Behavior of epithelial root sheath during tooth root formation in porcine molars: TUNEL, TEM, and immunohistochemical studies.

The behavior of epithelial cells during tooth root formation in porcine molars was investigated using TUNEL, immunohistochemical and transmission electron microscopic methods. Cells of Hertwig's epithelial root sheath (HERS) expressed bcl-2, but were TUNEL-negative. Epithelial cells of HERS were tightly connected and surrounded by a continuous basement membrane. When dental papilla cells were attached to the inner HERS, they became positive for osteopontin (OPN). The inner basement membrane was intermittent immediately after those cells started to synthesize dentin matrix, and the intercellular spaces of the HERS became wider. Dental sac cells then migrated between the fragmented HERS and were directly connected to epithelial cells by desmosome-like structures. After the cementum was synthesized by these mesenchymal cells, epithelial cells aggregated to form cell clusters, which were again surrounded by a continuous basement membrane. These results suggest that fragmentation of the HERS is due to the invasion of dental sac cells, but is not related to cell death. Furthermore, the differentiation of dental papilla cells into odontoblasts occurred following their connection to the HERS through the basement membrane. Moreover, it is suggested that the differentiation of dental sac cells into cementoblasts occurred upon their direct connection to epithelial cells of the HERS.