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Bioactive properties of milk proteins with particular focus on anticariogenesis.

Beyond nutrition, there is an increasing amount of data and information to demonstrate a bioactive role for dairy components in adults including a role in prevention of dental caries. Specifically, the casein fraction and hydrolysates thereof have been the focus of researchers investigating cariogenicity prevention. Tooth enamel is a polymeric substance consisting of crystalline calcium phosphate embedded in a protein matrix. Dental caries develop by acidic demineralization (calcium and phosphorus solubilization) of tooth enamel. Demineralization occurs directly (acidic food consumption) or indirectly (by fermentation products of dental plaque odontopathogenic bacteria growing on residual food particles between teeth or adhering to the plaque). Research efforts with milk derived bioactive peptides have focused on inhibition of cariogenic, plaque-forming bacteria, inhibition of tooth enamel demineralization, and subsequent enamel remineralization. Caseinophosphopeptides (CPP) and glycomacropeptide (GMP) have been patented for use in common personal hygiene products to prevent dental caries. Research has shown CPP and GMP to be growth inhibitory to the cariogenic bacteria Streptococcus mutans and other species. Additionally, CPP forms nanoclusters with amorphous calcium phosphate (AMP) at the tooth surface to provide a reservoir of calcium and phosphate ions to maintain a state of super saturation with respect to tooth enamel. This would buffer plaque pH, and also provide ions for tooth enamel remineralization. Glycosidic structures attached to GMP are important to numerous bioactive properties of the peptide including anticariogenicity. Like CPP, GMP has shown inhibitory activity to enamel demineralization and promotes tooth enamel remineralization.

Amino Acid Sequence↗

Explorer probing of root caries lesions: an in vitro study.

Probing with a dental explorer is the preferred method of root caries diagnoses. However, studies suggest that use of a dental explorer in coronal caries detection may damage tooth structure. This pilot study investigated the effect of probing on remineralization of root surface lesions in vitro. Root-surface lesions were created on 10 extracted teeth by exposing them to a demineralizing solution for 21 days. One side of each lesion was then randomly selected and probed with an explorer, while the other side was not probed. The teeth then were placed in a remineralizing solution containing 10 parts per million (ppm) fluoride for 21 days. Of the 9 teeth with usable sections, all showed frank evidence of defects on the probed halves. Sites of remineralization were seen in the non-probed halves and adjacent to the probed defects, but not within or at the base of these defects. These results suggest that probing of root surfaces may create defects that do not fully remineralize.

Cariostatic Agents↗

The role of salivary peptides in dental caries.

Dental caries is a complex disease, characterized by demineralization of tooth structure. With a protective role, several salivary phosphopeptides appear to be involved in remineralization processes, delaying the loss of tooth structure. In this work we have correlated peptide saliva composition with dental caries susceptibility through the analysis of saliva and hydroxyapatite-adsorbed salivary peptides samples. Saliva samples were obtained from two groups, a caries-free and a cariessusceptible group, and were analysed using HPLC-MS and a sequential extraction with 6 m of guanidine followed by tri fluoroacetate. Data analysis has allowed us to verify a strong correlation between large amounts phosphopeptides (PRP1/3, histatin 1 and statherin), and the absence of dental caries, which reinforces the importance of these peptides in the maintenance of tooth integrity. In addition, in the caries-susceptible group a high number of peptide fragments was observed, suggesting a high proteolytic activity.

Chromatography, High Pressure Liquid↗

Toothbrush resistance and fluoride retention of sound, etched, fluoridated, and remineralized bovine enamel.

The wear resistance of tooth enamel that had undergone eight different treatments regarding the bonding procedure has been investigated. In comparison to sound enamel, the outer 3 microns of the etched-surface enamel show a decreased wear resistance against toothbrushing in vitro. Remineralization, either direct or preceded by acidified phosphated fluoride (APF) application, did not improve the wear resistance. The prophylactic effect of the APF treatment of etched enamel is considerable and is not lost after the soft, outer surface layer has worn off.

Acid Etching, Dental↗

Prevention and reversal of dental caries: role of low level fluoride.

Dental caries is a bacterially based disease that progresses when acid produced by bacterial action on dietary fermentable carbohydrates diffuses into the tooth and dissolves the mineral, that is, demineralization. Pathological factors including acidogenic bacteria (mutans streptococci and lactobacilli), salivary dysfunction, and dietary carbohydrates are related to caries progression. Protective factors which include salivary calcium, phosphate and proteins, salivary flow, and fluoride in saliva can balance, prevent or reverse dental caries. Fluoride works primarily via topical mechanisms which include (1) inhibition of demineralization at the crystal surfaces inside the tooth, (2) enhancement of remineralization at the crystal surfaces (the resulting remineralized layer is very resistant to acid attack), and (3) inhibition of bacterial enzymes. Fluoride in drinking water and in fluoride-containing products reduces tooth decay via these mechanisms. Low but slightly elevated levels of fluoride in saliva and plaque provided from these sources help prevent and reverse caries by inhibiting demineralization and enhancing remineralization. The level of fluoride incorporated into dental mineral by systemic ingestion is insufficient to play a significant role in caries prevention. The effect of systemically ingested fluoride on caries is minimal. Fluoride "supplements" can be best used as a topical delivery system by sucking or chewing tablets or lozenges prior to ingestion.

Adolescent↗

The effect of clodronate (dichloromethylene diphosphonate) on the mineral concentration of dental enamel and bone of the miniature swine.

Diphosphonates are compounds which prevent the crystallisation of calcium phosphates from solution. Our aim was to study the effects of clodronate (dichloromethylene diphosphonate), one of the diphosphonates, on the mineral concentrations of dental enamel and bone. In a subchronic experiment the effects of this compound were investigated. The drug was administered orally in three doses (100, 300, 1000 mg/kg) per day to 32 Göttingen miniature swines for six months. From the acid etch biopsy of enamel the amount of dissolved hydroxyapatite and the concentrations of fluoride and magnesium were measured. The calcium, phosphorus, fluoride, magnesium, carbonate and citrate concentrations were determined from the cancellous bone sample. All values were calculated for dry bone. Calcium/phosphorus and calcium/magnesium ratios were calculated for both enamel and bone. In tooth enamel the magnesium concentration showed a significant dose dependent decrease and calcium/magnesium ratio a significant dose dependent rise. Calcium phosphorus/ratio exhibited a dose dependent increase in surface enamel. Bone fluoride concentration showed a highly significant and calcium/magnesium ratio a significant dose dependent rise. Clodronate seems to interfere with the remineralization process of tooth enamel, through its effect on magnesium. Incorporation of the drug also appears to exhibit a marked action on the fluoride metabolism of bone. This may require attention in certain long term treatment situations with clodronate in high fluoride areas.

Animals↗

[De- and remineralization processes of the surface layer of the enamel in intact and pulp-extirpated teeth].

Clinico-laboratory investigation of 61 intact teeth and 61 teeth after pulp extraction was performed, including in vivo acid enamel biopsy and intravital staining with 2% methylene blue solution. The regularities were established governing Ca and P efflux from superficial enamel layer of intact and pulp-extracted teeth into the acid solution with respect to age, group of the tooth and pulp extraction terms. The pulp extraction factor was investigated with special reference to the tooth acid resistance and its remineralization rate.

Adolescent↗

[Effects of fluoride in drinking water on health of deciduous teeth].

INTRODUCTION: High incidence of decayed deciduous teeth, as well as lack of adequate therapy, makes tooth decay prevention very important. One of the simplest ways to reduce tooth decay is fluoridation of drinking water. The optimal concentration of fluoride in drinking water is 1 ppm/l, and many waters naturally contain this quantity. Waters in Vojvodina are mainly poor in fluoride, except in a few regions. It has long been postulated that fluoride has a prophyilactic effect during intrauterine life. Today a theory of greater local impact of fluoride has been accepted, as well as its role in de- and remineralization of solid tooth tissue. MATERIAL AND METHOD: This epidemiological study was performed in the area of Vojvodina, in places with various fluoride concentrations in drinking water (0.18-1.04 ppm/l). Dental examination was performed among 145 children, 6 years of age. For tooth decay detection DMF index was used. RESULTS AND DISCUSSION: In places with low and optimal fluoride concentration in drinking water the percentage of children with decayed teeth is different, but without statistical significance. The percentage of affected deciduous teeth is high in all places. In places with low fluoride concentration it is 24.2-32.3%, in places with optimal concentration is it 27-32%. The average value of DMF in all places is between 4.7-6.4. These results show that the optimal fluoride concentration in drinking water decreases the incidence of tooth decay, but this difference is not significant. CONCLUSION: Presence of fluoride in drinking water doesn't affect health of deciduous teeth. Decreased incidence of decayed deciduous teeth can be achieved only with combined usage of fluoride (local and systemic), as well as with an intensive health education program.

Child↗

[The functions of saliva].

Saliva is crucial in maintaining the oral health. The watery, viscous phase of saliva solubilizes the various substances and prevents the oral surfaces against desiccation. An agent is tasted only when present in a soluble state. The electrolytes of the saliva are essential as a buffer (bicarbonate and phosphate) and they are involved in the remineralization of the tooth enamel.

Humans↗

Integration of a laser fluorescence caries detection device in dental hygiene practice.

Research has shown that most initial carious lesions are found on occlusal surfaces. In addition, the widespread use of fluoride has resulted in harder enamel surfaces. Recent studies have focused on carious lesions, called hidden caries, that are difficult to detect by conventional visual, tactile, and radiographic techniques. The newly developed DIAGNOdent, a diode-laser caries detector, can be used to determine the soundness of tooth structure on occlusal surfaces, which will aid clinicians in determining whether a tooth should receive remineralization therapy, a sealant, or would benefit by more invasive restorative procedures. It is estimated that 30% to 50% more occlusal caries will be detected at earlier stages with this device. Dental hygienists perform patient-assessment procedures on a regular basis and will find the DIAGNOdent an ideal patient-assessment tool.

Dental Caries↗

In vitro evaluation of different remineralization periods in improving the resistance of previously eroded bovine dentine against tooth-brushing abrasion.

One dentine specimen was prepared from each of 90 bovine incisors. The samples were then evenly distributed among nine groups (A-I) and submitted to 10 alternating de- and re-mineralization cycles, including abrasion by tooth brushing. Each cycle started with a demineralization using the erosive soft drink Sprite Light for 1 min, followed by storing the samples in pooled human saliva for a total of 240 min. The specimens were removed from the saliva at different intervals (group A, 0 min; B, 15 min; C, 30 min; D, 45 min; E, 60 min; F, 90 min; G, 120 min) and brushed in an automatic brushing machine. Groups H (erosion, but no brushing) and I (no erosion, but brushing), which were also stored in saliva for 240 min, served as controls. After these cycles, loss of dentine was determined by profilometry, producing the following values (mean+/-S.D.), which were analysed statistically (P< or = 0.05): group A (5.03+/-1.49 microm), B (4.44+/-1.09 microm), C (4.91+/-0.95 microm), D (5.47+/-1.52 microm), E (5.29+/-1.45 microm), F (4.76+/-0.74 microm), G (5.16+/-0.71 microm), H (2.61+/-1.31), I (1.11+/-0.39). Groups A-G had no significant differences, but they showed a significantly greater loss of dentine than groups H and I. It is concluded that the abrasion resistance of eroded bovine dentine is still decreased after a remineralization period of 120 min.

Analysis of Variance↗

Oral effects of sugars and sweeteners.

Increasingly, sweeteners are being used in the diets of caries-prone individuals to reduce sugar intake. Such substitution deprives the bacteria in the dental plaque of the sugars many of them use to produce the acids that cause demineralization of tooth tissue and development of the caries lesion. Sweeteners are particularly effective replacements for dietary sugars because they also stimulate the flow of saliva which can, through several mechanisms, prevent demineralization and even bring about remineralization of already demineralized enamel, dentine or cementum. More saliva means that more of its nitrogen-containing substances will reach the dental plaque where they can be degraded by plaque bacteria and thus produce base and the alkaline conditions that are conducive to a shift from tooth demineralization to tooth remineralization. At the same time, the additional saliva brings to the plaque more calcium and phosphate ions, the necessary ingredients for the remineralization process. At alkaline pH, saliva also provides a source of easily solubilized calcium phosphate that, in association with salivary carbohydrate protein, becomes part of the dental plaque. Because it is dissolved by acid more easily than is the calcium phosphate of the tooth, plaque calcium phosphate acts as a substitute for the tooth tissues when periods of acid attack occur following sugar ingestion. It is proposed that diagnostic tests be developed that can conveniently determine the acid-base and demineralization-remineralization potentials of different dental plaques and that these tests be used to determine the extent to which sugars in the diets of caries-active individuals be replaced with sweeteners.

Bacteria↗

Dependence of in vitro demineralization of apatite and remineralization of dental enamel on fluoride concentration.

The anti-caries activity of fluoride is contributed to in several ways. Two major aspects of fluoride action are (i) the inhibition of demineralization at the crystal surfaces within the tooth, and (ii) the enhancement of subsurface remineralization resulting in arrestment or reversal of caries lesions. Fluoride present in the aqueous phase at the apatite crystal surface may play a determining role in the inhibition of enamel or dentin demineralization. In one part of the present study, the initial dissolution rate of synthetic carbonated-apatite in acetate buffers was measured with fluoride present in the buffer in the 0-2.6 mmol/L (0-50 ppm) range. Inhibition of demineralization was shown to be a logarithmic function of the fluoride concentration in solution. In the second part of the present study, an in vitro pH-cycling model was used for determination of the effect on net de/remineralization of enamel by treatment solutions containing fluoride in the 0-26 mmol/L (0-500 ppm) range. The net mineral loss was shown to be negatively related to the logarithm of the fluoride concentration. These studies have demonstrated an exponential quantitative relationship between fluoride concentration and inhibition of apatite demineralization or enhancement of remineralization. The clinical implications are (i) that simply increasing fluoride concentration may not necessarily give increased cariostatic benefit, and (ii) that improving the means of delivery of relatively low fluoride concentrations for longer times should be more appropriate for enhancing clinical efficacy.

Apatites↗

Anticariogenic complexes of amorphous calcium phosphate stabilized by casein phosphopeptides: a review.

Using laboratory, animal, and human in situ caries models, investigators have shown that casein phosphopeptide amorphous calcium phosphate complexes (CPP-ACP) exhibit an anticariogenic activity. The casein phosphopeptides (CPP) are produced from a tryptic digest of the milk protein casein by aggregation with calcium phosphate and purification by ultrafiltration. The CPP have a remarkable ability to stabilize calcium phosphate in solution and substantially increase the level of calcium phosphate in dental plaque. Through their multiple phosphoseryl residues, the CPP bind to forming clusters of amorphous calcium phosphate (ACP) in metastable solution, preventing their growth to the critical size required for nucleation and precipitation. The proposed mechanism of anticariogenicity for the CPP-ACP is that they localize ACP in dental plaque, which buffers the free calcium and phosphate lon activities, thereby helping to maintain a state of supersaturation with respect to tooth enamel depressing demineralization and enhancing remineralization. The CPP-ACP, unlike fluoride, can be added to sugar-containing foods and therefore have commercial potential as an additive to foods as well as to toothpastes and mouthwashes for the control of dental caries.

Amino Acid Sequence↗