title: Piezo-power microdissection of mature human dental tissue.
abstract: Isolation of sufficient quantities of pure populations of odontoblasts from healthy and diseased teeth will facilitate our understanding of dentinogenesis during development and repair. Here we describe a novel Piezo-power microdissection (PPMD) technique for the isolation of pure populations of odontoblasts and pulpal tissue from formalin-fixed, paraffin-embedded, mature, healthy and carious human teeth. Odontoblasts and pulpal tissue gene expression were subsequently studied in ribonucleic acid isolated from PPMD preparations using a semi-quantitative reverse transcription polymerase chain reaction approach. Data confirmed that the genes for dentine sialophosphoprotein and Nestin are preferentially expressed in odontoblasts, whilst the genes for both collagen-1alpha and collagen-3alpha were expressed preferentially in pulpal tissue, particularly in carious samples. PPMD provides a novel and powerful approach to isolate pure populations of dental tissues and cells from fixed specimens for subsequent downstream molecular analyses.
authors.affiliation: Oral Biology, School of Dentistry, University of Birmingham, St. Chads Queensway, Birmingham B4 6NN, UK.
authors.forename: Julia L
authors.initials: JL
authors.lastname: McLachlan
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Smith
authors.forename: Paul R
authors.initials: PR
authors.lastname: Cooper
mesh_terms: D001483:Base Sequence; D024042:Collagen Type I / Q000032:analysis / Q000235:genetics; D024061:Collagen Type III / Q000032:analysis / Q000235:genetics; D003731:Dental Caries / Q000378:metabolism / Q000473:pathology / Q000601:surgery*; D003782:Dental Pulp / Q000378:metabolism* / Q000473:pathology; D003804:Dentin / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007381:Intermediate Filament Proteins / Q000032:analysis / Q000235:genetics; D042282:Microdissection / Q000379:methods*; D009419:Nerve Tissue Proteins; D064231:Nestin; D009804:Odontoblasts / Q000378:metabolism* / Q000473:pathology; D012313:RNA / Q000032:analysis; D012795:Sialoglycoproteins / Q000032:analysis
chemical_list: D024042:Collagen Type I; D024061:Collagen Type III; D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; D012795:Sialoglycoproteins; D012313:RNA