Hidenori Hamba

Effects of CPP-ACP with sodium fluoride on inhibition of bovine enamel demineralization: A quantitative assessment using micro-computed tomography

OBJECTIVES The present study evaluated the effects of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) and CPP-ACP with 900 ppm fluoride (CPP-ACPF) pastes on inhibition of enamel demineralization over time, using polychromatic micro-computed tomography (micro-CT). METHODS Enamel blocks were prepared from bovine teeth. The specimens were each treated by one of the following agents, 30 min daily for 7 days: deionized water (negative control); CPP-ACP paste; CPP-ACPF paste; and NaF solutions (positive controls) (90, 900, and 9000 ppm F). After treatment, the specimens were immersed in a demineralizing solution (pH 4.5) for 24, 72, and 120 h. Mean mineral loss (ML) and lesion depth (LD) after each period were determined from mineral density profiles obtained using micro-CT. RESULTS ML values in all the treatment groups were significantly smaller than those in the control group after 72 and 120 h of demineralization (p < 0.05, two-way ANOVA and t-test with Bonferroni correction). ML values in CPP-ACPF and NaF solution groups were significantly smaller compared to CPP-ACP group after 72 h (p < 0.05). LD values in the CPP-ACPF and all the NaF solutions groups were significantly smaller compared to the control group after 120 h (p < 0.05). The 9000 ppm F group showed the lowest nominal ML and LD values. CONCLUSIONS The application of CPP-ACP or CPP-ACPF pastes to sound enamel surfaces resulted in inhibition of enamel demineralization, and a better effect was noted for the latter paste. Quantitative assessment using polychromatic micro-CT demonstrated to be useful for detecting mineral density changes.

Enamel Lesion Parameter Correlations between Polychromatic Micro-CT and TMR

Transverse microradiography (TMR) is considered as the gold standard technique for the evaluation of enamel lesions. Micro-computed tomography (µCT) has the advantage of non-destructive measurements, but the beam-hardening effect with polychromatic x-rays is a major drawback. To date, no study has validated µCT against TMR. The objective of this study was to validate µCT measurements of enamel lesions under various x-ray conditions and software beam-hardening correction (BHC) against TMR. Human molars with natural white-spot lesions were scanned for 5 min by µCT at 100 kV in different conditions: 50 µA (0.5-mm Al filter), 165 µA (0.5-mm Al/0.3-mm Cu), and 200 µA (0.5-mm Al/0.4-mm Cu), with or without BHC. Grayscale values were converted into mineral density values using phantoms. Thin sections at the same positions were then prepared for TMR. Lesion depth (LD; µm) and mineral loss (ΔZ; vol%µm) were compared between µCT and TMR by Pearson’s correlations. µCT measurements correlated well with TMR under all conditions (p < 0.001, r > 0.86 for LD and ΔZ), except for 0.5-mm Al without BHC (p > 0.05). Even without BHC, combined Al/Cu filters successfully reduced the beam-hardening effect. µCT can be used as a non-destructive alternative to TMR with comparable parameters for the study of enamel lesions.

Gum Containing Calcium Fluoride Reinforces Enamel Subsurface Lesions in situ

The aim of this study was to assess the effect of chewing gum containing phosphoryl oligosaccharides of calcium (POs-Ca) and a low concentration of fluoride (F) on the hardness of enamel subsurface lesions, utilizing a double-blind, randomized, and controlled in situ model. Fifteen individuals wore removable lingual appliances with 3 bovine-enamel insets containing subsurface demineralized lesions. Three times a day for 14 days, they chewed one of the 3 chewing gums (placebo, POs-Ca, POs-Ca+F). After the treatment period, cross-sectional mineral content, nanoindentation hardness, and fluoride ion mapping by time-of-flight secondary ion mass spectrometry (TOF-SIMS) were evaluated. Although there were no statistical differences in overall mineral content and hardness recovery rates between POs-Ca and POs-Ca+F subsurface lesions (p > 0.05), nanoindentation at 1-μm distance increments from the surface showed statistical differences in hardness recovery rate between POs-Ca and POs-Ca+F in the superficial 20-μm region (p < 0.05). Fluoride mapping revealed distribution of the ion up to 20 μm from the surface in the POs-Ca+F group. Nanoindentation and TOF-SIMS results highlighted the benefits of bioavailability of fluoride ion on reinforcement of the superficial zone of subsurface lesions in situ (NCT01377493).

Non-destructive characterization of voids in six flowable composites using swept-source optical coherence tomography

OBJECTIVE The aim of this study was to evaluate the void frequency (V(F)) and void volume (V(V)) in different flowable composites using swept-source optical coherence tomography (SS-OCT). METHODS Standard class I cavities were prepared and filled with six different flowable composites: Clearfil Majesty LV (MJ; Kuraray), MI Flow (MW; GC), MI Fil (ML; GC), Beautifil flow plus (BF; Shofu), Palfique Estelite low flow (EL; Tokuyama) and Surefil SDR flow (SF; Dentsply). The restorations were scanned under OCT. The OCT tomograms were analyzed and average V(F) and V(V) per restoration for each composite were calculated. Scanning electron microscope (SEM) was used to observe the structure of each composite. In addition, the flowability of the materials was evaluated measuring the displacement of each material placed up-right on a glass slide. V(F) and V(V) obtained by OCT were also compared to those calculated using micro-computed tomography (micro-CT). RESULTS Kruskal-Wallis ANOVA and Mann-Whitney U tests revealed significantly different V(F) and V(V) values (p<0.05) among the composites. Voids ranging from 35 to 785 μm in diameter were detected in OCT tomograms. MJ showed highest V(F) and V(V) values followed by MW, but ML, BF, EL and SF showed no significant difference. Filler volume in composites showed a positive correlation with void formation, but flowability did not show a specific trend. Micro-CT evaluation validated the V(F) and V(V) calculation by OCT, with a significant correlation in void size (p<0.001, r=0.94). CONCLUSION The results of this study indicate the reliability of SS-OCT for real-time void characterization of composite materials and restorations. Void formation in flowable composites is material dependent.

Effects of a chewing gum containing phosphoryl oligosaccharides of calcium (POs-Ca) and fluoride on remineralization and crystallization of enamel subsurface lesions in situ

OBJECTIVES Manufacturers are adding fluoride (F) to calcium-containing chewing gums to further promote enamel remineralization. The aim of this study was to assess the effect of a chewing gum containing phosphoryl oligosaccharides of calcium (POs-Ca) and fluoride on remineralization of enamel subsurface lesions, in a double-blind, randomized controlled in situ trial. METHODS Thirty-six volunteer subjects wore removable buccal appliances with three different insets of bovine enamel with subsurface demineralized lesions. For 14 days the subjects chewed one of the three chewing gums (placebo, POs-Ca, POs-Ca+F), three times a day. After each treatment period, the insets were removed from the appliance, embedded, sectioned, polished and then subjected to laboratory tests; mineral level was determined by transverse microradiography (TMR; n=36), and hydroxyapatite (HAp) crystallites were assessed by synchrotron radiation wide-angle X-ray diffraction (WAXRD; n=13). Data were analysed by t-test or Wilcoxon rank-sum test with Bonferroni corrections at 0.05 significance level. RESULTS Chewing POs-Ca and POs-Ca+F gums resulted in 21.9±10.6 and 26.3±9.4 (mean±SD) percentage mineral recovery, which was significantly higher than that of placebo gum (15.0±11.4) (p<0.05). Chewing POs-Ca+F gum resulted in 24.9±5.4 (mean±SD) percentage HAp crystallites recovery, which was significantly higher compared to POs-Ca (16.0±4.1%) or placebo (11.1±4.8%) gums (p<0.05). CONCLUSIONS Addition of POs-Ca to the chewing gum resulted in significant remineralization of enamel subsurface lesions. Although POs-Ca+F gum was not superior in TMR recovery rate when compared with POs-Ca gum, WAXRD results highlighted the importance of fluoride ion bioavailability in the formation of HAp crystallites in enamel subsurface lesions in situ (NCT01377493).

Assessment of natural enamel lesions with optical coherence tomography in comparison with microfocus x-ray computed tomography

Abstract. A technology to characterize early enamel lesions is needed in dentistry. Optical coherence tomography (OCT) is a noninvasive method that provides high-resolution cross-sectional images. The aim of this study is to compare OCT with microfocus x-ray computed tomography (μCT) for assessment of natural enamel lesions in vitro. Ten human teeth with visible white spot-like changes on the enamel smooth surface and no cavitation (ICDAS code 2) were subjected to imaging by μCT (SMX-100CT, Shimadzu) and 1300-nm swept-source OCT (Dental SS-OCT, Panasonic Health Care). In μCT, the lesions appeared as radiolucent dark areas, while in SS-OCT, they appeared as areas of increased signal intensity beneath the surface. An SS-OCT attenuation coefficient based on Beer–Lambert law could discriminate lesions from sound enamel. Lesion depth ranged from 175 to 606  μm in SS-OCT. A correlation between μCT and SS-OCT was found regarding lesion depth (R=0.81, p<0.001) and also surface layer thickness (R=0.76, p<0.005). The images obtained clinically in real time using the dental SS-OCT system are suitable for the assessment of natural subsurface lesions and their surface layer, providing comparable images to a laboratory high-resolution μCT without the use of x-ray.

Sodium fluoride mouthrinse used twice daily increased incipient caries lesion remineralization in an in situ model

OBJECTIVES To investigate the remineralizing effects of fluoride mouthrinses used at different times and frequency in addition to fluoride toothpaste. METHODS A randomized crossover single blinded study comprised 4 experimental phases of 21 days each. Twelve orthodontic volunteers were fixed with an orthodontic bracket containing an artificial carious enamel slab, which was from the same tooth in all 4 phases, and were randomly assigned to the following groups: (1) brushing with F toothpaste 2× per day (F- brush), (2) F- brush+rinsing with 0.05% NaF (F- rinse) after lunch, (3) F- brush+F-rinse before bedtime, and (4) F- brush+F- rinse 2× per day. Mean mineral gain after each phase was determined from mineral density profiles obtained using Micro-CT. RESULTS The mean mineral gain in all treatments with F- brush and F-rinse were significantly greater than those in F- brush (p<0.05). Moreover F- rinse 2× per day increased lesion remineralization more than F- rinse once a day. CONCLUSIONS The twice-daily use of 0.05% NaF mouthrinse combined with twice-daily regular use of fluoride toothpaste resulted in the greatest remineralization of incipient caries. These data indicate that rinsing frequency is a factor affecting the effectiveness of fluoride mouthrinse. CLINICAL SIGNIFICANCE The rinsing frequency of NaF mouthrinse, when used with fluoride toothpaste, also affects the remineralization. This finding, if confirmed by a clinical study, would lead to a new recommendation for fluoride mouthrinse used in high caries risk patients who could benefit from using it twice a day.

Morphological Evaluation of the Adhesive/Enamel interfaces of Two-step Self-etching Adhesives and Multimode One-bottle Self-etching Adhesives.

PURPOSE To evaluate the acid-base resistant zone (ABRZ) at the adhesive/enamel interface of self-etching adhesives with or without prior phosphoric acid etching. MATERIALS AND METHODS Four adhesives were used in 8 groups: Clearfil SE Bond (SEB), Optibond XTR (XTR), Scotchbond Universal Adhesive (SBU), and Clearfil BOND SE ONE (ONE) without prior phosphoric-acid etching, and each adhesive with phosphoric acid etching for 10 s (P-SEB, P-XTR, P-SBU and P-ONE, respectively). After application of self-etching adhesives on ground enamel surfaces of human teeth, a flowable composite was placed. For observation of the acid-base resistant zone (ABRZ), the bonded interface was exposed to demineralizing solution (pH 4.5) for 4.5 h, followed by 5% NaOCl with ultrasonication for 20 min. After the acid-base challenge, morphological attributes of the interface were observed using SEM. RESULTS ABRZ formation was confirmed in all groups. The funnel-shaped erosion beneath the interface was present in SBU and ONE, where nearly 10 to 15 μm of enamel was dissolved. With phosphoric acid etching, the ABRZs were obviously thicker compared with no phosphoric acid etching. CONCLUSION Enamel beneath the bonding interface was more susceptible to acid dissolution in SBU and ONE. In the case of the one-bottle self-etching adhesives and universal adhesives that intrinsically have higher pH values, enamel etching should be recommended to improve the interfacial quality.

Effect of different desensitizers on inhibition of bovine dentin demineralization: micro-computed tomography assessment

This study evaluated the effect of two desensitizers on inhibition of dentin demineralization, after immersion in artificial saliva using micro-computed tomography (μCT). Dentin blocks cut from bovine incisors were treated with deionized water (DW, a negative control) or one of three desensitizers: a fluoride varnish (Duraphat, a positive control), a calcium phosphate desensitizer (Teethmate Desensitizer), and a fluoro-alumino-calcium silicate-based desensitizer (Nanoseal). After each treatment, the specimens in Duraphat, Nanoseal, and Teethmate Desensitizer groups were pre-immersed in artificial saliva (pH 6.5) for either 1 d or 1 wk. The mineral loss of the specimens after demineralization (pH 5.0, 3 h) was evaluated by μCT. The treated surface was investigated with scanning electron microscopy. Mineral loss in all treatment groups was significantly lower than that in DW. Duraphat was the most effective treatment against demineralization, followed by Nanoseal. Nanoseal showed significantly better reduction in mineral loss following immersion for 1 wk in artificial saliva than for 1 d. However, Teethmate Desensitizer and Duraphat did not exhibit enhanced inhibition of demineralization over a longer period of immersion in artificial saliva. Scanning electron microscopy images showed deposition of particles on the dentin in both Teethmate Desensitizer. The application of Teethmate Desensitizer and Nanoseal to the exposed dentin surface resulted in inhibition of demineralization, with Nanoseal resulting in improved inhibition after prolonged immersion in artificial saliva.

Effects of experimental pastes containing surface pre-reacted glass ionomer fillers on inhibition of enamel demineralization

This study aimed to evaluate the inhibitory effect of experimental pastes containing surface pre-reacted glass ionomer (S-PRG) fillers on enamel demineralization. Bovine blocks were treated twice a day for 4 days by 7 groups; experimental pastes containing 0-30 wt% S-PRG filler (S00, S01, S05, S10, and S30), deionized water (DW) as negative control, and NaF paste (MP) as positive control. The surfaces were demineralized by acetic acid for 3 days. Mineral loss (ML) was calculated by micro-computed X-ray tomography. The treated surface was finally investigated with scanning electron microscope (SEM) and micro-focused particle induced X-ray emission (micro-PIXE). S05, S10 and S30 demonstrated significantly lower ML than S00, S01 and DW (p<0.05). S10 showed the greatest inhibitory effect, which was significantly greater than MP. The S-PRG filler containing experimental pastes demonstrated a potential to inhibit enamel demineralization. Sr ion incorporation was confirmed on the enamel surface with the experimental pastes.