Erythropoietin and Cytoprotective Cytokines in Experimental Traumatic Brain Injury - Tissue-Protective Cytokines: Methods and Protocols - page 149

Table 2
Phases in the “Mild head injury complicated by hemorrhagic shock” model

Phase of study

Injury + hemorrhage Pre-hospital phase De fi nitive care

Ventilation Room air Room air 100% O 2

Injury Mild injury (3 m/s,

2.5 mm deformation)

- -

Hemorrhage Withdraw blood slowly till

MAP < 40 mmHg

- -

Resuscitation - Initial resuscitation with

saline to bring
MAP > 50 mmHg

Reinfuse shed blood and
additional saline to keep
MAP > 60 mmHg

0.3 ml/h. It is important to expel the air bubbles in the infusion
tubing.
6. Antibiotics and Analgesics: Enrofloxacin (available as 100 mg/
ml) is administered in the dosage of 0.05 ml/kg for both rats
and mice by subcutaneous route once daily starting on the day
of surgery for 3 days. Buprenorphine analgesic (available as
0.3 mg/ml) is given in the dosage of 0.1 mg/kg (rats) and
1 mg/kg (mice) by subcutaneous route twice daily for 3 days
starting on the day of surgery.
7. The study drugs (e.g., Epo, 5,000 U/kg) are administered by
intraperitoneal route or subcutaneous routes. The rats are
restrained in a polythene bag and the drug is injected into the
peritoneum. The mouse is held with one hand and stabilized
between the palm and fingers and then the drug is injected
into the peritoneum with the other hand.
8. TBI model complicated by hemorrhagic shock: After mild
TBI, blood is withdrawn into a syringe with citrate phosphate
dextrose to slowly drop the mean arterial pressure (MAP) to
40 mmHg. The blood volume withdrawn is usually 2 ml/100 g
weight of rat. Blood is slowly withdrawn (50% over first 5 min,
25% over next 5 min and 25% over the next 5 min) to mimic
the situation of traumatic blood loss in the battle field. The
shed blood is stored at 4°C. The physiological parameters are
monitored for 60 min during which anesthesia is maintained
with isoflurane and room air only. Following this hypotensive
period, lactated Ringer solution is infused at the rate of 1 ml/
min to maintain a MAP of 50 mm of Hg. This period mimics
the pre-hospital phase where resuscitation is with intravenous
fluids. The stored blood is rewarmed to 37°C, reperfused and
100% oxygen is restarted. The stages in this mild TBI with
hemorrhagic hypotension model are shown in Table 2 .