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The Generation of Closed Femoral Fractures in Mice: A Model to Study Bone Healing
Published on: August 16, 2018
Altered expression of microRNA during fracture healing in diabetic rats
S Takahara1, S Y Lee2, T Iwakura1
1Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine, 650-0017 Kobe, Japan.
Objectives:
Diabetes mellitus (DM) is known to impair fracture healing. Increasing evidence suggests that some microRNA (miRNA) is involved in the pathophysiology of diabetes and its complications. We hypothesized that the functions of miRNA and changes to their patterns of expression may be implicated in the pathogenesis of impaired fracture healing in DM.
Methods:
Closed transverse fractures were created in the femurs of 116 rats, with half assigned to the DM group and half assigned to the control group. Rats with DM were induced by a single intraperitoneal injection of streptozotocin. At post-fracture days five, seven, 11, 14, 21, and 28, miRNA was extracted from the newly generated tissue at the fracture site. Microarray analysis was performed with miRNA samples from each group on post-fracture days five and 11. For further analysis, real-time polymerase chain reaction (PCR) analysis was performed at each timepoint.
Results:
Microarray analysis showed that there were 14 miRNAs at day five and 17 miRNAs at day 11, with a greater than twofold change in the DM group compared with the control group. Among these types of miRNA, five were selected based on a comparative and extended literature review. Real-time PCR analysis revealed that five types of miRNA (miR-140-3p, miR-140-5p, miR-181a-1-3p, miR-210-3p, and miR-222-3p) were differentially expressed with changing patterns of expression during fracture healing in diabetic rats compared with controls.
Conclusions:
Our findings provide information to further understand the pathology of impaired fracture healing in a diabetic rat model. These results may allow the potential development of molecular therapy using miRNA for the treatment of impaired fracture healing in patients with DM.Cite this article: S. Takahara, S. Y. Lee, T. Iwakura, K. Oe, T. Fukui, E. Okumachi, T. Waki, M. Arakura, Y. Sakai, K. Nishida, R. Kuroda, T. Niikura. Altered expression of microRNA during fracture healing in diabetic rats. Bone Joint Res 2018;7:139-147. DOI: 10.1302/2046-3758.72.BJR-2017-0082.R1.
Insights
Diabetic rats show altered microRNA (miRNA) expression patterns during fracture healing. These findings may lead to new molecular therapies for impaired bone healing in diabetes mellitus.
Area of Science:
- Biomedical Science
- Molecular Biology
- Orthopedics
Background:
- Diabetes mellitus (DM) is known to impair fracture healing.
- MicroRNAs (miRNAs) are increasingly implicated in diabetes pathophysiology and complications.
- The role of miRNA in impaired fracture healing in DM requires further investigation.
Purpose of the Study:
- To investigate the role of microRNA (miRNA) in impaired fracture healing in a diabetic rat model.
- To identify specific miRNAs and their expression patterns during the fracture healing process in diabetic conditions.
Main Methods:
- A diabetic rat model was established using streptozotocin.
- Closed transverse femur fractures were created in diabetic and control rats.
- miRNA was extracted from fracture sites at multiple time points and analyzed using microarray and real-time PCR.
Main Results:
- Microarray analysis identified differentially expressed miRNAs at days five and 11 post-fracture in diabetic rats.
- Real-time PCR confirmed differential expression of five specific miRNAs (miR-140-3p, miR-140-5p, miR-181a-1-3p, miR-210-3p, miR-222-3p).
- These miRNAs exhibited altered expression patterns throughout the fracture healing process in diabetic rats compared to controls.
Conclusions:
- The study identifies specific miRNAs with altered expression during fracture healing in diabetic rats.
- These findings contribute to understanding the molecular mechanisms of impaired fracture healing in DM.
- The identified miRNAs may serve as potential targets for novel molecular therapies to treat impaired fracture healing in diabetic patients.
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