title: Purmorphamine increased adhesion, proliferation and expression of osteoblast phenotype markers of human dental pulp stem cells cultured on beta-tricalcium phosphate.
abstract: OBJECTIVE
  Growth factors play a significant role in cell proliferation and differentiation during different stages of the bone repair. However, several limitations have been brought researchers attention to an osteoinductive small molecule including Purmorphamine. In this study, we aimed to evaluate the effect of Purmorphamine on adhesion, proliferation and differentiation of human dental pulp stem cells (hDPSCs) seaded on beta-tricalcium phosphate (β-TCP) granules.
  
  
  METHODS
  hDPSCs were established from extracted wisdom teeth of healthy volenteers. Cells at passage 3 were seeded on β-TCP in the presence or absence of Purmorphamine. Cell adhesion and proliferation were assessed using scanning electeron microscopy (SEM) and DNA counting assay, respectively, after 1, 3 and 5days. Then, hDPSCs seeded on β-TCP were subjected to osteogenic medium with or without Purmorphamine. After 7 and 14days osteogenic diffrentiation capability of hDPSCs were determined using real-time RT-PCR and alkaline phosphatase (ALP) activity assay.
  
  
  RESULTS
  The significant increase in amount of DNA was observed at day 3 and 5 in the presence of Purmorphamine. SEM imaging also was confirmed the DNA counting assay; in all given time points, hDPSC attachment and growth was significantly higher in the presence of Purmorphamine. ALP activity was increased by Purmorphamine at both 7 and 14days of induction. Purmorphamine showed to effect on osteopontin expression at earlier stage of osteogenic differentiation, whereas for osteocalcin expression, this effect was more evident at later stage of differentiation.
  
  
  CONCLUSIONS
  Purmorphamine had a promotive effect on adhesion, proliferation and osteogenic differentiation of hDPSCs cultured on β-TCP. The outcome of the current study would help in development of in vitro culture conditions for better osteogenic differentiation of hDPSCs prior to transplantation.
authors.affiliation: Research Institute of Dental Sciences, Dental Research Center, Dental School, Shahid Beheshti University of Medical Sciences, Tehran 19839, Iran. Electronic address: m.rezai.rad@gmail.com.
authors.forename: Maryam
authors.initials: M
authors.lastname: Rezia Rad
authors.affiliation: Research Institute of Dental Sciences, Dental Research Center, Dental School, Shahid Beheshti University of Medical Sciences, Tehran 19839, Iran. Electronic address: moein.khojaste@yahoo.com.
authors.forename: Moein
authors.initials: M
authors.lastname: Khojaste
authors.affiliation: Research Institute of Dental Sciences, Dental Research Center, Dental School, Shahid Beheshti University of Medical Sciences, Tehran 19839, Iran. Electronic address: shahriarim@yahoo.com.
authors.forename: Mehrnoosh
authors.initials: M
authors.lastname: Hasan Shahriari
authors.affiliation: Iranian Center of Endodontic Research, Dental Research Center, Dental School, Shahid Beheshti University of Medical Sciences, Tehran 19839, Iran; Department of Endodontics, Dental School, Shahid Beheshti University of Medical Sciences, Tehran 19839, Iran. Electronic address: saasgar@yahoo.com.
authors.forename: Saeed
authors.initials: S
authors.lastname: Asgary
authors.affiliation: Research Institute of Dental Sciences, Dental Research Center, Dental School, Shahid Beheshti University of Medical Sciences, Tehran 19839, Iran; Department of Oral and Maxillofacial Surgery, Dental School, Shahid Beheshti University of Medical Sciences, Tehran 19839, Iran; School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran 19839, Iran. Electronic address: arashkhojasteh@yahoo.com.
authors.forename: Arash
authors.initials: A
authors.lastname: Khojasteh
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism*; D002130:Calcium Phosphates / Q000494:pharmacology*; D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003782:Dental Pulp / Q000166:cytology*; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D009025:Morpholines / Q000494:pharmacology*; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D015675:Osteocalcin / Q000235:genetics / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects; D053495:Osteopontin / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D011687:Purines / Q000494:pharmacology*; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure
chemical_list: D015415:Biomarkers; D002130:Calcium Phosphates; D009025:Morpholines; D011687:Purines; C485817:beta-tricalcium phosphate; D015675:Osteocalcin; D053495:Osteopontin; D000469:Alkaline Phosphatase; C470893:purmorphamine
keywords: Adhesion; Differentiation; Human dental pulp stem cells; Osteoinductive small molecule; Proliferation; Purmorphamine; β-TCP