DLX5 regulates the osteogenic differentiation of spinal ligaments cells derived from ossification of the posterior longitudinal ligament patients via NOTCH signaling
Abstract Background Ossification of the posterior longitudinal ligaments (OPLL) is common disorder characterized by heterotopic ossification of the spinal ligaments. Mechanical stimulation (MS) plays an important role in OPLL. DLX5 is an essential transcription factor required for osteoblast differe...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2023-06-01
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| Series: | JOR Spine |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/jsp2.1247 |
| _version_ | 1827919039331041280 |
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| author | Tao Tang Zhengya Zhu Zhongyuan He Fuan Wang Hongkun Chen Shengkai Liu Mingbin Zhan Jianmin Wang Wei Tian Dafu Chen Xinbao Wu Xizhe Liu Zhiyu Zhou Shaoyu Liu |
| author_facet | Tao Tang Zhengya Zhu Zhongyuan He Fuan Wang Hongkun Chen Shengkai Liu Mingbin Zhan Jianmin Wang Wei Tian Dafu Chen Xinbao Wu Xizhe Liu Zhiyu Zhou Shaoyu Liu |
| author_sort | Tao Tang |
| collection | DOAJ |
| description | Abstract Background Ossification of the posterior longitudinal ligaments (OPLL) is common disorder characterized by heterotopic ossification of the spinal ligaments. Mechanical stimulation (MS) plays an important role in OPLL. DLX5 is an essential transcription factor required for osteoblast differentiation. However, the role of DLX5 during in OPLL is unclear. This study aims to investigate whether DLX5 is associated with OPLL progression under MS. Methods Stretch stimulation was applied to spinal ligaments cells derived from OPLL (OPLL cells) and non‐OPLL (non‐OPLL cells) patients. Expression of DLX5 and osteogenesis‐related genes were determined by quantitative real‐time polymerase chain reaction and Western blot. The osteogenic differentiation ability of the cells was measured using alkaline phosphatase (ALP) staining and alizarin red staining. The protein expression of DLX5 in the tissues and the nuclear translocation of NOTCH intracellular domain (NICD) was examined by immunofluorescence. Results Compared with non‐OPLL cells, OPLL cells expressed higher levels of DLX5 in vitro and vivo (p < 0.01). Upregulated expression of DLX5 and osteogenesis‐related genes (OSX, RUNX2, and OCN) were observed in OPLL cells induced with stretch stimulation and osteogenic medium, whereas there was no change in the non‐OPLL cells (p < 0.01). Cytoplasmic NICD protein translocated from the cytoplasm to the nucleus inducing DLX5 under stretch stimulation, which was reduced by the NOTCH signaling inhibitors (DAPT) (p < 0.01). Conclusions These data suggest that DLX5 play a critical role in MS‐induced progression of OPLL through NOTCH signaling, which provides a new insight into the pathogenesis of OPLL. |
| first_indexed | 2024-03-13T03:52:07Z |
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| institution | Directory Open Access Journal |
| issn | 2572-1143 |
| language | English |
| last_indexed | 2024-03-13T03:52:07Z |
| publishDate | 2023-06-01 |
| publisher | Wiley |
| record_format | Article |
| series | JOR Spine |
| spelling | doaj.art-bc7476dfb1e548bbab0b67c215ca79412023-06-22T10:18:22ZengWileyJOR Spine2572-11432023-06-0162n/an/a10.1002/jsp2.1247DLX5 regulates the osteogenic differentiation of spinal ligaments cells derived from ossification of the posterior longitudinal ligament patients via NOTCH signalingTao Tang0Zhengya Zhu1Zhongyuan He2Fuan Wang3Hongkun Chen4Shengkai Liu5Mingbin Zhan6Jianmin Wang7Wei Tian8Dafu Chen9Xinbao Wu10Xizhe Liu11Zhiyu Zhou12Shaoyu Liu13Innovation Platform of Regeneration and Repair of Spinal Cord and Nerve Injury, Department of Orthopaedic Surgery, The Seventh Affiliated Hospital Sun Yat‐sen University Shenzhen ChinaInnovation Platform of Regeneration and Repair of Spinal Cord and Nerve Injury, Department of Orthopaedic Surgery, The Seventh Affiliated Hospital Sun Yat‐sen University Shenzhen ChinaInnovation Platform of Regeneration and Repair of Spinal Cord and Nerve Injury, Department of Orthopaedic Surgery, The Seventh Affiliated Hospital Sun Yat‐sen University Shenzhen ChinaInnovation Platform of Regeneration and Repair of Spinal Cord and Nerve Injury, Department of Orthopaedic Surgery, The Seventh Affiliated Hospital Sun Yat‐sen University Shenzhen ChinaInnovation Platform of Regeneration and Repair of Spinal Cord and Nerve Injury, Department of Orthopaedic Surgery, The Seventh Affiliated Hospital Sun Yat‐sen University Shenzhen ChinaGuangdong Provincial Key Laboratory of Orthopedics and Traumatology, Orthopaedic Research Institute/Department of Spinal Surgery The First Affiliated Hospital of Sun Yat‐sen University Guangzhou ChinaGuangdong Provincial Key Laboratory of Orthopedics and Traumatology, Orthopaedic Research Institute/Department of Spinal Surgery The First Affiliated Hospital of Sun Yat‐sen University Guangzhou ChinaInnovation Platform of Regeneration and Repair of Spinal Cord and Nerve Injury, Department of Orthopaedic Surgery, The Seventh Affiliated Hospital Sun Yat‐sen University Shenzhen ChinaLaboratory of Bone Tissue Engineering, Beijing Laboratory of Biomedical Materials Beijing Research Institute of Orthopaedics and Traumatology, Beijing Jishuitan Hospital Beijing ChinaLaboratory of Bone Tissue Engineering, Beijing Laboratory of Biomedical Materials Beijing Research Institute of Orthopaedics and Traumatology, Beijing Jishuitan Hospital Beijing ChinaLaboratory of Bone Tissue Engineering, Beijing Laboratory of Biomedical Materials Beijing Research Institute of Orthopaedics and Traumatology, Beijing Jishuitan Hospital Beijing ChinaGuangdong Provincial Key Laboratory of Orthopedics and Traumatology, Orthopaedic Research Institute/Department of Spinal Surgery The First Affiliated Hospital of Sun Yat‐sen University Guangzhou ChinaInnovation Platform of Regeneration and Repair of Spinal Cord and Nerve Injury, Department of Orthopaedic Surgery, The Seventh Affiliated Hospital Sun Yat‐sen University Shenzhen ChinaInnovation Platform of Regeneration and Repair of Spinal Cord and Nerve Injury, Department of Orthopaedic Surgery, The Seventh Affiliated Hospital Sun Yat‐sen University Shenzhen ChinaAbstract Background Ossification of the posterior longitudinal ligaments (OPLL) is common disorder characterized by heterotopic ossification of the spinal ligaments. Mechanical stimulation (MS) plays an important role in OPLL. DLX5 is an essential transcription factor required for osteoblast differentiation. However, the role of DLX5 during in OPLL is unclear. This study aims to investigate whether DLX5 is associated with OPLL progression under MS. Methods Stretch stimulation was applied to spinal ligaments cells derived from OPLL (OPLL cells) and non‐OPLL (non‐OPLL cells) patients. Expression of DLX5 and osteogenesis‐related genes were determined by quantitative real‐time polymerase chain reaction and Western blot. The osteogenic differentiation ability of the cells was measured using alkaline phosphatase (ALP) staining and alizarin red staining. The protein expression of DLX5 in the tissues and the nuclear translocation of NOTCH intracellular domain (NICD) was examined by immunofluorescence. Results Compared with non‐OPLL cells, OPLL cells expressed higher levels of DLX5 in vitro and vivo (p < 0.01). Upregulated expression of DLX5 and osteogenesis‐related genes (OSX, RUNX2, and OCN) were observed in OPLL cells induced with stretch stimulation and osteogenic medium, whereas there was no change in the non‐OPLL cells (p < 0.01). Cytoplasmic NICD protein translocated from the cytoplasm to the nucleus inducing DLX5 under stretch stimulation, which was reduced by the NOTCH signaling inhibitors (DAPT) (p < 0.01). Conclusions These data suggest that DLX5 play a critical role in MS‐induced progression of OPLL through NOTCH signaling, which provides a new insight into the pathogenesis of OPLL.https://doi.org/10.1002/jsp2.1247cyclic stretchDLX5mechanical stimulationNOTCH signalingOPLLosteogenic differentiation |
| spellingShingle | Tao Tang Zhengya Zhu Zhongyuan He Fuan Wang Hongkun Chen Shengkai Liu Mingbin Zhan Jianmin Wang Wei Tian Dafu Chen Xinbao Wu Xizhe Liu Zhiyu Zhou Shaoyu Liu DLX5 regulates the osteogenic differentiation of spinal ligaments cells derived from ossification of the posterior longitudinal ligament patients via NOTCH signaling JOR Spine cyclic stretch DLX5 mechanical stimulation NOTCH signaling OPLL osteogenic differentiation |
| title | DLX5 regulates the osteogenic differentiation of spinal ligaments cells derived from ossification of the posterior longitudinal ligament patients via NOTCH signaling |
| title_full | DLX5 regulates the osteogenic differentiation of spinal ligaments cells derived from ossification of the posterior longitudinal ligament patients via NOTCH signaling |
| title_fullStr | DLX5 regulates the osteogenic differentiation of spinal ligaments cells derived from ossification of the posterior longitudinal ligament patients via NOTCH signaling |
| title_full_unstemmed | DLX5 regulates the osteogenic differentiation of spinal ligaments cells derived from ossification of the posterior longitudinal ligament patients via NOTCH signaling |
| title_short | DLX5 regulates the osteogenic differentiation of spinal ligaments cells derived from ossification of the posterior longitudinal ligament patients via NOTCH signaling |
| title_sort | dlx5 regulates the osteogenic differentiation of spinal ligaments cells derived from ossification of the posterior longitudinal ligament patients via notch signaling |
| topic | cyclic stretch DLX5 mechanical stimulation NOTCH signaling OPLL osteogenic differentiation |
| url | https://doi.org/10.1002/jsp2.1247 |
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