Background: Bone sialoprotein (BSP) and osteopontin (OPN), two major noncollagenous proteins (NCPs) in collagen-based mineralized tissues, have been implicated in mineral deposition and cell- and matrix-matrix interactions during root development. However, their role in cementogenesis is still a subject of debate. Since distribution of proteins is indicative of function, we have analyzed their temporo-spatial appearance in relation to that of cementum collagen.
Methods: Human premolars and rat molars at various stages of root development characterized by differing rates of formation were fixed in aldehyde and embedded in epoxy and LR White resin. Sections were processed for ultrastructural analysis and postembedding colloidal gold (immuno)cytochemistry.
Results: Incubations with antibodies against BSP and OPN and with lectins recognizing prominent sugars in these proteins generally revealed similar labeling patterns in both human and rat teeth, with gold particles accumulating mainly in the interfibrillar spaces. The lectin Helix pomatia, specific for N-acetyl-D-galactosamine, was distinctive in that it consistently reacted with human cementum, but only sporadically labeled rat cementum. Regardless of both the species and the stage of root development, mineralization initiated in mantle predentin in association with distinct foci immunoreactive for BSP and OPN. In human teeth, the deposition of cementum collagen began before the start of dentin mineralization and thus prior to any detectable labeling for BSP and OPN. However, at early stages of root formation in the rat, cementum collagen appeared after BSP and OPN accumulated on the root surface, whereas at advanced stages the deposition of cementum collagen, BSP and OPN coincided.
Conclusions: The temporo-spatial differences in the appearance of BSP and OPN relative to cementum collagen correlate well with known differences in the speed of root elongation and explain the variable appearance of the dentino-cemental junction. The data reveal no causal relationship between BSP and OPN and the differentiation of cementoprogenitor cells and indicate that the distribution of collagen fibrils ultimately determines the amount and pattern of accumulation of these NCPs. There also is no consistent planar accumulation of BSP and OPN between dentin and cementum such as the cement lines found between "old" and "new" bone. It is concluded that the interlacement of collagen fibrils at the dentino-cemental junction, across which mineralization spreads, represents the primary attachment mechanism between cementum and dentin.