Anabolic role of lysyl oxidase like-2 in cartilage of knee and temporomandibular joints with osteoarthritis
Abstract Background Lysyl oxidase like-2 (LOXL2) is a copper-dependent amine oxidase. Our previous studies showed that LOXL2 is elevated during mouse fracture healing. The goal of this study was to evaluate the potential of LOXL2 to act as an anabolic agent in cartilage affected by osteoarthritis (O...
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| Format: | Article |
| Language: | English |
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BMC
2017-08-01
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| Series: | Arthritis Research & Therapy |
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| Online Access: | http://link.springer.com/article/10.1186/s13075-017-1388-8 |
| _version_ | 1818906404443914240 |
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| author | Weam Alshenibr Mustafa M. Tashkandi Saqer F. Alsaqer Yazeed Alkheriji Amelia Wise Sadanand Fulzele Pushkar Mehra Mary B. Goldring Louis C. Gerstenfeld Manish V. Bais |
| author_facet | Weam Alshenibr Mustafa M. Tashkandi Saqer F. Alsaqer Yazeed Alkheriji Amelia Wise Sadanand Fulzele Pushkar Mehra Mary B. Goldring Louis C. Gerstenfeld Manish V. Bais |
| author_sort | Weam Alshenibr |
| collection | DOAJ |
| description | Abstract Background Lysyl oxidase like-2 (LOXL2) is a copper-dependent amine oxidase. Our previous studies showed that LOXL2 is elevated during mouse fracture healing. The goal of this study was to evaluate the potential of LOXL2 to act as an anabolic agent in cartilage affected by osteoarthritis (OA). Methods LOXL2 was visualized in tissues from human knee and hip joints and temporomandibular joints (TMJ) by immunofluorescence. The activity of LOXL2 in human articular and TMJ chondrocytes was assessed by cell-based assays, microarray analysis, and RT-qPCR, and LOXL2-mediated activation of NF-κB and extracellular signal-related kinase (ERK) signaling pathways was measured by western blotting. To examine LOXL2-induced effect in vivo, we implanted Matrigel-imbedded human chondrocytes into nude mice and exposed them to exogenous LOXL2 for 6 weeks. Finally, LOXL2-induced effects on collagen type 2 α1 (COL2A1) and phospho-SMAD2/3 were evaluated by immunofluorescence analysis. Results LOXL2 staining was detected in damaged regions of human TMJ, hip and knee joints affected by OA. Stimulation with transforming growth factor (TGF)-β1 upregulated LOXL2 expression, while pro-inflammatory cytokines IL-1β and TNF-α downregulated LOXL2, in human chondrocytes. Viral transduction of LOXL2 in OA chondrocytes increased the mRNA levels of chondroitin sulfate proteoglycan (CSPG4), aggrecan (ACAN), sex determining region Y-box containing gene 9 (SOX9), and COL2A1 but reduced the levels of extracellular matrix (ECM)-degrading enzymes matrix metalloproteinase (MMP)1, MMP3, and MMP13. Further, forced expression of LOXL2 promoted chondrogenic lineage-specific gene expression, increased the expression of COL2A1 in the presence of TNF-α, and inhibited chondrocyte apoptosis. LOXL2 expression also inhibited IL-1β-induced phospho-NF-κB/p65 and TGF-β1-induced ERK1/2 phosphorylation. Matrigel constructs of human chondrocytes from the knee joint and TMJ implanted in nude mice showed anabolic responses after LOXL2 transduction, including increased expression of SOX9, ACAN, and COL2A1. Finally, immunofluorescence staining revealed co-localization of LOXL2 with SOX9 in the nuclei of cells in the implants, decreased phospho-SMAD2/3, and increased COL2A1 staining. Conclusion Our results suggest that although LOXL2 is upregulated in cartilage affected by OA, this may be a protective response that promotes anabolism while inhibiting specific catabolic responses in the pathophysiology of OA. |
| first_indexed | 2024-12-19T21:38:41Z |
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| language | English |
| last_indexed | 2024-12-19T21:38:41Z |
| publishDate | 2017-08-01 |
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| series | Arthritis Research & Therapy |
| spelling | doaj.art-06803ddf1f124c34beddad45be728b082022-12-21T20:04:43ZengBMCArthritis Research & Therapy1478-63622017-08-0119111510.1186/s13075-017-1388-8Anabolic role of lysyl oxidase like-2 in cartilage of knee and temporomandibular joints with osteoarthritisWeam Alshenibr0Mustafa M. Tashkandi1Saqer F. Alsaqer2Yazeed Alkheriji3Amelia Wise4Sadanand Fulzele5Pushkar Mehra6Mary B. Goldring7Louis C. Gerstenfeld8Manish V. Bais9Department of Molecular and Cell Biology, Boston University Henry M. Goldman School of Dental MedicineDepartment of Molecular and Cell Biology, Boston University Henry M. Goldman School of Dental MedicineDepartment of Molecular and Cell Biology, Boston University Henry M. Goldman School of Dental MedicineDepartment of Molecular and Cell Biology, Boston University Henry M. Goldman School of Dental MedicineDepartment of Orthopaedic Surgery, School of Medicine, Boston UniversityDepartment of Orthopaedic Surgery and Institute of Regenerative and Reparative Medicine, Georgia Regents UniversityDepartment of Oral and Maxillofacial Surgery, Boston University Henry M. Goldman School of Dental MedicineHospital for Special Surgery Research Institute, Weill Cornell Medical CollegeDepartment of Orthopaedic Surgery, School of Medicine, Boston UniversityDepartment of Molecular and Cell Biology, Boston University Henry M. Goldman School of Dental MedicineAbstract Background Lysyl oxidase like-2 (LOXL2) is a copper-dependent amine oxidase. Our previous studies showed that LOXL2 is elevated during mouse fracture healing. The goal of this study was to evaluate the potential of LOXL2 to act as an anabolic agent in cartilage affected by osteoarthritis (OA). Methods LOXL2 was visualized in tissues from human knee and hip joints and temporomandibular joints (TMJ) by immunofluorescence. The activity of LOXL2 in human articular and TMJ chondrocytes was assessed by cell-based assays, microarray analysis, and RT-qPCR, and LOXL2-mediated activation of NF-κB and extracellular signal-related kinase (ERK) signaling pathways was measured by western blotting. To examine LOXL2-induced effect in vivo, we implanted Matrigel-imbedded human chondrocytes into nude mice and exposed them to exogenous LOXL2 for 6 weeks. Finally, LOXL2-induced effects on collagen type 2 α1 (COL2A1) and phospho-SMAD2/3 were evaluated by immunofluorescence analysis. Results LOXL2 staining was detected in damaged regions of human TMJ, hip and knee joints affected by OA. Stimulation with transforming growth factor (TGF)-β1 upregulated LOXL2 expression, while pro-inflammatory cytokines IL-1β and TNF-α downregulated LOXL2, in human chondrocytes. Viral transduction of LOXL2 in OA chondrocytes increased the mRNA levels of chondroitin sulfate proteoglycan (CSPG4), aggrecan (ACAN), sex determining region Y-box containing gene 9 (SOX9), and COL2A1 but reduced the levels of extracellular matrix (ECM)-degrading enzymes matrix metalloproteinase (MMP)1, MMP3, and MMP13. Further, forced expression of LOXL2 promoted chondrogenic lineage-specific gene expression, increased the expression of COL2A1 in the presence of TNF-α, and inhibited chondrocyte apoptosis. LOXL2 expression also inhibited IL-1β-induced phospho-NF-κB/p65 and TGF-β1-induced ERK1/2 phosphorylation. Matrigel constructs of human chondrocytes from the knee joint and TMJ implanted in nude mice showed anabolic responses after LOXL2 transduction, including increased expression of SOX9, ACAN, and COL2A1. Finally, immunofluorescence staining revealed co-localization of LOXL2 with SOX9 in the nuclei of cells in the implants, decreased phospho-SMAD2/3, and increased COL2A1 staining. Conclusion Our results suggest that although LOXL2 is upregulated in cartilage affected by OA, this may be a protective response that promotes anabolism while inhibiting specific catabolic responses in the pathophysiology of OA.http://link.springer.com/article/10.1186/s13075-017-1388-8LOXL2Anabolic responseCartilageOsteoarthritis |
| spellingShingle | Weam Alshenibr Mustafa M. Tashkandi Saqer F. Alsaqer Yazeed Alkheriji Amelia Wise Sadanand Fulzele Pushkar Mehra Mary B. Goldring Louis C. Gerstenfeld Manish V. Bais Anabolic role of lysyl oxidase like-2 in cartilage of knee and temporomandibular joints with osteoarthritis Arthritis Research & Therapy LOXL2 Anabolic response Cartilage Osteoarthritis |
| title | Anabolic role of lysyl oxidase like-2 in cartilage of knee and temporomandibular joints with osteoarthritis |
| title_full | Anabolic role of lysyl oxidase like-2 in cartilage of knee and temporomandibular joints with osteoarthritis |
| title_fullStr | Anabolic role of lysyl oxidase like-2 in cartilage of knee and temporomandibular joints with osteoarthritis |
| title_full_unstemmed | Anabolic role of lysyl oxidase like-2 in cartilage of knee and temporomandibular joints with osteoarthritis |
| title_short | Anabolic role of lysyl oxidase like-2 in cartilage of knee and temporomandibular joints with osteoarthritis |
| title_sort | anabolic role of lysyl oxidase like 2 in cartilage of knee and temporomandibular joints with osteoarthritis |
| topic | LOXL2 Anabolic response Cartilage Osteoarthritis |
| url | http://link.springer.com/article/10.1186/s13075-017-1388-8 |
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