HIF-1alpha and VEGF Expression in Fracture Healing / 대한정형외과학회잡지
The Journal of the Korean Orthopaedic Association
;
: 479-487, 2008.
Article
in Korean
| 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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Osteoblasts
/
Bony Callus
/
RNA
/
RNA, Messenger
/
Immunohistochemistry
/
Proteins
/
Blotting, Western
/
Polymerase Chain Reaction
/
Fracture Healing
/
Cell Culture Techniques
Type of study:
Prognostic study
Limits:
Animals
/
Humans
Language:
Korean
Journal:
The Journal of the Korean Orthopaedic Association
Year:
2008
Type:
Article
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