) and anti-TRPV1 (cat# ACC-030), anti-TRPV2 (cat# ACC-039), and anti-TRPA1 (cat# ACC-037) from Alomone Labs. Species-appropriate secondary antibodies (goat, rabbit, and mouse) were obtained from Jackson ImmunoResearch.

Techniques: Western Blot, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: A novel role for TRPV1 in macrophage giant cell formation

doi: 10.64898/2026.05.11.724406

Figure Lengend Snippet: (A) Representative Giemsa-stained images of multinucleated FBGCs in WT BMDMs left untreated or stimulated with IL-4 plus GMCSF (25 ng/ml, 96 h), in the presence or absence of the TRPV1 antagonist AMG. (B-D) Quantitative analysis of FBGC formation from (A): (B) number of FBGCs per high-power field, (C) percentage of fused BMDMs, and (D) average FBGC size. Data represent n = 3 biological replicates with 5 images per group; scale bar, 100 μm; Student’s t-test, ***p < 0.001, ****p < 0.0001. (E) Representative immunofluorescence images of WT BMDMs transfected with scramble or TRPV1-targeting siRNA, stained for TRPV1 (red) using anti-TRPV1 IgG (original magnification, 60x; scale bar, 2 μm). (F) Quantification of TRPV1 fluorescence intensity (n = 10 cells per condition; Student’s t-test, ***p < 0.001). (G) Immunoblot showing TRPV1 expression in WT BMDMs 48 h after transfection with scramble or TRPV1 siRNA. (H-J) Quantification of FBGC formation following TRPV1 knockdown: (H) number of FBGCs per high-power field, (I) percentage of fused BMDMs, and (J) average FBGC size. Data represent n = 3 biological replicates with 5 images per group; Student’s t-test, ***p < 0.001.

Article Snippet: Primary antibodies comprised anti-actin (cat# 4970S; Cell Signaling Technology, Danvers, MA) and anti-TRPV1 (cat# ACC-030), anti-TRPV2 (cat# ACC-039), and anti-TRPA1 (cat# ACC-037) from Alomone Labs. Species-appropriate secondary antibodies (goat, rabbit, and mouse) were obtained from Jackson ImmunoResearch.

Techniques: Staining, Immunofluorescence, Transfection, Fluorescence, Western Blot, Expressing, Knockdown