1.0 INTRODUCTION:
Inflammation is a local response of living mammalian tissues to the injury. It is a body defense reaction in order to eliminate or limit the spread of injurious agents. There are various components to an inflammatory reaction that can contribute to the associated symptoms and tissue injury. Edema formation, leukocyte infiltration and granuloma formation represent such components of inflammation. However, it is related to infection caused by microorganisms, and various pathological changes are associated with it.
2.0 TEST SYSTEM DETAILS:
Species : Mus musculus
Strain : CS1; Swiss; C57;Balb/c
Age : 8-10 weeks
Body Wight : 25-30 g
Sex : Male or Female
No. of animals : 8 /Group
3.0 ALLOCATION OF GROUPS:
|
Groups |
Treatment |
Dose; ROA |
No. of Animals |
|
G1 |
Normal
Control |
Normal
saline or 0.25% Na-CMC |
8 |
|
G2 |
Disease
Control |
Normal
saline or 0.25% Na-CMC |
8 |
|
G3 |
Reference
Drug- Dexamethasone |
5
mpk; p.o. |
8 |
|
G4 |
Plant
Extract-1 |
X
mpk; p.o. |
8 |
|
G5 |
Plant
Extract-2 |
XX
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.
· To induce inflammation the mice will be implanted subcutaneously a sterilized cotton pellet weighing 10 ± 1mg into the groin region under anesthetic condition.
· The treatment schedule will be as follows:
· Group G1 animal will be treated as normal control and treated with normal saline or Na- CMC.
· Animals of group G2 will be treated as disease control and administered with normal saline or Na- CMC.
· Animals of group G3 will be treated orally with dexamethasone at the dose of 5mg/kg.
· Group G4 and G5 animals will be treated with test compound at different dose levels.
· All drug or vehicle and test compound will be administered to animals after cotton pallet implantation (Day 0) and continued for seven consecutive days.
· Animals will be sacrificed on the 8th day; the pellets surrounded by granuloma tissue will be dissected out carefully.
· The weight of wet cotton pellets will be noted and thereafter the cotton pellets were dried in oven at 60˚C for 24 h to obtain a dry cotton pellet weight, the mean weight of granuloma tissue formed around each pellet was obtained and the percentage inhibition was determined.
Percentage
inhibition (%) = (Control –Treated/Control) x 100
5.0 END POINT PARAMETER(S):
· % Anti-inflammatory activity (% Inhibition)
· Dry & wet weight of cotton pallet
6.0 REFERENCE(S):
6.1 Afsar SK, et al., Assessment of anti-inflammatory activity of Artemisia vulgaris leaves by cotton pellet granuloma method in Wistar albino rats, Journal of Pharmacy Research (2013), http://dx.doi.org/10.1016/ j.jopr.2013.04.056.
6.2 P. C. Lalawmpuiia*, c. Malsawmtluangia, r. Vanlalruataa, b. B. Kakoti. Evaluation of anti-inflammatory activity of helicia nilagirica bedd on cotton pellet-induced granuloma in rats. Int j pharm pharm sci, vol 8, issue 7, 455-456.
6.3 Kalpesh R. Patil, Umesh B. Mahajan, Banappa S. Unger, Sameer N. Goyal, Sateesh Belemkar, Sanjay J. Surana, Shreesh Ojha, and Chandragouda R. Patil. Animal Models of Inflammation for Screening of Anti-inflammatory Drugs: Implications for the Discovery and Development of Phytopharmaceuticals. Int. J. Mol. Sci. 2019, 20, 4367; doi:10.3390/ijms20184367
6.4 Tapas Kumar Sur, Biswajit Auddy, Susil Kumar Mitra, Dipak Kumar Sarkar, and Dipankar Bhattacharyya. Anti-inflammatory Activity of Detoxified Bacterial Strains in Wistar Rats. Natural Product Sciences. Vol. 16, No. 3, 2010 161
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