The in vitro and in vivo reproducibility of a video-based digital imaging system for tooth colour measurement.

OBJECTIVES: To assess the robustness of a new custom built video-based digital imaging system (VDIS) for measuring tooth colour and whiteness under in vitro and in vivo conditions. METHODS: The VDIS imaging system was developed for tooth colour measurement and evaluated in vitro and in vivo. The in vitro validation used extracted human teeth (HT, n=14) stored in water and VITA Classical shade guide tabs (SG, n=16). These were measured by the VDIS at baseline, 5min, 2h, 1 week and 2 weeks to evaluate the system repeatability. For in vivo validation, adult volunteers (male/female, n=34) with two natural, unrestored central incisors had their teeth imaged using the VDIS at baseline, 5min and 2h (3 images each) by two different operators to evaluate time and operator effects. Between taking individual images, subjects moved from the imaging-frame to assess the effect of re-positioning on reproducibility. From the in vitro and in vivo images, the average tooth RGB values were obtained, and the CIELAB values and a tooth whiteness index WIO value were calculated. Repeatability and reproducibility of VDIS imaging system was assessed using appropriate repeated measurement analysis techniques and ANOVA. RESULTS: The measurement variations in vitro were between 1 and 2 units of ΔWIO and the average colour differences were less than 1 ΔE*ab unit. For the in vivo study, analysis of the CIELAB parameters and WIO showed that subject variability accounted for between 82 and 99% of the observed variability in the measurement process. The operator variability was less than 0.5% and the overall measurement error was found to be only 0.3% for WIO. Across assessment times the variability was less than 0.5%. CONCLUSIONS: The dental imaging system V-DIS was shown to be a highly reproducible means for tooth colour and whiteness measurement. CLINICAL SIGNIFICANCE: Digital imaging based techniques gives a highly reproducible approach to measuring tooth colour.

In vitro and clinical evaluation of optical tooth whitening toothpastes.

OBJECTIVES: To measure tooth whitening effects delivered immediately after brushing with silica-based toothpastes containing either blue covarine or a combination of blue covarine and FD&C Blue No. 1 in vitro and in vivo. METHODS: Salivary pellicle coated human extracted teeth were brushed with either a slurry of a toothpaste containing blue covarine (BC) or a formulation containing a matched level of blue covarine and FD&C Blue No.1 (BC+D). The colour of the specimens were measured in vitro using a colorimeter, before and after brushing and changes in CIELAB and tooth Whiteness Index (WIO) values calculated. In a double-blind cross-over clinical study, subjects brushed with BC toothpaste, a toothpaste containing increased levels of blue covarine (BC+) or BC+D toothpaste and tooth colour changes were measured with a digital image analysis system. RESULTS: The in vitro study demonstrated that BC+D gave a significantly (p=0.002) greater change in WIO value than BC. Clinical results showed that BC, BC+ and BC+D gave a significant increase in WIO (p<0.0001) from baseline. The WIO change was significantly greater when brushing with BC+D toothpaste than with either toothpaste BC (p<0.0001) or BC+ (p<0.05). CONCLUSIONS: Toothpastes containing blue covarine or a combination of blue covarine and FD&C Blue No. 1 gave a statistically significant improvement in tooth whiteness immediately after brushing in both in vitro and clinical studies. In addition, the toothpaste containing both blue covarine and FD&C Blue No. 1 gave statistically significant greater tooth whitening from baseline than the blue covarine containing toothpastes. CLINICAL SIGNIFICANCE: The silica-based toothpastes containing blue covarine or a combination of blue covarine and FD&C Blue No. 1 evaluated in the current study gave significant tooth whitening benefits immediately after one brush.

Tooth whitening evaluation of blue covarine containing toothpastes.

OBJECTIVES: To measure the tooth whitening effects delivered immediately after brushing with silica-based toothpastes containing blue covarine in vitro and in vivo. METHODS: Salivary pellicle coated human extracted teeth were brushed with either a slurry of a toothpaste containing blue covarine (BC), a formulation containing an increased level of blue covarine (BC+) or a negative control toothpaste containing no blue covarine. The colour of the specimens were measured in vitro using either a Minolta chromameter or a VITA Easyshade spectrophotometer, before and after brushing and changes in CIELAB values and tooth Whiteness Index (WIO) values calculated. In a double-blind cross-over clinical study, subjects brushed with either BC or BC+ toothpaste and tooth colour changes were measured with a digital image analysis system. RESULTS: The in vitro studies demonstrated that toothpastes containing blue covarine gave a significantly (p<0.05) greater change in b* and WIO values than the negative control toothpaste; the BC+ toothpaste gave a significantly greater increase in b* and WIO values than the BC toothpaste, and BC+ gave a significant increase in shade change versus the negative control. Clinical results showed that BC and BC+ gave a significant reduction in b* (p<0.0001) and increase in WIO (p<0.0001) from baseline indicating significant tooth whitening had occurred. The parameter changes were significantly greater when brushing with the BC+ toothpaste than with the BC toothpaste (WIO p=0.006; b* p=0.013). CONCLUSIONS: Toothpastes containing blue covarine gave a statistically significant reduction in tooth yellowness and improvement in tooth whiteness immediately after brushing in both in vitro and clinical studies. In addition, the higher concentration blue covarine toothpaste gave statistically significant greater tooth whitening benefits than the lower concentration blue covarine toothpaste. CLINICAL SIGNIFICANCE: The silica-based toothpastes containing blue covarine evaluated in the current study gave tooth whitening benefits immediately after one brush.

Mode of action studies on the formation of enamel minerals from a novel toothpaste containing calcium silicate and sodium phosphate salts.

OBJECTIVES: To investigate in vitro and in situ the deposition and formation of hydroxyapatite (HAP) on enamel surfaces following brushing with a novel toothpaste containing calcium silicate (CaSi), sodium phosphate salts and fluoride. METHODS: Polished enamel blocks were brushed in vitro with a slurry of the CaSi toothpaste. After one brush and four weeks simulated brushing the enamel surfaces were analysed. In an in situ protocol, enamel blocks were attached to first or second molar teeth of healthy subjects, exposed to 4 weeks twice per day brushing with the CaSi toothpaste and then analysed. The surface deposits were analysed using scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), transmission electron microscopy (TEM) and selected area electron diffraction (SAED). In addition, the CaSi toothpaste was slurried in simulated oral fluid (SOF) over a 3 hour period and the solids were isolated and analysed by Fourier transform infrared spectroscopy (FTIR). RESULTS: The FTIR study demonstrated that calcium phosphate phases had formed and these became increasingly crystalline over 3 hours. CaSi was deposited onto enamel surfaces following one brushing with the toothpaste in vitro.The deposited particles showed evidence of HAP crystalline phases associated with the CaSi. Following 4 weeks brushing in vitro, the deposition increased and analyses showed that the deposited material was HAP. These results were confirmed by the in situ study. CONCLUSIONS: Calcium silicate can be deposited onto enamel surfaces from a novel toothpaste formulation where it can form the enamel mineral HAP. CLINICAL SIGNIFICANCE: A novel toothpaste formulation containing CaSi can form HAP on enamel surfaces. The potential of this technology is for a novel approach to the repair of demineralised enamel and the protection of enamel during acid exposure.

Candida albicans cell wall ssa proteins bind and facilitate import of salivary histatin 5 required for toxicity.

Fungicidal activity of Hst 5 is initiated by binding to cell surface proteins on Candida albicans, followed by intracellular transport to cytoplasmic effectors leading to cell death. As we identified heat shock 70 proteins (Ssa1p and/or Ssa2p) from C. albicans lysates that bind Hst 5, direct interactions between purified recombinant Ssa proteins and Hst 5 were tested by pull-down and yeast two-hybrid assays. Pulldown of both native complexes and those stabilized by cross-linking demonstrated higher affinity of Hst 5 for Ssa2p than for Ssa1p, in agreement with higher levels of interactions between Ssa2p and Hst 5 measured by yeast two-hybrid analyses. C. albicans ssa1Delta and ssa2Delta mutants were constructed to examine Hst 5 binding, translocation, and candidacidal activities. Both ssa1Delta and ssa2Delta mutants were indistinguishable from wild-type cells in growth and hyphal formation. However, C. albicans ssa2Delta mutants were highly resistant to the candidacidal activity of Hst 5, although the ssa1Delta mutant did not have any significant reduction in killing by Hst 5. Total cellular binding of 125I-Hst 5 in the ssa2Delta mutant was reduced to one-third that of wild-type cells, in contrast to the ssa1Delta mutant whose total cellular binding of Hst 5 was similar to the wild-type strain. Intracellular transport of Hst 5 was significantly impaired in the ssa2Delta mutant strain, but only mildly so in the ssa1Delta mutant. Thus, C. albicans Ssa2p facilitates fungicidal activity of Hst 5 in binding and intracellular translocation, whereas Ssa1p appears to have a lesser functional role in Hst 5 toxicity.