Co-culture of osteochondral explants and synovial membrane as in vitro model for osteoarthritis.
Animals
Chondrocytes
/ cytology
Coculture Techniques
Collagen Type I
/ genetics
Horses
Interleukin-1beta
/ pharmacology
Interleukin-6
/ genetics
Matrix Metalloproteinase 1
/ genetics
Models, Biological
Nitric Oxide
/ metabolism
Osteoarthritis
/ pathology
Synovial Membrane
/ cytology
Tumor Necrosis Factor-alpha
/ pharmacology
Urea
/ metabolism
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2019
2019
Historique:
received:
03
04
2018
accepted:
16
03
2019
entrez:
3
4
2019
pubmed:
3
4
2019
medline:
20
12
2019
Statut:
epublish
Résumé
The purpose of the current study was to establish an in vitro model for osteoarthritis (OA) by co-culture of osteochondral and synovial membrane explants. Osteochondral explants were cultured alone (control-1) or in co-culture with synovial membrane explants (control-2) in standard culture medium or with interleukin-1β (IL1β) and tumor necrosis factor (TNFα) added to the culture medium (OA-model-1 = osteochondral explant; OA-model-2 = osteochondroal-synovial explant). In addition, in OA-model groups a 2-mm partial-thickness defect was created in the centre of the cartilage explant. Changes in the expression of extracellular matrix (ECM) genes (collagen type-1 (Col1), Col2, Col10 and aggrecan) as well as presence and quantity of inflammatory marker genes (IL6, matrix metalloproteinase-1 (MMP1), MMP3, MMP13, a disintegrin and metalloproteinase with-thrombospondin-motif-5 (ADAMTS5) were analysed by immunohistochemistry, qPCR and ELISA. To monitor the activity of classically-activated pro-inflammatory (M1) versus alternatively-activated anti-inflammatory/repair (M2) synovial macrophages, the nitric oxide/urea ratio in the supernatant of osteochondral-synovial explant co-cultures was determined. In both OA-model groups immunohistochemistry and qPCR showed a significantly increased expression of MMPs and IL6 compared to their respective control group. ELISA results confirmed a statistically significant increase in MMP1and MMP3 production over the culturing period. In the osteochondral-synovial explant co-culture OA-model the nitric oxide/urea ratio was increased compared to the control group, indicating a shift toward M1 synovial macrophages. In summary, chemical damage (TNFα, IL1β) in combination with a partial-thickness cartilage defect elicits an inflammatory response similar to naturally occurring OA in osteochondral explants with and without osteochondral-synovial explant co-cultures and OA-model-2 showing a closer approximation of OA due to the additional shift of synovial macrophages toward the pro-inflammatory M1 phenotype.
Identifiants
pubmed: 30939166
doi: 10.1371/journal.pone.0214709
pii: PONE-D-18-09968
pmc: PMC6445514
doi:
Substances chimiques
Collagen Type I
0
Interleukin-1beta
0
Interleukin-6
0
Tumor Necrosis Factor-alpha
0
Nitric Oxide
31C4KY9ESH
Urea
8W8T17847W
Matrix Metalloproteinase 1
EC 3.4.24.7
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0214709Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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