title: Ascorbic acid induces osteoblast differentiation of human suspension mononuclear cells.
abstract: OBJECTIVE
  Suspension mononuclear cells (MNCs) can be differentiated into osteoblasts with the induction of ascorbic acid and β-glycerophosphate. The aim of this study was to determine the ability of suspension MNCs to differentiate into osteoblasts using ascorbic acid only.
  
  
  METHODS
  Suspension MNCs were obtained by a combination of gradient centrifugation and culture selection. Suspension MNCs were subjected to differentiation assay by culturing them inside proliferation medium supplemented with 10 μg/mL, 30 μg/mL, 50 μg/mL, 60 μg/mL, 90 μg/mL and 500 μg/mL of ascorbic acid. Proliferation medium supplemented with 50 μg/mL ascorbic acid and 10 mmol/L β-glycerophosphate was used as a positive control for osteoblast induction, and proliferation medium without ascorbic acid was used as a negative control. Differentiation analysis was performed using alkaline phosphatase (ALP) assay, von Kossa staining and expression of osteoblast-related genes.
  
  
  RESULTS
  With all concentrations of ascorbic acid used, there was a significant increase (P < 0.05) in ALP-specific activity and mineralized nodule formation throughout the differentiation course compared with negative control. Ascorbic acid was also able to activate the expression of osteopontin (SPP1), osteonectin (SPARC) and runt-related transcription factor 2 (RUNX2) messenger RNA in positive control and ascorbic acid-induced MNCs (30 μg/mL and 90 μg/mL) but not in negative control.
  
  
  CONCLUSIONS
  Ascorbic acid alone was sufficient to induce osteoblast differentiation from suspension MNCs; 30-90 μg/mL of ascorbic acid was found to be the optimal concentration. Ascorbic acid can be used as a nutritional supplement for cellular therapy of bone-related disease.
authors.affiliation: School of Biosciences & Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Bangi, Selangor, Malaysia.
authors.forename: Siti Norhaiza
authors.initials: SN
authors.lastname: Hadzir
authors.affiliation: School of Biosciences & Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Bangi, Selangor, Malaysia.
authors.forename: Siti Norsaidah
authors.initials: SN
authors.lastname: Ibrahim
authors.affiliation: Department of Orthodontics, Faculty of Dentistry, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.
authors.forename: Rohaya Megat
authors.initials: RM
authors.lastname: Abdul Wahab
authors.affiliation: School of Biosciences & Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Bangi, Selangor, Malaysia.
authors.forename: Intan Zarina
authors.initials: IZ
authors.lastname: Zainol Abidin
authors.affiliation: School of Biosciences & Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Bangi, Selangor, Malaysia.
authors.forename: Sahidan
authors.initials: S
authors.lastname: Senafi
authors.affiliation: School of Biology, Faculty of Applied Science, Universiti Teknologi MARA, Shah Alam, Selangor, Malaysia. Electronic address: hisham@ukm.my.
authors.forename: Zaidah Zainal
authors.initials: ZZ
authors.lastname: Ariffin
authors.affiliation: Allianze University College of Medical Sciences, Waziria Medical Square, Kepala Batas, Penang, Malaysia.
authors.forename: Mohamad
authors.initials: M
authors.lastname: Abdul Razak
authors.affiliation: School of Biosciences & Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Bangi, Selangor, Malaysia.
authors.forename: Shahrul Hisham
authors.initials: SH
authors.lastname: Zainal Ariffin
mesh_terms: D001205:Ascorbic Acid / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000187:drug effects*; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects; D010012:Osteogenesis / Q000187:drug effects; D015676:Osteonectin / Q000235:genetics; D053495:Osteopontin / Q000235:genetics
chemical_list: D050659:Core Binding Factor Alpha 1 Subunit; D015676:Osteonectin; D053495:Osteopontin; D001205:Ascorbic Acid
keywords: alkaline phosphatase; ascorbic acid; mineralization; osteoblasts; suspension mononuclear cell