Carbamylated erythropoietin is neuroprotective in an experimental model of traumatic brain injury.
OBJECTIVE
The well-documented neuroprotective effects of recombinant human erythropoietin (rhEPO) are commonly associated with untoward erythrocyte-stimulating effects (polycythemia), with subsequent risk of thromboembolic complications. A carbamylated-rhEPO (CEPO) derivative, which is neuroprotective but lacks hematopoietic activity, has been recently developed. In this study, we evaluated the neuroprotective capability of CEPO in an in vitro model of cerebral trauma in which rhEPO was previously shown to reduce posttraumatic cell death.


METHODS
Prospective, controlled experiment.


METHODS
Animal, basic science laboratory.


METHODS
Wistar rats, 8 days old.


METHODS
Organotypic hippocampal slices, obtained from rat brains, were subjected to a well-characterized model of mechanical injury followed by addition of 10 IU/mL rhEPO, 10-100 IU/mL CEPO, or vehicle (injured control) to the incubation medium at different times to assess the temporal window of therapeutic neuroprotection.


RESULTS
Posttraumatic cell death was quantified at 12, 24, or 48 hrs after injury by measuring propidium iodide fluorescence in the selectively vulnerable CA1 hippocampal area. Posttraumatic injury, observed in injured, vehicle-treated hippocampal slices, was significantly attenuated by addition of either 10 IU/mL rhEPO or 10 IU/mL CEPO. The neuroprotective efficacy of 10 IU/mL rhEPO or CEPO remained intact even when administration was delayed 1 hr after trauma. Qualitative microscopy in semithin sections showed that both rhEPO and CEPO exerted a marked pyramidal neuron-sparing effect.


CONCLUSIONS
Our study shows that 10 IU/mL CEPO exerts neuroprotective effects comparable with those of rhEPO in an in vitro model of mechanical cerebral trauma. Because CEPO lacks hematopoietic effects and seems to possess a prolonged therapeutic time window, this erythropoietin derivative may represent an exciting new pharmacologic tool in treating patients with mechanical injury to the brain.
Adembri
Chiara
C
Section of Anesthesiology and Intensive Care, Department of Critical Care Medicine, University of Florence, Florence, Italy. chiara.adembri@unifi.it
Massagrande
Alessandra
A
Tani
Alessia
A
Miranda
Marco
M
Margheri
Martina
M
De Gaudio
Raffaele
R
Pellegrini-Giampietro
Domenico E
DE
D000818:Animals; D001930:Brain Injuries / Q000517:prevention & control*; D004921:Erythropoietin / Q000031:analogs & derivatives* / Q000627:therapeutic use; D066298:In Vitro Techniques; D018696:Neuroprotective Agents / Q000627:therapeutic use*; D051381:Rats; D017208:Rats, Wistar
D018696:Neuroprotective Agents; C488171:carbamylated erythropoietin; D004921:Erythropoietin