Abstract
Objective: To develop a novel mandible slice organ culture model to investigate the effects of externally applied force on the dentine-pulp complex.
Design: In vitro organ culture.
Setting: School of Dentistry, Birmingham, UK.
Materials and methods: Transverse 2 mm thick sections were cut from the mandibles of five 28-day-old male Wistar rats. Serial sections were used for control and test pairs. Springs made from 0.016-inch and 0.019 x 0.025-inch stainless steel wires were used to apply a 50 g tensile or compressive force, respectively, to test specimens. Control and test specimens were cultured for 5 days in a humidified incubator with 5% CO2 at 37 degrees C and processed for routine histological investigation. Nine more rats were used to provide control and compression test pairs where the pulps were extirpated after 3 days culture and total RNA isolated for gene expression analysis by reverse transcriptase polymerase chain reaction (RT-PCR).
Results: Histology showed the dental and supporting tissues maintained a healthy appearance in the control cultures after culture. Histomorphometric analysis revealed a 20-27% increase in pulp fibroblast density in test specimens compared with controls. Gene expression analyses revealed up-regulation in the test groups of PCNA, c-Myc, Collagen 1 alpha, TGF-beta 1 and alkaline phosphatase, whilst expression of osteocalcin was reduced.
Conclusions: The results demonstrated that the present organ culture technique provides a valuable in vitro experimental model for studying the effects of externally applied forces. These forces stimulated a cellular response in the pulp chamber characterized by altered gene expression and proliferation of fibroblasts; the latter being unaffected by the nature of the force in terms of compression or tension.