Glenn D. Walker

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.

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.

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.

Increased remineralization of tooth enamel by milk containing added casein phosphopeptide-amorphous calcium phosphate

Casein phosphopeptide amorphous calcium phosphate nano-complexes (CPP-ACP) in chewing gum, lozenges and mouthrinses have been shown to re-mineralize enamel subsurface lesions in human in situ experiments. The aim of this double-blind, randomized clinical study was to investigate the capacity of CPP-ACP added to bovine milk to re-mineralize enamel subsurface lesions in situ. Ten subjects drank milk containing either 2.0 or 5.0 g CPP-ACP/l or a control milk whilst wearing removable appliances with enamel slabs containing subsurface demineralized lesions. Each 200 ml milk sample was consumed once a day for each weekday over three consecutive weeks. After each treatment and one weeks rest the subjects crossed over to the other treatments. At the completion of the treatments the enamel slabs were removed and remineralization determined using microradiography and microdensitometry. The results demonstrated that all three milk samples re-mineralized enamel subsurface lesions. However, the milk samples containing CPP-ACP produced significantly greater remineralization than the control milk. The re-mineralizing effect of CPP-ACP in milk was dose-dependent with 2.0 and 5.0 g CPP-ACP/l producing an increase in mineral content of 70 and 148%, respectively, relative to the control milk. The differences in remineralization following exposure to the three milk samples were all statistically significant (P<0.001). In conclusion, this study shows that the addition of 2.0-5.0 g CPP-ACP/l to milk substantially increases its ability to re-mineralize enamel subsurface lesions.

Porphyromonas gingivalis RgpA-Kgp Proteinase-Adhesin Complexes Penetrate Gingival Tissue and Induce Proinflammatory Cytokines or Apoptosis in a Concentration-Dependent Manner

ABSTRACT The RgpA-Kgp proteinase-adhesin complexes of Porphyromonas gingivalis were observed, using immunostaining, in human gingival tissue associated with periodontitis but not in healthy tissue. The staining pattern suggested a concentration gradient from the subgingival plaque into the subjacent gingival connective tissue. Intense immunostaining was observed in areas displaying gross disturbance of tissue architecture. P. gingivalis cells and the RgpA-Kgp complexes at low concentrations were shown to stimulate secretory intercellular adhesion molecule 1, interleukin-8 (IL-8), IL-6, and macrophage chemoattractant protein secretion from cultured human epithelial (KB) and fibroblast (MRC-5) cells. However, at high concentrations a reduction in the level of these mediators was observed. In contrast, macrophage inflammatory protein 1α and IL-1α were stimulated only at high P. gingivalis cell concentrations. P. gingivalis cells and the RgpA-Kgp complexes were shown to induce apoptosis in KB and MRC-5 cells in a time- and dose-dependent manner. These data suggest that the RgpA-Kgp complexes penetrate the gingival connective tissue; at low concentrations distal from the plaque the complexes stimulate the secretion of proinflammatory mediators, while at high concentrations proximal to the plaque they induce apoptosis and attenuate the secretion of proinflammatory mediators.

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 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.

Effect of added calcium phosphate on enamel remineralization by fluoride in a randomized controlled in situ trial.

UNLABELLED Dental products containing calcium phosphate and fluoride are claimed to enhance enamel remineralization over fluoride products. OBJECTIVES To compare remineralization of enamel subsurface lesions by dental products with added calcium phosphate in a double-blind, randomized, cross-over in situ study. METHODS Human enamel specimens with subsurface lesions were prepared and inserted into intra-oral appliances worn by volunteers. A slurry (1g product plus 4 ml H(2)O) of each product was rinsed for 60s, 4 times per day for 10 days. Six products were tested (i) placebo, (ii) 1000 ppm F, (iii) 5000 ppm F, (iv) Tooth Mousse (TM), (v) TM plus 900 ppm F (TMP) and (vi) Clinpro with 950 ppm F. Calcium, inorganic phosphate and fluoride levels were measured in post-rinse/saliva samples using ion chromatography. Mineral content was measured using transverse microradiography. RESULTS Only TM and TMP significantly increased salivary calcium and phosphate levels. The products produced remineralization in the following order from lowest to highest: placebo<1000 ppm F=Clinpro<5000 ppm F<TM<TMP. CONCLUSION Clinpro was not significantly different to 1000 ppm F whereas TM and TMP were superior to 5000 ppm F with TMP producing the highest level of enamel lesion remineralization.

Consumption of milk with added casein phosphopeptide-amorphous calcium phosphate remineralizes enamel subsurface lesions in situ.

BACKGROUND Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) is an anticariogenic agent that is suitable to be added to foods. The aim of this double-blind, three-way crossover randomized study was to investigate the capacity of CPP-ACP, when added to bovine milk, to remineralize enamel subsurface lesions in situ. METHODS Ten subjects drank 100 mL of bovine milk containing no added CPP-ACP (control milk), 0.2% (w/v) CPP-ACP or 0.3% (w/v) CPP-ACP, for 30 seconds once daily for 15 days, whilst wearing removable appliances with attached slabs of enamel containing subsurface enamel lesions. After each treatment and a one-week washout period, subjects crossed over to another treatment and this was repeated until they had consumed each of the three milk products. At the completion of each treatment the enamel slabs were removed and remineralization was determined using microradiography. RESULTS The results demonstrated that all three milk samples remineralized enamel subsurface lesions in situ. However, the two milk samples containing added CPP-ACP each produced significantly greater remineralization than the control milk. CONCLUSIONS The remineralizing effect of CPP-ACP in milk was dose-dependent with milk containing 0.2% CPP-ACP and 0.3% CPP-ACP producing an increase in mineral content of 81% and 164%, respectively, relative to the control milk.

Effect of casein phosphopeptide-amorphous calcium phosphate added to acidic beverages on enamel erosion in vitro.

BACKGROUND To investigate, in vitro, the effect on enamel erosion of the addition of 0.2% w/v casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) to four commercially-available soft drinks, two of which were carbonated. METHODS Enamel specimens (n=27) were sectioned from sound extracted human third molar teeth and polished to a mirror finish. Exposed enamel windows of 1 mm2 were created by painting the surface with acid-resistant nail varnish. Four citric flavoured soft drinks (pH range 2.2 to 2.4) and distilled deionized water (DDW) were tested. Each drink was tested with and without 0.2% CPP-ACP w/v. The specimens were placed into 50 mL of solution at 37 °C for 30 minutes, rinsed and varnish removed. The samples were profiled with a white light profilometer and erosive depths recorded. RESULTS All soft drinks tested caused enamel erosion but adding 0.2% w/v CPP-ACP significantly reduced (p<.05) erosive depth in all test solutions in comparison with the solutions without CPP-ACP. The erosive depths for all solutions with 0.2% CPP-ACP did not differ significantly from those of DDW. CONCLUSIONS Adding CPP-ACP at 0.2% w/v significantly decreased the erosivity of all four soft drinks. The erosivity of the soft drinks with 0.2% CPP-ACP added did not differ significantly from that of distilled water.