Journal: Cell Proliferation

Article Title: Inhibition of Fibroblast Activation Protein‐α Ameliorates Intervertebral Disc Degeneration via Reduced Vascular Invasion in Cartilage Endplate

doi: 10.1111/cpr.70162

Figure Lengend Snippet: FAP‐α is highly expressed in degenerative cartilage endplate (CEP) tissues. (A) Bioinformatics analysis of the expression profiles from patients with degenerative CEP and healthy controls. Heatmap showing the top differentially expressed genes (DEGs). (B) Volcano plot of DEGs indicating significant changes in mRNA expression ( p < 0.05). Green dots represent upregulated DEGs, while orange dots represent downregulated DEGs. (C) MRI images and tissue photos of intervertebral discs at different stages of degeneration. (D) Immunohistochemical staining of human CEP tissue for FAP‐α, MMP3, CD31, VEGFA, Collagen X, and Collagen II. Black scale bar = 100 μm. Data are presented as means ± SE, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The 3‐month‐old rats were randomly assigned to three groups ( n = 6 each): the sham operation group, the LSI model group and the LSI + FAP‐α suppression treatment group [ ] (5 mg/kg talabostat orally once daily, HY‐13233B, MedChemExpress).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Cell Proliferation

Article Title: Inhibition of Fibroblast Activation Protein‐α Ameliorates Intervertebral Disc Degeneration via Reduced Vascular Invasion in Cartilage Endplate

doi: 10.1111/cpr.70162

Figure Lengend Snippet: FAP‐α is upregulated in the CEP tissues of aged rats. (A) Representative Safranin O‐Fast Green staining images for each group (female rats, n = 6/group). Black scale bar = 200 μm, yellow scale bar = 40 μm. (B) Histological scores of intervertebral discs from rats of different ages. (C, D) IHC detection and quantitative analysis of FAP‐α, MMP3, CD31, VEGFA, Collagen X, and Collagen II. Black scale bar = 200 μm, yellow scale bar = 40 μm. Data are presented as means ± SE, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. AF: annulus fibrosus, CEP: cartilage endplate, IVD: intervertebral disc, NP: nucleus pulposus.

Article Snippet: The 3‐month‐old rats were randomly assigned to three groups ( n = 6 each): the sham operation group, the LSI model group and the LSI + FAP‐α suppression treatment group [ ] (5 mg/kg talabostat orally once daily, HY‐13233B, MedChemExpress).

Techniques: Staining

Journal: Cell Proliferation

Article Title: Inhibition of Fibroblast Activation Protein‐α Ameliorates Intervertebral Disc Degeneration via Reduced Vascular Invasion in Cartilage Endplate

doi: 10.1111/cpr.70162

Figure Lengend Snippet: Talabostat ameliorates IDD in the rat lumbar spine instability model 3 months post‐surgery. (A, B) Representative X‐ray images and disc height indices (DHI) of rat intervertebral discs. DHI% was calculated as: DHI% = (A0 + B0 + C0) × 100% / (A0 + B0 + C0 + A1 + B1 + C1). White scale bar = 200 μm, n = 8. (C, D) T2‐weighted MRI images of rat lumbar spines from different experimental groups. Grey‐scale values of regions of interest (red irregular circle) in the intervertebral disc (IVD) are shown. White scale bar = 200 μm, n = 8. (E, F) Representative Safranin O‐Fast Green and H&E staining of disc samples from different experimental groups with corresponding histological grades. Black scale bar = 200 μm, yellow scale bar = 40 μm. (G, H) Representative immunofluorescence images of FAP‐α, Collagen II, MMP3, and CD31 in intervertebral discs from different experimental groups. White scale bar = 200 μm, n = 8. Semi‐quantitative analysis of fluorescence intensity. Averages ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. CEP: cartilage endplate, IVD: intervertebral disc.

Article Snippet: The 3‐month‐old rats were randomly assigned to three groups ( n = 6 each): the sham operation group, the LSI model group and the LSI + FAP‐α suppression treatment group [ ] (5 mg/kg talabostat orally once daily, HY‐13233B, MedChemExpress).

Techniques: Staining, Immunofluorescence, Fluorescence

Journal: Cell Proliferation

Article Title: Inhibition of Fibroblast Activation Protein‐α Ameliorates Intervertebral Disc Degeneration via Reduced Vascular Invasion in Cartilage Endplate

doi: 10.1111/cpr.70162

Figure Lengend Snippet: Lumbar intervertebral discs of wild‐type (WT) and FAP KO mice in the ageing model. (A) Representative Safranin O‐Fast Green and H&E staining images of different groups of aged mice. Black scale bar = 200 μm, red scale bar = 40 μm, n = 8. (B) Histological scores of intervertebral discs in different groups of aged mice. (C, D) Representative immunofluorescence images of FAP‐α, Collagen II, Aggrecan, MMP3, H2AX, and CD31 in disc samples. White scale bar = 200 μm, yellow scale bar = 40 μm, n = 8. Semi‐quantitative analysis of fluorescence intensity. Averages ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. AF: annulus fibrosus, CEP: cartilage endplate, IVD: intervertebral disc, NP: nucleus pulposus.

Article Snippet: The 3‐month‐old rats were randomly assigned to three groups ( n = 6 each): the sham operation group, the LSI model group and the LSI + FAP‐α suppression treatment group [ ] (5 mg/kg talabostat orally once daily, HY‐13233B, MedChemExpress).

Techniques: Staining, Immunofluorescence, Fluorescence

Journal: Cell Proliferation

Article Title: Inhibition of Fibroblast Activation Protein‐α Ameliorates Intervertebral Disc Degeneration via Reduced Vascular Invasion in Cartilage Endplate

doi: 10.1111/cpr.70162

Figure Lengend Snippet: Effects of FAP‐α on blood vessel function. (A‐C) Optical images showing the effect of FAP‐α knockdown or overexpression in conditioned media on HUVEC migration at 24 and 48 h (scale bar = 200 μm). Quantification of migration area (%) by ImageJ. (D‐E) Schematic of the co‐culture experiment of HUVECs and CEP cells, and image of crystal violet‐stained migrating HUVECs (scale bar = 200 μm). (F‐H) Representative images of the tube formation assay on Matrigel in HUVECs treated with FAP‐α conditioned medium (scale bar = 200 μm). Quantitative analysis of the number of branch points (%) and tube length (%). (I‐L) Representative immunofluorescence images of p‐p65 and MMP3 in CEP cells, and semi‐quantitative analysis of fluorescence intensity. Scale bar = 50 μm. Averages ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The 3‐month‐old rats were randomly assigned to three groups ( n = 6 each): the sham operation group, the LSI model group and the LSI + FAP‐α suppression treatment group [ ] (5 mg/kg talabostat orally once daily, HY‐13233B, MedChemExpress).

Techniques: Knockdown, Over Expression, Migration, Co-Culture Assay, Staining, Tube Formation Assay, Immunofluorescence, Fluorescence

Journal: Cell Proliferation

Article Title: Inhibition of Fibroblast Activation Protein‐α Ameliorates Intervertebral Disc Degeneration via Reduced Vascular Invasion in Cartilage Endplate

doi: 10.1111/cpr.70162

Figure Lengend Snippet: FAP‐α activates the PI3K/Akt/HIF‐1α/VEGFA pathway, increases the release of inflammatory cytokines, and promotes Collagen II degradation. (A) Volcano plot of gene differences between CEP cells with siFAP and the control group. (B) KEGG analysis of differentially expressed gene pathways. (C) GO analysis of differentially expressed gene pathways. (D, E) Western blot results show the protein levels of FAP‐α, HIF‐1α, p‐PI3K, PI3K, p‐AKT, AKT, and VEGFA in the control group and the siFAP‐α group. (F, G) Western blot results show the protein levels of FAP‐α, HIF‐1α, p‐PI3K, PI3K, p‐AKT, AKT, and VEGFA in the Vector group and the overexpression of FAP‐α group. (H, I) Representative immunofluorescence images of Collagen II in CEP cells and semi‐quantitative analysis of fluorescence intensity. Scale bar = 50 μm. (J–L) The levels of IL‐1β, IL‐6, and TNF‐α in supernate of each group were measured using ELSIA. Averages ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The 3‐month‐old rats were randomly assigned to three groups ( n = 6 each): the sham operation group, the LSI model group and the LSI + FAP‐α suppression treatment group [ ] (5 mg/kg talabostat orally once daily, HY‐13233B, MedChemExpress).

Techniques: Control, Western Blot, Plasmid Preparation, Over Expression, Immunofluorescence, Fluorescence