Quantitative assessment of dentine mineralization and tubule occlusion by NovaMin and stannous fluoride using serial block face scanning electron microscopy.

Dentine hypersensitivity (DH) is one of the most common dental conditions affecting most adults during their lifetime. Tubule occlusion is a widely accepted method for treating DH. Current in-vitro techniques such as focused ion beam, scanning electron microscopy (SEM), or hydraulic conductance that are used to determine tubule occlusion do not provide the depth of occlusion, are time-consuming, expensive and the volume of dentine tested is limited. The presented study aimed to assess the ability of serial block-face SEM (SBF-SEM) to section dentine, to quantify the number of occluded tubules including the depth of penetration by NovaMin and stannous fluoride (SnF2 ) and to compare mineral density between the control and treated dentine. Results demonstrated that NovaMin provided a better occlusion with 100% of the tubules blocked at the surface compared to 83% for SnF2 . The grayscale value (230.42) was significantly higher (p ≤ 0.05) after treatment with NovaMin compared to SnF2 (222.06) and the control (196.37), indicating increased mineral density and dentine mineralization. SBF-SEM has the potential to be used for large volume analysis of bone-like materials at high resolution with minimal sample preparation over a short period. It can be significantly useful in the development and research of new biomaterials.

An in vitro evaluation of a novel high fluoride daily mouthrinse using a combination of microindentation, 3D profilometry and DSIMS.

OBJECTIVES: Firstly, to evaluate the in vitro anti-erosion efficacy of a new mouthrinse formulation containing 450 ppm fluoride using profilometry and microindentation. Secondly, to compare fluoride uptake by erosive lesions from two mouthrinses containing different fluoride sources using dynamic secondary ion mass spectrometry (DSIMS). METHODS: Sound human enamel was treated (60s) with mouthrinses containing different fluoride concentrations, then immersed in 1.0% citric acid pH 3.8 for either 300 s or 30 min (Studies 1 & 2 respectively). Surface roughness and erosion depth were determined profilometrically in Study 1, and surface microhardness monitored as a function of time in Study 2. Lesion rehardening was monitored following a 60 s rinse and immersion in artificial saliva for 48 h (Study 3), whilst Study 4 employed DSIMS to quantify fluoride uptake by lesions treated (60s) with rinses containing either sodium fluoride (NaF) or a NaF/Olaflur/stannous chloride combination. RESULTS: The test rinse (450 ppm fluoride) suppressed surface roughening and bulk tissue loss versus all comparators (p< 0.0001), except in the latter measure for the rinse containing 112 ppm fluoride. The test rinse significantly inhibited enamel surface softening versus the three rinses containing ≤112 ppm fluoride (as NaF) at 30 min (p<0.05), but was not statistically significantly different from the 225 ppm fluoride rinse. The test rinse conferred statistically superior lesion rehardening versus all comparators at both 24 and 48 h (p< 0.0001). DSIMS demonstrated statistically significantly higher fluoride uptake by incipient erosive lesions treated with the test rinse versus the NaF/Olaflur/stannous rinse. CONCLUSIONS: Anti-erosion efficacy was positively correlated with fluoride concentration. DSIMS showed significantly higher levels of fluoride uptake by incipient erosive lesions treated with the 450 ppm fluoride rinse versus the NaF/Olaflur/stannous rinse.

Studies on a novel combination polymer system: in vitro erosion prevention and promotion of fluoride uptake in human enamel.

OBJECTIVES: Firstly, determine the effect of pre-treating sound human enamel with a hydrosoluble combination polymer system (TriHydra™) comprising 0.20% carboxymethylcellulose, 0.010% xanthan gum and 0.75% copovidone, alone or in combination with fluoride, on in vitro erosion by citric acid. Secondly, investigate the effect of the polymers on fluoride uptake by incipient erosive lesions. METHODS: Study 1: Sound enamel specimens were treated (60s, 20°C, 150 rpm) with either (i) deionised water, (ii) polymers in deionised water, (iii) 300 mg/L fluoride or (iv) polymers in 300 mg/L fluoride. Specimen groups (n=5) were then immersed in 1.0% citric acid (pH 3.8, 300 s, 20°C, 50 rpm) and non-contact profilometry was used to determine surface roughness (Sa) and bulk tissue loss. Study 2: Incipient erosive lesions were similarly treated with (i)-(iv). Dynamic Secondary Ion Mass Spectrometry (DSIMS) was then used to determine the fluoride depth-distribution. RESULTS: Study 1: Mean±SD Sa and erosion depths for treatment groups (i)-(iv) were (a)657±243, (b)358±50, (c)206±72, (d)79±16 nm and (a)19.73±8.70, (b)2.52±1.34, (b)0.49±0.34 and (b)0.31±0.21 mm respectively (matching superscripts denote statistically equivalent groups). Study 2: Lesions treated with (iii) and (iv) exhibited similar fluoride penetration depths (∼ 60 µm). Mean fluoride intensity ratios based on F/(F+P) at 1 µm for treatment groups (i)-(iv) were (a)0.010±0.004, (a)0.011±0.004, (b)0.803±0.148 and (c)0.994±0.004 respectively. CONCLUSIONS: The combination polymer system exhibited anti-erosion efficacy in its own right. The polymer/fluoride admixture statistically significantly reduced Sa, however suppression of bulk tissue loss was not statistically significantly different versus either treatment alone. The presence of polymer appears to promote fluoride uptake by erosive lesions most noticeably in the first 6 µm.

In vitro microhardness studies on a new anti-erosion desensitizing toothpaste.

OBJECTIVE: To evaluate in vitro the efficacy of a new anti-erosion desensitizing toothpaste to inhibit enamel surface softening by a dietary acid, and promote re-hardening of artificial erosive lesions. METHODOLOGY: The ability of the toothpaste to inhibit formation and promote repair of erosive lesions in human enamel has been investigated. In an enamel surface softening study, sound human enamel was pre-treated with one of four toothpaste slurries for two minutes, before exposure to 1.0% citric acid, pH 3.8, for a total of 30 minutes. The surface microhardness (SMH) of the specimens was determined at baseline and at 10-minute intervals using a Struers Duramin-1 microindentor. In an enamel re-hardening study, the erosive lesions were prepared by exposure of the specimens to 1.0% citric acid, pH 3.8, for 30 minutes. After two minutes treatment with a toothpaste slurry, lesion repair was monitored by SMH after 4, 24, and 48 hours incubation in artificial saliva. This remineralizing phase was modified by the addition of an aliquot of the relevant toothpaste slurry, to mimic in vivo carryover of the formulation. RESULTS: The new test formulation, Elmex Sensitive, and Colgate Sensitive exhibited statistically significant inhibition of citric acid-mediated enamel surface softening versus a fluoride-free placebo at all time points. The test toothpaste gave statistically superior protection against the erosive challenge compared to Elmex Sensitive and Colgate Sensitive after 20- and 30-minute exposures. In the remineralization studies, erosive lesions treated with the test toothpaste exhibited statistically superior re-hardening versus lesions treated with Elmex Sensitive and Colgate Sensitive after 24- and 48-hour incubation in the artificial saliva. Lesions treated with Elmex Sensitive re-hardened to a statistically significant extent versus the fluoride-free placebo toothpaste. A re-hardening study, in which a series of the new toothpaste-base formulations containing increasing concentrations of NaF were evaluated, showed a clear fluoride dose response. CONCLUSION: The present microhardness studies show that treatment with fluoride-containing toothpastes helps protect sound enamel from acid-mediated surface softening, and promotes re-hardening of erosive lesions. The new test toothpaste exhibited statistically superior efficacy to Elmex Sensitive and Colgate Sensitive in both in vitro models.

Review of the use of polymers in saliva substitutes for symptomatic relief of xerostomia.

It is clear from the many clinical studies carried out on currently available saliva substitutes, that the main factors limiting the success of polymers in treating xerostomia is their short duration of action and/or their lack of acceptability by users. The most effective formulations for severe sufferers of xerostomia contain relatively viscous, thixotropic polymers, which mild to moderate sufferers in particular tend to dislike. Relief can also be extended by increasing concentrations of non-thixotropic polymers; however, this also has to be balanced by the decrease in palatability resulting from higher polymer levels. In addition, gels rather than solutions tend to result from higher polymer concentrations, which are only employed by the most severe sufferers of xerostomia, and in fact, have been suggested to offer little improvement in efficacy over oral sprays. It would be highly advantageous to improve the duration of relief without impacting negatively on the sensory profile of the product. This would provide a longer lasting effect that is acceptable to all sufferers of xerostomia. The essential role of polymers in saliva substitutes is to provide lubrication, hydration, and protection of the oral mucosa, providing symptomatic relief to sufferers of xerostomia. However, saliva substitutes should also be of neutral or near neutral pH, to prevent demineralization of the oral hard tissues. These products should always be used in conjunction with a thorough oral health regimen, but may also contain appropriate levels of electrolytes for remineralization of enamel and dentine.

Co-operative mineralization and protein self-assembly in amelogenesis: silica mineralization and assembly of recombinant amelogenins in vitro.

An amorphous silica mineralization technique was used to produce inorganic/protein composites to elucidate the structure and mechanism of formation of amelogenin assemblies, which may play an important role in regulating enamel structure during the initial stages of amelogenesis. Full-length recombinant amelogenins from mouse (rM179) and pig (rP172) were investigated along with key degradation products (rM166 and native P148) lacking the hydrophilic C terminus found in parent molecules. The resulting products were examined using transmission electron microscopy and/or small-angle X-ray scattering. Using protein concentrations of 0.1-3 mg ml-1, large monodisperse spheres of remarkably similar mean diameters were observed using rM179 (124+/-4 nm) and rP172 (126+/-7 nm). These spheres also exhibited 'internal structure', comprising nearly spherical monodisperse particles of approximately 20 nm in diameter. In the presence of rM166, P148, and bovine serum albumin (control), large unstructured and randomly shaped particles (250-1000 nm) were observed. Without added protein, large dense spherical particles of silica (mean approximately 500 nm) lacking internal structure were produced. These findings demonstrate that full-length amelogenins have the ability to form higher-order structures, whereas amelogenins that lack the hydrophilic C terminus do not. The results also suggest that full-length amelogenin can guide the formation of organized mineralized structures through co-operative interactions between assembling protein and forming mineral.