Investigation of MicroRNA Biomarkers in Equine Distal Interphalangeal Joint Osteoarthritis
Abstract
:1. Introduction
2. Results and Discussion
2.1. Samples
2.2. Small RNA Sequencing Analysis
2.3. Validation of Small RNA Sequencing by RT-qPCR
2.4. Gain and Loss Function Studies for miR-92a
Discussion
3. Methods and Materials
3.1. Sample Collection
3.2. Macroscopic and Microscopic Cartilage Assessment
3.3. Total RNA Extraction, Library Preparation and Small RNA Sequencing
3.4. Small RNA Sequencing Data Analysis
3.5. Small RNA Sequencing Validation
3.6. Primary Chondrocyte Collection, Isolation and Cell Culture
3.7. miRNA Cell Transfection
3.8. RNA Extraction, Reverse Transcription and RT-qPCR of Equine Chondrocytes
3.9. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Mild OA Group | Severe OA Group | |
---|---|---|
Sex | Three geldings Two mares | Six geldings Six mares |
Mean weight | 550 ± 70 kg | 440 ± 70 kg |
Mean age | 9 ± 3 years | 18 ± 9 years |
Breeds | Highland Pony, Dutch Warmblood, Belgian Warmblood and Thoroughbred | Irish Draught, Welsh Pony, Icelandic Pony, Irish Sports Horse, Cob, Cross-breed pony, Thoroughbred, Sports Horse and Connemara Pony |
Reasons for euthanasia | Colic, lameness, cervical vertebral malformation and one horse had a complete open third metacarpal bone fracture | Colic, lameness, incisional hernia, congestive heart failure, severe behavioural problems and advanced dental disease |
Horse Number | Breed | Age (Years) | Sex | Weight (kg) | Reason for Euthanasia | Group |
---|---|---|---|---|---|---|
H3 | Irish Draught | 28 | G | 500 kg | Lameness | Severe OA |
H16 | Cross-breed horse | 22 | G | 400 kg | Age-related concerns | Severe OA |
H17 | Cob-cross | 26 | G | 400 kg | Age-related concerns | Severe OA |
H26 | Irish Sports Horse | 5 | M | 500 kg | Multi-limb lameness | Mild OA |
H27 | Cob | 7 | M | 500 kg | Grass sickness | Mild OA |
H28 | Irish Draught | 12 | G | 600 kg | Lameness | Severe OA |
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Baker, M.E.; Lee, S.; Clinton, M.; Hackl, M.; Castanheira, C.; Peffers, M.J.; Taylor, S.E. Investigation of MicroRNA Biomarkers in Equine Distal Interphalangeal Joint Osteoarthritis. Int. J. Mol. Sci. 2022, 23, 15526. https://doi.org/10.3390/ijms232415526
Baker ME, Lee S, Clinton M, Hackl M, Castanheira C, Peffers MJ, Taylor SE. Investigation of MicroRNA Biomarkers in Equine Distal Interphalangeal Joint Osteoarthritis. International Journal of Molecular Sciences. 2022; 23(24):15526. https://doi.org/10.3390/ijms232415526
Chicago/Turabian StyleBaker, Melissa E., Seungmee Lee, Michael Clinton, Matthias Hackl, Catarina Castanheira, Mandy J. Peffers, and Sarah E. Taylor. 2022. "Investigation of MicroRNA Biomarkers in Equine Distal Interphalangeal Joint Osteoarthritis" International Journal of Molecular Sciences 23, no. 24: 15526. https://doi.org/10.3390/ijms232415526