2.3. Animals and experimental design
The Graduate Studies and the Ethical Committees of the College of Pharmacy, University of Baghdad, approved the study protocol.
Wistar Albino experimental rats of both sexes aged six weeks with an average weight of 150gm were utilized in this study; since the animals were acquired and maintained in the College of Pharmacy Experimental Animal House, University of Baghdad, Iraq; in addition, the experimental rats were acclimatized for one week prior to starting the experiment; the rats were housed under controlled conditions of a light/dark cycle (12hours), temperature at (23±2°C) and humidity (50±5%); and had free access to a standard commercial diet, which was purchased from the local market, and tap water ad libitum .
The experimental animals (32 Rats) were randomly assigned into four groups (n=8) as follows:
Group I : Each rat was orally administered vehicle only (5% tween in DDW) via oral gavage for 14 consecutive days. Then a single dose of normal saline (NaCl) (0.9%) was IP injected 1 hour after the last vehicle administration on day 14. This group served as the control group.
Group II : Each rat was orally administered DMF only (15mg/kg/day) for 14 consecutive days.
Group III : Each rat was orally-administered vehicle (5% tween in DDW) only via oral gavage for 14 consecutive days. Then a single dose of DOX (90mg/kg) was IP injected 1hour after the last vehicle administration on day 14. This group served as the model group.
Group IV : Each rat was orally administered DMF (15mg/kg/day) for 14 consecutive days, then a single dose of DOX (90mg/kg) was IP -injected 1 hour after the last DMF treatment on day 14.
Twenty-four hours after DOX injection (i.e., at day 15), rats were anaesthetised using diethyl ether, blood samples were collected from the Jugular vein in non-heparinized tubes and were left to clot at room temperature, then centrifuged for 20 min at 4000 rpm to obtain serum and stored at -20°C for biochemical analysis; the animals were sacrificed by cervical dislocation, and the rats’ femoral bone marrow (BM) was harvested and processed for genotoxicity evaluations (17).