Immunohistochemical characterization of noncollagenous matrix molecules on the alveolar bone surface at the initial principal fiber attachment in rat molars

Abstract

This study was designed to immunodetect proteoglycans (PGs) and the noncollagenous glycoproteins, bone sialoprotein (BSP) and osteopontin (OPN) on developing alveolar bone surface in rat molars by the indirect immunoperoxidase method, and to discuss the roles of these molecules at the initial principal fiber (PF) attachment. To characterize PGs, antibodies against five species of glycosaminoglycans (GAGs), chondroitin-4-sulfate (C4S), chondroitin-6-sulfate (C6S), unsulfated chondroitin (C0S), dermatan sulfate (DS), and keratan sulfate (KS) were used. Maxillary alveolar bone facing the distal root of the second molar was examined in 20- and 25-day-old male Wistar rats. Routine histological staining was also used. A hematoxylin-stained, fibril-poor layer always appeared on the alveolar bone surface just prior to the initial PF organization. This layer was strongly immunoreactive for C4S, C0S, OPN, and BSP, and weakly for C6S, but not for DS and KS. Then the initial PFs were attached to this layer. When new bone containing Sharpey's fibers covered this layer, it remained as a hematoxylin-stained, fibril-poor layer between Sharpey's fiber-containing and -lacking bone. The layer was consistently immunoreactive for OPN and BSP but had no immunoreactivity for GAGs. The results suggest that the accumulation of C4S-, C0S-, and C6S-carrying PGs, and of BSP and OPN is a primary event at the initial PF attachment, and is involved in the adhesion of PFs and mineralization of the initial attachment layer. The BSP and OPN act to maintain the interface integrity between Sharpey's fiber-containing and Sharpey's fiber-lacking alveolar bone after the PF attachment is established.