The metabolic dynamics of cartilage explants over a long-term culture period

Moo, Eng Kuan; Osman, Noor Azuan Abu; Pingguan‐Murphy, Belinda

doi:10.1590/s1807-59322011000800021

Cited by 16 publications

(13 citation statements)

References 28 publications

(31 reference statements)

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“…Gene expression of cartilage relevant ECM marker genes like collagen II and aggrecan stayed relatively stable for 14 days before abruptly decreasing. Similar genetic dynamics have been observed beforehand in explant culture models which used immobilized, 6-mm-diameter bovine cartilage discs (25). These results indicate, that osteochondral live slices do not additional treatment like immobilization in agarose or strong perfusion pressures for nutrition and retaining their tissue speci c genetic phenotype -effectively reducing the handling necessities by a great deal.…”

Section: Discussionsupporting

confidence: 74%

Osteochondral Live Slices as Physiological in-vitro Model of the Human Joint for High-throughput Applications

Spinnen

Rendenbach

Kühl

et al. 2020

Preprint

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Background: Current advances in musculoskeletal research yielded numerous new insights and therapy options for diseases of the joints but preclinical in-vitro testing is currently limited to self-manufactured 2D and 3D cell cultures only remotely mimicking the in-vivo situation in regards of tissue composition and cell configuration. While in-vivo animal models are a lot more lifelike, they are inherently connected with ethical concerns, translation to the human situation, logistic efforts and high costs. Here we explored the use of native human life slice cultures from explanted tibial plateaus upon their feasibility to serve as a highly lifelike and easy-to-handle model of the human joint which can be obtained in masses from few tissue samples.Methods: Osteochondral slices (1cm x 1cm x 500µm) were prepared with a special microtome from 23 human tibial plateaus and subsequently cultivated in hanging inserts over the course of 7 or 21 days. Short-term cultivated slices were stimulated with either 800 pM/1.2 nM TNF-α or 800 pM TGF-β3. During cultivation, viability of tissue cells was assessed via laser microscopy / resazurin assay. After cultivation, gene expression of cartilage ECM Marker proteins was quantified with RT-PCR. TNF-α stimulated slices and their controls were stained with Safranin-O and analyzed via histomorphometry to quantify tissue proteoglycan content. Results: Tissue cell viability remained >90% over the three weeks. Laser scanning microscopy revealed highly conserved spatial alignment of cells inside the cartilage. Incubation of the slice cultures with TNF-α showed a significant, dose dependent decrease in mRNA expression of cartilage proteins collagen II, cartilage oligomeric matrix protein and aggrecan while incubation with TGF-β3 caused a significant increase of early bone formation proteins collagen I and also cartilage oligomeric matrix protein. Histologically, TNF-α incubation caused a 32% reduction of proteoglycans, detachment of collagen fibers and cell swelling.Conclusion: In summary, native osteochondral slice cultures provide a stable and manipulable, physiological model of human joint bone and cartilage biology. The setup is easy to produce and handle, scalable, and contributes to 3R principles in biomedical research, suggesting itself as a platform for preclinical testing especially regarding high-throughput applications.

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Section: Discussionsupporting

confidence: 74%

Osteochondral Live Slices as Physiological in-vitro Model of the Human Joint for High-throughput Applications

Spinnen

Rendenbach

Kühl

et al. 2020

Preprint

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“…It is quite difficult to control the response from individual samples. However, this model more closely represents the physiological environment than does the monolayer culture model and this model minimizes the presence of the complicated systems found in the animal model study [18, 19]. The experiment of this study was designed to mimic the microenvironment surrounding the inflammatory joint found in RA.…”

Section: Discussionmentioning

confidence: 99%

“…Nevertheless, animal experiments have a serious limitation due to the differences in biological responses; this is due to the differences between species. Furthermore, the model is made even more difficult because of the complicated biological systems involved as revealed in in vivo studies [18, 19]. In order to avoid such systemic and environmental complications in the animal model, the cartilage explant culture model has been used.…”

Section: Introductionmentioning

confidence: 99%

“…In order to avoid such systemic and environmental complications in the animal model, the cartilage explant culture model has been used. It is considered the most effective choice due to its ability to closely represent the physiological situation, while at the same time, it downplays any unexpected biological responses outside of the condition of interest found in in vivo studies [19]. …”

Section: Introductionmentioning

confidence: 99%

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Ex vivo model exhibits protective effects of sesamin against destruction of cartilage induced with a combination of tumor necrosis factor-alpha and oncostatin M

Khansai

Boonmaleerat

Pothacharoen

et al. 2016

BMC Complement Altern Med

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BackgroundRheumatoid arthritis (RA) is an autoimmune disease associated with chronic inflammatory arthritis. TNF-α and OSM are pro-inflammatory cytokines that play a key role in RA progression. Thus, reducing the effects of both cytokines is practical in order to relieve the progression of the disease. This current study is interested in sesamin, an active compound in sesame seeds. Sesamin has been shown to be a chondroprotective agent in osteoarthritis models. Here, we have evaluated a porcine cartilage explant as a cartilage degradation model related to RA induced by TNF-α and/or OSM in order to investigate the effects of sesamin on TNF-α and OSM in the cartilage degradation model.MethodsA porcine cartilage explant was induced with a combination of TNF-α and OSM (test group) or IL-1β and OSM (control group) followed by a co-treatment of sesamin over a long-term period (35 days). After which, the tested explants were analyzed for indications of both the remaining and the degradation aspects using glycosaminoglycan and collagen as an indicator.ResultsThe combination of TNF-α and OSM promoted cartilage degradation more than either TNF-α or OSM alone and was comparable with the combination of IL-1β and OSM. Sesamin could be offering protection against cartilage degradation by reducing GAGs and collagen turnover in the generated model.ConclusionsSesamin might be a promising agent as an alternative treatment for RA patients. Furthermore, the generated model revealed itself to be an impressive test model for the analysis of phytochemical substances against the cartilage degradation model for RA. The model could be used to test for the prevention of cartilage degradation in other biological agents induced with TNF-α and OSM as well.

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“…In the present study, we hypothesized that the paracrine chondroprotective effects of ASC on the OA cartilage would be advantageous due to the recovery of MMP/TIMP homeostasis. To avoid the dedifferentiation of chondrocytes, cultured in monolayer, or the synthesis of cartilage extracellular matrix (ECM) de novo in pellet cultures, we have chosen a model of human OA cartilage explants (CE), containing viable cells, readily surrounded by native OA cartilage ECM environment [ 10 ]. CE were cocultured or not with ASC, and the alterations in MMP and TIMP secretion by CE in response to ASC cocultures were analysed.…”

Section: Introductionmentioning

confidence: 99%

Paracrine Potential of the Human Adipose Tissue-Derived Stem Cells to Modulate Balance between Matrix Metalloproteinases and Their Inhibitors in the Osteoarthritic Cartilage In Vitro

Denkovskij

Bagdonas

Kusleviciute

et al. 2017

Stem Cells International

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Adipose tissue represents an abundant source of stem cells. Along with anti-inflammatory effects, ASC secrete various factors that may modulate metabolism of extracellular matrix in osteoarthritic (OA) cartilage, suggesting that the presence of ASC could be advantageous for OA cartilage due to the recovery of homeostasis between matrix metalloproteinases (MMPs) and their tissue inhibitors of metalloproteinases (TIMPs). To evaluate these effects, cartilage explants (CE) were cocultured with ASC for 3 and 7 days under stimulation with or without IL-1β. The pattern of gene expression in CE was modified by ASC, including the upregulation of COL1A1 and COL3A1 and the downregulation of MMP13 and COL10A1. The production of MMP-1, MMP-3, and MMP-13 by ASC was not significant; moreover, cocultures with ASC reduced MMP-13 production in CE. In conclusion, active production of TIMP-1, TIMP-2, TIMP-3, IL-6, IL-8, and gelatinases MMP-2 and MMP-9 by ASC may be involved in the extracellular matrix remodelling, as indicated by the altered expression of collagens, the downregulated production of MMP-13, and the reduced chondrocyte apoptosis in the cocultured CE. These data suggest that ASC modulated homeostasis of MMPs/TIMPs in degenerated OA cartilage in vitro and might be favourable in case of the intra-articular application of ASC therapy for the treatment of OA.

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The metabolic dynamics of cartilage explants over a long-term culture period

Cited by 16 publications

References 28 publications

Osteochondral Live Slices as Physiological in-vitro Model of the Human Joint for High-throughput Applications

Osteochondral Live Slices as Physiological in-vitro Model of the Human Joint for High-throughput Applications

Ex vivo model exhibits protective effects of sesamin against destruction of cartilage induced with a combination of tumor necrosis factor-alpha and oncostatin M

Paracrine Potential of the Human Adipose Tissue-Derived Stem Cells to Modulate Balance between Matrix Metalloproteinases and Their Inhibitors in the Osteoarthritic Cartilage In Vitro

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