Acid resistance of enamel subsurface lesions remineralized by a sugar-free chewing gum containing casein phosphopeptide-amorphous calcium phosphate

The aim of this clinical study was to investigate the acid resistance of enamel lesions remineralized in situ by a sugar-free chewing gum containing casein phosphopeptide-amorphous calcium phosphate nanocomplexes (CPP-ACP: Recaldent). The study utilized a double-blind, randomized, crossover design with two treatments: (i) sugar-free gum containing 18.8 mg of CPP-ACP, and (ii) sugar-free gum not containing CPP-ACP as control. Subjects wore removable palatal appliances with insets of human enamel containing demineralized subsurface lesions and chewed the gum for 20 min 4 times per day for 14 days. After each treatment the enamel slabs were removed and half of each lesion challenged with acid in vitro for 8 or 16 h. The level of remineralization was determined using microradiography. The gum containing CPP-ACP produced approximately twice the level of remineralization as the control sugar-free gum. The 8- and 16-hour acid challenge of the lesions remineralized with the control gum resulted in 65.4 and 88.0% reductions, respectively, of deposited mineral, while for the CPP-ACP-remineralized lesions the corresponding reductions were 30.5 and 41.8%. The acid challenge after in situ remineralization for both control and CPP-ACP-treated lesions resulted in demineralization underneath the remineralized zone, indicating that the remineralized mineral was more resistant to subsequent acid challenge. The results show that sugar-free gum containing CPP-ACP is superior to an equivalent gum not containing CPP-ACP in remineralization of enamel subsurface lesions in situ with mineral that is more resistant to subsequent acid challenge.     

Effect of addition of citric acid and casein phosphopeptide-amorphous calcium phosphate to a sugar-free chewing gum on enamel remineralization in situ

Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) has been shown to remineralize enamel subsurface lesions in situ. The aim of this study was to investigate the effects of CPP-ACP in a fruit-flavoured sugar-free chewing gum containing citric acid on enamel remineralization, and acid resistance of the remineralized enamel, using an in situ remineralization model. The study utilized a double-blind, randomized, crossover design with three treatments: (i) sugar-free gum (2 pellets) containing 20 mg citric acid and 18.8 mg CPP-ACP, (ii) sugar-free gum containing 20 mg citric acid alone, (iii) sugar-free gum not containing CPP-ACP or citric acid. Ten subjects were instructed to wear removable palatal appliances, with 4 half-slab insets of human enamel containing demineralized subsurface lesions and to chew gum (2 pellets) for 20 min 4 times per day for 14 days. At the completion of each treatment the enamel half-slabs were removed and half of the remineralized lesion treated with demineralization buffer for 16 h in vitro. The enamel slabs (remineralized, acid-challenged and control) were then embedded, sectioned and subjected to microradiography to determine the level of remineralization. Chewing with gum containing citric acid and CPP-ACP resulted in significantly higher remineralization (13.0 +/- 2.2%) than chewing with either gum containing no CPP-ACP or citric acid (9.4 +/- 1.2%) or gum containing citric acid alone (2.6 +/- 1.3%). The acid challenge of the remineralized lesions showed that the level of mineral after acid challenge was significantly greater for the lesions exposed to the gum containing CPP-ACP.     

Remineralization of enamel subsurface lesions in situ by sugar-free lozenges containing casein phosphopeptide-amorphous calcium phosphate

BACKGROUND: The anticariogenic potential of casein phosphopeptide-amorphous calcium phosphate nanocomplexes (CPP-ACP) has been demonstrated using laboratory, animal and human in situ caries models. The aim of this study was to determine the effect of CPP-ACP incorporation into a sugar-free lozenge (pressed mint tablet) on enamel remineralization in a human in situ model. METHODS: The study utilized a double-blind, randomized, cross-over design with four treatments: (i) a lozenge containing 56.4mg (3 per cent w/w) CPP-ACP; (ii) a lozenge containing 18.8mg (1 per cent w/w) CPP-ACP; (iii) a lozenge not containing CPP-ACP; and (iv) a no lozenge nil-treatment control. Ten subjects wore removable palatal appliances with four, human-enamel, half-slab insets containing subsurface lesions. Lozenges were consumed, without chewing, four times per day for 14 days duration. After each treatment period the enamel slabs were removed, paired with their respective demineralized control, embedded, sectioned and subjected to microradiography and computer-assisted densitometric image analysis to determine the level of remineralization. RESULTS: The incorporation of CPP-ACP into the lozenge significantly increased enamel subsurface lesion remineralization with 18.8 and 56.4mg of CPP-ACP increasing remineralization by 78 and 176 per cent respectively, relative to the control sugarfree lozenge. CONCLUSION: This study demonstrates that lozenges are a suitable vehicle for the delivery of CPP-ACP to promote enamel remineralization.     

Remineralization of enamel subsurface lesions in situ by the use of three commercially available sugar-free gums

Background.  Commercially available sugar-free chewing gums have been claimed to provide oral health benefits.
Aim.  The aim of this randomized, double-blind crossover in situ study was to compare the efficacy of three commercially available sugar-free chewing gums: Trident White, Orbit, and Orbit Professional, in remineralizing enamel subsurface lesions in situ .
Design.  Specimens containing enamel subsurface lesions were sectioned into test and control half-slabs with the test half-slabs inserted into removable palatal appliances. For each test chewing period, subjects were randomly allocated one of three test gums. Subjects ( n  = 10) chewed the randomly allocated gum for a 20-min period four times per day for 14 days. Each subject chewed all three test gums, with a 7-day washout period between crossovers. After each 14-day cycle, test and control half-slabs were paired, embedded in resin, sectioned, and subjected to microradiography to determine remineralization.
Results.  The gum TW produced significantly greater remineralization (18.4 ± 0.9%) than Orbit (8.9 ± 0.5%) and Orbit Professional (10.5 ± 0.9%).
Conclusion.  The superior remineralization activity of the TW gum in situ was attributed to the presence of casein phosphopeptide–amorphous calcium phosphate nanocomplexes.     

Remineralization of enamel subsurface lesions by sugar-free chewing gum containing casein phosphopeptide-amorphous calcium phosphate.

Casein phosphopeptide-amorphous calcium phosphate nanocomplexes (CPP-ACP) exhibit anticariogenic potential in laboratory, animal, and human in situ experiments. The aim of this study was to determine the ability of CPP-ACP in sugar-free chewing gum to remineralize enamel subsurface lesions in a human in situ model. Thirty subjects in randomized, cross-over, double-blind studies wore removable palatal appliances with six human-enamel half-slabs inset containing sub-surface demineralized lesions. The appliances were inserted immediately before gum-chewing for 20 min and then retained for another 20 min. This was performed four times per day for 14 days. At the completion of each treatment, the enamel half-slabs were paired with their respective demineralized control half-slabs, embedded, sectioned, and subjected to microradiography and densitometric image analysis, for measurement of the level of remineralization. The addition of CPP-ACP to either sorbitol- or xylitol-based gum resulted in a dose-related increase in enamel remineralization, with 0.19, 10.0, 18.8, and 56.4 mg of CPP-ACP producing an increase in enamel remineralization of 9, 63, 102, and 152%, respectively, relative to the control gum, independent of gum weight or type.     

Fluoride and casein phosphopeptide-amorphous calcium phosphate

Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) slows the progression of caries and remineralizes enamel subsurface lesions. The aim of this study was to determine the ability of CPP-ACP to increase the incorporation of fluoride into plaque and to promote enamel remineralization in situ. Randomized, double-blind, cross-over studies involved mouthrinses and dentifrices containing CPP-ACP and fluoride. The mouthrinses were used for 60 sec, three times/day for 5 days, and supragingival plaque was collected and analyzed for F. The dentifrices were rinsed as a water slurry for 60 sec four times/day for 14 days in an in situ model. The addition of 2% CPP-ACP to the 450-ppm-F mouthrinse significantly increased the incorporation of fluoride into plaque. The dentifrice containing 2% CPP-ACP produced a level of remineralization similar to that achieved with a dentifrice containing 2800 ppm F. The dentifrice containing 2% CPP-ACP plus 1100 ppm F was superior to all other formulations.     

Retention in plaque and remineralization of enamel lesions by various forms of calcium in a mouthrinse or sugar-free chewing gum

Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) nanocomplexes incorporated into sugar-free chewing gum have been shown to remineralize enamel subsurface lesions in situ. The aim of this study was to compare the ability of CPP-ACP, with that of other forms of calcium, to be retained in supragingival plaque and remineralize enamel subsurface lesions in situ when delivered in a mouthrinse or sugar-free gum in randomized, double-blind trials. In the mouthrinse study, only the CPP-ACP-containing mouthrinse significantly increased plaque calcium and inorganic phosphate levels, and the CPP were immunolocalized to the surfaces of bacterial cells as well as the intercellular matrix. In the chewing gum studies, the gum containing the CPP-ACP, although not containing the most calcium per piece of gum, produced the highest level of enamel remineralization independent of gum-chewing frequency and duration. The CPP could be detected in plaque extracts 3 hrs after subjects chewed the CPP-ACP-containing gum. The results showed that CPP-ACP were superior to other forms of calcium in remineralizing enamel subsurface lesions.     

Effect of casein phosphopeptide - amorphous calcium phosphate containing chewing gum on salivary concentration of calcium and phosphorus: An in-vivo study

Aim: Caries clinical trials of sugar-free chewing gum have shown that the gum is noncariogenic and in fact has anticariogenic effect through the stimulation of saliva. Sugar-free gums, therefore, may be an excellent delivery vehicle for safe and effective additive, capable of promoting enamel remineralization. Casein phosphopeptide - amorphous calcium phosphate (CPP-ACP) nanocomplexes incorporated into sugar-free chewing gum have shown to remineralize enamel subsurface lesions in situ. So this study was conducted to evaluate the effect of CPP-ACP containing sugar-free chewing gum on salivary concentration of calcium and phosphorous. Materials and Methods : Unstimulated saliva from each 24 selected subjects was collected. Then each subject was given two pellets of chewing gum containing CPP-ACP and asked to chew for a period of 20 min, after which saliva samples were collected from each individual. Once all the samples were collected they were assessed for calcium and phosphorous concentration using affiliated reagent kits and photometer. Statistical Analysis Used: Data obtained were analyzed using student's paired t test. Results: Significant difference was found in the calcium and phosphorus concentration of saliva before and after chewing CPP-ACP containing chewing gum. Conclusions: Chewing of CPP-ACP containing chewing gum showed a significant increase in the salivary concentration of calcium for a prolonged period of time hence it may help in the remineralization of tooth surfaces.     

Effects of Isomalt on enamel de- and remineralization,a combined in vitro pH-cycling model and in situ study

Isomalt is a non-cariogenic sweetener, which is widely used in sugar-free candy and chewing gum. Little is known about the effects of Isomalt on de- and remineralization. Binding between calcium and Isomalt has been reported, which could affect the mineral balance. The objective of this study was to examine the effects of Isomalt on de- and remineralization of bovine enamel lesions, both in vitro and in situ. In in vitro study, subsurface enamel lesions were subjected to 3-weeks pH-cycling. Treatments were 5-min rinses with 10% Isomalt solutions daily and 10% Isomalt additions to re- or demineralizing solutions. Standard pH-cycling conditions were used with a 0.2 ppm fluoride background during the remineralization phase. In in situ study, subsurface lesions were exposed 2 months in vivo and brushed three times daily with 10% Isomalt containing toothpaste. Treatment effects were assessed by chemical analysis of the solutions (in vitro) and transversal microradiography (in vitro and in situ). In in vitro study, while 5-min rinses with 10% Isomalt gave slightly increased remineralization, continuous presence of 10% Isomalt (in re- or demineralizing solutions) inhibited both de- and/or remineralization. This lead to significantly smaller overall mineral loss when Isomalt was added during demineralization. In in situ study, remineralization enhancement during short Isomalt treatments was confirmed. Isomalt had a positive effect on the de/remineralization balance when given under conditions relevant to practical use.     

Remineralization of enamel subsurface lesions by chewing gum with added calcium

Objectives

Chewing sugar-free gum has been shown to promote enamel remineralization. Manufacturers are now adding calcium to the gum in an approach to further promote enamel remineralization. The aim of this study was to compare the remineralization efficacy of four sugar-free chewing gums, two containing added calcium, utilizing a double-blind, randomized, crossover in situ model.

Methods

The sugar-free gums were: Trident Xtra Care, Orbit Professional, Orbit and Extra. Ten subjects wore removable palatal appliances with four human-enamel half-slab insets containing subsurface demineralized lesions. For four times a day for 14 consecutive days subjects chewed one of the chewing gums for 20 min. After each treatment the enamel slabs were removed, paired with their respective demineralized control slabs, embedded, sectioned and mineral level determined by microradiography. After 1-week rest the subjects chewed another of the four gums and this was repeated until each subject had used the four gum products.

Results

Chewing with Trident Xtra Care resulted in significantly higher remineralization (20.67 ± 1.05%) than chewing with Orbit Professional (12.43 ± 0.64%), Orbit (9.27 ± 0.59%) or Extra (9.32 ± 0.35%). The form of added calcium in Trident Xtra Care was CPP–ACP and that in Orbit Professional calcium carbonate with added citric acid/citrate for increased calcium solubility.

Conclusions

Although saliva analysis confirmed release of the citrate and calcium from the Orbit Professional gum the released calcium did not result in increased enamel remineralization over the normal sugar-free gums. These results highlight the importance of calcium ion bioavailability in the remineralization of enamel subsurface lesions in situ.     

Enamel Demineralization Prevention during Fixed Orthodontic Treatment and Remineralization Strategies

Preventing enamel demineralization and remineralization of orthodontic patients is important and it di-ffers from other dental patients. Oral hygiene education and oral examination is essential to prevent enamel demi-neralization, and dietary education or control of sugar intake should not be ignored during fixed orthodontic treat-ment. Laser irradiation is a noteworthy method to prevent enamel mineralization. Products containing fluoride andfluoride released not only can be used to prevent demineralization but also have the ability of remineralization during orthodontic treatment. Oral hygiene products containing casein-phosphopeptide amorphous calcium phosphate(CPP-ACP) have been demonstrated to have the ability of remineralization such as sugar-free chewing gum added CPP-ACP, lozenges containing CPP-ACP and milk protein casein stabilized by phosphopeptides.     

Comparison of the remineralization potential of CPP-ACP and CPP-ACP with 900 ppm fluoride on eroded human enamel: An in situ study

Objective

The aim of this in situ study was to compare the remineralization potential of pastes containing CPP-ACP and CPP-ACP with 900 ppm fluoride on human enamel softened by a cola drink.

Design

Forty-five enamel specimens obtained from human third molar teeth were eroded in a cola drink for 8 min and then attached to intra-oral devices worn by five volunteers. The specimens were subjected to three different in situ remineralization protocols using: (1) CPP-ACP (Group I), (2) CPP-ACP with 900 ppm fluoride (Group II), and (3) saliva (Group III, control). Vickers microhardness measurements were obtained at baseline followed by demineralization and remineralization stages.

Results

The CPP-ACP, CPP-ACP with 900 ppm fluoride and saliva controls resulted in 46.24%, 64.25% and 2.98% increase in post-erosion microhardness values, respectively. One-way ANOVA revealed statistically significant differences in the mean microhardness values between pastes containing CPP-ACP and CPP-ACP with 900 ppm fluoride.

Conclusions

Both CPP-ACP and CPP-ACP with 900 ppm fluoride substantially remineralized the softened enamel, with the CPP-ACP and fluoride combination showing higher remineralization potential than CPP-ACP. This study confirmed the synergistic effect of fluoride with CPP-ACP on remineralization of eroded enamel.     

Incorporation of casein phosphopeptide-amorphous calcium phosphate into a glass-ionomer cement

Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) nanocomplexes have been shown to prevent demineralization and promote remineralization of enamel subsurface lesions in animal and in situ caries models. The aim of this study was to determine the effect of incorporating CPP-ACP into a self-cured glass-ionomer cement (GIC). Incorporation of 1.56% w/w CPP-ACP into the GIC significantly increased microtensile bond strength (33%) and compressive strength (23%) and significantly enhanced the release of calcium, phosphate, and fluoride ions at neutral and acidic pH. MALDI mass spectrometry also showed casein phosphopeptides from the CPP-ACP nanocomplexes to be released. The release of CPP-ACP and fluoride from the CPP-ACP-containing GIC was associated with enhanced protection of the adjacent dentin during acid challenge in vitro.     

The effect of adding calcium lactate to xylitol chewing gum on remineralization of enamel lesions

The purpose of the study was to determine whether adding calcium lactate to chewing gum containing xylitol enhances remineralization of enamel surfaces using an early caries lesion model. Enamel slabs were cut from human extracted sound teeth and artificial subsurface lesions created within each. Half the enamel slabs were used as controls and stored in a humidifier while half were mounted into oral appliances worn by 10 volunteers (22-27 years old, 2 males and 8 females) in a three-leg trial, during which they wore the appliance without chewing gum, chewed gum containing xylitol + calcium lactate or chewed gum containing only xylitol 4 times a day for 2 weeks. Calcium concentrations in the enamel surfaces of control and test slabs were measured by X-ray spectrometry and degrees of remineralization were calculated. The mean degree of remineralization was greater after chewing xylitol-Ca gum (0.46 +/- 0.10) than after no gum (0.16 +/- 0.14) or after chewing xylitol gum (0.33 +/- 0.10) (p < 0.01). In conclusion, chewing gum containing xylitol + calcium lactate could enhance remineralization of enamel surface compared to chewing gum containing only xylitol or no gum chewing.     

CPP-ACP与不同浓度氟促进釉质再矿化的研究

目的：探讨酪蛋白磷酸多肽-无定形磷酸钙(casein phosphopeptide-amorphous calcium phosphate,CPP-ACP) 与不同浓度氟离子联合应用对脱矿釉质再矿化的作用。方法：取正畸拔除的健康前磨牙30颗,乳酸凝胶法制备人工早期釉质脱矿后, 随机分为对照组(5% CPP-ACP组)、实验组1(5% CPP-ACP+500 mg/L F-组)和实验组2(5% CPP-ACP+900 mg/L F-组),采用MH-5型显微硬度仪测量处理前、后釉质标本的显微硬度,采用SPSS 18.0软件包对数据进行统计学分析。结果：显微硬度测量显示,实验组2牙釉质的表面显微硬度变化显著高于实验组1(P<0.05)及对照组(P<0.01)。结论：CPP-ACP能够促进脱矿釉质再矿化,氟对其有加强协同作用。在一定范围内,氟浓度越高,协同作用越好。     

Remineralisation by chewing sugar-free gums in a randomised, controlled in situ trial including dietary intake and gauze to promote plaque formation

Remineralisation has been shown to be an effective mechanism of preventing the progression of enamel caries. The aim of this double-blind, randomised, cross-over in situ study was to compare enamel remineralisation by chewing sugar-free gum with or without casein phosphopeptide amorphous calcium phosphate (CPP-ACP) where the enamel lesions were exposed to dietary intake and some were covered with gauze to promote plaque formation. Participants wore removable palatal appliances containing 3 recessed enamel half-slabs with subsurface lesions covered with gauze and 3 without gauze. Mineral content was measured by transverse microradiography, and plaque composition was analysed by real-time polymerase chain reaction. For both the gauze-free and gauze-covered lesions, the greatest amount of remineralisation was produced by the CPP-ACP sugar-free gum, followed by the gum without CPP-ACP and then the no-gum control. Recessing the enamel in the appliance allowed plaque accumulation without the need for gauze. There was a trend of less remineralisation and greater variation in mineral content for the gauze-covered lesions. The cell numbers of total bacteria and streptococci were slightly higher in the plaque from the gauze-covered enamel for 2 of the 3 treatment legs; however, there was no significant difference in Streptococcus mutans cell numbers. In conclusion, chewing sugar-free gum containing CPP-ACP promoted greater levels of remineralisation than a sugar-free gum without CPP-ACP or a no-gum control using an in situ remineralisation model including dietary intake irrespective of whether gauze was used to promote plaque formation or not.     

不同再矿化处理对漂白后釉质表面形貌及钙磷比的影响

目的 探究不同再矿化处理对漂白后釉质表面形貌及钙磷比的影响。方法 制备牛牙釉质样本,漂白处理后,随机分为5组：去离子水(DW)组;2%氟化钠(NaF)组;10%原花青素(PC)组;酪蛋白磷酸肽-无定形磷酸钙(CPP-ACP)组;PC+CPP-ACP组。用色差仪、扫描电镜、能谱仪和原子力显微镜分别检测样本漂白前、漂白后即刻及再矿化1周的颜色、表面形貌、元素和粗糙度变化。结果 各组间色差值无统计学意义(P>0.05)。扫描电镜显示,再矿化后各组均可见程度不一的矿物质沉积,PC+CPP-ACP组沉积物更明显。能谱仪显示,漂白后釉质表面钙磷比降低,再矿化后均有升高,NaF及PC+CPP-ACP组更高(P<0.05)。原子力显微镜显示,再矿化后各组均可见沉积物且粗糙度降低(P<0.05),PC+CPP-ACP组沉积物更明显、粗糙度更小(P<0.05)。结论 PC和CPP-ACP均能促进漂白后釉质再矿化,联合作用效果更佳。PC和CPP-ACP没有影响漂白效果。     

Influences of bicarbonate on processes of enamel subsurface remineralization and demineralization: assessment using micro‐Raman spectroscopy and transverse microradiography

In the present study, we investigated, using micro‐Raman spectroscopy (Raman) and transverse microradiography, the influence of bicarbonate [sodium hydrogen carbonate (NaHCO₃)] on the effects of carbonate ions in the mineral phase during demineralization (acid resistance test) of subsurface lesions. Baseline lesions were created by demineralizing bovine enamel, and specimens were then exposed to remineralization solutions containing 0, 5, or 50 mM bicarbonate. Acid resistance tests were performed on remineralized and sound enamel specimens. Raman spectra showed that carbonate and phosphate were incorporated into both surface layers and lesion bodies during remineralization in the presence of bicarbonate. Moreover, the presence of bicarbonate did not affect the rates of remineralization, although the average mineral profiles of remineralized enamel differed from those of sound enamel after acid resistance tests. Raman analyses enabled close evaluation of site‐specific characteristics of carbonate and phosphate in subsurface lesions. In conclusion, incorporation of carbonate and phosphate ions into enamel subsurface lesions during remineralization does not affect the magnitude of remineralization or acid resistance.     

Increased remineralization of subsurface enamel lesions with molybdenum treatment

The present study was conducted to determine the effect of high molybdenum (Mo) concentration on fluoride (F) uptake and remineralization of subsurface lesions utilizing acid cyclic methods in vitro. Bovine enamel with artificial subsurface lesions were treated with test solutions containing 1,000 ppm F (NaF) with and without 1,000 ppm Mo at pH 7.0. F uptake was analyzed using a microdrilling technique and remineralization was evaluated by polarized light microscopy. The results indicate that addition of 1,000 ppm Mo in a 1,000 ppm F solution modestly increases F uptake and promotes remineralization of subsurface lesions compared with 1,000 ppm F alone. These findings suggest that a high level of Mo may enhance the remineralizing activity of F in subsurface bovine enamel lesions in vitro.     

精氨酸牙膏对釉质早期龋再矿化作用的研究

目的 研究精氨酸牙膏对人牙釉质早期龋的再矿化效果。方法 制备人牙釉质龋标本,随机分为 3组：空白组（去离子水处理）,对照组（普通市售含氟牙膏处理,含质量分数 0.14%单氟磷酸钠）,实验组（精氨酸牙膏处理,含质量分数 0.14%单氟磷酸钠和 8%精氨酸）。经体外 pH循环 10 d,测量 pH循环前后各组样本的表面显微硬度,计算其差值;并通过偏光显微镜观察各组样本再矿化前后的形态学改变。pH循环结束后,脱矿处理 2h,测量其表面显微硬度,计算与 pH循环前的差值,评估再矿化处理对釉质抗酸能力的影响。采用酸蚀法测定各组的氟摄入量。结果 与对照组和空白组比较,实验组釉质表面的显微硬度、抗酸能力,以及氟摄入量均明显升高。偏光显微镜下观察,实验组病损深度明显变浅,且有表层下再矿化条带,而空白组与对照组病损无明显变化。结论 精氨酸牙膏能有效促进人牙釉质龋表层及表层下再矿化,在早期龋的防治领域具有较大的临床应用前景。