title: VEGF and odontoblast-like cells: stimulation by low frequency ultrasound.
abstract: OBJECTIVE
  Vascular endothelial growth factor (VEGF) has been implicated in the regulation of dental pulp and dentine repair. Therapeutic ultrasound was shown to be effective for fracture repair. We investigated whether low frequency ultrasound influences the production of VEGF by odontoblast-like cells. Moreover, we examined the direct effects of VEGF on odontoblast-like cell proliferation.
  
  
  METHODS
  MDPC-23, an established odontoblast-like cell line, was exposed to increasing intensities of 30kHz ultrasound using an ultrasonic tip probe.
  
  
  RESULTS
  After 24h cell culture, WST-1 analysis of cell viability and number showed a dose-dependent decrease in the number of viable cells with increasing ultrasound power. However, the relative concentration of VEGF as analysed by ELISA and normalised to cell number was significantly increased in the culture supernatants indicating an ultrasound-induced stimulation of odontoblastic VEGF secretion. Analysis of VEGF gene expression by sqRT-PCR revealed the expression of the main VEGF isoforms in the MDPC-23 cells, i.e. VEGF(120) and VEGF(164) as well as to a minor extent VEGF(188). Low power ultrasound increased gene expression of all VEGF isoforms. Addition of recombinant VEGF to the cell cultures significantly stimulated cell proliferation. Gene expression of the VEGF receptors Flt1/VEGFR1 and KDR/VEGFR2 was detected in the MDPC-23, suggesting the possibility that VEGF may act on the odontoblast-like cells in an autocrine manner.
  
  
  CONCLUSIONS
  Our results indicate that ultrasound promoted VEGF expression and production by odontoblast-like cells and that VEGF may have autocrine effects on these cells. It is proposed that ultrasound may influence odontoblast activity and dentine repair by modulating production of endogenous growth factors in the dentine-pulp complex.
authors.affiliation: University of Birmingham School of Dentistry, St Chad's Queensway, Birmingham B46NN, UK. b.a.scheven@bham.ac.uk
authors.forename: B A
authors.initials: BA
authors.lastname: Scheven
authors.forename: J
authors.initials: J
authors.lastname: Man
authors.forename: J L
authors.initials: JL
authors.lastname: Millard
authors.forename: P R
authors.initials: PR
authors.lastname: Cooper
authors.forename: S C
authors.initials: SC
authors.lastname: Lea
authors.forename: A D
authors.initials: AD
authors.lastname: Walmsley
authors.forename: A J
authors.initials: AJ
authors.lastname: Smith
mesh_terms: D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D005786:Gene Expression Regulation; D051379:Mice; D009804:Odontoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011994:Recombinant Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014464:Ultrasonic Therapy; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology
chemical_list: D011994:Recombinant Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A