Limits...
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 Sterile saline is infused and withdrawn at 100 microl min-1 until a 250 microl sample is collected.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 perfusion needles and the perfusion system managing inflow and outflow from the knee joint space. A Watson-Marlow pump controlled the rate of saline infusion and sample extraction (100 μl min-1) from the joint. After the knee was secured to prevent movement of the limb, needles were inserted into the knee joint through the patella tendon.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC1919375&req=5

Figure 1: The perfusion needles and the perfusion system managing inflow and outflow from the knee joint space. A Watson-Marlow pump controlled the rate of saline infusion and sample extraction (100 μl min-1) from the joint. After the knee was secured to prevent movement of the limb, needles were inserted into the knee joint through the patella tendon.

Mentions: A needle perfusion system was constructed by binding a 25- and a 23-gauge needle together using epoxy putty, with the bevels of the needles positioned on the outside edges facing away from one another (see Figure 1). The tips of the needles were set 1–1.5 mm apart.


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 perfusion needles and the perfusion system managing inflow and outflow from the knee joint space. A Watson-Marlow pump controlled the rate of saline infusion and sample extraction (100 μl min-1) from the joint. After the knee was secured to prevent movement of the limb, needles were inserted into the knee joint through the patella tendon.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 1: The perfusion needles and the perfusion system managing inflow and outflow from the knee joint space. A Watson-Marlow pump controlled the rate of saline infusion and sample extraction (100 μl min-1) from the joint. After the knee was secured to prevent movement of the limb, needles were inserted into the knee joint through the patella tendon.
Mentions: A needle perfusion system was constructed by binding a 25- and a 23-gauge needle together using epoxy putty, with the bevels of the needles positioned on the outside edges facing away from one another (see Figure 1). The tips of the needles were set 1–1.5 mm apart.

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