Journal: Investigative Ophthalmology & Visual Science

Article Title: Corneal Nerves Promote Alkali Burn Repair by Modulating Macrophages and Neutrophils via Calcitonin Gene-Related Peptide

doi: 10.1167/iovs.67.4.60

Figure Lengend Snippet: CGRP suppresses corneal tissue inflammation following alkali burn. ( A ) The volcano plot displayed the upregulated and downregulated genes in the corneas of mice from the CGRP-treated group compared with the PBS-treated group at 7 days post-injury ( n = 3). ( B ) The heatmap showed the differentially expressed genes in the corneas of mice from the CGRP-treated group compared with the PBS-treated group at 7 days post-injury ( n = 3). ( C ) The mRNA expression of the inflammatory factors IL-1β, iNOS, MCP-1, MMP3, and MMP9 was detected by qPCR on day 7 post-injury following CGRP treatment ( n = 4). ( D, E ) Representative flow cytometry plots and bar charts of CD45+ cells in the cornea after 7 days of treatment with CGRP following injury. ( F, G ) Representative micrographs and bar charts of macrophages in the cornea after 7 days treatment with topical CGRP or PBS ( n = 4). ( H ) Corneal macrophages were determined by flow cytometry ( n = 4). ( I, J ) Representative flow cytometry plots and bar charts of CD206+ macrophages in the cornea of the CGRP treatment group ( n = 4). ( K, L ) Representative micrographs and bar charts of neutrophils in the cornea after 7 days treatment with topical CGRP or PBS ( n = 4). (M) Corneal neutrophils were determined by flow cytometry ( n = 4). Data are shown as mean ± SD. ns: not significant; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: We dissolved CGRP powder (HY-P0203; MedChenExpress) in sterile PBS to a final concentration of 50 μM, aliquoted it, and stored it at –80°C.

Techniques: Expressing, Flow Cytometry

Journal: Investigative Ophthalmology & Visual Science

Article Title: Corneal Nerves Promote Alkali Burn Repair by Modulating Macrophages and Neutrophils via Calcitonin Gene-Related Peptide

doi: 10.1167/iovs.67.4.60

Figure Lengend Snippet: Expression of the CGRP receptor in neutrophils and macrophages, and transcriptomic analysis of macrophages following CGRP treatment in vitro. ( A, B ) Light microscope image of bone marrow-derived macrophages and bone marrow-derived neutrophils ( n = 4). ( C ) CALCRL and RAMP1 expression was detected in bone marrow-derived macrophages and neutrophils using immunostaining. Green = CALCRL, red = RAMP1 ( n = 4). ( D ) Heatmap of selected significantly upregulated and downregulated genes depicting standardized gene expression values in CGRP-treated cells compared to PBS-treated cells. The colored circles next to the heatmap denote gene functions. ( E ) Volcano plot displaying upregulated and downregulated genes in macrophages from the CGRP-treated group versus the PBS-treated group. ( F ) GO enrichment analysis of significantly upregulated ( red ) and downregulated ( blue ) genes in CGRP-treated and saline-treated macrophages.

Article Snippet: We dissolved CGRP powder (HY-P0203; MedChenExpress) in sterile PBS to a final concentration of 50 μM, aliquoted it, and stored it at –80°C.

Techniques: Expressing, In Vitro, Light Microscopy, Derivative Assay, Immunostaining, Gene Expression, Saline

Journal: Investigative Ophthalmology & Visual Science

Article Title: Corneal Nerves Promote Alkali Burn Repair by Modulating Macrophages and Neutrophils via Calcitonin Gene-Related Peptide

doi: 10.1167/iovs.67.4.60

Figure Lengend Snippet: CGRP suppresses corneal tissue inflammation following alkali burn. ( A ) The volcano plot displayed the upregulated and downregulated genes in the corneas of mice from the CGRP-treated group compared with the PBS-treated group at 7 days post-injury ( n = 3). ( B ) The heatmap showed the differentially expressed genes in the corneas of mice from the CGRP-treated group compared with the PBS-treated group at 7 days post-injury ( n = 3). ( C ) The mRNA expression of the inflammatory factors IL-1β, iNOS, MCP-1, MMP3, and MMP9 was detected by qPCR on day 7 post-injury following CGRP treatment ( n = 4). ( D, E ) Representative flow cytometry plots and bar charts of CD45+ cells in the cornea after 7 days of treatment with CGRP following injury. ( F, G ) Representative micrographs and bar charts of macrophages in the cornea after 7 days treatment with topical CGRP or PBS ( n = 4). ( H ) Corneal macrophages were determined by flow cytometry ( n = 4). ( I, J ) Representative flow cytometry plots and bar charts of CD206+ macrophages in the cornea of the CGRP treatment group ( n = 4). ( K, L ) Representative micrographs and bar charts of neutrophils in the cornea after 7 days treatment with topical CGRP or PBS ( n = 4). (M) Corneal neutrophils were determined by flow cytometry ( n = 4). Data are shown as mean ± SD. ns: not significant; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: We dissolved CGRP powder (HY-P0203; MedChenExpress) in sterile PBS to a final concentration of 50 μM, aliquoted it, and stored it at –80°C.

Techniques: Expressing, Flow Cytometry

Journal: Investigative Ophthalmology & Visual Science

Article Title: Corneal Nerves Promote Alkali Burn Repair by Modulating Macrophages and Neutrophils via Calcitonin Gene-Related Peptide

doi: 10.1167/iovs.67.4.60

Figure Lengend Snippet: Expression of the CGRP receptor in neutrophils and macrophages, and transcriptomic analysis of macrophages following CGRP treatment in vitro. ( A, B ) Light microscope image of bone marrow-derived macrophages and bone marrow-derived neutrophils ( n = 4). ( C ) CALCRL and RAMP1 expression was detected in bone marrow-derived macrophages and neutrophils using immunostaining. Green = CALCRL, red = RAMP1 ( n = 4). ( D ) Heatmap of selected significantly upregulated and downregulated genes depicting standardized gene expression values in CGRP-treated cells compared to PBS-treated cells. The colored circles next to the heatmap denote gene functions. ( E ) Volcano plot displaying upregulated and downregulated genes in macrophages from the CGRP-treated group versus the PBS-treated group. ( F ) GO enrichment analysis of significantly upregulated ( red ) and downregulated ( blue ) genes in CGRP-treated and saline-treated macrophages.

Article Snippet: We dissolved CGRP powder (HY-P0203; MedChenExpress) in sterile PBS to a final concentration of 50 μM, aliquoted it, and stored it at –80°C.

Techniques: Expressing, In Vitro, Light Microscopy, Derivative Assay, Immunostaining, Gene Expression, Saline

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