title: Osteogenic potential of punica granatum through matrix mineralization, cell cycle progression and runx2 gene expression in primary rat osteoblasts.
abstract: BACKGROUND
  Osteoporosis is one of the prevalent diseases in ageing populations. Due to side effects of many chemotherapeutic agents, there is always a need to search for herbal products to treat the disorder. Punica granatum (PG) represent a potent fruit-bearing medicinal herb which exerted valuable anti-osteoporotic activities. The present study was carried out to validate the in vitro osteogenic effects of the PG seed extract in primary calvarial osteoblast cultures harvested from neonatal rats.
  
  
  METHODS
  The ethanolic extract of PG was subjected to evaluate cell proliferation, regeneration, mineralization and formation of collagen matrix using MTT, alkaline phosphatase, Alizarin Red-S staining and Sirius Red dye, respectively. Cell cycle progression and osteogenic gene Runx2 expression were carried out by flow cytometry and real time PCR, respectively.
  
  
  RESULTS
  Exposure of different concentrations (10-100 μg/ml) of the extract on osteoblastic cells showed characteristic morphological changes and increment in cell number. A significant growth in cell proliferation, ALP activity, collagen contents and matrix mineralization of osteoblasts in a dose dependent manner (p < 0.05), suggested that PG has a stimulatory effect on osteoblastic bone formation or potential activity against osteoporosis. In addition, PG extract also enhanced DNA content in S phase of cell cycle and Runx2 gene expression level in osteoblasts.
  
  
  CONCLUSIONS
  The data clearly indicated that PG promoting bone cell proliferation and differentiation in primary osteoblasts might be due to elevating the osteogenic gene Runx2 expression. The present study provides an evidence for PG could be a promising herbal medicinal candidate that able to develop drugs for osteoporosis.
authors.affiliation: Molecular Endocrinology Laboratory, Department of Zoology, University of Lucknow, Lucknow, 226007, India. sahabjadabiotech04@gmail.com.
authors.forename: Sahabjada
authors.initials: S
authors.lastname: Siddiqui
authors.affiliation: Molecular Endocrinology Laboratory, Department of Zoology, University of Lucknow, Lucknow, 226007, India. molendolab@gmail.com.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Arshad
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D050071:Bone Density Conservation Agents / Q000494:pharmacology*; D002113:Calcification, Physiologic / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism*; D004261:DNA Replication / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D029561:Lythraceae; D010006:Osteoblasts / Q000187:drug effects* / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects*; D008517:Phytotherapy; D010936:Plant Extracts / Q000494:pharmacology*; D010946:Plants, Medicinal; D061251:Primary Cell Culture; D051381:Rats; D016196:S Phase / Q000187:drug effects*; D012639:Seeds; D015854:Up-Regulation
chemical_list: D050071:Bone Density Conservation Agents; D050659:Core Binding Factor Alpha 1 Subunit; D010936:Plant Extracts; C493717:Runx2 protein, rat; D003094:Collagen; D000469:Alkaline Phosphatase