Recombinant human erythropoietin improves angiogenesis and wound healing in experimental burn wounds.
OBJECTIVE
Erythropoietin interacts with vascular endothelial growth factor (VEGF) and stimulates endothelial cell mitosis and motility; thus it may be of importance in the complex phenomenon of wound healing. The purpose of this study was to investigate the effect of recombinant human erythropoietin (rHuEPO) on experimental burn wounds.


METHODS
Randomized experiment.


METHODS
Research laboratory.


METHODS
C57BL/6 male mice weighing 25-30 g.


METHODS
Mice were immersed in 80 degrees C water for 10 secs to achieve a deep-dermal second degree burn. Animals were randomized to receive either rHuEPO (400 units/kg/day for 14 days in 100 microL subcutaneously) or its vehicle alone (100 microl/day distilled water for 14 days subcutaneously). On day 14 the animals were killed. Burn areas were used for histologic examination, evaluation of neoangiogenesis by immunohistochemistry, and expression (Western blot) of the specific endothelial marker CD31 as well as quantification of microvessel density, measurement of VEGF wound content (enzyme-linked immunosorbent assay), expression (Western blot) of endothelial and inducible nitric oxide synthases, and determination of wound nitric oxide (NO) products.


RESULTS
rHuEPO increased burn wound reepithelialization and reduced the time to final wound closure. These effects were completely abated by a passive immunization with specific antibodies against erythropoietin. rHuEPO improved healing of burn wound through increased epithelial proliferation, maturation of the extracellular matrix, and angiogenesis. The hematopoietic factor augmented neoangiogenesis as suggested by the marked increase in microvessel density and by the robust expression of the specific endothelial marker CD31. Furthermore, rHuEPO enhanced the wound content of VEGF caused a marked expression of endothelial and inducible nitric oxide synthases and increased wound content of nitric oxide products.


CONCLUSIONS
Our study suggests that rHuEPO may be an effective therapeutic approach to improve clinical outcomes after thermal injury.
Galeano
Mariarosaria
M
Department of Clinical and Experimental Medicine and Pharmacology, University of Messina, Italy.
Altavilla
Domenica
D
Bitto
Alessandra
A
Minutoli
Letteria
L
Calò
Margherita
M
Lo Cascio
Patrizia
P
Polito
Francesca
F
Giugliano
Giovanni
G
Squadrito
Giovanni
G
Mioni
Chiara
C
Giuliani
Daniela
D
Venuti
Francesco S
FS
Squadrito
Francesco
F
D000818:Animals; D002056:Burns / Q000188:drug therapy*; D004921:Erythropoietin / Q000627:therapeutic use*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D011994:Recombinant Proteins; D012867:Skin / Q000033:anatomy & histology / Q000098:blood supply*; D014945:Wound Healing / Q000187:drug effects*
D011994:Recombinant Proteins; D004921:Erythropoietin