Human amniotic membrane modulates Wnt/β-catenin and NF-κβ signaling pathways in articular chondrocytes in vitro
Objective: Inflammation, catabolism, and hypertrophy in chondrocytes play a central role in osteoarthritis (OA). The Wnt/β-catenin and NF-κβ pathways contribute to these degradative processes. This in vitro study evaluates the inhibitory effect of a novel therapeutic, micronized dehydrated human amn...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-12-01
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| Series: | Osteoarthritis and Cartilage Open |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2665913121000741 |
| _version_ | 1819099649327235072 |
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| author | Connie Chung Michelle Massee Thomas J. Koob |
| author_facet | Connie Chung Michelle Massee Thomas J. Koob |
| author_sort | Connie Chung |
| collection | DOAJ |
| description | Objective: Inflammation, catabolism, and hypertrophy in chondrocytes play a central role in osteoarthritis (OA). The Wnt/β-catenin and NF-κβ pathways contribute to these degradative processes. This in vitro study evaluates the inhibitory effect of a novel therapeutic, micronized dehydrated human amnion/chorion membrane (μdHACM), as a potential treatment to offset elevated Wnt/β-catenin and NF-κβ signaling. Design: Three-dimensional human articular chondrocyte pellets were stimulated with an inflammatory cocktail to induce a degenerative phenotype. Treatments included varying doses of μdHACM. Protein and gene expression were analyzed using qRT-PCR, immunoblotting, and immunofluorescence to assess changes in the major constituents of Wnt/β-catenin and NF-κβ signaling. Regulation of catabolic activity was evaluated using enzymatic assays that detect MMP-13 and aggrecanase-mediated degradation products in conditioned media. Results: Confirmation of the model was established through the expression of specific markers and extracellular matrix genes, verifying a chondrogenic phenotype was maintained. Inflammatory stimulation elicited a change in the chondrocyte proteome and secretome, elevating Wnt/β-catenin and NF-κβ signaling and downstream expression of inflammatory, proteolytic, and hypertrophic markers, while decreasing primary cartilage matrix components, ACAN and COL2A1. μdHACM reversed these inflammatory-induced changes, suppressing phospho-GSK-3β, β-catenin expression/nuclear localization of the Wnt signaling axis and inhibiting IKKβ, phospho-IκBα, and phospho-p65 in the NF-κβ signaling cascade. Additionally, μdHACM altered expression of direct downstream targets, namely MCP1, MMP3, MMP13, ADAMTS4, ADAMTS5, RUNX2 and COL10A1. Moreover, μdHACM reduced MMP-13 and aggrecanase-mediated substrate degradation. Conclusion: μdHACM ameloriated the effects of inflammatory-induced degeneration in chondrocytes through Wnt/β-catenin and NF-κβ inhibition, subsequently downregulating key inflammatory, hypertrophic and catabolic mediators in vitro. |
| first_indexed | 2024-12-22T00:50:14Z |
| format | Article |
| id | doaj.art-bb9cf1376908415f951568a77c7d3c87 |
| institution | Directory Open Access Journal |
| issn | 2665-9131 |
| language | English |
| last_indexed | 2024-12-22T00:50:14Z |
| publishDate | 2021-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Osteoarthritis and Cartilage Open |
| spelling | doaj.art-bb9cf1376908415f951568a77c7d3c872022-12-21T18:44:28ZengElsevierOsteoarthritis and Cartilage Open2665-91312021-12-0134100211Human amniotic membrane modulates Wnt/β-catenin and NF-κβ signaling pathways in articular chondrocytes in vitroConnie Chung0Michelle Massee1Thomas J. Koob2MiMedx Group, Inc. 1775 West Oak Commons Court NE, Marietta, GA, 30062, USACorresponding author.; MiMedx Group, Inc. 1775 West Oak Commons Court NE, Marietta, GA, 30062, USAMiMedx Group, Inc. 1775 West Oak Commons Court NE, Marietta, GA, 30062, USAObjective: Inflammation, catabolism, and hypertrophy in chondrocytes play a central role in osteoarthritis (OA). The Wnt/β-catenin and NF-κβ pathways contribute to these degradative processes. This in vitro study evaluates the inhibitory effect of a novel therapeutic, micronized dehydrated human amnion/chorion membrane (μdHACM), as a potential treatment to offset elevated Wnt/β-catenin and NF-κβ signaling. Design: Three-dimensional human articular chondrocyte pellets were stimulated with an inflammatory cocktail to induce a degenerative phenotype. Treatments included varying doses of μdHACM. Protein and gene expression were analyzed using qRT-PCR, immunoblotting, and immunofluorescence to assess changes in the major constituents of Wnt/β-catenin and NF-κβ signaling. Regulation of catabolic activity was evaluated using enzymatic assays that detect MMP-13 and aggrecanase-mediated degradation products in conditioned media. Results: Confirmation of the model was established through the expression of specific markers and extracellular matrix genes, verifying a chondrogenic phenotype was maintained. Inflammatory stimulation elicited a change in the chondrocyte proteome and secretome, elevating Wnt/β-catenin and NF-κβ signaling and downstream expression of inflammatory, proteolytic, and hypertrophic markers, while decreasing primary cartilage matrix components, ACAN and COL2A1. μdHACM reversed these inflammatory-induced changes, suppressing phospho-GSK-3β, β-catenin expression/nuclear localization of the Wnt signaling axis and inhibiting IKKβ, phospho-IκBα, and phospho-p65 in the NF-κβ signaling cascade. Additionally, μdHACM altered expression of direct downstream targets, namely MCP1, MMP3, MMP13, ADAMTS4, ADAMTS5, RUNX2 and COL10A1. Moreover, μdHACM reduced MMP-13 and aggrecanase-mediated substrate degradation. Conclusion: μdHACM ameloriated the effects of inflammatory-induced degeneration in chondrocytes through Wnt/β-catenin and NF-κβ inhibition, subsequently downregulating key inflammatory, hypertrophic and catabolic mediators in vitro.http://www.sciencedirect.com/science/article/pii/S2665913121000741WntNF-κβOsteoarthritisChondrocyteAmniotic membraneECM catabolism |
| spellingShingle | Connie Chung Michelle Massee Thomas J. Koob Human amniotic membrane modulates Wnt/β-catenin and NF-κβ signaling pathways in articular chondrocytes in vitro Osteoarthritis and Cartilage Open Wnt NF-κβ Osteoarthritis Chondrocyte Amniotic membrane ECM catabolism |
| title | Human amniotic membrane modulates Wnt/β-catenin and NF-κβ signaling pathways in articular chondrocytes in vitro |
| title_full | Human amniotic membrane modulates Wnt/β-catenin and NF-κβ signaling pathways in articular chondrocytes in vitro |
| title_fullStr | Human amniotic membrane modulates Wnt/β-catenin and NF-κβ signaling pathways in articular chondrocytes in vitro |
| title_full_unstemmed | Human amniotic membrane modulates Wnt/β-catenin and NF-κβ signaling pathways in articular chondrocytes in vitro |
| title_short | Human amniotic membrane modulates Wnt/β-catenin and NF-κβ signaling pathways in articular chondrocytes in vitro |
| title_sort | human amniotic membrane modulates wnt β catenin and nf κβ signaling pathways in articular chondrocytes in vitro |
| topic | Wnt NF-κβ Osteoarthritis Chondrocyte Amniotic membrane ECM catabolism |
| url | http://www.sciencedirect.com/science/article/pii/S2665913121000741 |
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