Proteomic analysis reveals dexamethasone rescues matrix breakdown but not anabolic dysregulation in a cartilage injury model
Summary: Objectives: In this exploratory study, we used discovery proteomics to follow the release of proteins from bovine knee articular cartilage in response to mechanical injury and cytokine treatment. We also studied the effect of the glucocorticoid dexamethasone (Dex) on these responses. Desig...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2020-12-01
|
| Series: | Osteoarthritis and Cartilage Open |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2665913120300935 |
| _version_ | 1819124368052060160 |
|---|---|
| author | Rebecca Mae Black Yang Wang André Struglics Pilar Lorenzo Viveka Tillgren Martin Rydén Alan J. Grodzinsky Patrik Önnerfjord |
| author_facet | Rebecca Mae Black Yang Wang André Struglics Pilar Lorenzo Viveka Tillgren Martin Rydén Alan J. Grodzinsky Patrik Önnerfjord |
| author_sort | Rebecca Mae Black |
| collection | DOAJ |
| description | Summary: Objectives: In this exploratory study, we used discovery proteomics to follow the release of proteins from bovine knee articular cartilage in response to mechanical injury and cytokine treatment. We also studied the effect of the glucocorticoid dexamethasone (Dex) on these responses. Design: Bovine cartilage explants were treated with either cytokines alone (10 ng/ml TNFα, 20 ng/ml IL-6, 100 ng/ml sIL-6R), a single compressive mechanical injury, cytokines and injury, or no treatment, and cultured in serum-free DMEM supplemented with 1% ITS for 22 days. All samples were incubated with or without addition of 100 nM Dex. Mass spectrometry and Western blot analyses were performed on medium samples for the identification and quantification of released proteins. Results: We identified 500 unique proteins present in all three biological replicates. Many proteins involved in the catabolic response of cartilage degradation had increased release after inflammatory stress. Dex rescued many of these catabolic effects. The release of some proteins involved in anabolic and chondroprotective processes was inconsistent, indicating differential effects on processes that may protect cartilage from injury. Dex restored only a small fraction of these to the control state, while others had their effects exacerbated by Dex exposure. Conclusions: We identified proteins that were released upon cytokine treatment which could be potential biomarkers of the inflammatory contribution to cartilage degradation. We also demonstrated the imperfect rescue of Dex on the effects of cartilage degradation, with many catabolic factors being reduced, while other anabolic or chondroprotective processes were not. |
| first_indexed | 2024-12-22T07:23:08Z |
| format | Article |
| id | doaj.art-15cb81ecb47d4e6cbfdb7673cf7b5853 |
| institution | Directory Open Access Journal |
| issn | 2665-9131 |
| language | English |
| last_indexed | 2024-12-22T07:23:08Z |
| publishDate | 2020-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Osteoarthritis and Cartilage Open |
| spelling | doaj.art-15cb81ecb47d4e6cbfdb7673cf7b58532022-12-21T18:34:13ZengElsevierOsteoarthritis and Cartilage Open2665-91312020-12-0124100099Proteomic analysis reveals dexamethasone rescues matrix breakdown but not anabolic dysregulation in a cartilage injury modelRebecca Mae Black0Yang Wang1André Struglics2Pilar Lorenzo3Viveka Tillgren4Martin Rydén5Alan J. Grodzinsky6Patrik Önnerfjord7Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USADepartment of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USAOrthopaedics, Department of Clinical Sciences Lund, Faculty of Medicine, Lund University, Lund, SwedenRheumatology and Molecular Skeletal Biology, Department of Clinical Sciences Lund, Faculty of Medicine, Lund University, Lund, SwedenRheumatology and Molecular Skeletal Biology, Department of Clinical Sciences Lund, Faculty of Medicine, Lund University, Lund, SwedenOrthopaedics, Department of Clinical Sciences Lund, Faculty of Medicine, Lund University, Lund, SwedenDepartment of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA; Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA; Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, MA, USARheumatology and Molecular Skeletal Biology, Department of Clinical Sciences Lund, Faculty of Medicine, Lund University, Lund, Sweden; Corresponding author. Rheumatology and Molecular Skeletal Biology, Department of Clinical Sciences, Lund University, BMC Plan C12, Lund, 22184, Sweden.Summary: Objectives: In this exploratory study, we used discovery proteomics to follow the release of proteins from bovine knee articular cartilage in response to mechanical injury and cytokine treatment. We also studied the effect of the glucocorticoid dexamethasone (Dex) on these responses. Design: Bovine cartilage explants were treated with either cytokines alone (10 ng/ml TNFα, 20 ng/ml IL-6, 100 ng/ml sIL-6R), a single compressive mechanical injury, cytokines and injury, or no treatment, and cultured in serum-free DMEM supplemented with 1% ITS for 22 days. All samples were incubated with or without addition of 100 nM Dex. Mass spectrometry and Western blot analyses were performed on medium samples for the identification and quantification of released proteins. Results: We identified 500 unique proteins present in all three biological replicates. Many proteins involved in the catabolic response of cartilage degradation had increased release after inflammatory stress. Dex rescued many of these catabolic effects. The release of some proteins involved in anabolic and chondroprotective processes was inconsistent, indicating differential effects on processes that may protect cartilage from injury. Dex restored only a small fraction of these to the control state, while others had their effects exacerbated by Dex exposure. Conclusions: We identified proteins that were released upon cytokine treatment which could be potential biomarkers of the inflammatory contribution to cartilage degradation. We also demonstrated the imperfect rescue of Dex on the effects of cartilage degradation, with many catabolic factors being reduced, while other anabolic or chondroprotective processes were not.http://www.sciencedirect.com/science/article/pii/S2665913120300935Post-traumatic osteoarthritisMass spectrometryCartilage matrixCytokinesProteomicsDexamethasone |
| spellingShingle | Rebecca Mae Black Yang Wang André Struglics Pilar Lorenzo Viveka Tillgren Martin Rydén Alan J. Grodzinsky Patrik Önnerfjord Proteomic analysis reveals dexamethasone rescues matrix breakdown but not anabolic dysregulation in a cartilage injury model Osteoarthritis and Cartilage Open Post-traumatic osteoarthritis Mass spectrometry Cartilage matrix Cytokines Proteomics Dexamethasone |
| title | Proteomic analysis reveals dexamethasone rescues matrix breakdown but not anabolic dysregulation in a cartilage injury model |
| title_full | Proteomic analysis reveals dexamethasone rescues matrix breakdown but not anabolic dysregulation in a cartilage injury model |
| title_fullStr | Proteomic analysis reveals dexamethasone rescues matrix breakdown but not anabolic dysregulation in a cartilage injury model |
| title_full_unstemmed | Proteomic analysis reveals dexamethasone rescues matrix breakdown but not anabolic dysregulation in a cartilage injury model |
| title_short | Proteomic analysis reveals dexamethasone rescues matrix breakdown but not anabolic dysregulation in a cartilage injury model |
| title_sort | proteomic analysis reveals dexamethasone rescues matrix breakdown but not anabolic dysregulation in a cartilage injury model |
| topic | Post-traumatic osteoarthritis Mass spectrometry Cartilage matrix Cytokines Proteomics Dexamethasone |
| url | http://www.sciencedirect.com/science/article/pii/S2665913120300935 |
| work_keys_str_mv | AT rebeccamaeblack proteomicanalysisrevealsdexamethasonerescuesmatrixbreakdownbutnotanabolicdysregulationinacartilageinjurymodel AT yangwang proteomicanalysisrevealsdexamethasonerescuesmatrixbreakdownbutnotanabolicdysregulationinacartilageinjurymodel AT andrestruglics proteomicanalysisrevealsdexamethasonerescuesmatrixbreakdownbutnotanabolicdysregulationinacartilageinjurymodel AT pilarlorenzo proteomicanalysisrevealsdexamethasonerescuesmatrixbreakdownbutnotanabolicdysregulationinacartilageinjurymodel AT vivekatillgren proteomicanalysisrevealsdexamethasonerescuesmatrixbreakdownbutnotanabolicdysregulationinacartilageinjurymodel AT martinryden proteomicanalysisrevealsdexamethasonerescuesmatrixbreakdownbutnotanabolicdysregulationinacartilageinjurymodel AT alanjgrodzinsky proteomicanalysisrevealsdexamethasonerescuesmatrixbreakdownbutnotanabolicdysregulationinacartilageinjurymodel AT patrikonnerfjord proteomicanalysisrevealsdexamethasonerescuesmatrixbreakdownbutnotanabolicdysregulationinacartilageinjurymodel |