Anticariogenic Activity Research Articles

Anticariogenic Activity and Haemolytic Study of Some Medicinal Plants Leaf Protein Extract against Six Oral pathogens in In vitro condition

Leaf extract of 12 medicinal plants were obtained by using 10mM sodium phosphate buffer and studied for their antibacterial activity against 6 cariogenic pathogens. Among them, the extract from Streblus asper showed the most intensive activity. Antibacterial activity of Streblus asper was investigated by the minimum inhibitory concentration (MIC) test and the viable cell count method. MIC of Streblus asper against Candida albicans causing dental caries was determined to be 2.5 μg/ml. The kinetic study showed that Streblus asper completely inactivated C. albicans at the concentration 50μg/ml in 15hours.Hemolytic study of the Streblus asper plant leaf protein gives HC50 value at a very high range than that of the MIC values against C. albicans of the organisms and thus flourish its application as successful pharmaceutical drug in practice. DOI: http://dx.doi.org/10.3126/ijasbt.v2i3.9344Int J Appl Sci Biotechnol, Vol. 2(3): 253-259

Phytopharmacological overview of Tribulus terrestris

Tribulus terrestris (family Zygophyllaceae), commonly known as Gokshur or Gokharu or puncture vine, has been used for a long time in both the Indian and Chinese systems of medicine for treatment of various kinds of diseases. Its various parts contain a variety of chemical constituents which are medicinally important, such as flavonoids, flavonol glycosides, steroidal saponins, and alkaloids. It has diuretic, aphrodisiac, antiurolithic, immunomodulatory, antidiabetic, absorption enhancing, hypolipidemic, cardiotonic, central nervous system, hepatoprotective, anti-inflammatory, analgesic, antispasmodic, anticancer, antibacterial, anthelmintic, larvicidal, and anticariogenic activities. For the last few decades or so, extensive research work has been done to prove its biological activities and the pharmacology of its extracts. The aim of this review is to create a database for further investigations of the discovered phytochemical and pharmacological properties of this plant to promote research. This will help in confirmation of its traditional use along with its value-added utility, eventually leading to higher revenues from the plant.

Anticariogenic activity and phytochemical studies of crude extract from some Indian plant leaves

Aim:The aim was to screen the selected Indian plants for their antibacterial efficacy against four cariogenic bacteria Lactobacillus acidophilus (LA)(Microbial Type Culture Collection [MTCC]-*447), Lactobacillus casei (LC) (MTCC-1423), Streptococcus mutans (SMU) (MTCC-890) and Staphylococcus aureus (MTCC-96). To identify and characterize active principle present in these plants for the treatment of dental caries.Materials and Methods:The dried plant leaves materials are extracted by cold extraction using hexane, ethyl acetate, methanol, and distilled water. The solvents were evaporated, and the dried masses were suspended in dimethyl sulfoxide and used for anticariogenic activity by agar well diffusion method. Minimum inhibitory concentration (MIC) was evaluated by two-fold serial broth dilution method. Preliminary phytochemical analysis of effective extract was carried out by thin-layer chromatography (TLC) and bioautography.Results:Ethyl acetate and hexane extract of Eucalyptus globules was found most effective against L. acidophilus with MIC value 31 μg/ml and 62 μg/ml, respectively. Ethyl acetate extracts of Acacia nilotica and methanolic extract of E. globules also exhibited antibacterial activity against SMU and L. casei with MIC value of 50 μg/ml. Qualitative analysis of E. globules revealed the presence of alkaloids, terpenoids, phenolic compounds, and cardiac glycosides. The active principle responsible for the anticariogenic activity from E. globules were separated by TLC and subjected to bioautography using SMU, LA and LC.Conclusion:Anticariogenic activity and preliminary phytochemical analysis revealed that E. globule have potential to treat dental caries.

Anticariogenic Activity of <i>Lagerstroemia speciosa</i> (L.)

Dental caries is the common infectious diseases of the oral cavity and is caused mainly by oral streptococci. The present study was carried out to investigate the anticariogenic activity of methanol extract of Lagerstroemia speciosa (L.) (Lythraceae) leaves. The inhibitory efficacy of methanol extract was tested against 12 oral isolates of Streptococcus mutans by Agar well diffusion method. The broth cultures of bacteria were swabbed uniformly on sterile Brain heart infusion agar plates and wells of 6mm were punched in the inoculated plates. Standard antibiotic and different concentrations of extract were transferred into labeled wells. Zone of inhibition was measured after incubation. The extract caused a concentration dependent inhibition of cariogenic isolates. Inhibition caused by standard antibiotic was higher than the methanol extract. Preliminary phytochemical analysis showed the presence of saponins, glycosides, tannins and terpenoids. The result of the present study reveals that methanol extract showed significant inhibitory activity against cariogenic isolates. The inhibitory efficacy of extract against cariogenic isolates could be due to the presence of these metabolites. In suitable form, the leaves could be used to treat dental caries.

Identification of Multiphosphorylated Peptides in Milk

Multiphosphorylated peptides endogenously present in milk exert anticariogenic activity due to their calcium binding capacity. This study performed comprehensive analysis of multiphosphorylated peptides in raw milk using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Since phosphopeptides are often negatively discriminated during ionization, putative phosphopeptides were identified by three different methods: (i) selective detection in 4-chloro-α-cyanocinnamic acid MALDI matrix compared to α-cyano-4-hydroxycinnamic acid; (ii) higher relative signal intensity in negative compared to positive ionization mode; and (iii) detection of signal pairs with mass differences of -80 Da or multiples thereof before and after enzymatic dephosphorylation. Thus, 18 putative phosphopeptides from raw milk were annotated. Peptide structures were then determined by product ion spectra from targeted liquid chromatography electrospray ionization tandem-MS analysis. Thus, β-casein33-48, β-casein29-48, β-casein1-21, β-casein2-25, β-casein1-25, β-casein1-27, β-casein1-28, β-casein1-29, β-casein1-32, αS2-casein1-21, and αS2-casein1-24 were revealed as major peptides with one or four phosphorylation sites in raw milk.

In Vitro Anticariogenic Effects of Drymocallis rupestris Extracts and Their Quality Evaluation by HPLC-DAD-MS3 Analysis

In this study, for the first time, we investigated in vitro inhibitory effects of Drymocallis rupestris extracts and their subfractions obtained with solvents of different polarity (aqueous, 50% ethanolic, diethyl ether, ethyl acetate and n-butanolic) against bacterial viability and caries virulence factors of Streptococcus spp. strains. The diethyl ether subfraction (PRU2) showed bacteriostatic and bactericidal activity against mutans streptococci, with minimum inhibitory concentrations (MICs) in the range of 0.75–1.5 mg/mL and minimum bactericidal concentrations (MBCs) in the range of 1.5–3 mg/mL. Furthermore, PRU2 inhibited biofilm formation by Streptococci in a dose-dependent manner. It was also found that all five D. rupestris preparations exhibited diverse inhibitory effects on de novo synthesis of water-insoluble and water-soluble α-d-glucans by glucosyltransferases of the mutans group streptococci. The phytochemical profile of investigated samples was determined by spectrophotometric and chromatographic (HPLC-DAD-MS3) methods. The high polyphenol (total phenol, phenolic acids, tannins, proantocyanidins, and flavonoids) contents were found which correlated with anticariogenic activity of the analyzed samples. The results demonstrate that D. rupestris extracts and their subfractions could become useful supplements for pharmaceutical products as a new anticariogenic agent in a wide range of oral care products. Further studies are necessary to clarify which phytoconstituents of D. rupestris are responsible for anticaries properties.

Anticariogenic Activity of Gnidia glauca (Fresen.) Gilg, Pothos scandens L. and Elaegnus kologa Schlecht

The present study was conducted to estimate total phenolic and flavonoid content and to determine anticariogenic efficacy of methanol extract of Gnidia glauca (Fresen.) Gilg (Thymelaeaceae), Pothos scandens L. (Araceae) and Elaegnus kologa Schlecht (Elaeagnaceae) leaves. Total phenolic and flavonoid content of leaf extracts was determined by Folin-Ciocalteau method and Aluminium chloride colorimetric estimation method respectively. Anticariogenic activity of leaf extracts was assessed against 13 clinical isolates of Streptococcus mutans by Agar well diffusion technique. The contents of total phenolics and flavonoids were higher in G. glauca followed by P. scandens and E. kologa. The extract of G. glauca showed greater inhibition of cariogenic isolates than P. scandens and E. kologa as revealed by wider inhibition zones. The study revealed a correlation between the amount of phenolics and flavonoids present in the extracts and the anticariogenic activity of the extracts. The plants used in this study could be used against dental caries.

Coffee melanoidins: structures, mechanisms of formation and potential health impacts

During the roasting process, coffee bean components undergo structural changes leading to the formation of melanoidins, which are defined as high molecular weight nitrogenous and brown-colored compounds. As coffee brew is one of the main sources of melanoidins in the human diet, their health implications are of great interest. In fact, several biological activities, such as antioxidant, antimicrobial, anticariogenic, anti-inflammatory, antihypertensive, and antiglycative activities, have been attributed to coffee melanoidins. To understand the potential of coffee melanoidin health benefits, it is essential to know their chemical structures. The studies undertaken to date dealing with the structural characterization of coffee melanoidins have shown that polysaccharides, proteins, and chlorogenic acids are involved in coffee melanoidin formation. However, exact structures of coffee melanoidins and mechanisms involved in their formation are far to be elucidated. This paper systematizes the available information and provides a critical overview of the knowledge obtained so far about the structure of coffee melanoidins, mechanisms of their formation, and their potential health implications.

용매 변화에 따른 버섯추출물의 항충치활성

버섯류는 기호식품으로서의 성격과 기능성식품으로서의 성격을 동시에 갖는 매우 독특한 식품소재로서 본 연구에서는 국내에서 자생하는 8종류의 버섯에서 항충치 활성성분을 조사해 보았으며, 더불어 높은 항충치활성을 가진 추출물을 얻고자 다양한 용매를 이용, 최적의 용매조성을 탐색해 보았다. Chloroform, methanol, acetone, ethyl acetate 및 증류수를 용매로 사용하였으며, 이들 추출물의 S. mutans에 대한 항균 효과와 glucosyltransferase (GTase) 활성 억제효과를 살펴보았다. 표고버섯, 영지버섯 및 만가닥버섯 추출물이 상대적으로 높은 항충치활성을 나타내었으며, 추출용매에 의한 항균효과를 조사해본 결과, chloroform, methanol, ethyl acetate로 추출한 버섯 추출물에서 상대적으로 높은 항균효과를 보여주었다. 8종의 버섯 중 항균효과가 뛰어나고 GTase 활성 억제효과가 높은 만가닥(Lyopyllum ulmarium) 버섯으로부터 chloroform, methanol 및 ethyl acetate 추출물을 얻어 항충치활성을 조사하였다. S. mutans에 대한 만가닥 버섯의 최대 항균효과를 얻기 위해 심플렉스 중심 합성 계획법(simplex centroid design)을 이용하여 최적의 추출용매조성을 살펴본 결과, chloroform, methanol 및 ethyl acetate 3가지 용매를 모두 동일한 비율(1:1:1)로 혼합하여 추출한 추출물에서 가장 높은 항균효과가 나타났다. 만가닥버섯추출물은 항충치 소재로서 중요한 역할을 할 수 있을 것으로 기대되며, 버섯추출물의 항충치 효과는 추출용매에 의해 영향을 받으므로 버섯에서 항충치활성물질을 얻고자 할 때에는 추출용매선택이 매우 중요할 것으로 사료된다. Various mushroom extracts were examined for their anti-Streptococcus mutans and anti-glucosyltransferase (GTase) activities. Mushrooms were extracted with chloroform, methanol, acetone, ethyl acetate, and distilled water. The chloroform, methanol, and ethyl acetate extracts showed higher antimicrobial activity than that of the others. The inhibitory effects of the chloroform, methanol, and ethyl acetate extracts of Lyopyllum ulmarium, which have high anti-microbial and GTase activities, on the growth of S. mutans were examined and the optimal ratio of solvents was also evaluated to investigate the influence of solvents using a simplex centroid design. The anti-cariogenic effects of the L. ulmarium extract on S. mutans was influenced by the solvent, and the optimum anti-cariogenic activity of the extract was obtained with the ratio of chloroform:methanol:ethyl acetate=1:1:1 (v/v/v). These results suggest that solvent selection is an important factor to extract anti-cariogenic materials effectively from mushrooms.

Antimicrobial activity of nanoemulsion on cariogenic planktonic and biofilm organisms

IntroductionNanoemulsions (NE) are a unique class of disinfectants produced by mixing a water immiscible liquid phase into an aqueous phase under high shear forces. NE have antimicrobial properties and are also effective anti-biofilm agents. Materials and methodsThe effectiveness of nanoemulsion and its components was determined against Streptococcus mutans and Lactobacillus casei by live/dead staining. In vitro antimicrobial effectiveness of nanoemulsion against planktonic S. mutans, L. casei, Actinomyces viscosus, Candida albicans and mixed culture was determined by a serial dilution technique to obtain minimum inhibitory concentration and minimum bactericidal concentration (MIC/MBC). In addition, efficacy was investigated by kinetics of killing, adherence and biofilm assays. ResultsCompared to its components, nanoemulsion showed notable antimicrobial activity against biofilm organisms, up to 83.0% kill within 1min. NE dilutions ranging from 243 to 19683 were effective against planktonic S. mutans, L. casei, A. viscosus, C. albicans and mixed culture of these four strains as shown through MIC/MBC assays. NE showed antimicrobial activity against planktonic cells at high dilutions, confirmed by time kill studies. The level of adhesion on glass surface was reduced by 94.2–99.5% in nanoemulsion treated groups (p<0.001). 4-Day-old S. mutans, L. casei, A. viscosus, C. albicans and mixed cultures biofilms treated with NE showed reductions of bacterial counts with decreasing dilutions (p<0.001). ConclusionThese results suggest that nanoemulsion has effective anti-cariogenic activity against cariogenic microorganisms and may be a useful medication in the prevention of caries.

Extracts of Eremophila longifolia inhibit the cariogenic activities of streptococcus mutans and streptococcus sobrinus

Plants belonging to the Eremophila genus are an integral part of the traditional medicine of indigenous Australian populations. Extracts of a number of Eremophilia species have demonstrated the presence of bioactivity, including inhibitory effects against Gram positive bacteria. The aim of the present study was to determine the activity of solvent and aqueous extracts of Eremophilia longifoliastem and leaves against Streptococcus mutans and Streptococcus sobrinus. A stem ethanol extract (SEE) demonstrated growth inhibition of the two cariogenic bacteria with a minimum inhibitory concentration (MIC) of 0.5% (w/v). Time-kill experiments indicated bactericidal activity by SEE within a 2 h period. Furthermore, this study assessed the anticariogenic activity of SEE in terms of its effect on glycolytic pH drop, viability of cells within an artificial biofilm and cell attachment to a membrane. In all experiments, SEE demonstrated significant anticariogenic activity compared with controls (p<0.05, Students t-test). SEE also compared favourably with commercial oral healthcare products. Preliminary phytochemical investigations suggested that the active components within SEE were phenolic compounds but unlikely to be flavonoids. Our study advocates SEE as a worthy candidate for further research into alternative chemotherapeutic approaches to dental caries. Key words: Eremophila longifolia, dental caries, biofilm, antibacterial.

Evaluation of ent-Kaurenoic Acid Derivatives for their Anticariogenic Activity

Ent-kaur-16(17)-en-19-oic acid (kaurenoic acid, KA) is a tetracyclic diterpene prototype for natural anticaries agents. Six KA derivatives were prepared and their antimicrobial activity against the main microorganisms involved in the caries process evaluated. The sodium salt of KA (KA-Na) was the most active, displaying very promising MIC values for most pathogens. Time-kill assays against the primary causative agent of caries (Streptococcus mutans) indicated that KA and KA-Na only inhibited growth in the first 12 h, suggesting a bacteriostatic effect. After this period (12-24 h), their bactericidal effect was clearly noted. KA and KA-Na showed no synergy when combined with the gold standard anticariogenic (chlorhexidine dihydrochloride, CHD) in the checkerboard assays against S. mutans.

Chemical composition, antimicrobial potential against cariogenic bacteria and cytotoxic activity of Tunisian Nigella sativa essential oil and thymoquinone

We investigate in this work the chemical composition by GC–EIMS, the antibacterial and the cytotoxic activities of Tunisian Nigella sativa essential oil and its bioactive compound, thymoquinone, were tested against various clinical cariogenic bacteria (n=30). Eighty-four compounds were identified in the essential oil. The major one was p-cymene (49.48%) whereas thymoquinone represented only 0.79%. The essential oil (2.43mg/disc) containing only 3.35μg of thymoquinone showed pronounced dose dependant antibacterial activity against Streptococcus mitis, Streptococcus mutans, Streptococcus constellatus and Gemella haemolysans (15.5±0.707mm). However, pure thymoquinone compound (150μg/disk) was active against all the studied strains especially S. mutans and S. mitis (24.5±0.71 and 22±1.41mm inhibition zones, respectively).Their MIC values are representatives of a good effect. So, the essential oil strongest activity was seen against S. mitis, S. mutans, S. constellatus and G. haemolysans (MIC 2.13mg/ml). However, thymoquinone was active against all the studied strains. The most important antibacterial activity was seen against S. constellatus (MIC 4μg/ml). The growth inhibition effects of thymoquinone on human epithelial cell lines (Hep-2) were significantly stronger than the essential oil. The IC50 of thymoquinone and essential oil were 19.25±1.6 and 55.2±2.1μg/ml, respectively.Our results demonstrate an important anticariogenic activity of the Tunisian N. sativa essential oil and thymoquinone. These effects further validate the traditional use of N. sativa in the folk medicine against various bacterial infections.

Antimicrobial activity of terpenoids from Copaifera langsdorffii Desf. against cariogenic bacteria

In the present work, the anticariogenic activities of nine labdane type-diterpenes and four sesquiterpenes were investigated. Among these metabolites, (-)-copalic acid (CA) was the most active compound displaying MIC values very promising (ranging from 2.0 to 6.0 µg/mL) against the main microorganisms responsible for dental caries: Streptococcus salivarius, S. sobrinus, S. mutans, S. mitis, S. sanguinis and Lactobacillus casei. Time kill assays performed with CA against the primary causative agent (S. mutans) revealed that, in the first 12 h, this compound only inhibits the growth of the inoculum (bacteriostatic effect). However, its bactericidal effect is clearly noted thereafter (between 12 and 24 h). Also, CA did not show a synergistic effect when combined with the anticariogenic gold standard (chlorhexidine, CHD) in the checkerboard assays against S. mutans. In conclusion, the results points out CA as an important metabolite in the search for new effective anticariogenic agents.

Anticariogenic Properties of ent-Pimarane Diterpenes Obtained by Microbial Transformation

In the present work, the anticariogenic activities of three pimarane-type diterpenes obtained by fungal biotransformation were investigated. Among these metabolites, ent-8(14),15-pimaradien-19-ol was the most active compound, displaying very promising MIC values (ranging from 1.5 to 4.0 μg mL−1) against the main microorganisms responsible for dental caries: Streptococcus salivarius, S. sobrinus, S. mutans, S. mitis, S. sanguinis, and Lactobacillus casei. Time kill assays performed with ent-8(14),15-pimaradien-19-ol against the primary causative agent S. mutans revealed that this compound only avoids growth of the inoculum in the first 12 h (bacteriostatic effect). However, its bactericidal effect is clearly noted thereafter (between 12 and 24 h). The curve profile obtained by combining ent-8(14),15-pimaradien-19-ol and chlorhexidine revealed a significant reduction in the time necessary for killing S. mutans compared with each of these two chemicals alone. However, no synergistic effect was observed using the same combination in the checkerboard assays against this microorganism. In conclusion, our results point out that ent-8(14),15-pimaradien-19-ol is an important metabolite in the search for new effective anticariogenic agents.

Anti-cariogenic and anti-biofilms activity of Tunisian propolis extract and its potential protective effect against cancer cells proliferation

Propolis is a multifunctional substance used by bees to maintain the safety of their hives. It is worldwide used for its potential therapeutic effects. In this study, Tunisian propolis ethanol extract (EEP) was tested for their anti-cariogenic, anti-biofilms and antiproliferative effects of many cell lines. The Tunisian EEP was evaluated in vitro against 33 oral pathogens including streptococci and enterococci using broth microdilution method. The anti-biofilms activity of EEP was assessed via Crystal Violet staining and MTT assays. The Tunisian EEP antiproliferative effect was evaluated on normal (MRC-5) and cancer cell lines (HT-29, A549, Hep-2, raw 264.7, Vero) by the ability of the cells to metabolically reduce MTT to a formazan dye. Our results revealed that Tunisian EEP possessed excellent protective effects against cariogenic and biofilms activity of oral streptococci. Furthermore, EEP showed a strong antiproliferative potencies against all studied cancer cell lines as judged by IC50 and its value ranges from 15.7 ± 3.4 to 200 ± 22.2 μg mL −1. These results suggest that EEP is able to inhibit cancer cell proliferation, cariogenic bacteria and oral biofilms formation. It could have a promising role in the future medicine and nutrition when used as antibiotic or food additive.

Antibacterial actvity and synergistic effect investigation of terpenoids from Copaifera langsdorffii Desf. against cariogenic bacteria

In the present work, the anticariogenic activities of nine labdane type-diterpenes and four sesquiterpenes were investigated (Figure). Among these metabolites, (–)- copalic acid (2) was the most active compound displaying very promising MIC values (ranging from 2.0 to 6.0µg mL-1) against the main microorganisms responsible for dental caries: Streptococcus salivarius, S. sobrinus, S. mutans, S. mitis, S. sanguinis and Lactobacillus casei [1]. Due to its highest anticariogenic activity, the synergistic effect of 2 combined with the anticariogenic gold standard (chlorhexidine, CHD) was investigated in the checkerboard assays [2] against the most important cariogenic microorganism (S. mutans). No synergistic effect was observed but the MIC values obtained allow us to conclude that 2 is an important metabolite in the search for new effective anticariogenic agents.

Anticariogenic and cytotoxic activity of clove essential oil (Eugenia caryophyllata) against a large number of oral pathogens

The occurrence of dental caries is mainly associated with oral pathogens, especially cariogenic bacteria. Numerous studies have validated the traditional use of medicinal plants by investigating the biological activity of essential oils. The Eugenia caryophyllata (clove) essential oil was tested in vitro against a large number of oral pathogens (114 streptococci and 46 yeast strains) using a disc diffusion method. The cytotoxicity assay of Eugenia caryophyllata essential oil on cancer cells (HT29, A549, Hep2, raw 264.7) and normal cells (MRC-5) was determined by the ability of the cells to metabolically reduce MTT to a formazan dye. Our results revealed that Eugenia essential oil possessed an excellent antibacterial activity against oral streptococci including the cariogenic bacteria as well as an excellent antifungal activity. Furthermore, the Eugenia caryophyllata essential oil showed significant cytotoxic effects against all studied cancer cell lines as judged by IC50 and its value ranges from 15.75 to 200 μg/ml. In conclusion, it is clear that clove oil shows powerful antibacterial and antifungal activity. The cytotoxic activity of the essential oil was dependent on the tested cell lines.

Variation in Total Polyphenolics Contents of Aerial Parts of Potentilla Species and Their Anticariogenic Activity

The aerial parts of selected Potentilla species (P. anserina, P. argentea, P. erecta, P. fruticosa, P. grandiflora, P. nepalensis, P. norvegica, P. pensylvanica, P. crantzii and P. thuringiaca) were investigated in order to determine their contents of polyphenolic compounds. The results showed that P. fruticosa has relatively high concentrations of tannins (167.3 ± 2.0 mg/g dw), proanthocyanidins (4.6 ± 0.2 mg/g dw) and phenolic acids (16.4 ± 0.8 mg/g dw), as well as flavonoids (7.0 ± 1.1 mg/g dw), calculated as quercetin. Furthermore, we investigated the in vitro inhibitory effects of aqueous extracts from these species against cariogenic Streptococcus spp. strains. It was found that the tested samples moderately inhibit the growth of oral streptococci. However, all the preparations exhibited inhibitory effects on water-insoluble α-(1→3)-, α-(1→6)-linked glucan (mutan) and artificial dental plaque formation. The extract from P. fruticosa showed the highest anti-biofilm activities, with minimum mutan and biofilm inhibition concentrations of 6.25–25 and 50–100 µg/mL, respectively. The results indicate that the studied Potentilla species could be a potential plant material for extracting biologically active compounds, and could become a useful supplement for pharmaceutical products as a new anticariogenic agent in a wide range of oral care products.

Casein Phosphopeptide-Amorphous Calcium Phosphate Incorporated into Sugar Confections Inhibits the Progression of Enamel Subsurface Lesions in situ

Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) has been demonstrated to exhibit anticariogenic activity in randomized, controlled clinical trials of sugar-free gum and a tooth cream. Two randomized, double-blind, crossover studies were conducted to investigate the potential of CPP-ACP added to hard candy confections to slow the progression of enamel subsurface lesions in an in situ model. The confections studied were: (1) control sugar (65% sucrose + 33% glucose syrup); (2) control sugar-free; (3) sugar + 0.5% (w/w) CPP-ACP; (4) sugar + 1.0% (w/w) CPP-ACP; (5) sugar-free + 0.5% (w/w) CPP-ACP. Participants (10 and 14 in study 1 and 2) wore a removable palatal appliance containing enamel half-slabs with subsurface lesions, except for meals and oral hygiene procedures, and consumed 1 confection 6 times a day for 10 days. The enamel half-slabs were inset to allow the development of plaque on the enamel surface. Participants rested for 1 week before crossing over to another confection. The appliances were stored in a humid container at 37°C when not in the mouth. After each treatment period, the enamel half-slabs were removed, paired with their demineralized control half-slabs, embedded, sectioned and then analysed using transverse microradiography. In both studies consumption of the control sugar confection resulted in significant demineralization (progression) of the enamel subsurface lesions. However, consumption of the sugar confections containing CPP-ACP did not result in lesion progression, but in fact in significant remineralization (regression) of the lesions. Remineralization by consumption of the sugar + 1.0% CPP-ACP confection was significantly greater than that obtained with the sugar-free confection.