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Studies of the effects of carbon dioxide (CO2) lasers on dental enamel have demonstrated that surface changes can be produced at low fluences (< 10 J/cm2) if wavelengths are used which are efficiently absorbed by the hard tissues. In this study, scanning electron microscopy (SEM) was used to characterize the wavelength dependence of surface changes in dental enamel after exposure to an extensive range of CO2 laser conditions. Bovine and human enamel were irradiated by a tunable, pulsed CO2 laser (9.3, 9.6, 10.3, 10.6 microns), with 5, 25, or 100 pulses, at absorbed fluences of 2, 5, 10, or 20 J/cm2, and pulse widths of 50, 100, 200, 500 microseconds. SEM micrographs revealed evidence of melting, crystal fusion, and exfoliation in a wavelength-dependent manner. Crystal fusion occurred at absorbed fluences as low as 5 J/cm2 per pulse at 9.3, 9.6, and 10.3 microns, in contrast to no crystal fusion at 10.6 microns (< or = 20 J/cm2). Longer pulses at constant fluence conditions decreased the extent of surface melting and crystal fusion. The total number of laser pulses delivered to the tissue did not significantly affect surface changes as long as a minimum of 5 to 10 pulses was used. Within the four easily accessible wavelengths of the CO2 laser, there are dramatic differences in the observed surface changes of dental hard tissue.

Scanning electron microscopy was used to compare the morphology, integrity and distribution of bacterial cells in a test plaque grown on the surface of enamel with that of the cell sediment plaque routinely used in a short-term intraoral caries model. Cultures of S. mutans IB-1600 or S. sobrinus 6715-13 were grown in complex media supplemented with either 2.0% sucrose (glucan plaque) or 0.2% glucose (non-glucan plaque). Cell sediment (CS) plaque was prepared by centrifuging the cultures after incubation, recovering the cell sediment, and spreading it on Metricel membrane filter paper. Surface grown (SG) plaque was prepared by suspending saliva-coated bovine enamel in the culture medium, incubating, and recovering the enamel assembly with bacterial accumulations. Cell morphology and integrity, as well as the appearance of glucan-like material produced by the cells, was similar in both CS and SG test plaques. The cell distribution however, varied in the SG plaque from extremes of all cells to all glucan, whereas the cell sediment plaque was more uniform in cell distribution. A highly standardized test plaque minimizes variability in the intraoral caries model. These findings support the contention that the bacterial cells in a cell sediment plaque are similar in morphology, integrity and glucan production to surface grown plaque, and have the added advantage of uniform distribution, which makes the cell sediment plaque more appropriate for intraoral caries model studies.

The formation of calcium (Ca) fluoride (CaF2) on bovine enamel blocks during clinically relevant treatment times using neutral fluoride (F) solutions (0.26 mol/l F) with and without 0.1 mol/l lactate was investigated. Uncoated and pellicle-coated blocks were evaluated for alkali-soluble (1 mol/l KOH, three consecutive 24-hour treatments) Ca, PO4, and F after treatment by the F solutions for 0, 5, 15, 30, and 60 min. There was an overall time-related increase in F recovery, while Ca tended to remain at baseline levels. Less F was recovered from the pellicle-coated blocks. The addition of lactate to the F treatment solution did not result in an overall increase in alkali-soluble F recovery, but did result in the formation of cuboidal shaped crystals which closely approached the morphology of pure CaF2. A 1:2 stoichiometric ratio Ca:2F (mol:mol) was not established based on chemical analyses. The ultrastructural and elemental composition of surface deposits on the samples, as determined using scanning electron microscopy, X-ray diffraction, and energy-dispersive spectroscopy, established the presence of CaF2 after 24-hour F treatments; however, it was not possible to directly demonstrate the formation of CaF2 after clinically relevant treatment times.

The intra-oral enamel demineralization test (IEDT) was introduced by Brudevold et al. (1984). This caries model involves human subjects wearing palatal appliances each holding eight bovine enamel blocks covered by a bacterial cell layer prepared by the harvesting of cultures of Streptococcus mutants (test plaque). The original model used the iodide permeability test for assessment of the extent of demineralization of bovine enamel blocks resulting from acid production by the test plaque after dietary substrate challenge. The IEDT model has been expanded and improved by us in the following ways: (1) Based on encouraging findings from an in vitro study (Zero et al., 1990), the surface microhardness test has been adopted to measure the extent of demineralization occurring at three sites on the enamel blocks corresponding to an area over which the effective plaque thickness is 0.5, 1.5, and 2.5 mm; (2) intra-oral pH of the test plaque is measured by means of a Beetrode miniature pH electrode at baseline, then at five, 10, 15, 30, and 45 min after the start of a test; (3) plaque samples are collected at the end of a test and analyzed for organic acid content by means of HPLC; (4) the bacterial test challenge has been expanded to include different cariogenic bacteria which are grown under various growth conditions. The improved model has the capability of studying fundamental aspects of the caries process, namely, the relationships among dietary substrate challenge, plaque pH change, plaque organic acid profiles, microbial virulence properties, and enamel demineralization. Furthermore, the model has the potential for use in more applied research on caries-preventive agents such as fluoride.

Modifications to the standard operating settings for accelerating voltage, condenser lens current, scan rate, working distance and tilt on the conventional scanning electron microscope (SEM) enabled non-metal coated dental hard tissues and synthetic apatite pellets to be viewed free of charging effects. Well-resolved images at magnifications as high as 35,000x were achieved using accelerating voltages less than 5 kV. The methodology detailed here allowed for serial SEM examination of the same sample at various points during an experimental procedure, and may be applied to other sample types. The procedure is non-destructive to the sample and requires no physical modification to the microscope.