title: Immunohistochemical identification of type I and type III collagen and chondroitin sulphate in human pre-dentine: a correlative FEI-SEM/TEM study.
abstract: OBJECTIVE
  To identify type I- (I-CF) and type III-collagen fibrils (III-CF) and chondroitin 4/6 sulphate (CS) within human pre-dentine by means of a correlative analysis under field emission in-lens-scanning electron microscopy (FEI-SEM) and transmission electron microscopy (TEM).
  
  
  METHODS
  Human-extracted teeth were obtained and submitted to either a pre-embedding or a post-embedding immunolabelling procedure using monoclonal primary antibodies anti-I-CF, anti-III-CF and anti-CS. Gold-conjugated secondary antibodies were coupled to primary antibodies to visualize labelling under the electron beam. Correlative labelling patterns were obtained for I-CF and CS under both FEI-SEM and TEM.
  
  
  RESULTS
  Field emission in lens-SEM analysis revealed an intricate three-dimensional network of I-CF and CS clarifying the intimate relationship between the two main components of the pre-dentine organic matrix. TEM analysis revealed odontoblasts exhibiting intracellular labelling for CS, which became more intense and diffuse over the pre-dentine organic matrix. The same diffuse immunoreaction was revealed for I-CF, whereas a weak immunolocalization of III-CF was found scattered throughout the pre-dentine layer and over the collagen fibrils.
  
  
  CONCLUSIONS
  Both the pre- and post-embedding immunohistochemical approaches have led to the visualization of CF- and CS-labelling distribution within the pre-dentine layer, adding further knowledge on the elucidation of collagen-proteoglycans interaction in the organic matrix of human dental roots.
authors.affiliation: Department of Biomorphology, University of Chieti-Pescara, Chieti-Pescara, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Orsini
authors.forename: A
authors.initials: A
authors.lastname: Ruggeri
authors.forename: A
authors.initials: A
authors.lastname: Mazzoni
authors.forename: V
authors.initials: V
authors.lastname: Papa
authors.forename: M
authors.initials: M
authors.lastname: Piccirilli
authors.forename: M
authors.initials: M
authors.lastname: Falconi
authors.forename: R
authors.initials: R
authors.lastname: Di Lenarda
authors.forename: L
authors.initials: L
authors.lastname: Breschi
mesh_terms: D002809:Chondroitin Sulfates / Q000032:analysis*; D024042:Collagen Type I / Q000032:analysis*; D024061:Collagen Type III / Q000032:analysis*; D003804:Dentin / Q000737:chemistry* / Q000648:ultrastructure; D006801:Humans; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D011509:Proteoglycans / Q000032:analysis
chemical_list: D024042:Collagen Type I; D024061:Collagen Type III; D011509:Proteoglycans; D002809:Chondroitin Sulfates