Synovial Fibroblast Extracellular Vesicles Induce Inflammation via Delivering miR-21-5p in Osteoarthritis

Small extracellular vesicles (sEVs) derived from different osteoarthritic (OA) tissues regulate OA-related biological processes through transporting their content (proteins, miRNAs, etc.) to recipient cells. This study aimed to characterize the miRNA profile of synovial fibroblasts-derived small EVs...

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Main Authors: Vasileios Konteles, Ioanna Papathanasiou, Maria Tzetis, Anastasios Kriebardis, Aspasia Tsezou
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Cells
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Online Access:https://www.mdpi.com/2073-4409/14/7/519
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author Vasileios Konteles
Ioanna Papathanasiou
Maria Tzetis
Anastasios Kriebardis
Aspasia Tsezou
author_facet Vasileios Konteles
Ioanna Papathanasiou
Maria Tzetis
Anastasios Kriebardis
Aspasia Tsezou
author_sort Vasileios Konteles
collection DOAJ
description Small extracellular vesicles (sEVs) derived from different osteoarthritic (OA) tissues regulate OA-related biological processes through transporting their content (proteins, miRNAs, etc.) to recipient cells. This study aimed to characterize the miRNA profile of synovial fibroblasts-derived small EVs (FS_OA_sEVs) and investigate their role in inflammation in chondrocytes. Chondrocytes were isolated from macroscopically preserved and lesioned OA cartilage (C_OAmin and C_OAmax) and synovial fibroblasts from OA synovium. Synovial fibroblasts-derived small EVs (FS_OA_sEVs) were characterized according to ISEV guidelines and used for miRNA profiling and bioinformatics analysis. miR-21-5p was identified as one of the most abundant, and its target genes, such as KLF6, were enriched in OA-related processes including inflammation. Treatment of C_OAmin chondrocytes with FS_OA_sEVs resulted in decreased expression of COL2A1 and ACAN and an increase in catabolic markers MMP-3 and MMP-13. Moreover, C-OAmin receiving FS_OA_sEVs exhibited increased levels of inflammatory markers and miR-21-5p expression, resembling chondrocytes’ phenotype from lesioned OA cartilage, whereas miR-21-5p inhibition reversed their expression of inflammatory markers and miR-21-5p. Compared to C_OA min, C_OAmax chondrocytes exhibited increased miR-21-5p and inflammatory markers expression and decreased KLF6 expression. miR-21-5p inhibition in C_OAmax led to KLF6 upregulation and suppression of inflammatory mediators, whereas co-treatment with siRNA against KLF6 negated this effect, confirming a potential direct regulatory relationship between miR-21-5p and KLF6. Our results provide novel insights into the FS_OA_sEV-mediated inflammation axis, highlighting FS_OA_sEV-derived miR-21-5p as a driver of OA progression via regulating inflammation in chondrocytes.
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spelling doaj-art-9e46c8e24d0c4a5686c712f239d0bcd92025-08-20T03:08:46ZengMDPI AGCells2073-44092025-03-0114751910.3390/cells14070519Synovial Fibroblast Extracellular Vesicles Induce Inflammation via Delivering miR-21-5p in OsteoarthritisVasileios Konteles0Ioanna Papathanasiou1Maria Tzetis2Anastasios Kriebardis3Aspasia Tsezou4Laboratory of Cytogenetics and Molecular Genetics, Faculty of Medicine, University of Thessaly, 41222 Larissa, GreeceLaboratory of Cytogenetics and Molecular Genetics, Faculty of Medicine, University of Thessaly, 41222 Larissa, GreeceChoremion Research Laboratory, Department of Medical Genetics, National and Kapodistrian University of Athens, 11527 Athens, GreeceLaboratory of Reliability and Quality Control in Laboratory Hematology (HemQcR), Department of Biomedical Sciences, School of Health & Caring Sciences, University of West Attica (UniWA), 12243 Egaleo, GreeceLaboratory of Cytogenetics and Molecular Genetics, Faculty of Medicine, University of Thessaly, 41222 Larissa, GreeceSmall extracellular vesicles (sEVs) derived from different osteoarthritic (OA) tissues regulate OA-related biological processes through transporting their content (proteins, miRNAs, etc.) to recipient cells. This study aimed to characterize the miRNA profile of synovial fibroblasts-derived small EVs (FS_OA_sEVs) and investigate their role in inflammation in chondrocytes. Chondrocytes were isolated from macroscopically preserved and lesioned OA cartilage (C_OAmin and C_OAmax) and synovial fibroblasts from OA synovium. Synovial fibroblasts-derived small EVs (FS_OA_sEVs) were characterized according to ISEV guidelines and used for miRNA profiling and bioinformatics analysis. miR-21-5p was identified as one of the most abundant, and its target genes, such as KLF6, were enriched in OA-related processes including inflammation. Treatment of C_OAmin chondrocytes with FS_OA_sEVs resulted in decreased expression of COL2A1 and ACAN and an increase in catabolic markers MMP-3 and MMP-13. Moreover, C-OAmin receiving FS_OA_sEVs exhibited increased levels of inflammatory markers and miR-21-5p expression, resembling chondrocytes’ phenotype from lesioned OA cartilage, whereas miR-21-5p inhibition reversed their expression of inflammatory markers and miR-21-5p. Compared to C_OA min, C_OAmax chondrocytes exhibited increased miR-21-5p and inflammatory markers expression and decreased KLF6 expression. miR-21-5p inhibition in C_OAmax led to KLF6 upregulation and suppression of inflammatory mediators, whereas co-treatment with siRNA against KLF6 negated this effect, confirming a potential direct regulatory relationship between miR-21-5p and KLF6. Our results provide novel insights into the FS_OA_sEV-mediated inflammation axis, highlighting FS_OA_sEV-derived miR-21-5p as a driver of OA progression via regulating inflammation in chondrocytes.https://www.mdpi.com/2073-4409/14/7/519extracellular vesiclesosteoarthritismiR-21-5pinflammation
spellingShingle Vasileios Konteles
Ioanna Papathanasiou
Maria Tzetis
Anastasios Kriebardis
Aspasia Tsezou
Synovial Fibroblast Extracellular Vesicles Induce Inflammation via Delivering miR-21-5p in Osteoarthritis
Cells
extracellular vesicles
osteoarthritis
miR-21-5p
inflammation
title Synovial Fibroblast Extracellular Vesicles Induce Inflammation via Delivering miR-21-5p in Osteoarthritis
title_full Synovial Fibroblast Extracellular Vesicles Induce Inflammation via Delivering miR-21-5p in Osteoarthritis
title_fullStr Synovial Fibroblast Extracellular Vesicles Induce Inflammation via Delivering miR-21-5p in Osteoarthritis
title_full_unstemmed Synovial Fibroblast Extracellular Vesicles Induce Inflammation via Delivering miR-21-5p in Osteoarthritis
title_short Synovial Fibroblast Extracellular Vesicles Induce Inflammation via Delivering miR-21-5p in Osteoarthritis
title_sort synovial fibroblast extracellular vesicles induce inflammation via delivering mir 21 5p in osteoarthritis
topic extracellular vesicles
osteoarthritis
miR-21-5p
inflammation
url https://www.mdpi.com/2073-4409/14/7/519
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AT ioannapapathanasiou synovialfibroblastextracellularvesiclesinduceinflammationviadeliveringmir215pinosteoarthritis
AT mariatzetis synovialfibroblastextracellularvesiclesinduceinflammationviadeliveringmir215pinosteoarthritis
AT anastasioskriebardis synovialfibroblastextracellularvesiclesinduceinflammationviadeliveringmir215pinosteoarthritis
AT aspasiatsezou synovialfibroblastextracellularvesiclesinduceinflammationviadeliveringmir215pinosteoarthritis