A study of the initial attachment and subsequent behavior of rat oral epithelial cells cultured on titanium.

BACKGROUND: The seal between an implant and the oral epithelium is an important factor for successful implant therapy. The purpose of this study was to evaluate the initial attachment and subsequent behavior of rat oral epithelial (OE) cells on pure titanium (Ti) used for dental implants. METHODS: OE cells derived from 4-day-old rats were cultured on Ti, polystyrene culture dishes, and glass coverslips. The number of attached cells, cell areas, number of colonies, and proliferation rates were measured. Additionally, immunostaining of vinculin and laminin-5 (LN5) was performed, and LN5-immunoreactive areas were measured. RESULTS: After 24 hours of culture, there were fewer cells attached to the Ti than to the polystyrene dishes or glass coverslips, and the area of cells was greater on the polystyrene than on the Ti or glass. OE cells reached their maximum proliferation rate after 48 hours of culture on the polystyrene and glass, and after 72 hours on Ti. LN5 was deposited behind cells as they migrated, and the LN5-immunoreactive area was smaller on Ti than on polystyrene after 96 hours of culture. CONCLUSIONS: The initial attachment of OE cells to Ti was inferior to that on polystyrene or glass, and the OE cell migration area indicated by the deposition of LN5 was smaller on Ti than on the other materials. Therefore, this study suggests that further improvement of Ti surface properties is needed for rapid attachment and spreading of oral epithelium to dental implants.

Difference in penetration of horseradish peroxidase tracer as a foreign substance into the peri-implant or junctional epithelium of rat gingivae.

Horseradish peroxidase (HRP) tracer was applied to the gingival sulcus of implants or natural teeth at 5, 25, or 50 mg/ml to investigate the sealing capacities of the peri-implant epithelium (PIE) and junctional epithelium (JE); the extent of HRP penetration was observed under electron microscopy. A Ti-6Al-4V implant was inserted either immediately (immediate implantation) or 2 weeks (delayed implantation) after extraction of the maxillary left first molar of rats. The JE of the right molar was used as a control. Although the whole PIE of undecalcified sections appeared to be attached to the implant surface, electron microscopically, the internal basement lamina (IBL) and hemidesmosomes were deficient in the coronal-middle region of the PIE. There were extensive extracellular deposits of HRP in the intercellular spaces between PIE cells, and HRP was blocked to some extent by the lamina lucida and lamina densa of the external basal lamina and basal cell junction. HRP was detected in the connective tissue under the PIE, but was not found in the connective tissue under the JE. Intracellularly, HRP was found in the vesicles and granules of PIE cells and JE cells. These were fewer in number in PIE cells than in JE cells. There were no differences between the findings for immediate and delayed implantation. The results indicate that a deficiency in the IBL permitted penetration of HRP from the gingival sulcus into the connective tissue under the PIE, and suggest that the endocytotic capacity of PIE cells is inferior to that of JE cells.