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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 Sterile saline is infused and withdrawn at 100 microl min-1 until a 250 microl sample is collected.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

The effects of 150 μg (low dose; n = 5) and 500 μg (high dose; n = 3) FCA on total cell count from joint perfusates. Naïve joints contained no cells (0), whereas all other joints contained increased levels, although only high dose ipsilateral joints proved to have significantly raised levels (P < 0.05, Mann Whitney); statistical significance donated by *.
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Figure 4: The effects of 150 μg (low dose; n = 5) and 500 μg (high dose; n = 3) FCA on total cell count from joint perfusates. Naïve joints contained no cells (0), whereas all other joints contained increased levels, although only high dose ipsilateral joints proved to have significantly raised levels (P < 0.05, Mann Whitney); statistical significance donated by *.

Mentions: Total inflammatory cell counts from normal animals (n = 5) and those injected with FCA (n = 8) 14 days prior to sampling are shown in Figure 4. Normal joints had no cells detectable, whereas all others samples had measurable levels. However, only the 500 μg FCA ipsilateral (n = 3) joints proved to have a significantly greater number of cells than normal joints (4.8 ± 0.06 × 106 cells ml-1; P < 0.05, Mann Whitney). A dose-response relationship was demonstrated by the total cell count in both ipsilateral and contralateral joints.


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)

The effects of 150 μg (low dose; n = 5) and 500 μg (high dose; n = 3) FCA on total cell count from joint perfusates. Naïve joints contained no cells (0), whereas all other joints contained increased levels, although only high dose ipsilateral joints proved to have significantly raised levels (P < 0.05, Mann Whitney); statistical significance donated by *.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: The effects of 150 μg (low dose; n = 5) and 500 μg (high dose; n = 3) FCA on total cell count from joint perfusates. Naïve joints contained no cells (0), whereas all other joints contained increased levels, although only high dose ipsilateral joints proved to have significantly raised levels (P < 0.05, Mann Whitney); statistical significance donated by *.
Mentions: Total inflammatory cell counts from normal animals (n = 5) and those injected with FCA (n = 8) 14 days prior to sampling are shown in Figure 4. Normal joints had no cells detectable, whereas all others samples had measurable levels. However, only the 500 μg FCA ipsilateral (n = 3) joints proved to have a significantly greater number of cells than normal joints (4.8 ± 0.06 × 106 cells ml-1; P < 0.05, Mann Whitney). A dose-response relationship was demonstrated by the total cell count in both ipsilateral and contralateral joints.

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