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4.3.3. Formalin-induced paw edema bioassay (sub-acute infla-
mmatory model)
measuring the inhibition of edema volume 3 h after formalin injec-
tion (2%, 0.1 mL).
This sub-acute inflammatory model was performed as previ-
ously described.35,36 Rats in the first experiment were given the
same test compounds at a dose of 20 mg/kg body weight daily
for 7 consecutive days. A solution of formalin (2%, 0.1 mL) was in-
jected into the subplantar region of the left hind paw under light
ether anesthesia 1 h after oral administration of the test com-
pound. A second injection of formalin (2%, 0.1 mL) was given on
the third day. The changes in the weight (g) of paw edema were
measured at the eighth day. Percentage inhibition of inflammation
was calculated according to the following equation:
4.3.8. Statistical analysis
Data are presented as means standard error of the mean
(SEM). The concentration-dependent effects of various drugs
in vitro were evaluated statistically by the randomized blocks de-
sign analysis of variance (ANOVA). Statistical differences between
various in vivo data sets were estimated by using the Analysis of
Variance (ANOVA) followed by Student–Newman–Keuls Multiple
Comparison Test. Statistical Analysis System (SAS) and SPSS ana-
lytical software were used. The difference in results was consid-
ered significant when P <0.05.
% Inhibition ¼ ðweight of paw edema of control
ꢁ weight of paw edema of treatedÞ=
Acknowledgments
weight of paw edema of control ꢀ 100
This research was supported by grants from the Jack Brown and
Family Alzheimer’s Disease Research Foundation and Natural Sci-
ences and Engineering Research Council of Canada. The authors
wish to thank Dr. Ed Neeland for helpful discussions and com-
ments on the manuscript.
4.3.4. Turpentine oil-induced granuloma pouch bioassay (sub-
acute inflammatory model)
This sub-acute inflammatory model was performed as previ-
ously described.36,37 Male albino rats weighing 120–150 g were
used throughout this assay. One group of five rats was kept as a
control and another group received the standard drug celecoxib
at a dose of 20 mg/kg body weight, po. Subcutaneous dorsal gran-
uloma pouch was made in ether-anesthetized rats by injecting
2 mL of air, followed by injection of 0.5 mL of turpentine oil into
it. All of the test compounds were administered orally at a dose
of 20 mg/kg body weight 1 h prior to turpentine oil injection and
continued for seven consecutive days. On the eighth day, the
paw was opened under anesthesia and the exudates were taken
out with a syringe. The volume (mL) of the exudates was measured
and the percentage inhibition of inflammation was determined as
follows:
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The selected compounds were further tested in rats at 5, 10, 20,
40 and 50 mg/kg body weight and the ED50 was determined by