Extracellular matrix in tooth cementum and mantle dentin: localization of osteopontin and other noncollagenous proteins, plasma proteins, and glycoconjugates by electron microscopy.

Abstract

Noncollagenous proteins (NCPs) are considered to have multiple functions related to the formation, turnover, and repair of the collagen-based mineralized tissues. Collectively, they comprise a class of generally acidic, mineral-binding proteins showing extensive posttranslational modifications, including glycosylation, phosphorylation, and sulfation. METHODS. We have used colloidal-gold immunocytochemistry and lectin-gold cytochemistry, together with transmission electron microscopy, to examine the organic matrix composition of tooth cementum and the subjacent mantle dentin in rodent molar teeth. Molars were processed for immunocytochemistry using antibodies against osteopontin (OPN), osteocalcin (OC), bone sialoprotein (BSP), bone acidic glycoprotein-75 (BAG-75), albumin (ALB), and alpha 2HS-glycoprotein (alpha 2HS-GP), or for glycoconjugate cytochemistry using lectin-gold complexes. Ultrastructurally, at the advancing root edge in developing molars, OPN and BSP initially were associated with small calcification foci in the mantle dentin. With progressing mineralization, OC and alpha 2HS-GP appeared diffusely distributed throughout the calcified mantle dentin, and diminished as a gradient toward the circumpulpal dentin. Immediately following disruption of Hertwig's epithelial root sheath, cementum deposition commenced at the root surface occasionally with the appearance of a cement line rich in OPN. Cementum matrix proper contained abundant OPN, BSP, OC, and alpha 2HS-GP, but no or little BAG-75 or ALB. Protein immunolabeling, as well as lectin labeling for beta-D-galactose and N-acetyl-neuraminic acid and/or N-glycolyl-neuraminic acid, both being prominent sugars of certain NCPs, was primarily concentrated between, and at the surface of, collagen fibrils in acellular extrinsic fiber cementum. OPN, BSP, OC, and alpha 2HS-GP were also prominent components of cellular cementum and of Sharpey's fibers. In cellular cementum, laminae limitantes sometimes present delimiting cementocyte lacunae and cell process-containing canaliculi were also rich in OPN. Along the root surface, occasional cementoblasts exhibited intracellular labeling for OPN over the Golgi apparatus and secretory granules. We have identified OPN, BSP, OC, and alpha 2HS-GP as being prominent organic constituents of both mantle dentin and acellular and cellular cementum, and, have elucidated the details of their distribution at the ultrastructural level. The temporal appearance and spatial distribution of these organic moieties in the teeth root are similar to those seen during bone formation and are consistent with proposals that certain NCPs may be involved in regulating calcification and/or participating in cell-matrix and matrix-matrix/mineral adhesion events.