HIF-1alpha and VEGF Expression in Fracture Healing / 대한정형외과학회잡지
The Journal of the Korean Orthopaedic Association
;
: 479-487, 2008.
Artigo
em Coreano
| WPRIM
| ID: wpr-652613
ABSTRACT
PURPOSE:
To elucidate the relation between fracture healing and angiogenesis, we checked expression of Hypoxia-inducible factor (HIF) and Vascular endothelial growth factor (VEGF) in hypoxic cell cultures and the callus from a rat femur fracture model. MATERIALS ANDMETHODS:
Human osteoblasts, chondrocytes, and rat ST2 cells were cultured in DME/F12 media with 10% FBS. Hypoxic DME/F12 media (PO2<60 mmHg) was generated by bubbling with 95% N2 and 5% CO2 and added to cells. After 2, 6, and 24 hours, RNA and proteins were collected for reverse transcription - polymerase chain reaction (RT-PCR) and Western blot. In addition, immunocytochemistry and siRNA treatment for HIF-1alpha were performed. Next, femurs from 9-week SD rats were fractured after fixation with needles. The rats were sacrificed at post-fracture day (PFD) 3, 5, 7, 10, 14, 21 and calluses were collected for RT-PCR and Western blot.RESULTS:
HIF-1alpha and HIF-2alpha expression were not increased in RT-PCR but protein levels were increased. VEGF expression in RT-PCR was increased. Treatment with siRNA directed towards HIF inhibited VEGF expression. In the rat fracture callus, HIF-1alpha and VEGF expression peaked between PFD 5 and 7 and decreased after PFD 10. In contrast to cell culture, mRNA expression of HIF-1alpha was increased at PFD 7.CONCLUSION:
HIF-1alpha and VEGF peaked early in fracture healing. With expression decreasing as O2 tension increased. Further study is needed to identify other factors affecting chondrogenic differentiation.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Osteoblastos
/
Calo Ósseo
/
RNA
/
RNA Mensageiro
/
Imuno-Histoquímica
/
Proteínas
/
Western Blotting
/
Reação em Cadeia da Polimerase
/
Consolidação da Fratura
/
Técnicas de Cultura de Células
Tipo de estudo:
Estudo prognóstico
Limite:
Animais
/
Humanos
Idioma:
Coreano
Revista:
The Journal of the Korean Orthopaedic Association
Ano de publicação:
2008
Tipo de documento:
Artigo
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