Journal: International Dental Journal

Article Title: NLRP12 Alleviated Periodontal Destruction via Suppressing Nuclear Factor Kappa-B Signalling Pathway

doi: 10.1016/j.identj.2026.109417

Figure Lengend Snippet: LPS triggered an inflammatory response and suppressed the osteogenic differentiation potential of PDLSCs. (A) The proliferative capacity of PDLSCs exposed to varying LPS concentrations over 2, 4, and 6 days was measured with CCK-8 assay. (B) The qRT-PCR analysis revealed elevated mRNA expression of inflammatory mediators IL-6, IL-8 , and IL-10 in PDLSCs following treatment with 10 µg/mL LPS. The qRT-PCR (C) and WB (D) analysis revealed significantly lower expression of osteogenic markers (COL1 and RUNX2) in PDLSCs exposed to 10 µg/mL LPS. The ALP (E) and alizarin red (F) staining assays showed that 10 µg/mL LPS significantly inhibited the osteogenic differentiation capacity of PDLSCs compared to the control group. Data were presented as mean ± SD ( n = 3). *** P < .001, **** P < .0001.

Article Snippet: After blocking with 10% normal goat serum (Beyotime), sections were covered overnight at 4°C with primary antibodies against IL-6 (#21865-1-AP; Proteintech), IL-8 (#94407; CST), IL-10 (#ab290735; Abcam), COL1 (#12256; CST) and RUNX2 (#8486; CST), then covered with secondary antibodies Goat Anti-Rabbit IgG (Zsbio), and then stained with DAB (Zsbio) for 20 to 60 seconds and counterstained with haematoxylin.

Techniques: CCK-8 Assay, Quantitative RT-PCR, Expressing, Staining, Control

Journal: International Dental Journal

Article Title: NLRP12 Alleviated Periodontal Destruction via Suppressing Nuclear Factor Kappa-B Signalling Pathway

doi: 10.1016/j.identj.2026.109417

Figure Lengend Snippet: NLRP12 overexpression reduced LPS-induced inflammatory response and reversed LPS-induced suppression of osteogenic differentiation in PDLSCs. (A and B) The qRT-PCR and WB data revealed that expression level of NLRP12 in oeNLRP12 group was increased significantly compared to oeNC group. (C) The qRT-PCR data revealed that mRNA expression of IL-6 and IL-8 was suppressed, while the mRNA expression of IL-10 was increased in oeNLRP12 + LPS group compared to oeNC + LPS group. (D) The expression of COL1 and RUNX2 was increased significantly detected by WB assays in oeNLRP12 + LPS group compared to oeNC + LPS group. (E) Representative pictures of ALP staining showed lower staining in oeNC + LPS group compared to oeNC group and higher staining in oeNLRP12 + LPS group compared to oeNC + LPS group (scale bar = 500 μm). (F) Representative pictures of alizarin red staining showed fewer mineralized nodules in oeNC + LPS group compared to oeNC group and more mineralized nodules in oeNLRP12 + LPS group compared to oeNC + LPS group (scale bar = 500 μm). oeNC: PDLSCs transfected via negative control lentiviral. oeNLRP12: PDLSCs transfected via lentiviral with overexpression- NLRP12 . oeNC + LPS: PDLSCs transfected via negative control lentiviral and subsequently cultured under 10 µg/mL LPS induction. oeNLRP12 + LPS: PDLSCs transfected via lentiviral with overexpression- NLRP12 and subsequently cultured under 10 µg/mL LPS induction. Data were presented as mean ± SD ( n = 3). ns, no significant difference, *** P < .001, **** P < .0001.

Article Snippet: After blocking with 10% normal goat serum (Beyotime), sections were covered overnight at 4°C with primary antibodies against IL-6 (#21865-1-AP; Proteintech), IL-8 (#94407; CST), IL-10 (#ab290735; Abcam), COL1 (#12256; CST) and RUNX2 (#8486; CST), then covered with secondary antibodies Goat Anti-Rabbit IgG (Zsbio), and then stained with DAB (Zsbio) for 20 to 60 seconds and counterstained with haematoxylin.

Techniques: Over Expression, Quantitative RT-PCR, Expressing, Staining, Transfection, Negative Control, Cell Culture

Journal: International Dental Journal

Article Title: NLRP12 Alleviated Periodontal Destruction via Suppressing Nuclear Factor Kappa-B Signalling Pathway

doi: 10.1016/j.identj.2026.109417

Figure Lengend Snippet: Overexpression of NLRP12 alleviated the inflammatory responses and osteogenic differentiation inhibition of PDLSCs by suppressing the NF-κB pathway. (A) WB results showing the changes of protein expression levels of p-p65, p65, p-IκBα, and IκBα in PDLSCs after NLRP12 overexpression. (B) WB results showing the changes of protein expression levels of p-p65 and p65 in PDLSCs overexpressing NLRP12 after PMA treatment. (C)The qRT-PCR results demonstrating the transcriptional expression levels of IL-6, IL-8 , and IL-10 in PDLSCs overexpressing NLRP12 after PMA treatment. (D)WB results demonstrating alterations in the protein expression levels of COL1 and RUNX2 in PDLSCs overexpressing NLRP12 after PMA treatment. (E) Representative pictures showing ALP staining (scale bar = 500 μm) (F) Representative pictures showing alizarin red staining (scale bar = 500 μm). oeNC: PDLSCs transfected via negative control lentiviral. oeNLRP12: PDLSCs transfected via lentiviral with overexpression- NLRP12 . oeNC + LPS: PDLSCs transfected via negative control lentiviral and subsequently cultured under 10 µg/mL LPS. oeNLRP12 + LPS: PDLSCs transfected via lentiviral with overexpression- NLRP12 and subsequently cultured under 10 µg/mL LPS. Data were presented as mean ± SD ( n = 3). ns, no significant difference, * P < .05, ** P < .01, *** P < .001, **** P < .0001.

Article Snippet: After blocking with 10% normal goat serum (Beyotime), sections were covered overnight at 4°C with primary antibodies against IL-6 (#21865-1-AP; Proteintech), IL-8 (#94407; CST), IL-10 (#ab290735; Abcam), COL1 (#12256; CST) and RUNX2 (#8486; CST), then covered with secondary antibodies Goat Anti-Rabbit IgG (Zsbio), and then stained with DAB (Zsbio) for 20 to 60 seconds and counterstained with haematoxylin.

Techniques: Over Expression, Inhibition, Expressing, Quantitative RT-PCR, Staining, Transfection, Negative Control, Cell Culture

Journal: International Dental Journal

Article Title: NLRP12 Alleviated Periodontal Destruction via Suppressing Nuclear Factor Kappa-B Signalling Pathway

doi: 10.1016/j.identj.2026.109417

Figure Lengend Snippet: Overexpression of NLRP12 in PDLSCs alleviated the inflammatory responses and promoted periodontal regeneration in periodontitis rats. (A) The timeline demonstrated the experimental procedures. (B) The micro-CT scanning results. (C) Representative images of H&E staining (the black scale bars and white scale bars are 200 and 100 μm, respectively). (D) Representative images of Masson trichrome staining (the black scale bars and white scale bars are 200 and 100 μm, respectively). (E) Cementoenamel junction (CEJ)–alveolar bone crest (ABC) distance. (F) bone volume/total volume (BV/TV). (G) Loss of attachment in H&E staining. (H) Semiquantitative analysis of Masson trichrome staining. (I) Representative immunohistochemical images showing the expression level of inflammatory factor IL-6, IL-8, and IL-10, osteogenic factor COL1, and RUNX2, and TRAP staining showing the number of osteoclasts. Scale bar = 100 μm. (J) Quantitative analysis of IL-6-positive cells, IL-8-positive cells, IL-10-positive cells, COL1-positive cells, and RUNX2-positive cells. (K) Quantitative result of TRAP+ cells. Control group: without treatment. Periodontitis group: ligature-induced experimental periodontitis model. oeNC group: periodontitis group treated with oeNC PDLSCs. oeNLRP12 group: periodontitis group treated with oeNLRP12 PDLSCs. IHC: immunohistochemistry. IOD, integrated option density. The data are presented as mean ± SD ( n = 6 rats per group). *P < .1 , **P < .01 , ***P < .001 , ****P < .0001 .

Article Snippet: After blocking with 10% normal goat serum (Beyotime), sections were covered overnight at 4°C with primary antibodies against IL-6 (#21865-1-AP; Proteintech), IL-8 (#94407; CST), IL-10 (#ab290735; Abcam), COL1 (#12256; CST) and RUNX2 (#8486; CST), then covered with secondary antibodies Goat Anti-Rabbit IgG (Zsbio), and then stained with DAB (Zsbio) for 20 to 60 seconds and counterstained with haematoxylin.

Techniques: Over Expression, Micro-CT, Staining, Immunohistochemical staining, Expressing, Control, Immunohistochemistry

Journal: Scientific Reports

Article Title: IL-8 contributes to postoperative adhesion formation through the crosstalk of neutrophils and mesothelial cells

doi: 10.1038/s41598-026-42159-x

Figure Lengend Snippet: Evaluation of functional differences between IL-8 and CXCL1 in neutrophils. ( A ) Evaluation of IL-8 and CXCL1 activity in the U2OS-CXCR1 reporter cells ( left ) and CHO-K1-CXCR2 reporter cells ( right ). ( B ) Flow cytometric analysis of CXCR1 and CXCR2 surface expression on neutrophils. ( C ) Neutrophil chemotaxis assay toward IL-8 or CXCL1. RLU, relative luminescence units. Data in ( A ) and ( C ) are presented as the mean ± SD (n = 3).

Article Snippet: The concentrations of IL-8 in the tissue lysates were measured by sandwich immunoassay using two noncompetitive anti-human IL-8 antibodies (Chugai Pharmaceutical Co., Ltd.).

Techniques: Functional Assay, Activity Assay, Expressing, Chemotaxis Assay

Journal: Scientific Reports

Article Title: IL-8 contributes to postoperative adhesion formation through the crosstalk of neutrophils and mesothelial cells

doi: 10.1038/s41598-026-42159-x

Figure Lengend Snippet: Reciprocal amplification of IL-8 and TNF-α in MeT-5A cells. ( A , B ) Flow cytometric analysis of CXCR1 ( A ) and CXCR2 ( B ) surface expression on MeT-5A cells. Left: representative overlaid histograms. Right: quantification of MFI across biological replicates. P values were determined using Student’s t test. *** indicates P < 0.001. ( C ) Evaluation of TNF-α concentrations in culture medium. P values were determined using Tukey’s multiple comparison test. *** indicates P < 0.001. ( D ) Evaluation of IL-8 concentrations in culture medium. P values were determined using Williams’ test. * indicates P < 0.025. Quantitative data are presented as individual data points with mean ± SD. (n = 3).

Article Snippet: The concentrations of IL-8 in the tissue lysates were measured by sandwich immunoassay using two noncompetitive anti-human IL-8 antibodies (Chugai Pharmaceutical Co., Ltd.).

Techniques: Amplification, Expressing, Comparison

Journal: Scientific Reports

Article Title: IL-8 contributes to postoperative adhesion formation through the crosstalk of neutrophils and mesothelial cells

doi: 10.1038/s41598-026-42159-x

Figure Lengend Snippet: Fibrotic reactions in neutrophils and MeT-5A cells. ( A ) RNA expression change of TGFB1 in neutrophils stimulated with IL-8, IL-8 + AMY109, or TNF-α. ( B ) RNA expression change of TGFB1 , COL1A1 , FN1 , CCN2 ( CTGF ), SERPINE1 (PAI-1) and VIM in MeT-5A cells stimulated with TGF-β1 (0.4, 2, 10 ng/mL), TNF-α or IL-8. Data are presented as individual data points with mean ± SD. (n = 3) P values were determined using Dunnett’s test. * and *** indicate P < 0.05 and P < 0.001, respectively.

Article Snippet: The concentrations of IL-8 in the tissue lysates were measured by sandwich immunoassay using two noncompetitive anti-human IL-8 antibodies (Chugai Pharmaceutical Co., Ltd.).

Techniques: RNA Expression

Journal: Scientific Reports

Article Title: IL-8 contributes to postoperative adhesion formation through the crosstalk of neutrophils and mesothelial cells

doi: 10.1038/s41598-026-42159-x

Figure Lengend Snippet: Evaluation of IL-8 protein expression in injured abdominal walls from monkeys following cesarean sections (C-sections) and in postoperative adhesions (PA) model. ( A ) IL-8 concentrations in injured abdominal walls from monkeys following C-sections. Data are presented as individual data points with mean values indicated (n = 10 lesions for 0 h, 2 lesions for 6 h, Day 1, 3, 7, and 1 month). ( B ) IL-8 concentrations in injured abdominal walls from monkey PA models. Data are presented as individual data points with mean ± SD. (n = 6 lesions for 0 h, 8 lesions for 6 h, 4 lesions for Day 7) P values were determined using Steel’s test. * and ** indicate P < 0.05 and P < 0.01, respectively.

Article Snippet: The concentrations of IL-8 in the tissue lysates were measured by sandwich immunoassay using two noncompetitive anti-human IL-8 antibodies (Chugai Pharmaceutical Co., Ltd.).

Techniques: Expressing

Journal: Scientific Reports

Article Title: IL-8 contributes to postoperative adhesion formation through the crosstalk of neutrophils and mesothelial cells

doi: 10.1038/s41598-026-42159-x

Figure Lengend Snippet: Possible role of IL-8 and AMY109 in postoperative adhesion formation. IL-8 promotes neutrophil infiltration and forms a reciprocal amplification with TNF-α in mesothelial cells. Elevated TNF-α upregulates TGF-β1 in neutrophils, driving adhesion formation. Neutralization of IL-8 by AMY109 blocks neutrophil recruitment and subsequent inflammatory responses and fibrotic reactions, thereby preventing adhesion formation.

Article Snippet: The concentrations of IL-8 in the tissue lysates were measured by sandwich immunoassay using two noncompetitive anti-human IL-8 antibodies (Chugai Pharmaceutical Co., Ltd.).

Techniques: Amplification, Neutralization