miR-1323 suppresses bone mesenchymal stromal cell osteogenesis and fracture healing via inhibiting BMP4/SMAD4 signaling
Abstract Background Atrophic non-union fractures show no radiological evidence of callus formation within 3 months of fracture. microRNA dysregulation may underlie the dysfunctional osteogenesis in atrophic non-union fractures. Here, we aimed to analyze miR-1323 expression in human atrophic non-unio...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2020-06-01
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| Series: | Journal of Orthopaedic Surgery and Research |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13018-020-01685-8 |
| _version_ | 1811298070451716096 |
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| author | Hui Xie Ming Liu Yaofeng Jin Haiqing Lin Yushan Zhang Song Zheng |
| author_facet | Hui Xie Ming Liu Yaofeng Jin Haiqing Lin Yushan Zhang Song Zheng |
| author_sort | Hui Xie |
| collection | DOAJ |
| description | Abstract Background Atrophic non-union fractures show no radiological evidence of callus formation within 3 months of fracture. microRNA dysregulation may underlie the dysfunctional osteogenesis in atrophic non-union fractures. Here, we aimed to analyze miR-1323 expression in human atrophic non-union fractures and examine miR-1323’s underlying mechanism of action in human mesenchymal stromal cells. Methods Human atrophic non-union and standard healing fracture specimens were examined using H&E and Alcian Blue staining, immunohistochemistry, qRT-PCR, immunoblotting, and ALP activity assays. The effects of miR-1323 mimics or inhibition on BMP4, SMAD4, osteogenesis-related proteins, ALP activity, and bone mineralization were analyzed in human mesenchymal stromal cells. Luciferase reporter assays were utilized to assay miR-1323’s binding to the 3'UTRs of BMP4 and SMAD4. The effects of miR-1323, BMP4, and SMAD4 were analyzed by siRNA and overexpression vectors. A rat femur fracture model was established to analyze the in vivo effects of antagomiR-1323 treatment. Results miR-1323 was upregulated in human atrophic non-union fractures. Atrophic non-union was associated with downregulation of BMP4 and SMAD4 as well as the osteogenic markers ALP, collagen I, and RUNX2. In vitro, miR-1323 suppressed BMP4 and SMAD4 expression by binding to the 3'UTRs of BMP4 and SMAD4. Moreover, miR-1323’s inhibition of BMP4 and SMAD4 inhibited mesenchymal stromal cell osteogenic differentiation via modulating the nuclear translocation of the transcriptional co-activator TAZ. In vivo, antagomiR-1323 therapy facilitated the healing of fractures in a rat model of femoral fracture. Conclusions This evidence supports the miR-1323/BMP4 and miR-1323/SMAD4 axes as novel therapeutic targets for atrophic non-union fractures. |
| first_indexed | 2024-04-13T06:14:20Z |
| format | Article |
| id | doaj.art-43a4fc5717cd4b77a48681c7bcda1389 |
| institution | Directory Open Access Journal |
| issn | 1749-799X |
| language | English |
| last_indexed | 2024-04-13T06:14:20Z |
| publishDate | 2020-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Orthopaedic Surgery and Research |
| spelling | doaj.art-43a4fc5717cd4b77a48681c7bcda13892022-12-22T02:58:54ZengBMCJournal of Orthopaedic Surgery and Research1749-799X2020-06-0115111310.1186/s13018-020-01685-8miR-1323 suppresses bone mesenchymal stromal cell osteogenesis and fracture healing via inhibiting BMP4/SMAD4 signalingHui Xie0Ming Liu1Yaofeng Jin2Haiqing Lin3Yushan Zhang4Song Zheng5Department of Orthopedics, The Second Affiliated Hospital of Jiaxing UniversityDepartment of Orthopedics, The Second Affiliated Hospital of Jiaxing UniversityDepartment of Orthopedics, The Second Affiliated Hospital of Jiaxing UniversityDepartment of Orthopedics, The Second Affiliated Hospital of Jiaxing UniversityDepartment of Orthopedics, The Second Affiliated Hospital of Jiaxing UniversityDepartment of Orthopedics, The Second Affiliated Hospital of Jiaxing UniversityAbstract Background Atrophic non-union fractures show no radiological evidence of callus formation within 3 months of fracture. microRNA dysregulation may underlie the dysfunctional osteogenesis in atrophic non-union fractures. Here, we aimed to analyze miR-1323 expression in human atrophic non-union fractures and examine miR-1323’s underlying mechanism of action in human mesenchymal stromal cells. Methods Human atrophic non-union and standard healing fracture specimens were examined using H&E and Alcian Blue staining, immunohistochemistry, qRT-PCR, immunoblotting, and ALP activity assays. The effects of miR-1323 mimics or inhibition on BMP4, SMAD4, osteogenesis-related proteins, ALP activity, and bone mineralization were analyzed in human mesenchymal stromal cells. Luciferase reporter assays were utilized to assay miR-1323’s binding to the 3'UTRs of BMP4 and SMAD4. The effects of miR-1323, BMP4, and SMAD4 were analyzed by siRNA and overexpression vectors. A rat femur fracture model was established to analyze the in vivo effects of antagomiR-1323 treatment. Results miR-1323 was upregulated in human atrophic non-union fractures. Atrophic non-union was associated with downregulation of BMP4 and SMAD4 as well as the osteogenic markers ALP, collagen I, and RUNX2. In vitro, miR-1323 suppressed BMP4 and SMAD4 expression by binding to the 3'UTRs of BMP4 and SMAD4. Moreover, miR-1323’s inhibition of BMP4 and SMAD4 inhibited mesenchymal stromal cell osteogenic differentiation via modulating the nuclear translocation of the transcriptional co-activator TAZ. In vivo, antagomiR-1323 therapy facilitated the healing of fractures in a rat model of femoral fracture. Conclusions This evidence supports the miR-1323/BMP4 and miR-1323/SMAD4 axes as novel therapeutic targets for atrophic non-union fractures.http://link.springer.com/article/10.1186/s13018-020-01685-8FractureAtrophic non-unionBMP4SMAD4miR-1323 |
| spellingShingle | Hui Xie Ming Liu Yaofeng Jin Haiqing Lin Yushan Zhang Song Zheng miR-1323 suppresses bone mesenchymal stromal cell osteogenesis and fracture healing via inhibiting BMP4/SMAD4 signaling Journal of Orthopaedic Surgery and Research Fracture Atrophic non-union BMP4 SMAD4 miR-1323 |
| title | miR-1323 suppresses bone mesenchymal stromal cell osteogenesis and fracture healing via inhibiting BMP4/SMAD4 signaling |
| title_full | miR-1323 suppresses bone mesenchymal stromal cell osteogenesis and fracture healing via inhibiting BMP4/SMAD4 signaling |
| title_fullStr | miR-1323 suppresses bone mesenchymal stromal cell osteogenesis and fracture healing via inhibiting BMP4/SMAD4 signaling |
| title_full_unstemmed | miR-1323 suppresses bone mesenchymal stromal cell osteogenesis and fracture healing via inhibiting BMP4/SMAD4 signaling |
| title_short | miR-1323 suppresses bone mesenchymal stromal cell osteogenesis and fracture healing via inhibiting BMP4/SMAD4 signaling |
| title_sort | mir 1323 suppresses bone mesenchymal stromal cell osteogenesis and fracture healing via inhibiting bmp4 smad4 signaling |
| topic | Fracture Atrophic non-union BMP4 SMAD4 miR-1323 |
| url | http://link.springer.com/article/10.1186/s13018-020-01685-8 |
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