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Cytokine profiles in the joint depend on pathology, but are different between synovial fluid, cartilage tissue and cultured chondrocytes.

Tsuchida AI, Beekhuizen M, 't Hart MC, Radstake TR, Dhert WJ, Saris DB, van Osch GJ, Creemers LB - Arthritis Res. Ther. (2014)

Bottom Line: Cartilage formation was lower by healthy unexpanded chondrocytes than by osteoarthritic or defect chondrocytes.Several pro-inflammatory, pro-angiogenic and pro-repair cytokines were elevated in joints with symptomatic cartilage defects and/or osteoarthritis, although different cytokines were elevated in synovial fluid compared to tissue or cells.These alterations did not affect in vitro tissue formation with these chondrocytes, as this was at least as effective or even better compared to healthy chondrocytes.

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: This study aimed to evaluate whether profiles of several soluble mediators in synovial fluid and cartilage tissue are pathology-dependent and how their production is related to in vitro tissue formation by chondrocytes from diseased and healthy tissue.

Methods: Samples were obtained from donors without joint pathology (n = 39), with focal defects (n = 65) and osteoarthritis (n = 61). A multiplex bead assay (Luminex) was performed measuring up to 21 cytokines: Interleukin (IL)-1α, IL-1β, IL-1RA, IL-4, IL-6, IL-6Rα, IL-7, IL-8, IL-10, IL-13, tumor necrosis factor (TNF)α, Interferon (IFN)γ, oncostatin M (OSM), leukemia inhibitory factor (LIF), adiponectin, leptin, monocyte chemotactic factor (MCP)1, RANTES, basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), vascular growth factor (VEGF).

Results: In synovial fluid of patients with cartilage pathology, IL-6, IL-13, IFNγ and OSM levels were higher than in donors without joint pathology (P ≤ 0.001). IL-13, IFNγ and OSM were also different between donors with cartilage defects and OA (P < 0.05). In cartilage tissue from debrided defects, VEGF was higher than in non-pathological or osteoarthritic joints (P ≤ 0.001). IL-1α, IL-6, TNFα and OSM concentrations (in ng/ml) were markedly higher in cartilage tissue than in synovial fluid (P <0.01). Culture of chondrocytes generally led to a massive induction of most cytokines (P < 0.001). Although the release of inflammatory cytokines was also here dependent on the pathological condition (P < 0.001) the actual profiles were different from tissue or synovial fluid and between non-expanded and expanded chondrocytes. Cartilage formation was lower by healthy unexpanded chondrocytes than by osteoarthritic or defect chondrocytes.

Conclusions: Several pro-inflammatory, pro-angiogenic and pro-repair cytokines were elevated in joints with symptomatic cartilage defects and/or osteoarthritis, although different cytokines were elevated in synovial fluid compared to tissue or cells. Hence a clear molecular profile was evident dependent on disease status of the joint, which however changed in composition depending on the biological sample analysed. These alterations did not affect in vitro tissue formation with these chondrocytes, as this was at least as effective or even better compared to healthy chondrocytes.

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Safranin O staining of newly formed tissue by healthy, cartilage defect and osteoarthritis chondrocytes. Safranin O staining of newly formed tissue after redifferentiation of chondrocytes on type II collagen-coated filters after 28 days of culture. (A) Nonexpanded healthy chondrocytes, (B) nonexpanded cartilage defect chondrocytes, (C) nonexpanded osteoarthritic chondrocytes, (D) expanded healthy chondrocytes, (E) expanded cartilage defect chondrocytes, and (F) expanded osteoarthritic chondrocytes.
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Fig2: Safranin O staining of newly formed tissue by healthy, cartilage defect and osteoarthritis chondrocytes. Safranin O staining of newly formed tissue after redifferentiation of chondrocytes on type II collagen-coated filters after 28 days of culture. (A) Nonexpanded healthy chondrocytes, (B) nonexpanded cartilage defect chondrocytes, (C) nonexpanded osteoarthritic chondrocytes, (D) expanded healthy chondrocytes, (E) expanded cartilage defect chondrocytes, and (F) expanded osteoarthritic chondrocytes.

Mentions: Total GAG production per DNA was not different between healthy, cartilage defect and OA nonexpanded chondrocytes (Figure 1C). However, most of the GAG produced during culture was not retained in the newly formed tissue but released in the medium (Figure 1B). Remarkably, unexpanded chondrocytes from defect cartilage (GAG/DNA, 19.6 ± 15.1 μg/μg) and OA chondrocytes (GAG/DNA, 22.8 ± 17.1 μg/μg) retained more of the produced GAG in the matrix than isolated unexpanded chondrocytes from healthy cartilage (GAG/DNA, 9.4 ± 4.4 μg/μg; P <0.001) (Figure 1A). However, we did not see this same difference in safranin-O staining intensity (Figure 2). Nonquantitative evaluation of collagen II deposition showed clear collagen II staining without apparent differences between donor types (Figure 3).Figure 1


Cytokine profiles in the joint depend on pathology, but are different between synovial fluid, cartilage tissue and cultured chondrocytes.

Tsuchida AI, Beekhuizen M, 't Hart MC, Radstake TR, Dhert WJ, Saris DB, van Osch GJ, Creemers LB - Arthritis Res. Ther. (2014)

Safranin O staining of newly formed tissue by healthy, cartilage defect and osteoarthritis chondrocytes. Safranin O staining of newly formed tissue after redifferentiation of chondrocytes on type II collagen-coated filters after 28 days of culture. (A) Nonexpanded healthy chondrocytes, (B) nonexpanded cartilage defect chondrocytes, (C) nonexpanded osteoarthritic chondrocytes, (D) expanded healthy chondrocytes, (E) expanded cartilage defect chondrocytes, and (F) expanded osteoarthritic chondrocytes.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4201683&req=5

Fig2: Safranin O staining of newly formed tissue by healthy, cartilage defect and osteoarthritis chondrocytes. Safranin O staining of newly formed tissue after redifferentiation of chondrocytes on type II collagen-coated filters after 28 days of culture. (A) Nonexpanded healthy chondrocytes, (B) nonexpanded cartilage defect chondrocytes, (C) nonexpanded osteoarthritic chondrocytes, (D) expanded healthy chondrocytes, (E) expanded cartilage defect chondrocytes, and (F) expanded osteoarthritic chondrocytes.
Mentions: Total GAG production per DNA was not different between healthy, cartilage defect and OA nonexpanded chondrocytes (Figure 1C). However, most of the GAG produced during culture was not retained in the newly formed tissue but released in the medium (Figure 1B). Remarkably, unexpanded chondrocytes from defect cartilage (GAG/DNA, 19.6 ± 15.1 μg/μg) and OA chondrocytes (GAG/DNA, 22.8 ± 17.1 μg/μg) retained more of the produced GAG in the matrix than isolated unexpanded chondrocytes from healthy cartilage (GAG/DNA, 9.4 ± 4.4 μg/μg; P <0.001) (Figure 1A). However, we did not see this same difference in safranin-O staining intensity (Figure 2). Nonquantitative evaluation of collagen II deposition showed clear collagen II staining without apparent differences between donor types (Figure 3).Figure 1

Bottom Line: Cartilage formation was lower by healthy unexpanded chondrocytes than by osteoarthritic or defect chondrocytes.Several pro-inflammatory, pro-angiogenic and pro-repair cytokines were elevated in joints with symptomatic cartilage defects and/or osteoarthritis, although different cytokines were elevated in synovial fluid compared to tissue or cells.These alterations did not affect in vitro tissue formation with these chondrocytes, as this was at least as effective or even better compared to healthy chondrocytes.

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: This study aimed to evaluate whether profiles of several soluble mediators in synovial fluid and cartilage tissue are pathology-dependent and how their production is related to in vitro tissue formation by chondrocytes from diseased and healthy tissue.

Methods: Samples were obtained from donors without joint pathology (n = 39), with focal defects (n = 65) and osteoarthritis (n = 61). A multiplex bead assay (Luminex) was performed measuring up to 21 cytokines: Interleukin (IL)-1α, IL-1β, IL-1RA, IL-4, IL-6, IL-6Rα, IL-7, IL-8, IL-10, IL-13, tumor necrosis factor (TNF)α, Interferon (IFN)γ, oncostatin M (OSM), leukemia inhibitory factor (LIF), adiponectin, leptin, monocyte chemotactic factor (MCP)1, RANTES, basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), vascular growth factor (VEGF).

Results: In synovial fluid of patients with cartilage pathology, IL-6, IL-13, IFNγ and OSM levels were higher than in donors without joint pathology (P ≤ 0.001). IL-13, IFNγ and OSM were also different between donors with cartilage defects and OA (P < 0.05). In cartilage tissue from debrided defects, VEGF was higher than in non-pathological or osteoarthritic joints (P ≤ 0.001). IL-1α, IL-6, TNFα and OSM concentrations (in ng/ml) were markedly higher in cartilage tissue than in synovial fluid (P <0.01). Culture of chondrocytes generally led to a massive induction of most cytokines (P < 0.001). Although the release of inflammatory cytokines was also here dependent on the pathological condition (P < 0.001) the actual profiles were different from tissue or synovial fluid and between non-expanded and expanded chondrocytes. Cartilage formation was lower by healthy unexpanded chondrocytes than by osteoarthritic or defect chondrocytes.

Conclusions: Several pro-inflammatory, pro-angiogenic and pro-repair cytokines were elevated in joints with symptomatic cartilage defects and/or osteoarthritis, although different cytokines were elevated in synovial fluid compared to tissue or cells. Hence a clear molecular profile was evident dependent on disease status of the joint, which however changed in composition depending on the biological sample analysed. These alterations did not affect in vitro tissue formation with these chondrocytes, as this was at least as effective or even better compared to healthy chondrocytes.

Show MeSH
Related in: MedlinePlus