title: Pueraria mirifica extract and puerarin enhance proliferation and expression of alkaline phosphatase and type I collagen in primary baboon osteoblasts.
abstract: Phytoestrogen-rich Pueraria mirifica (PM) tuberous extract is a promising candidate for the development of anti-osteoporosis drugs for postmenopausal women, but its action has never been validated in humans or in non-human primates, which are more closely related to humans than rodents. In vitro study of non-human primate osteoblasts is thus fundamental to prepare for in vivo studies of phytoestrogen effects on primate bone. This study aimed to establish a culture system of baboon primary osteoblasts and to investigate the effects of PM extract and its phytoestrogens on these cells. Primary osteoblasts from adult baboon fibulae exhibited osteoblast characteristics in regard to proliferation, differentiation, mineralization, and estrogen receptor expression. They responded to 17β-estradiol by increased proliferation rate and mRNA levels of alkaline phosphatase (ALP), type I collagen, and osteocalcin. After being exposed for 48 h to 100 μg/ml PM extract, 1000 nM genistein, or 1000 nM puerarin, primary baboon osteoblasts markedly increased the rate of proliferation and mRNA levels of ALP and type I collagen without changes in Runx2, osterix, or osteocalcin expression. PM extract, genistein, and puerarin also decreased the RANKL/OPG ratio, suggesting that they could decrease osteoclast-mediated bone resorption. However, neither PM extract nor its phytoestrogens altered calcium deposition in osteoblast culture. In conclusion, we have established baboon primary osteoblast culture, which is a new tool for bone research and drug discovery. Furthermore, the present results provide substantial support for the potential of PM extract and its phytoestrogens to be developed as therapeutic agents against bone fragility.
authors.affiliation: Biological Sciences Program, Faculty of Science, Chulalongkorn University, Bangkok, Thailand; Department of Biology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.
authors.forename: Wacharaporn
authors.initials: W
authors.lastname: Tiyasatkulkovit
authors.affiliation: Department of Biology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand. Electronic address: Suchinda.M@chula.ac.th.
authors.forename: Suchinda
authors.initials: S
authors.lastname: Malaivijitnond
authors.affiliation: Department of Physiology, Faculty of Science, Mahidol University, Bangkok, Thailand; Center of Calcium and Bone Research (COCAB), Faculty of Science, Mahidol University, Bangkok, Thailand.
authors.forename: Narattaphol
authors.initials: N
authors.lastname: Charoenphandhu
authors.affiliation: Southwest National Primate Research Center and Department of Genetics, Texas Biomedical Research Institute, San Antonio, TX 78245, USA.
authors.forename: Lorena M
authors.initials: LM
authors.lastname: Havill
authors.affiliation: Southwest National Primate Research Center and Department of Genetics, Texas Biomedical Research Institute, San Antonio, TX 78245, USA.
authors.forename: Allen L
authors.initials: AL
authors.lastname: Ford
authors.affiliation: Southwest National Primate Research Center and Department of Genetics, Texas Biomedical Research Institute, San Antonio, TX 78245, USA.
authors.forename: John L
authors.initials: JL
authors.lastname: VandeBerg
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism*; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D024042:Collagen Type I / Q000378:metabolism*; D005260:Female; D007529:Isoflavones / Q000494:pharmacology*; D010006:Osteoblasts / Q000187:drug effects*; D010215:Papio; D048789:Phytoestrogens / Q000494:pharmacology*; D010936:Plant Extracts / Q000494:pharmacology*; D061251:Primary Cell Culture; D029908:Pueraria / Q000737:chemistry; D011960:Receptors, Estrogen / Q000378:metabolism
chemical_list: D024042:Collagen Type I; D007529:Isoflavones; D048789:Phytoestrogens; D010936:Plant Extracts; D011960:Receptors, Estrogen; D000469:Alkaline Phosphatase; C033607:puerarin
keywords: Baboon; Genistein; Osteoblasts; Osteoporosis; Pueraria mirifica; Puerarin