EVALUATION OF ANTI-INFLAMMATORY POTENTIAL OF TEST SAMPLE IN CARRAGEENAN INDUCED RAT PAW EDEMA (ACUTE MODEL)

1.0 INTRODUCTION:

Among many methods aimed at screening of anti-inflammatory drugs, one of the most useful techniques is based upon ability of such agents to inhibit edema produced in rat hind paw after injection of 1 % carrageenan. It has been widely used as a simple and reliable model to assess anti-inflammatory activity of various agents. Also, inhibition of carrageenan-induced inflammation has been shown to be highly predictive of anti-inflammatory drug activity in human inflammatory disease and doses of NSAIDs in this model correlate well with effective dose in patients. In addition, this model allows detecting orally active anti-inflammatory agent particularly in acute phase of inflammation. The paw volume is measured before and after injection of carrageenan and the paw volume of treated animals is compared to the controls. Any substance having ability to reduce paw volume in this model potentially acts as an anti-inflammatory agent by inhibiting synthesis of release of inflammatory mediators.

2.0 TEST SYSTEM DETAILS:

Species : Rattus norvegicus (Rat)

Strain : Wistar or Sprague Dawley

Age : 8-10 weeks

Body Wight : 160-180 g

Sex : Male or Female

No. of animals : 8 /Group

3.0 ALLOCATION OF GROUPS:

Groups	Treatment	Dose; ROA	No. of Animals
G1	Disease Control	Normal saline or 0.25% Na-CMC	8
G2	Reference Drug- Indomethacin	10 mpk; p.o.	8
G3	Plant Extract-1	X mpk; p.o.	8
G4	Plant Extract-2	XX mpk; p.o.	8
G5	Plant Extract-3	XXX mpk; p.o.	8

*The dose and ROA (Routes of administration) will be decided based on the type of reference drug

4.0 METHODOLOGY:

· The study protocol (Form B) shall be approved from the IAEC before commencing the experiment.

· Animals shall be procured from the CPCSEA authorized vendor.

· Animals shall be quarantined for 1 week as per the in house SOP.

· Healthy animals will be selected, randomized based on body weight and divided into five different groups consisting of 8 animals each.

· Group G1 animal will be treated as disease control and treated with normal saline or Na- CMC.

· Animals of group G2 will be treated orally with Indomethacin at the dose of 10mpk.

· Group G3, G4 and G5 animals will be treated with test compound at different dose levels.

· The rat paw will be marked with ink at the level of lateral malleolus.

· After one hour of drug treatment, all the animals will be challenged with subcutaneous (s.c.) injection of carrageenan (0.1 ml of 1 % solution in normal saline) into the plantar side of the left hind paw.

· The paw volume will be measured up to the mark using digital plethysmometer before (−1 h) and at 1h, 2h, 3h, 4h, and 5 h after injection of carrageenan for all the animals.

· Paw edema will be calculated by subtracting paw volume before carrageenan challenge (−1h) from the respective paw volumes at 1h, 2h, 3h, 4h, and 5h.

· Similarly, percentage of anti-inflammatory activity will be calculated for each animal using the following formula:

Paw volume of control (mL) − Paw volume of test (mL)/Paw volume of control (mL) ×100 %

5.0 END POINT PARAMETER(S):

· Paw Edema Volume

· % Anti-inflammatory activity (% Inhibition)

6.0 REFERENCE(S):

6.1 Fernandes J, H. Spindola, V. de Sousa, et al. 2010. Antiinflammatory activity of chitooligosaccharides in vivo. Marine Drugs 8: 1763–1768

6.2 Gursoy, A., L. Eroglu, S. Ulutin, et al. 1989. Evaluation of indomethacin nanocapsules for their physical stability and inhibitory activity on inflammation and platelet aggregation. International Journal of Pharmaceutics 52: 101–108.

6.3 Otterness, I.G., E.H. Wiseman, and D. Gans. 1979. A comparison of the carrageenan edema test and the ultraviolet light-induced erythema test as predictors of the clinical dose in rheumatoid arthritis. Agents and Actions 9: 177–183.

6.4 Di Rosa, M., J.P. Giroud, and D.A. Willoughdy. 1971. Studies of the mediators of the acute inflammatory response induced in rats in different sites by carrageenan and turpentine. The Journal of Pathology 104: 15–29.

6.5 Varpe SS, Juvekar AR, Bidikar MP, Juvekar PR. Evaluation of anti-inflammatory activity of Typha angustifolia pollen grains extracts in experimental animals. Indian J Pharmacol 2012;44:788-91

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