Identification of the pyroptosis, apoptosis, and necroptosis (PANoptosis) involved in osteogenic differentiation inhibition impaired by tumor necrosis factor-α

Abstract Background Inflammatory diseases of the bones often result in excessive release of inflammatory cytokines and impaired osteogenic differentiation. Tumor necrosis factor‐α (TNF-α) is an important inflammatory cytokine in disease, and implicated in developing PANoptosis. Although TNF-α has ca...

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Main Authors: Chang-liang Xia, Fei Liang, Jia-xuan Li, Yudun Qu, Wei Zhang, Suanji Ou, Yang Yang, Yong Qi, Chang-peng Xu
Format: Article
Language:English
Published: BMC 2025-07-01
Series:European Journal of Medical Research
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Online Access:https://doi.org/10.1186/s40001-025-02702-4
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author Chang-liang Xia
Fei Liang
Jia-xuan Li
Yudun Qu
Wei Zhang
Suanji Ou
Yang Yang
Yong Qi
Chang-peng Xu
author_facet Chang-liang Xia
Fei Liang
Jia-xuan Li
Yudun Qu
Wei Zhang
Suanji Ou
Yang Yang
Yong Qi
Chang-peng Xu
author_sort Chang-liang Xia
collection DOAJ
description Abstract Background Inflammatory diseases of the bones often result in excessive release of inflammatory cytokines and impaired osteogenic differentiation. Tumor necrosis factor‐α (TNF-α) is an important inflammatory cytokine in disease, and implicated in developing PANoptosis. Although TNF-α has captured much attention in PANoptosis, their relationship in osteogenic differentiation remains elusive. Therefore, it is necessary to explore how TNF-α inhibits osteogenic differentiation and the mechanism involved. Methods RNA sequencing was used to sequence normal and TNF-α-treated cells and screen for differentially expressed 43 genes of PANotosis. PCR and Western blotting were performed to validate sequencing results consistent. The degree of osteoblast differentiation was determined by alkaline phosphatase and calcium ion quantification. The extent of cell death was evaluated by cell death staining, lactate dehydrogenase quantification, and propidium iodide staining. Results Calcium ion quantification, RUNX2 and ALP confirmed that osteogenic differentiation was inhibited, and related to the cell death increase. More importantly, scanning electron microscopy revealed that multiple PANotosis morphologies appeared in the same field of view. We also demonstrated that by interdict NLRP3, cells could be rescued from PANoptosis and the inhibition of osteogenic differentiation caused by TNF-α. Further, immunofluorescence and coimmunoprecipitation of several closely related proteins were performed to verify the possible presence of PANoptosome. Conclusion In summary, inhibition PANoptosis may serve as a new therapeutic target for bone infection and may plays an important role in osteogenic differentiation and inflammatory cell death in bone infection.
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spelling doaj-art-bd0c0c3c5e564dc4a87201bcd6d89aff2025-08-20T04:01:25ZengBMCEuropean Journal of Medical Research2047-783X2025-07-0130111510.1186/s40001-025-02702-4Identification of the pyroptosis, apoptosis, and necroptosis (PANoptosis) involved in osteogenic differentiation inhibition impaired by tumor necrosis factor-αChang-liang Xia0Fei Liang1Jia-xuan Li2Yudun Qu3Wei Zhang4Suanji Ou5Yang Yang6Yong Qi7Chang-peng Xu8Department of Orthopaedics, The Affiliated Guangdong Second Provincial General Hospital of Jinan UniversityDepartment of Anaesthesiology, Guangdong Second Provincial General HospitalDepartment of Orthopaedics, The Affiliated Guangdong Second Provincial General Hospital of Jinan UniversityDepartment of Orthopaedics, The Affiliated Guangdong Second Provincial General Hospital of Jinan UniversityDepartment of Orthopaedics, The Affiliated Guangdong Second Provincial General Hospital of Jinan UniversityDepartment of Orthopaedics, The Affiliated Guangdong Second Provincial General Hospital of Jinan UniversityDepartment of Orthopaedics, The Affiliated Guangdong Second Provincial General Hospital of Jinan UniversityDepartment of Orthopaedics, The Affiliated Guangdong Second Provincial General Hospital of Jinan UniversityDepartment of Orthopaedics, The Affiliated Guangdong Second Provincial General Hospital of Jinan UniversityAbstract Background Inflammatory diseases of the bones often result in excessive release of inflammatory cytokines and impaired osteogenic differentiation. Tumor necrosis factor‐α (TNF-α) is an important inflammatory cytokine in disease, and implicated in developing PANoptosis. Although TNF-α has captured much attention in PANoptosis, their relationship in osteogenic differentiation remains elusive. Therefore, it is necessary to explore how TNF-α inhibits osteogenic differentiation and the mechanism involved. Methods RNA sequencing was used to sequence normal and TNF-α-treated cells and screen for differentially expressed 43 genes of PANotosis. PCR and Western blotting were performed to validate sequencing results consistent. The degree of osteoblast differentiation was determined by alkaline phosphatase and calcium ion quantification. The extent of cell death was evaluated by cell death staining, lactate dehydrogenase quantification, and propidium iodide staining. Results Calcium ion quantification, RUNX2 and ALP confirmed that osteogenic differentiation was inhibited, and related to the cell death increase. More importantly, scanning electron microscopy revealed that multiple PANotosis morphologies appeared in the same field of view. We also demonstrated that by interdict NLRP3, cells could be rescued from PANoptosis and the inhibition of osteogenic differentiation caused by TNF-α. Further, immunofluorescence and coimmunoprecipitation of several closely related proteins were performed to verify the possible presence of PANoptosome. Conclusion In summary, inhibition PANoptosis may serve as a new therapeutic target for bone infection and may plays an important role in osteogenic differentiation and inflammatory cell death in bone infection.https://doi.org/10.1186/s40001-025-02702-4TNF-αPANoptosisInflammatory bone diseaseNLRP3
spellingShingle Chang-liang Xia
Fei Liang
Jia-xuan Li
Yudun Qu
Wei Zhang
Suanji Ou
Yang Yang
Yong Qi
Chang-peng Xu
Identification of the pyroptosis, apoptosis, and necroptosis (PANoptosis) involved in osteogenic differentiation inhibition impaired by tumor necrosis factor-α
European Journal of Medical Research
TNF-α
PANoptosis
Inflammatory bone disease
NLRP3
title Identification of the pyroptosis, apoptosis, and necroptosis (PANoptosis) involved in osteogenic differentiation inhibition impaired by tumor necrosis factor-α
title_full Identification of the pyroptosis, apoptosis, and necroptosis (PANoptosis) involved in osteogenic differentiation inhibition impaired by tumor necrosis factor-α
title_fullStr Identification of the pyroptosis, apoptosis, and necroptosis (PANoptosis) involved in osteogenic differentiation inhibition impaired by tumor necrosis factor-α
title_full_unstemmed Identification of the pyroptosis, apoptosis, and necroptosis (PANoptosis) involved in osteogenic differentiation inhibition impaired by tumor necrosis factor-α
title_short Identification of the pyroptosis, apoptosis, and necroptosis (PANoptosis) involved in osteogenic differentiation inhibition impaired by tumor necrosis factor-α
title_sort identification of the pyroptosis apoptosis and necroptosis panoptosis involved in osteogenic differentiation inhibition impaired by tumor necrosis factor α
topic TNF-α
PANoptosis
Inflammatory bone disease
NLRP3
url https://doi.org/10.1186/s40001-025-02702-4
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