IN VIVO ANTI INFLAMMATORY EVALUATION OF DRUGS

1. Presented by:
Anushma Chorsiya
M.Pharm
(Pharmacognosy & Phytochemistry)
DELHI PHARMACEUTICAL SCIENCES AND
RESEARCH UNIVERSITY (DPSRU)
IN-VIVO EVALUATION OF
ANTI-INFLAMMATORY DRUGS

2. Inflammation is local defensive response of living body
against injury or chemical agents.
A drug or substance that reduces inflammation i.e. redness
swelling and pain in the body are anti inflammatory agents.
These agents blocks certain substances in the body that
causes inflammation.
INTRODUCTION

3. Lack of
Nutrients
Bad
Diet
Stress
Pathogen
Exposure
Obesity
Enviromental
Toxins
FACTORS

4. TYPES
• SHORT TERM PROCESS
• 5 CARDINAL SIGNS: PAIN,
REDNESS, IMMOBILITY,
SWELLING AND HEAT
ACUTE
INFLAMMATION
• SLOW LONG TERM PROCESS
• SIGNS: FATIGUE, FEVER,
MOUTH SORES, RASHES,
ABDOMINAL PAIN AND
CHEST PAIN
CHRONIC
INFLAMMATION

5. MODELS
GRANULOMA
POUCH TECHNIQUE
COTTON WOOL
GRANULOMA
UV- ERYTHEMA
IN GUINEA PIGS
PLEURISY TESTS
PAW EDEMA
IN RATS
OXAZOLONE
INDUCED EAR
EDEMA IN MICE
SPONGE
IMPLANTATION
TECHNIQUE
VASCULAR
PERMEABILITY
CROTON OIL
EAR EDEMA IN
RATS AND MICE
GLASS ROD
GRANULOMA

6. VASCULAR
PERMEABILITY
PG’s,
histamine,
leucotrienes
release
Arterioles
and venules
dilation
Vascular
permeability
increased
Plasma
protein and
fluids
extravagated
Edema
produced
Animal Model: Albino Wistar Rat
Inducer: 0.25 ml; 0.6% v/v solution of acetic acid intraperitoneally
Dye: 10ml/kg; 10% w/v Evan Blue i.v. via tail vein
Absorbance: Measured at 590 nm

7. UV ERYTHEMA IN
GUINEA PIG
Exposure to
280-320 nm UV
radiations
During 24 hrs
Prostaglandin E
increased
Animal Model: Albino Guinea Pig (approx 350 gm)
Chemical Depilatory: Suspension of Barium Sulphide
Drug Administration: Gavages 10mg/kg
Leather cuff Hole: 1.5 * 2.5 cm
Assesment: By scoring erythema from 0 to 5

8. Croton oil ear edema in rat and mice
Croton Oil
(Irritant
agents)
Activates
PK-C, MAPK
AND PLA-2
Release PAF
Stimulates vasodilation,
vascular permeability,
polymer phonuclear,
leukocytes migration etc.
Skin
Inflammation
(Ear edema)
Aim: Study anti inflammatory activity of steroids.
Irritants: 12-o-tetracanoliphorbol-13-acetate (TPA) and other
phorbol esters
Dose: 15ml acetone solution of 75µg of croton oil.
Standard: Indomethacin

9. Oxazolon induced ear edema in mice and
rat
Aim: To quantify tropical and systemic drug anti- inflammatory activity.
Inducer: 10ml, 0.5% oxazolone in acetone thrice a week for 8 days, later
0.01ml of 2% oxazolone solution.
Oxazolone
repeated
dose
Increased
level of Th2
cytokines
Decreased
Th1
cytokines
Th2 especially IL4 plays role in
development of Dermatitis

10. Pleurisy test
•Irritants: Histamine, Bradykinin, Prostaglandins, Mast cell degranulators, Dextran,
Enzymes, Antigens, Microbes.
•Non-specific Irritants: Turpentine, Carrageenan
Route of Administration: Intra plural injections (0.1 ml of 1% Carrageenan)
Determination: Myeloperoxidase, Adenosine-deaminase activities, Nitric oxide levels, Total
and Differential leukocytes counts and ED50 value for different doses.

11. PAW EDEMA IN RATS
Animal Model: Sprague–Dawley rat (100-150 gms)
Inducer: Carrageenin, Brewer’s yeast, Formaldehyde, Dextran,
Egg albumin, Kaolin, Zymosan, Histamine, Serotonin, Cobra
Venom
Standard: Ibuprofen
FIRST
PHASE
SECOND
PHASE
THIRD
PHASE
• Mediated by mast
cell degranulation.
• Histamine and
serotonin release
• 1 hr
• Bradykinin release
and pain.
• 1-2.5 hrs
• Eicosanoid
production.
• 3-4 hrs

12.  Animal Model: Swiss Albino Mice (25-30 gm)
 Cage: Polypropylene
 Temperature: 25±2ºC
 Relative Humidity: 60-70%
 Inducer: 0.1 ml Carrageenan 1% w/v by sub plantar injection
 Test Drug: 200 and 400 mg/kg oral administration of Ficus
virens ethanolic extract
 Standard Drug: 40 mg/kg Ibuprofen oral administration
 Control: 10ml/kg Saline oral administration
 Instrument: Plethysmometer
Evaluation of carrageenan induced anti-inflammatory
activity of ethanolic extract of bark of ficus virens L. In
swiss albino mice

13. PLETHYSMOMETER

14.  Animal Models: Swiss albino mice, 20-25 gms (Chronic)
Wistar albino rat, 150-200 gms (Acute)
 Study: Carrageenan induced rat paw odema and Cotton pellet induced granuloma
 Standard Drug: Diclofenan sodium (25 mg/kg- Acute model and 10 mg/kg – Chronic
model)
 Test Extract: Ethanolic, Chloroform and n- Hexane extract of Bryophyllum calycinum
each of dose 100 mg/kg, 200 mg/kg and 400 mg/kg for acute model. Ethanolic extract of
Bryophyllum calycinum of dose 100 mg/kg, 200 mg/kg and 400 mg/kg for chronic
model.
EVALUATION OF ANTI INFLAMMATORY ACTIVITY
OF Bryophyllum calycinum (CRASSULACEAE )
ON ACUTE AND CHRONIC INFLAMMATION
MODELS

15. REFERNCES
• Azza, Z., & Oudghiri, M. (2015). In vivo anti-inflammatory and antiarthritic activities of aqueous extracts
from Thymelaea hirsuta. Pharmacognosy research, 7(2), 213.
• Wang, Y., Li, G. H., Liu, X. Y., Xu, L., Wang, S. S., & Zhang, X. M. (2017). In vivo anti-inflammatory effects of
taraxasterol against animal models. African Journal of Traditional, Complementary and Alternative
Medicines, 14(1), 43-51.
• Phanse, M. A., Patil, M. J., Abbulu, K., Chaudhari, P. D., & Patel, B. (2012). In-vivo and in-vitro screening of
medicinal plants for their anti-inflammatory activity: an overview. J. Appl. Pharm. Sci, 2(6), 19-33.
• Pandurangan, A., Kaur, A., Kumar, M., Kaushik, V., & Arunachalam, G. (2019). Evaluation of Anti-
inflammatory Activity of Bryophyllum calycinum (Crassulaceae) on Acute and Chronic Inflammation
Models. Journal of Pharmaceutical Sciences and Research, 11(1), 263-266.
• Hafeez, A., Jain, U., Sajwan, P., Srivastava, S., & Thakur, A. (2013). Evaluation of Carrageenan induced anti-
inflammatory activity of ethanolic extract of bark of Ficus virens Linn. in swiss albino mice. J
Phytopharmacol, 2, 39-43.