title: Removal of dentin matrix proteoglycans by trypsin digestion and its effect on dentin bonding.
abstract: The aim of this study was to investigate the effect of trypsin digestion on removal of chondroitin sulfate proteoglycans (CS-PGs) in demineralized dentin, and subsequent dentin bonding. Bovine dentin fragments were demineralized, treated with or without trypsin, stained with cupromeronic blue, and observed under transmission electron microscopy. Demineralized sections with or without trypsin digestion were also subjected to immunohistochemical analysis with anti-chondroitin-4-sulfate (C4S) monoclonal antibody, 2-B-6. The presence of galactosamine and glucosamine in the trypsin digest was confirmed by amino acid analysis. Bond strength testing was performed on trypsin treated and control specimens where samples were either kept moist or dried and re-wet, then bonded. Bond strength significantly decreased after trypsin treatment (p < 0.05). TEM, immunohistochemical, and amino acid analyses demonstrated that trypsin digestion efficiently removed C4S-PGs from demineralized dentin matrix. This study indicates that the detrimental effects observed on dentin bonding by trypsinization may be due in part to the removal/cleavage of the C4S-PGs, and further underscore the importance of C4S-PGs on dentin bonding.
authors.affiliation: Department of Restorative Dentistry, University of Illinois at Chicago, Illinois, USA.
authors.forename: Ana Karina B
authors.initials: AK
authors.lastname: Bedran-Russo
authors.forename: Patricia N R
authors.initials: PN
authors.lastname: Pereira
authors.forename: Wagner R
authors.initials: WR
authors.lastname: Duarte
authors.forename: Katsushi
authors.initials: K
authors.lastname: Okuyama
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Yamauchi
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D011508:Chondroitin Sulfate Proteoglycans / Q000235:genetics / Q000378:metabolism*; D001840:Dental Bonding; D003804:Dentin / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D008422:Materials Testing; D013718:Tensile Strength; D014357:Trypsin / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D011508:Chondroitin Sulfate Proteoglycans; C093073:Demineralized Dentin Matrix; D014357:Trypsin