title: The proliferation, differentiation, and mineralization effects of puerarin on osteoblasts in vitro.
abstract: OBJECTIVE
  Osteoblasts are key functional cells in the process of bone metabolic balance. Phytoestrogens have an important influence on the proliferation and differentiation of osteoblasts. Puerarin, a plant estrogen, has a wide range concentration in vitro on the function of osteoblasts. The current study investigates the effect of the phytoestrogen puerarin on the proliferation, differentiation, and mineralization of osteoblasts in vitro.
  
  
  METHODS
  The calvaria bone of eight-ten Wistar rats which were born within 24 h were obtained in aseptic condition. After enzyme digestion, isolation, purified osteoblasts of rats were cultured for further study. The cells of the first to third generation were divided into a control group and a puerarin-treated group with 10(-3)-10(-10) mol·L(-1) puerarin. The cells were exposed to the medium containing a low level of carbohydrates, 10% (V/V) FBS for 24 h. After 1 to 4 days of culture, the OD values on the proliferation of osteoblasts in each group were determined by microplate reader. The cells were cultured in the medium containing 50 μg·mL(-1) vitamin C, 10(-2) mol·L(-1) sodium glycerophosphate, 10% FBS and the medium was changed every 3 to 4 days. After 2 to 8 days of culture, expression of alkaline phosphatase were tested and compared by microplate reader. The mineral nodes of osteoblasts were dyed using alizarin red or improved Von Kossa way after four weeks.
  
  
  RESULTS
  Compared with those in the 10(-5)-10(-9) mol·L(-1) puerarin, the proliferation of osteoblasts, the expression of alkaline phosphatase, and the number of mineral nodes of osteoblasts were significantly decreased in the control group. The increase was the fastest in the third day, while on the fourth day it was decreased, and arrived at statistical significance compared with the alkaline phosphatase activities and control group. The 10(-6) mol·L(-1) group was the most distinct, and formed the most mineralized nodule. Compared with the 10(-3) mol·L(-1) puerarin group, those changes were markedly increased in the control group.
  
  
  CONCLUSIONS
  Puerarin has proliferation, differentiation, and mineralization effects on osteoblasts in a dose-dependent manner, and has a double-way effect on the osteoblasts in vitro. A low-dose showed positive effects on the development of osteoblasts, and high-dose puerarin could inhibit the formation of bone.
authors: The Affiliated Guang-ren Hospital of Xi'an Jiaotong University Medical School, Xi'an No. 4 Hospital, Xi'an 710004, China.
authors: Chang
authors: C
authors: Wang
authors: The Affiliated Guang-ren Hospital of Xi'an Jiaotong University Medical School, Xi'an No. 4 Hospital, Xi'an 710004, China.
authors: Mei-Xia
authors: MX
authors: Meng
authors: Department of Endocrinology, First Affiliated Hospital of Lanzhou University, Lanzhou 730000, China.
authors: Xu-Lei
authors: XL
authors: Tang
authors: Institute of Orthopaedics, Lanzhou General Hospital, Lanzhou Command, PLA, Lanzhou 730050, China.
authors: Ke-Ming
authors: KM
authors: Chen
authors: The Affiliated Guang-ren Hospital of Xi'an Jiaotong University Medical School, Xi'an No. 4 Hospital, Xi'an 710004, China. Electronic address: sdd1962@126.com.
authors: Li
authors: L
authors: Zhang
authors: Department of Medicine, Chinese Medicine Hospital of the Baqiao District of the Xi'an, Xi'an 710038, China.
authors: Wei-Ning
authors: WN
authors: Liu
authors: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an 710069, China. Electronic address: zyy@nwu.edu.cn.
authors: Ying-Yong
authors: YY
authors: Zhao
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D015519:Bone Density; D001842:Bone and Bones / Q000166:cytology / Q000187:drug effects*; D002113:Calcification, Physiologic / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005260:Female; D066298:In Vitro Techniques; D007529:Isoflavones / Q000494:pharmacology*; D010006:Osteoblasts / Q000187:drug effects*; D048789:Phytoestrogens / Q000494:pharmacology*; D017208:Rats, Wistar
chemical_list: D007529:Isoflavones; D048789:Phytoestrogens; D000469:Alkaline Phosphatase; C033607:puerarin
keywords: Differentiation; Mineralization; Osteoblast; Phytoestrogen; Proliferation; Puerarin