[Icariin enhances differentiation and maturation of rat calvarial osteoblasts in collagen hydrogel three-dimensional culture].
Zhejiang Da Xue Xue Bao Yi Xue Ban
; 44(3): 301-7, 2015 05.
Article
em Zh
| MEDLINE
| ID: mdl-26350011
ABSTRACT
OBJECTIVE:
To investigate the effects of icariin on the differentiation and maturation of rat calvarial osteoblasts(ROB) in collagen hydrogel three-dimensional culture.METHODS:
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 × 10â»4, 1 × 10â»5, 1 × 10â»6 and 1 × 10â»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.RESULTS:
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 × 10â»5, 1 × 10â»6 and 1×10â»7 mol/L all enhanced the activity of ALP of collagen hydrogel three-dimensional cultured ROB, and 1 × 10â»6 mol/L was the optimal concentration. Besides, icariin (1 × 10â»6 mol/L) increased mRNA and protein expression of BMP-2ãRUNX-2 and Osterix compared to control group.CONCLUSION:
Icariin can enhance the expression of osteogenic markers of ROB in collagen hydrogel three-dimensional culture significantly.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Osteoblastos
/
Flavonoides
/
Colágeno
/
Hidrogéis
Limite:
Animals
Idioma:
Zh
Ano de publicação:
2015
Tipo de documento:
Article