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Demonstration of a novel technique to quantitatively assess inflammatory mediators and cells in rat knee joints.

Barton NJ, Stevens DA, Hughes JP, Rossi AG, Chessell IP, Reeve AJ, McQueen DS - J Inflamm (Lond) (2007)

Bottom Line: Our results demonstrate up to 125 fold increases in synovial IL1alpha and IL1beta concentrations, approximately 30 fold increases in levels of IL6 and IL10 and a 200-300 fold elevation in synovial concentrations of TNFalpha during FCA-induced experimental arthritis.Finally, this novel technique has demonstrated a dose-response relationship between FCA and the total cell counts of synovial perfusates.In summary, this new technique provides a robust method for quantifying inflammatory mediators and cells from the synovial cavity itself, thereby detailing the inflammatory processes from within the capsule and excluding those processes occurring in other tissues surrounding the entire articulation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Neuroscience, University of Edinburgh, Medical College, 1 George Sq, Edinburgh, EH8 9JZ, UK. N.J.Barton@sms.ed.ac.uk

ABSTRACT

Background: The inflammation that accompanies the pain and swelling associated with osteo- and rheumatoid arthritis is mediated by complex interactions of inflammatory mediators. Cytokines play a pivotal role in orchestrating many of these processes, including inflammatory cell recruitment, adhesion and activation. In addition, prostaglandins are secreted into the synovial cavity and are involved in perpetuation of local inflammation, vasodilatation and vasoconstriction, and also with bone resorption. Pre-clinical models have been developed in order to correlate to the human disease and principle among these is the adjuvant-induced arthritis model in the rat.

Methods: We have developed a technique to quantitatively assess the contents of synovial fluid samples from rat joints. Two needles joined together are inserted into the knee joint of anaesthetised rats and connected to a Watson-Marlow perfusion pump. Sterile saline is infused and withdrawn at 100 microl min-1 until a 250 microl sample is collected.

Results: Our results demonstrate up to 125 fold increases in synovial IL1alpha and IL1beta concentrations, approximately 30 fold increases in levels of IL6 and IL10 and a 200-300 fold elevation in synovial concentrations of TNFalpha during FCA-induced experimental arthritis. Finally, this novel technique has demonstrated a dose-response relationship between FCA and the total cell counts of synovial perfusates.

Conclusion: In summary, this new technique provides a robust method for quantifying inflammatory mediators and cells from the synovial cavity itself, thereby detailing the inflammatory processes from within the capsule and excluding those processes occurring in other tissues surrounding the entire articulation.

No MeSH data available.


Related in: MedlinePlus

Levels of IL1α, IL1β, IL6, IL10 and TNFα in (a) ipsilateral and (b) contralateral joints of normal rats and those injected with low (150 μg; n = 8) and high (500 μg; n = 3) dose FCA 14 days earlier. There were negligible levels of any of the mediators measure in naïve joints (n = 10), but a significant increase in the expression of IL1α, IL1β, IL6 and TNFα was seen in all ipsilateral inflamed joints and in contralateral joints of rats injected with the high dose FCA (P < 0.05, Two-way ANOVA; compared with normal joints); statistical significance represented by *.
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Figure 3: Levels of IL1α, IL1β, IL6, IL10 and TNFα in (a) ipsilateral and (b) contralateral joints of normal rats and those injected with low (150 μg; n = 8) and high (500 μg; n = 3) dose FCA 14 days earlier. There were negligible levels of any of the mediators measure in naïve joints (n = 10), but a significant increase in the expression of IL1α, IL1β, IL6 and TNFα was seen in all ipsilateral inflamed joints and in contralateral joints of rats injected with the high dose FCA (P < 0.05, Two-way ANOVA; compared with normal joints); statistical significance represented by *.

Mentions: Fourteen days after rats received 150 μg or 500 μg FCA i.art (n = 8 and 3 respectively), the ipsilateral joint contained significantly higher levels of IL1α, IL1β, IL6 and TNFα compared with samples from naïve joints (n = 10), as measured by the Luminex assay (P < 0.05, Two-way ANOVA; see Figure 3a). The contralateral joints of rats injected with 500 μg FCA also contained significantly higher levels of IL1α, IL1β, IL6 and TNFα (P < 0.05, Two-way ANOVA; see Figure 3b).


Demonstration of a novel technique to quantitatively assess inflammatory mediators and cells in rat knee joints.

Barton NJ, Stevens DA, Hughes JP, Rossi AG, Chessell IP, Reeve AJ, McQueen DS - J Inflamm (Lond) (2007)

Levels of IL1α, IL1β, IL6, IL10 and TNFα in (a) ipsilateral and (b) contralateral joints of normal rats and those injected with low (150 μg; n = 8) and high (500 μg; n = 3) dose FCA 14 days earlier. There were negligible levels of any of the mediators measure in naïve joints (n = 10), but a significant increase in the expression of IL1α, IL1β, IL6 and TNFα was seen in all ipsilateral inflamed joints and in contralateral joints of rats injected with the high dose FCA (P < 0.05, Two-way ANOVA; compared with normal joints); statistical significance represented by *.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC1919375&req=5

Figure 3: Levels of IL1α, IL1β, IL6, IL10 and TNFα in (a) ipsilateral and (b) contralateral joints of normal rats and those injected with low (150 μg; n = 8) and high (500 μg; n = 3) dose FCA 14 days earlier. There were negligible levels of any of the mediators measure in naïve joints (n = 10), but a significant increase in the expression of IL1α, IL1β, IL6 and TNFα was seen in all ipsilateral inflamed joints and in contralateral joints of rats injected with the high dose FCA (P < 0.05, Two-way ANOVA; compared with normal joints); statistical significance represented by *.
Mentions: Fourteen days after rats received 150 μg or 500 μg FCA i.art (n = 8 and 3 respectively), the ipsilateral joint contained significantly higher levels of IL1α, IL1β, IL6 and TNFα compared with samples from naïve joints (n = 10), as measured by the Luminex assay (P < 0.05, Two-way ANOVA; see Figure 3a). The contralateral joints of rats injected with 500 μg FCA also contained significantly higher levels of IL1α, IL1β, IL6 and TNFα (P < 0.05, Two-way ANOVA; see Figure 3b).

Bottom Line: Our results demonstrate up to 125 fold increases in synovial IL1alpha and IL1beta concentrations, approximately 30 fold increases in levels of IL6 and IL10 and a 200-300 fold elevation in synovial concentrations of TNFalpha during FCA-induced experimental arthritis.Finally, this novel technique has demonstrated a dose-response relationship between FCA and the total cell counts of synovial perfusates.In summary, this new technique provides a robust method for quantifying inflammatory mediators and cells from the synovial cavity itself, thereby detailing the inflammatory processes from within the capsule and excluding those processes occurring in other tissues surrounding the entire articulation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Neuroscience, University of Edinburgh, Medical College, 1 George Sq, Edinburgh, EH8 9JZ, UK. N.J.Barton@sms.ed.ac.uk

ABSTRACT

Background: The inflammation that accompanies the pain and swelling associated with osteo- and rheumatoid arthritis is mediated by complex interactions of inflammatory mediators. Cytokines play a pivotal role in orchestrating many of these processes, including inflammatory cell recruitment, adhesion and activation. In addition, prostaglandins are secreted into the synovial cavity and are involved in perpetuation of local inflammation, vasodilatation and vasoconstriction, and also with bone resorption. Pre-clinical models have been developed in order to correlate to the human disease and principle among these is the adjuvant-induced arthritis model in the rat.

Methods: We have developed a technique to quantitatively assess the contents of synovial fluid samples from rat joints. Two needles joined together are inserted into the knee joint of anaesthetised rats and connected to a Watson-Marlow perfusion pump. Sterile saline is infused and withdrawn at 100 microl min-1 until a 250 microl sample is collected.

Results: Our results demonstrate up to 125 fold increases in synovial IL1alpha and IL1beta concentrations, approximately 30 fold increases in levels of IL6 and IL10 and a 200-300 fold elevation in synovial concentrations of TNFalpha during FCA-induced experimental arthritis. Finally, this novel technique has demonstrated a dose-response relationship between FCA and the total cell counts of synovial perfusates.

Conclusion: In summary, this new technique provides a robust method for quantifying inflammatory mediators and cells from the synovial cavity itself, thereby detailing the inflammatory processes from within the capsule and excluding those processes occurring in other tissues surrounding the entire articulation.

No MeSH data available.


Related in: MedlinePlus