Piezo-power microdissection of mature human dental tissue.
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.
McLachlan
Julia L
JL
Oral Biology, School of Dentistry, University of Birmingham, St. Chads Queensway, Birmingham B4 6NN, UK.
Smith
Anthony J
AJ
Cooper
Paul R
PR
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
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