Immuno-scanning electron microscope characterization of large tubules in human deciduous dentin

This study was undertaken to elucidate the type and origin of collagen fibrils which construct the large tubules in deciduous coronal dentin by scanning electron microscope and anti-types I and III collagen antibody procedures.

The studies were performed on human deciduous teeth. The teeth were fixed in 4% paraformaldehyde solution and then fractured either mesio-distally parallel to the long axis of the tooth or transversely perpendicular to the long axis of the tooth crown. The specimens were three-dimensionally observed employing the scanning electron microscope to distinguish the content of large tubules. Polyclonal antibodies of anti-type I and anti-type III collagen with 20 nm colloidal gold, and secondary electron imaging and backscatter electron imaging of high-resolution field emission scanning electron microscopy were used to examine the types of collagen fibrils.

The large tubules extended from the vicinity of the incisal edge of the dentino-enamel junction to the pulp cavity. Inside the large tubules, fibers in compact bundles run parallel to the longitudinal axis of the tubules. The fiber bundles consisted of collagen fibrils which were 50-150 nm in diameter with typical cross striation. Immuno-scanning electron microscopy showed type I collagen-labelling gold particles and type III collagen-labelling gold particles to be abundant on the fibrils. Types I and III collagen-labelling gold particles were present on the banded collagen fibrils regardless of their diameter.

It was found that type III collagen is present together with type I collagen on the fibrils constructing the large tubules of the human deciduous dentin. This immunohistochemical study suggested that the fibrils constructing the large tubules were derived from the von Korff fibers, and types I and III collagens formed copolymers.