Icariin enhances differentiation and maturation of rat calvarial osteoblasts in collagen hydrogel three-dimensional culture / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
;
(6): 301-307, 2015.
Article
in Chinese
| WPRIM
| ID: wpr-255195
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of icariin on the differentiation and maturation of rat calvarial osteoblasts(ROB) in collagen hydrogel three-dimensional culture.</p><p><b>METHODS</b>ROB were obtained by enzyme digestion from the segregated neonatal SD rats skull and were embedded in 2 mg/mL rat tail collagen for three-dimensional culture. The growth state of ROB was observed by FDA/PI staining, HE staining and scanning electron microscopy. ROB were treated with icariin at the concentration of 1 × 10⁻⁴, 1 × 10⁻⁵, 1 × 10⁻⁶ and 1 × 10⁻⁷ mol/L respectively. The activity of alkaline phosphatase(ALP) was detected after 3, 6, 9 d of icariin treatment. Three-dimensional cultured ROB were treated with optimal concentration icariin for 12, 24, 36, 48 h and total RNA was extracted and the mRNA expressions of bone morphogenetic protein-2 (BMP-2), Runt-related transcription factor 2 (RUNX-2) and Osterix were detected by real time RT-PCR. The protein expression of BMP-2, RUNX-2 and Osterix were examined by Western-blotting.</p><p><b>RESULTS</b>ROB were cultured in collagen hydrogel successfully. FDA/PI staining, HE staining, and scanning electron microscopy showed that ROB adhered with collagen tightly and distributed homogeneously. Icariin at final concentration of 1 × 10⁻⁵, 1 × 10⁻⁶ and 1×10⁻⁷ mol/L all enhanced the activity of ALP of collagen hydrogel three-dimensional cultured ROB, and 1 × 10⁻⁶ mol/L was the optimal concentration. Besides, icariin (1 × 10⁻⁶ mol/L) increased mRNA and protein expression of BMP-2、RUNX-2 and Osterix compared to control group.</p><p><b>CONCLUSION</b>Icariin can enhance the expression of osteogenic markers of ROB in collagen hydrogel three-dimensional culture significantly.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Osteoblasts
/
Pharmacology
/
Skull
/
Transcription Factors
/
Flavonoids
/
Drugs, Chinese Herbal
/
Cell Differentiation
/
Cells, Cultured
/
Chemistry
/
Collagen
Limits:
Animals
Language:
Chinese
Journal:
Journal of Zhejiang University. Medical sciences
Year:
2015
Type:
Article
Similar
MEDLINE
...
LILACS
LIS