Potential for Drug Repositioning of Midazolam as an Inhibitor of Inflammatory Bone Resorption
Abstract
:1. Introduction
2. Results
2.1. Effect of MDZ on the Differentiation of RAW264 Cells into Osteoclasts
2.2. Effect of MDZ on the Expression of Osteoclast Differentiation Marker Genes
2.3. Effect of MDZ on Osteoclast-Mediated Bone Resorption
2.4. Animal Experiments
2.4.1. Morphological Assessment of the Effect of MDZ on Lipopolysaccharide (LPS)-Induced Calvarial Mouse
2.4.2. Validation of Suture Width and Bone Matrix Volume on Bone Destruction in LPS-Induced Calvarial Mice
2.5. Elucidation of the Mechanism of MDZ-Induced Inhibition of Osteoclast Differentiation
3. Discussion
4. Materials and Methods
4.1. Biochemical and Cell Biology Experiments
4.1.1. Preparation of Mouse Macrophage-like Cell Line (RAW264 Cells)
4.1.2. TRAP Assay
4.1.3. TRAP Staining of RAW264 Cells
4.1.4. Pit Formation Assay
4.1.5. Quantitative Polymerase Chain Reaction Analysis
4.2. Animal Experiments
4.2.1. Establishment of Lipopolysaccharide-Induced Calvarial Model Mouse and TRAP Staining
4.2.2. Micro-Computed Tomography (Micro-CT) and Quantitative Analysis of Suture
4.3. Measurement of Reactive Oxygen Species
4.4. Western Blot Analysis
4.5. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
EC50 | Half-maximal (50%) effective concentration |
PBS | Phosphate-buffered saline |
mRNA | Messenger ribonucleic acid |
qPCR | Quantitative polymerase chain reaction |
WHO | World Health Organization |
NADPH | Nicotinamide adenine dinucleotide phosphate reduced form |
Na2HPO4 | Disodium hydrogen phosphate |
RIPA | Radioimmunoprecipitation assay |
HRP | Horseradish peroxidase |
ECL | Enhanced chemiluminescence |
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Harigaya, H.; Chiba-Ohkuma, R.; Karakida, T.; Yamamoto, R.; Fujii-Abe, K.; Kawahara, H.; Yamakoshi, Y. Potential for Drug Repositioning of Midazolam as an Inhibitor of Inflammatory Bone Resorption. Int. J. Mol. Sci. 2024, 25, 7651. https://doi.org/10.3390/ijms25147651
Harigaya H, Chiba-Ohkuma R, Karakida T, Yamamoto R, Fujii-Abe K, Kawahara H, Yamakoshi Y. Potential for Drug Repositioning of Midazolam as an Inhibitor of Inflammatory Bone Resorption. International Journal of Molecular Sciences. 2024; 25(14):7651. https://doi.org/10.3390/ijms25147651
Chicago/Turabian StyleHarigaya, Hiroko, Risako Chiba-Ohkuma, Takeo Karakida, Ryuji Yamamoto, Keiko Fujii-Abe, Hiroshi Kawahara, and Yasuo Yamakoshi. 2024. "Potential for Drug Repositioning of Midazolam as an Inhibitor of Inflammatory Bone Resorption" International Journal of Molecular Sciences 25, no. 14: 7651. https://doi.org/10.3390/ijms25147651