[Icariin enhances differentiation and maturation of rat calvarial osteoblasts in collagen hydrogel three-dimensional culture].
To investigate the effects of icariin on the differentiation and maturation of rat calvarial osteoblasts(ROB) in collagen hydrogel three-dimensional culture.



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.



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.



Icariin can enhance the expression of osteogenic markers of ROB in collagen hydrogel three-dimensional culture significantly.
School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China;2. Institute of Orthopedics, Lanzhou CPLC Command General Hospital, Lanzhou 730050, China.
Yan-fang
YF
Xie
School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China;2. Institute of Orthopedics, Lanzhou CPLC Command General Hospital, Lanzhou 730050, China.
Ming-gang
MG
Wang
School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China;2. Institute of Orthopedics, Lanzhou CPLC Command General Hospital, Lanzhou 730050, China.
Ke-ming
KM
Chen
School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China;2. Institute of Orthopedics, Lanzhou CPLC Command General Hospital, Lanzhou 730050, China.
Wen-gui
WG
Shi
School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China;2. Institute of Orthopedics, Lanzhou CPLC Command General Hospital, Lanzhou 730050, China.
Jian
J
Zhou
School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China;2. Institute of Orthopedics, Lanzhou CPLC Command General Hospital, Lanzhou 730050, China.
Yu-hai
YH
Gao
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000378:metabolism; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry*; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism; D004365:Drugs, Chinese Herbal; D005419:Flavonoids / Q000494:pharmacology*; D020100:Hydrogels / Q000737:chemistry*; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D012886:Skull / Q000166:cytology; D014157:Transcription Factors / Q000378:metabolism
C525720:Bmp2 protein, rat; D055396:Bone Morphogenetic Protein 2; D050659:Core Binding Factor Alpha 1 Subunit; D004365:Drugs, Chinese Herbal; D005419:Flavonoids; D020100:Hydrogels; C493717:Runx2 protein, rat; C495181:Sp7 protein, rat; D014157:Transcription Factors; D003094:Collagen; D000469:Alkaline Phosphatase; C056599:icariin