Induction of odontoblast-like cell differentiation in dog dental pulps after in vivo implantation of dentine matrix components.

The effects of dentine extracellular matrix components on dental mesenchymal cells were studied by light and transmission electron microscopy after their implantation at central sites of mechanically exposed pulps in dog molar teeth. The implants were Millipore filters that had been soaked with solutions containing 30 or 300 micrograms/ml of an EDTA-soluble fraction of rabbit incisor dentine. Control filters were soaked with dog albumin or phosphate buffered saline. Columnar, polarized cells were consistently seen after 8 days in close proximity to the filters coated with both concentrations of dentine matrix components. Characteristic features of these polarized cells included widened cisternae of the rough endoplasmic reticulum, a rich microfilamentous network in the long cytoplasmic extensions invading the filter pores and numerous cytoplasmic bodies. These cells also showed evidence of functional as well as cytological differentiation. Polarized processing of secretory granules could be observed after 8 days' implantation, and also the presence of matrix vesicles and deposition of a fine, collagenous matrix into the filters apically to the distal end of the cytoplasmic processes. After 24 days' implantation, secretion of a tubular matrix could be consistently seen in association with the odontoblast-like cells. No changes in cell organization or matrix synthesis were seen after implantation of control filters. These studies demonstrate that bioactive components present in the EDTA-soluble dentine matrix fraction are able to directly induce cell polarization and apical secretion of tubular matrix when implanted in contact with dental pulp cells at sites remote from the odontoblast layer.

Immunolocalization of fibronectin during the early response of dog dental pulp to demineralized dentine or calcium hydroxide-containing cement.

The role of fibronectin during the events initiating the post-developmental histogenesis of dentine was investigated by exposing the pulp to implants of autogenous demineralized dentine or calcium hydroxide-containing cement for short periods. Implants exposed for 3 days were processed for immunoelectron-microscopic analysis of fibronectin adsorption on to their surfaces. The localization of fibronectin in the critical area of interaction was examined by immunofluorescence 6, 14 and 21 days after implantation. Heavy adsorption of fibronectin on to the dentine implants and the crystalline structures that had been deposited on the cement implants was demonstrated. Positive fluorescence was consistently seen around dentine implants. Strongly immunopositive fibroblast-like cells and weakly reactive, differentiating odontoblast-like cells were found in association with the implanted matrix. Uncalcified matrix secreted by the polarized or non-polarized cells was consistently rich in fibronectin. Fibroblast-like cells exhibiting intense immunoreaction only at 14 and 21 days were mainly associated with the crystalline precipitates on the cement surfaces or within the surrounding pulp. The findings indicate that the specific inductive effects of demineralized dentine on pulp cells are initiated by exposure of the pulp to a fibronectin-containing surface; adhesion of pulp cells and synthesis of a fibronectin-rich matrix characterize the development of new dentine. The reparative response to non-specific inductive influences such as calcium hydroxide seems to be mediated by progressive enhancement of fibronectin synthesis in pulp cells.