Redifferentiation of dedifferentiated human articular chondrocytes: comparison of 2D and 3D cultures

Caron, M.M.; Emans, Pieter J.; Coolsen, Mariëlle M.E.; Voss, L.; Surtel, D.A.; Cremers, A.; Rhijn, Lodewijk W. van; Welting, Tim J. M.

doi:10.1016/j.joca.2012.06.016

Cited by 410 publications

(384 citation statements)

References 53 publications

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“…RNA isolation, RNA quantification by ultraviolet (UV) spectrometry (Biodrop; Isogen Life Sciences, Utrecht, the Netherlands) and cDNA synthesis were performed as described before (Caron et al, 2012a;Welting et al, 2011). Real time quantitative PCR (RT-qPCR) was performed using MESAGREEN qPCR MasterMix Plus for SYBR ® Assay (Eurogentec, Seraing, Belgium).…”

Section: Cox-2 Inhibition Impairs Chondrogenic Differentiationmentioning

confidence: 99%

Impairment of the chondrogenic phase of endochondral ossification in vivo by inhibition of cyclooxygenase-2

Janssen

Caron

Rietbergen

et al. 2017

eCM

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Many studies have reported on the effects of cyclooxygenase-2 (COX-2) inhibition on osteogenesis. However, far less is known about the effects of COX-2 inhibition on chondrogenic differentiation. Previous studies conducted by our group show that COX-2 inhibition influences in vitro chondrogenic differentiation. Importantly, this might have consequences on endochondral ossification processes occurring in vivo, such as bone fracture healing, growth plate development and ectopic generation of cartilage. The goal of our study was to investigate, in vivo, the effect of COX-2 inhibition by celecoxib on the cartilaginous phase of three different endochondral ossification scenarios.10 mg/kg/d celecoxib or placebo were orally administered for 25 d to skeletally-immature New Zealand White rabbits (n = 6 per group). Endochondral ossification during fracture healing of a non-critical size defect in the ulna, femoral growth plate and ectopically-induced cartilaginous tissue were examined by radiography, micro-computed tomography (µ-CT), histology and gene expression analysis.Celecoxib treatment resulted in delayed bone fracture healing, alterations in growth plate development and progression of mineralisation. In addition, chondrogenic differentiation of ectopically-induced cartilaginous tissue was severely impaired by celecoxib. In conclusion, we found that celecoxib impaired the chondrogenic phase of endochondral ossification.

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Section: Cox-2 Inhibition Impairs Chondrogenic Differentiationmentioning

confidence: 99%

Impairment of the chondrogenic phase of endochondral ossification in vivo by inhibition of cyclooxygenase-2

Janssen

Caron

Rietbergen

et al. 2017

eCM

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“…While several previous OA models have employed 2D monolayers, [19][20][21][22][23] the literature reveals that cell behavior in 2D is often not representative of cell responses observed in physiologically relevant 3D contexts. [24][25][26][27] Indeed, an increasing number of studies recently reporting in vitro OA models have cultured chondrocytes within 3D microenvironments. [28][29][30][31][32] However, important auxiliary cell types such as macrophages have continued to be cultured in 2D, 28,31,33 despite the primary location of the macrophages in the knee joint being within the synovial membrane.…”

Section: Introductionmentioning

confidence: 99%

A Three-Dimensional Chondrocyte-Macrophage Coculture System to Probe Inflammation in Experimental Osteoarthritis

Samavedi

Díaz‐Rodríguez

Erndt-Marino

et al. 2017

Tissue Engineering Part A

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The goal of the present study was to develop a fully three-dimensional (3D) coculture system that would allow for systematic evaluation of the interplay between activated macrophages (AMs) and chondrocytes in osteoarthritic disease progression and treatment. Toward this end, our coculture system was first validated against existing in vitro osteoarthritis models, which have generally cultured healthy normal chondrocytes (NCs)-in two-dimensional (2D) or 3D-with proinflammatory AMs in 2D. In this work, NCs and AMs were both encapsulated within poly(ethylene glycol) diacrylate hydrogels to mimic the native 3D environments of both cell types within the osteoarthritic joint. As with previous studies, increases in matrix metalloproteinases (MMPs) and proinflammatory cytokines associated with the early stages of osteoarthritis were observed during NC-AM coculture, as were decreases in protein-level Aggrecan and collagen II. Thereafter, the coculture system was extended to osteoarthritic chondrocytes (OACs) and AMs to evaluate the potential effects of AMs on pre-existing osteoarthritic phenotypes. OACs in coculture with AMs expressed significantly higher levels of MMP-1, MMP-3, MMP-9, MMP-13, IL-1b, TNF-a, IL-6, IL-8, and IFN-g compared to OACs in mono-culture, indicating that proinflammatory macrophages may intensify the abnormal matrix degradation and cytokine secretion already associated with OACs. Likewise, AMs cocultured with OACs expressed significantly more IL-1b and VEGF-A compared to AM mono-culture controls, suggesting that OACs may intensify abnormal macrophage activation. Finally, OACs cultured in the presence of nonactivated macrophages produced lower levels of MMP-9 and proinflammatory cytokines IL-1b, TNF-a, and IFN-g compared to OACs in the OAC-AM system, results that are consistent with anti-inflammatory agents temporarily reducing certain OA symptoms. In summary, the 3D coculture system developed herein captures several key features of inflammatory OA and may prove useful in future screening of therapeutic agents and/or assessment of disease progression mechanisms.

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“…Articular chondrocytes will display phenotypic or morphological changes and lose their ability of cartilage tissue regeneration after the fourth passage due to rapid dedifferentiation and senescence, (Kang et al 2007;Caron et al 2012;Cha et al 2013;Ashraf et al 2016). Therefore, chondrocytes that passaged within three generations (P0, P1, and P2) were sub-cultured in vitro in this study.…”

Section: Discussionmentioning

confidence: 99%

“…Cartilage is a unique avascular, aneural, and alymphatic tissue that lacks resident progenitor stem cells, and thus has little self-healing potential once injured (Goessler et al 2005;Candela et al 2014). Chondrocytes, the primary component of articular cartilage, are essential for the production of extracellular matrix (ECM) and maintenance of cartilage structure and function (Caron et al 2012). In addition, chondrocytes are recently demonstrated to be important in preventing articular degeneration (Tang et al 2017).…”

Section: Introductionmentioning

confidence: 99%

MicroRNA-483-5p Modulates the Expression of Cartilage-Related Genes in Human Chondrocytes through Down-Regulating TGF-<i>β</i>1 Expression

Wei

et al. 2017

Tohoku J. Exp. Med.

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Transforming growth factor-β1 (TGF-β1) has been reported to improve chondrocytes phenotype and function. The expression levels of microRNA-483-5p (miR-483-5p), a potential regulator of TGF-β signaling pathway, were elevated in chondrocytes of patients with osteoarthritis. In this study, we aimed to explore the role of miR-483-5p for the expression of cartilage-related genes in chondrocytes. Human chondrocytes were harvested from femoral condyle and tibial plateau of different donors (n = 10) following amputation due to sarcomas not involving the joint space. The expression levels of miR-483-5p and TGF-β1 mRNA were measured by qRT-PCR. Runt-related transcription factor 2 (RUNX2) is a transcription factor involved in chondrocyte differentiation, and type II collagen-degrading matrix metalloproteinase13 (MMP13) contributes to cartilage degradation. qRT-PCR was also used to measure the levels of RUNX2 and MMP-13 mRNAs, as well as type II collagen alpha 1 (COL2A1) and aggrecan mRNAs. COL2A1 and aggrecan are major cartilage extracellular matrix proteins that are essential for normal cartilage function. The expression levels of miR-483-5p were significantly increased in chondrocytes from Passage 0 to 2, whereas the expression levels of TGF-β1 mRNA were marginally decreased. Passage 1 chondrocytes were employed for following experiments. MiR-483-5p overexpression reduced TGF-β1 expression, while miR-483-5p knockdown dramatically elevated TGF-β 1 expression both at mRNA and protein levels. Further, miR-483-5p overexpression significantly decreased the levels of COL2A1 and aggrecan mRNAs, and increased those of RUNX2 and MMP13 mRNAs, by down-regulating TGF-β1 expression. These findings suggest that modulating miR-483-5p expression may contribute to maintaining the cartilage tissues.

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Redifferentiation of dedifferentiated human articular chondrocytes: comparison of 2D and 3D cultures

Cited by 410 publications

References 53 publications

Impairment of the chondrogenic phase of endochondral ossification in vivo by inhibition of cyclooxygenase-2

Impairment of the chondrogenic phase of endochondral ossification in vivo by inhibition of cyclooxygenase-2

A Three-Dimensional Chondrocyte-Macrophage Coculture System to Probe Inflammation in Experimental Osteoarthritis

MicroRNA-483-5p Modulates the Expression of Cartilage-Related Genes in Human Chondrocytes through Down-Regulating TGF-<i>β</i>1 Expression

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