Study on the effects of spermidine on LPS-induced inflammatory osteolysis in mouse calvaria
Objective·To investigate the inhibitory effects of spermidine (SPD) on inflammatory osteolysis both in vivo and in vitro.Methods·The CCK-8 assay was used to assess the viability of RAW264.7 macrophages treated with various concentrations of SPD. The levels of intracellular reactive oxygen species (R...
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Editorial Office of Journal of Shanghai Jiao Tong University (Medical Science)
2025-06-01
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| Series: | Shanghai Jiaotong Daxue xuebao. Yixue ban |
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| Online Access: | https://xuebao.shsmu.edu.cn/article/2025/1674-8115/1674-8115-2025-45-6-673.shtml |
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| author | ZHAO Xinyu ZHANG Wenchao CHEN Xuzhuo SONG Jiaqi HUANG Hui ZHANG Shanyong |
| author_facet | ZHAO Xinyu ZHANG Wenchao CHEN Xuzhuo SONG Jiaqi HUANG Hui ZHANG Shanyong |
| author_sort | ZHAO Xinyu |
| collection | DOAJ |
| description | Objective·To investigate the inhibitory effects of spermidine (SPD) on inflammatory osteolysis both in vivo and in vitro.Methods·The CCK-8 assay was used to assess the viability of RAW264.7 macrophages treated with various concentrations of SPD. The levels of intracellular reactive oxygen species (ROS) in lipopolysaccharide (LPS)-activated RAW264.7 cells were evaluated by staining with dichlorodihydrofluorescein diacetate (DCFH-DA) and dihydroethidium (DHE), respectively. Reverse transcription real-time fluorescence quantitative PCR (RT-qPCR) was utilized to determine the effects of SPD on the expression of pro-inflammatory genes in LPS-activated RAW264.7 cells. Tartrate-resistant acid phosphatase (TRAP) staining was used to evaluate the effect of SPD on the differentiation of mouse primary bone marrow-derived macrophages (BMMs) into osteoclasts. RT-qPCR was employed to further analyze the effect of SPD on the expression of genes related to osteoclast differentiation and functions after BMM-induced differentiation. An LPS-induced mouse calvarial osteolysis model was constructed, and the therapeutic efficacy of SPD on inflammatory osteolysis was assessed using Micro-CT analysis, hematoxylin-eosin (H-E) staining and TRAP staining of histological sections.Results·The CCK-8 assay showed that SPD, even at a concentration of 1 000 μmol/L, exhibited no significant cytotoxicity to RAW264.7 cells. ROS analysis revealed that SPD markedly inhibited LPS-induced elevation of intracellular ROS levels in macrophages. RT-qPCR results indicated that SPD suppressed the expression of pro-inflammatory genes induced by LPS. Both TRAP staining and RT-qPCR demonstrated that SPD effectively inhibited the differentiation of BMMs into osteoclasts induced by receptor activator of nuclear factor-κB ligand (RANKL) and reduced the expression of genes associated with osteoclast differentiation and function. In the mouse calvarial osteolysis model, Micro-CT analysis showed that the bone volume fraction and bone mineral density in the SPD-treated groups were significantly higher than those in the LPS group. Histological staining revealed that SPD treatment reduced inflammatory cell infiltration, decreased osteoclast numbers, and alleviated tissue damage.Conclusion·SPD inhibits macrophage inflammatory responses and RANKL-induced osteoclast differentiation in vitro; in vivo, it alleviates LPS-induced inflammatory calvarial osteolysis in mice. |
| format | Article |
| id | doaj-art-caa47acb80e842a0a03f31f67d5608e3 |
| institution | DOAJ |
| issn | 1674-8115 |
| language | zho |
| publishDate | 2025-06-01 |
| publisher | Editorial Office of Journal of Shanghai Jiao Tong University (Medical Science) |
| record_format | Article |
| series | Shanghai Jiaotong Daxue xuebao. Yixue ban |
| spelling | doaj-art-caa47acb80e842a0a03f31f67d5608e32025-08-20T03:15:19ZzhoEditorial Office of Journal of Shanghai Jiao Tong University (Medical Science)Shanghai Jiaotong Daxue xuebao. Yixue ban1674-81152025-06-0145667368310.3969/j.issn.1674-8115.2025.06.0021674-8115（2025）06-0673-11Study on the effects of spermidine on LPS-induced inflammatory osteolysis in mouse calvariaZHAO Xinyu0ZHANG Wenchao1CHEN Xuzhuo2SONG Jiaqi3HUANG Hui4ZHANG Shanyong5School of Stomatology, Shandong Second Medical University, Weifang 261053, ChinaSchool of Stomatology, Shandong Second Medical University, Weifang 261053, ChinaDepartment of Oral Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology, Shanghai Research Institute of Stomatology, Shanghai 200011, ChinaDepartment of Stomatology, Shenzhen Hospital of Peking University, Shenzhen 518000, ChinaDepartment of Oral Imageology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, ChinaDepartment of Oral Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology, Shanghai Research Institute of Stomatology, Shanghai 200011, ChinaObjective·To investigate the inhibitory effects of spermidine (SPD) on inflammatory osteolysis both in vivo and in vitro.Methods·The CCK-8 assay was used to assess the viability of RAW264.7 macrophages treated with various concentrations of SPD. The levels of intracellular reactive oxygen species (ROS) in lipopolysaccharide (LPS)-activated RAW264.7 cells were evaluated by staining with dichlorodihydrofluorescein diacetate (DCFH-DA) and dihydroethidium (DHE), respectively. Reverse transcription real-time fluorescence quantitative PCR (RT-qPCR) was utilized to determine the effects of SPD on the expression of pro-inflammatory genes in LPS-activated RAW264.7 cells. Tartrate-resistant acid phosphatase (TRAP) staining was used to evaluate the effect of SPD on the differentiation of mouse primary bone marrow-derived macrophages (BMMs) into osteoclasts. RT-qPCR was employed to further analyze the effect of SPD on the expression of genes related to osteoclast differentiation and functions after BMM-induced differentiation. An LPS-induced mouse calvarial osteolysis model was constructed, and the therapeutic efficacy of SPD on inflammatory osteolysis was assessed using Micro-CT analysis, hematoxylin-eosin (H-E) staining and TRAP staining of histological sections.Results·The CCK-8 assay showed that SPD, even at a concentration of 1 000 μmol/L, exhibited no significant cytotoxicity to RAW264.7 cells. ROS analysis revealed that SPD markedly inhibited LPS-induced elevation of intracellular ROS levels in macrophages. RT-qPCR results indicated that SPD suppressed the expression of pro-inflammatory genes induced by LPS. Both TRAP staining and RT-qPCR demonstrated that SPD effectively inhibited the differentiation of BMMs into osteoclasts induced by receptor activator of nuclear factor-κB ligand (RANKL) and reduced the expression of genes associated with osteoclast differentiation and function. In the mouse calvarial osteolysis model, Micro-CT analysis showed that the bone volume fraction and bone mineral density in the SPD-treated groups were significantly higher than those in the LPS group. Histological staining revealed that SPD treatment reduced inflammatory cell infiltration, decreased osteoclast numbers, and alleviated tissue damage.Conclusion·SPD inhibits macrophage inflammatory responses and RANKL-induced osteoclast differentiation in vitro; in vivo, it alleviates LPS-induced inflammatory calvarial osteolysis in mice.https://xuebao.shsmu.edu.cn/article/2025/1674-8115/1674-8115-2025-45-6-673.shtmlspermidine (spd)osteolysisosteoclastinflammation |
| spellingShingle | ZHAO Xinyu ZHANG Wenchao CHEN Xuzhuo SONG Jiaqi HUANG Hui ZHANG Shanyong Study on the effects of spermidine on LPS-induced inflammatory osteolysis in mouse calvaria Shanghai Jiaotong Daxue xuebao. Yixue ban spermidine (spd) osteolysis osteoclast inflammation |
| title | Study on the effects of spermidine on LPS-induced inflammatory osteolysis in mouse calvaria |
| title_full | Study on the effects of spermidine on LPS-induced inflammatory osteolysis in mouse calvaria |
| title_fullStr | Study on the effects of spermidine on LPS-induced inflammatory osteolysis in mouse calvaria |
| title_full_unstemmed | Study on the effects of spermidine on LPS-induced inflammatory osteolysis in mouse calvaria |
| title_short | Study on the effects of spermidine on LPS-induced inflammatory osteolysis in mouse calvaria |
| title_sort | study on the effects of spermidine on lps induced inflammatory osteolysis in mouse calvaria |
| topic | spermidine (spd) osteolysis osteoclast inflammation |
| url | https://xuebao.shsmu.edu.cn/article/2025/1674-8115/1674-8115-2025-45-6-673.shtml |
| work_keys_str_mv | AT zhaoxinyu studyontheeffectsofspermidineonlpsinducedinflammatoryosteolysisinmousecalvaria AT zhangwenchao studyontheeffectsofspermidineonlpsinducedinflammatoryosteolysisinmousecalvaria AT chenxuzhuo studyontheeffectsofspermidineonlpsinducedinflammatoryosteolysisinmousecalvaria AT songjiaqi studyontheeffectsofspermidineonlpsinducedinflammatoryosteolysisinmousecalvaria AT huanghui studyontheeffectsofspermidineonlpsinducedinflammatoryosteolysisinmousecalvaria AT zhangshanyong studyontheeffectsofspermidineonlpsinducedinflammatoryosteolysisinmousecalvaria |