Pharmacological Evaluation in Male Mice

Male inbred Swiss mice of NRC breeding colony (5–6 weeks, 25–30 g) were used. Animals were housed in plastic cages (4 per cage), maintained in controlled laboratory conditions (23–26 °C, 30–50% relative humidity, 12 h light/dark cycle, lights on 6:00 a.m.) and kept on standard diet and tap water ad libitum. Animals were acclimatized for a week to the experimental room, where behavioral experiments were carried out. All experimental procedures were conducted in accordance with the National Institutes of Health guide for the care and use of Laboratory animals (NIH Publications No. 8023, revised 1978) and were reviewed and approved by the Institutional Animal Care and Use Committee of NRC. Reporting of experimental data was in accordance with the ARRIVE guidelines.
Tramadol HCl was generously provided from ADWIA, Egypt. Tramadol was dissolved in sterile water (50 mg/ml). Different powdered composites were dispersed by tween 80 in sterile water to make 10 mg/ml solution. Diazepam (2 mg/ml vial, Valium, Egypt) was diluted in sterile water (1 mg/ml). Sodium Pentobarbital (50 mg/ml, Egypt) was applied for anesthesia purposes.