This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Research article
Dongxu Liu
Shandong University School of Medicine: Shandong University Cheeloo College of Medicine
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
Biomechanical forces are vital for the regulation of skeletal tissue. Mechanical stretch plays a vital role in osteogenic differentiation of periodontal ligament stem cells (PDLSCs) during orthodontic treatment. Cyclic mechanical stretch may trigger the up-regulated production of reactive oxygen species (ROS). ROS has a critical effect on bone cell function and the pathophysiology of bone diseases. N-acetylcysteine (NAC), a ROS scavenger, possesses powerful antioxidant capacity. The aim of this study was to determine the role of ROS and NAC in PDLSCs during osteogenic differentiation under cyclic mechanical stretch. We further investigated that the therapeutic potential of NAC to improve the changes of the microstructure of alveolar bone during orthodontic tooth movement in rats by micro-CT system.
Methods
The expression of COL1 (collagen type I), RUNX2 (runt-related transcription factor 2) and OPN (osteopontin) by qRT-PCR and Western blot experiments, and alkaline phosphatase (ALP) staining as well as ALP activity tests were used to examine osteogenic differentiation tendency of PDLSCs subjected to cyclic mechanical stretch of 10% and 0.5Hz deformation induced by the Flexcell tension system. ROS production in PDLSCs were measured under cyclic mechanical stretch by Flow Cytometry. The levels of reduced glutathione (GSH), oxidized GSH (GSSG) and the GSH/GSSG ratio with or without NAC treatment were analyzed. And we evaluated the changes of the microstructure of alveolar bone during orthodontic tooth movement in rats employing micro-CT system.
Results
NAC treatment could promote the osteogenic differentiation of PDLSCs under cyclic mechanical stretch. Down-regulated ROS generation and the up-regulated level of GSH and the ratio of GSH/GSSG in PDLSCs treated with NAC were observed in response to cyclic mechanical stretch. NAC improved the microstructure of alveolar bone, including BV/TV (bone volume/total volume), Tb.Th (trabecular thickness), Tb.Sp (trabecular separation) and SMI (microstructure model index), during orthodontic tooth movement in rats.
Conclusion
These results revealed that NAC might be a potential therapeutic approach for the remodeling of the alveolar bone during orthodontic tooth movement.
Keywords
Cyclic mechanical stretch, Periodontal ligament stem cells, Osteogenic differentiation, Reactive oxygen species, N-acetylcysteine, Orthodontic tooth movement
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
This is a list of supplementary files associated with this preprint. Click to download.
- Table1NAC.xls
Table 1 The primer sequences used in this study
- Additionalfile1FigureS1..tif
Additional file 1: Figure S1. The direction of orthodontic tooth movement and ROI. The arrow showed the direction of orthodontic tooth movement. The red area showed the ROI for the alveolar bone microstructural parameter analysis at the tension side. (.tif)
- Additionalfile2TableS1.xls
Additional file 2: Table S1. The effect of NAC on microstructure of alveolar bone during orthodontic tooth movement in rats. Data are expressed as means ± standard deviation, n = 5 specimens/group. *P < 0.05 vs. the control group, respectively. (.xls)
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This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Research article
Dongxu Liu
Shandong University School of Medicine: Shandong University Cheeloo College of Medicine
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 22 Mar, 2021
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Orthopedic Surgery
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
Biomechanical forces are vital for the regulation of skeletal tissue. Mechanical stretch plays a vital role in osteogenic differentiation of periodontal ligament stem cells (PDLSCs) during orthodontic treatment. Cyclic mechanical stretch may trigger the up-regulated production of reactive oxygen species (ROS). ROS has a critical effect on bone cell function and the pathophysiology of bone diseases. N-acetylcysteine (NAC), a ROS scavenger, possesses powerful antioxidant capacity. The aim of this study was to determine the role of ROS and NAC in PDLSCs during osteogenic differentiation under cyclic mechanical stretch. We further investigated that the therapeutic potential of NAC to improve the changes of the microstructure of alveolar bone during orthodontic tooth movement in rats by micro-CT system.
Methods
The expression of COL1 (collagen type I), RUNX2 (runt-related transcription factor 2) and OPN (osteopontin) by qRT-PCR and Western blot experiments, and alkaline phosphatase (ALP) staining as well as ALP activity tests were used to examine osteogenic differentiation tendency of PDLSCs subjected to cyclic mechanical stretch of 10% and 0.5Hz deformation induced by the Flexcell tension system. ROS production in PDLSCs were measured under cyclic mechanical stretch by Flow Cytometry. The levels of reduced glutathione (GSH), oxidized GSH (GSSG) and the GSH/GSSG ratio with or without NAC treatment were analyzed. And we evaluated the changes of the microstructure of alveolar bone during orthodontic tooth movement in rats employing micro-CT system.
Results
NAC treatment could promote the osteogenic differentiation of PDLSCs under cyclic mechanical stretch. Down-regulated ROS generation and the up-regulated level of GSH and the ratio of GSH/GSSG in PDLSCs treated with NAC were observed in response to cyclic mechanical stretch. NAC improved the microstructure of alveolar bone, including BV/TV (bone volume/total volume), Tb.Th (trabecular thickness), Tb.Sp (trabecular separation) and SMI (microstructure model index), during orthodontic tooth movement in rats.
Conclusion
These results revealed that NAC might be a potential therapeutic approach for the remodeling of the alveolar bone during orthodontic tooth movement.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
This is a list of supplementary files associated with this preprint. Click to download.
- Table1NAC.xls
Table 1 The primer sequences used in this study
- Additionalfile1FigureS1..tif
Additional file 1: Figure S1. The direction of orthodontic tooth movement and ROI. The arrow showed the direction of orthodontic tooth movement. The red area showed the ROI for the alveolar bone microstructural parameter analysis at the tension side. (.tif)
- Additionalfile2TableS1.xls
Additional file 2: Table S1. The effect of NAC on microstructure of alveolar bone during orthodontic tooth movement in rats. Data are expressed as means ± standard deviation, n = 5 specimens/group. *P < 0.05 vs. the control group, respectively. (.xls)
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