Physical properties and enamel remineralization potential of arginine-fluoride varnishes.

OBJECTIVE: The study objectives were to examine the physical properties and enamel remineralization potential of fluoride (F) varnishes incorporated with arginine (Arg). METHODS: Four commercial F varnishes: 1) Duraphat®; 2) Flúor Protector®, 3) Fluor Protector S®, and 4) Fluorimax™ were supplemented with 2% w/v. Arg. The control/experimental varnishes underwent rheometric analysis to assess varnish density (δ), velocity (ν), and associated viscosity, both quantitatively (ν/δ) and qualitatively based on determined mass, volume, distance flow, and time under experimentation. The varnish wet/dry weights (at 2 h) were also analysed. Further, sound enamel specimens (T0) with artificial incipient caries-like lesions (T1) were treated with control/experimental varnishes and subjected to remineralization assay with artificial saliva for 6 h. Thereafter (T2), the specimens were characterized to estimate precipitated Ca and net enamel F uptake. Additionally, mineral density (MD) was assessed using micro-CT at T0, T1, and T2 to derive mineral gain (MG) and % remineralization for the treatment groups. RESULTS: When Arg is incorporated, the physical properties of the F-containing varnishes undergo a significant transformation, resulting in higher density, varnish weight, dry varnish weight, and viscosity compared to their respective control varnishes (p<0.05). Incorporating Arg in Duraphat®, Fluor Protector S®, and Fluorimax™ significantly improved both enamel Ca precipitation and F uptake compared to the respective controls (p<0.05). Additionally, the enamel F uptake was significantly higher with all the tested varnishes when enriched with Arg (p<0.05). The combined data for MD, MG, and % remineralization suggests that the remineralization potential of F-varnishes significantly increased when enriched with Arg (p<0.05). CONCLUSION: Incorporating Arg in inorganic F varnishes improves their physical properties and enhances the enamel remineralization potential of the varnishes. CLINICAL SIGNIFICANCE: This study highlights the possibility of incorporating Arg in distinct F-source varnishes. The synergism between active components (Arg-F) aids in enhanced remineralization and superior varnish physical properties, demonstrating a promising approach for high caries-risk patients.

Fluoride release potential of arginine-incorporated fluoride varnishes.

The study aimed to examine the fluoride (F) release potential of arginine (Arg)-incorporated F varnishes. Four commercially available F varnishes were included in the study: Duraphat® (5% NaF), Flúor Protector® (0.9% SiH2F2), Fluor Protector S® (NH4F), and Fluorimax™ (2.5% NaF). L-arginine (2% w/v.) was incorporated in these varnishes to estimate F release at 1 h, 4 h, 6 h, 24 h, 3 days, and 7 days using an F-ion selective electrode. The media pH of eluded varnishes was estimated and primary inorganic F extraction was performed. The main effects pH, F release, and computed integrated mean/cumulative F release for experimental groups were significantly higher than the controls (p<0.01). The primary extracted F concentrations for the Arg-containing groups were significantly lower than the control groups (p<0.001) demonstrating a chemical interplay with Arg incorporation. To conclude, irrespective of the inorganic F content, incorporating Arg in F-containing varnishes increases their F release potential.

Effect of arginine-fluoride varnish on preventing enamel erosion by paediatric liquid medicaments.

BACKGROUND: The study objective was to examine the effect of arginine-sodium fluoride (Arg-NaF) varnish on preventing enamel erosion by acidic paediatric liquid medicaments (PLM). METHODS: The treatment groups were: 1) 2% Arg-NaF; 2) 4% Arg-NaF; 3) 8% Arg-NaF; 4) NaF; 5) MI (CPP-ACFP) varnishes; and 6) no varnish. The pH of PLM (paracetamol and chlorpheniramine) was measured at baseline and after immersing the Perspex® blocks coated with varnishes at 0 min, 30 min, 1 h, and 4 h. Seventy-two enamel specimens (n = 72) were randomly divided into 2 groups by PLM and further by treatment groups. Then, the specimens were pre-treated with varnishes and subjected to erosive cycles (5 min, 2×/day for 4 days) by PLM. After each erosive challenge, the specimens were stored in artificial saliva. At baseline and after 4 days, the specimens were assessed for surface roughness (Ra) using 2D-surface profilometric analysis (SPA) and atomic force microscopy (AFM). Additionally, the Ca/P ratio was determined using scanning electron microscopy with energy-dispersive X-ray spectroscopy. Paired samples dependent t-test, 1-way ANOVA and 2-way ANOVA with Bonferroni post-hoc tests were used to analyse data with the level of significance set at p < 0.05. RESULTS: The pH of PLM with 8% Arg-NaF was significantly higher than the other groups at 30 min and 4 h (p < 0.05). With paracetamol, no significant difference was observed between the baseline and post-erosive cycle measured enamel Ra (by SPA/AFM) and Ca/P ratio for all treatment groups (p > 0.05). The Ra determined by AFM, at the post-erosive cycle with chlorpheniramine, when treated with 4 and 8% Arg-NaF was significantly lower than the other groups (p < 0.05); except CPP-ACFP (p > 0.05). With the chlorpheniramine post-erosive cycle, the Ca/P ratio for 4, 8% Arg-NaF and CPP-ACFP treated specimens was significantly higher than the baseline Ca/P (p < 0.05). CONCLUSION: The 4%/8% Arg-NaF and MI varnish® application exhibit an enhanced preventive effect against low pH (pH < 3.0) PLM-mediated enamel erosive challenges compared to 5% NaF varnish.

Porphyromonas gingivalis can degrade dental zirconia.

OBJECTIVES: The aim of present study was to examine the effect of Porphyromonas gingivalis (P.g.) adhesion on dental zirconia by characterizing the physical and chemical properties. METHODS: Eighty polished-sintered zirconia discs were prepared and randomly distributed to 5 groups (n = 16): Zirconia cultured with - Group 1: broth containing P.g. for - 3 days; Group 2: 7 days; Group 3: broth (alone) for - 3 days; Group 4: 7 days; and Group 5: dry discs (negative control). After experimental period, broths were analyzed for pH and Zr release with inductively coupled plasma-optical emission spectroscopy (ICP-OES). The zirconia surface was evaluated by scanning electron microscope (SEM), atomic force microscopy (AFM), X-ray diffraction (XRD), water contact angle (WCA), and biaxial flexural strength (BFS). RESULTS: The mean pH with zirconia adhesion to P.g. group was significantly higher than the broth control (p < 0.05). As per ICP-OES, Zr ion/particulate release with P.g. adhesion to zirconia were significantly higher than the controls (p < 0.05). Post-experimental incubation, no defects were found on zirconia surfaces; tetragonal phase remained constant with no transformation to monoclinic phase but lower peak intensities were identified in experimental groups. WCA of zirconia surfaces with P.g. bacteria for 3 days (12.04° ± 2.05°) and 7 days (15.09° ± 2.95°) were significantly higher than zirconia surfaces immersed with broth (only) for 3 days (7.17° ± 1.09°) and 7 days (7.55° ± 0.65°), respectively (p < 0.05). BFS values of zirconia with P.g. for 3 days (632.57 ± 119.96 MPa) and 7 days (656.17 ± 100.29 MPa) were significantly lower than zirconia incubated in broth alone (765.01 ± 20.12 MPa) conditions (p < 0.05). SIGNIFICANCE: Under the conditions of present study, it can be concluded that P.g. adhesion on zirconia leads to structural alterations of dental zirconia further contributing to zirconia degradation.

Enamel remineralization potential and antimicrobial effect of a fluoride varnish containing calcium strontium silicate.

OBJECTIVES: To investigate enamel remineralization and antimicrobial effect of sodium fluoride (NaF) varnish containing calcium strontium silicate (CSR). METHODS: CSR was synthesized by sol-gel process and incorporated in 5 % NaF varnish at three different concentrations (1 %, 2 %, and 4 % w/v). The treatment/control groups were: 1 % CSR+NaF, 2 % CSR+NaF, 4 % CSR+NaF, NaF, and no treatment. Strontium and fluoride release from the varnishes was evaluated. Sound enamel specimens (n = 6) were demineralized, varnish-treated, and subjected to remineralization cycle. Mineral density of enamel specimens was evaluated using micro-CT. Antimicrobial effect of the varnishes on Streptococcus mutans and Lactobacillus acidophilus biofilms was assessed using confocal laser scanning microscopy. The HGF-1 cytotoxicity of the varnishes was examined using CCK-8 assay. RESULTS: Both 2 % and 4 % CSR+NaF varnishes showed significantly higher F release and remineralization potential than NaF varnish (p < 0.05). Dead bacterial proportion of 4 % CSR+NaF varnish was significantly higher than NaF varnish (p < 0.05). The CFUs values of both S. mutans and L. acidophilus were significantly lower in 4 % CSR+NaF group than NaF group (p < 0.05). No significant difference in cell viability was observed among the groups (p > 0.05). CONCLUSIONS: Incorporation of 4 % CSR in a NaF varnish significantly enhanced its enamel remineralization and antimicrobial potential with no cytotoxic effect. CLINICAL SIGNIFICANCE: Dental caries is a major public health problem globally. The study highlights the great potential of CSR-doped NaF varnish as a novel anti-caries agent with synergistic remineralizing and antimicrobial properties to combat early enamel caries lesions in the general population.

The effect of synbiotic-fluoride therapy on multi-species biofilm.

OBJECTIVES: The study objective was to examine the effect of synbiotic-fluoride (SF) therapy within a multi-species cariogenic biofilm model system comprising of S. mutans, S. sanguinis, and S. gordonii. METHODS: The SF therapy was prepared using 2% L-arginine (Arg), 0.2% NaF and probiotic L. rhamnosus GG (LRG). The 8 treatment groups were: Group 1: No treatment, Group 2: 2% Arg, Group 3: 0.2% NaF, Group 4: LRG, Group 5: 2% Arg+0.2% NaF, Group 6: 2% Arg+LRG, Group 7: 0.2% NaF+LRG, and Group 8: SF therapy (2% Arg+0.2% NaF +LRG). Multi-species biofilm model over 96 h comprising Streptococcus mutans, Streptococcus sanguinis, and Streptococcus gordonii was utilized. The biofilms received cariogenic challenge and SF therapy 2 × /day. The extracellular matrix components were analyzed for carbohydrates, proteins, and extra-cellular DNA (eDNA). The live/dead cells were imaged and quantified using confocal microscopy. The viable/dead bacterial concentrations were estimated using propidium monoazide-quantitative polymerase chain reaction (PMA-qPCR). The gene expressions for gtfB, sagP, arcA, argG, and argH were measured using real-time reverse transcriptase qPCR. RESULTS: Carbohydrates and protein content with SF therapy were higher than non-LRG containing groups, while eDNA content was lower than other groups (p<0.05). Live bacterial proportions determined using confocal imaging with SF therapy were the lowest (p<0.05). The 2% Arg+LRG and SF therapy showed higher viable L. rhamnosus GG than 0.2% NaF+LRG (p<0.05). The dead S. mutans with SF therapy were higher than the other groups (p<0.05) with no difference from 2% Arg+0.2% NaF and 2% Arg+LRG (p>0.05). The SF therapy significantly downregulates gtfB and upregulates sagP, arcA, argG, argH gene expression (p<0.05). CONCLUSION: Synbiotic-fluoride therapy effectuates multi-fold changes in the multi-species biofilm matrix and cellular components leading to superior ecological homeostasis than its individual contents, prebiotics (arginine), probiotic (L. rhamnosus GG), and fluorides (NaF). CLINICAL SIGNIFICANCE: The ecological-based synbiotic-fluoride caries-preventive therapy aids in maintaining biofilm homeostasis to preempt/restore dysbiosis thereby sustaining dynamic-diverse health-associated microbial stability significant as a preventive regimen for high caries-risk patients.

Synbiotic-fluoride synergism on enamel remineralization, cytotoxicity and genotoxicity.

OBJECTIVE(S): The objectives of the present study were to examine the - a) enamel remineralization potential of synbiotic-fluoride (SF) therapy using a multi-species bacterial pH-cycling model; and b) cytotoxic and genotoxic effects of SF therapy extracts. MATERIALS AND METHODS: The SF therapy group comprised of 2% arginine (Arg), 0.2% NaF, and a probiotic Lactobacillus rhamnosus GG (LRG). The intervention groups studied were: 1) No treatment; 2) 2% Arg; 3) 0.2% NaF; 4) LRG; 5) 2% Arg+0.2% NaF; 6) 2% Arg+LRG; 7) 0.2% NaF+LRG; and 8) 2% Arg+0.2% NaF+LRG (SF therapy). The enamel remineralization potential of SF therapy was investigated under cariogenic biofilm challenge; while the cytotoxicity and genotoxicity of SF therapy extracts were examined on HGF-1 and Chinese hamster fibroblast V79, respectively. To determine the remineralization effect, the specimens were subjected to mineral density (MD) assessment using micro-CT, Ca/P molar ratio with SEM-EDX, and enamel fluoride uptake (EFU) estimates. The HGF-1 proliferation assessment was quantified using MTT/CCK-8 assays with qualitative analysis by nuclei staining Hoechst-based fluorescence imaging. The genotoxicity was determined by micronuclei formation test. RESULTS: Mineral gain and %remineralization derived from MD assessment for the SF therapy were significantly higher than the other groups (p<0.05). The %ΔCa/P for the SF and 2% Arg+0.2% NaF were significantly higher than the other groups (p<0.05). The SF and 2% Arg+0.2% NaF groups had the highest EFU compared to the other groups (p<0.05). No significant difference in the %viable HGF-1 cells were observed between the treatment interventions and no treatment group (p>0.05). Compared to the EMS-positive control, the micronuclei formation for all the intervention groups was significantly lower (p<0.05), with no significant difference among the treatment groups (p>0.05). CONCLUSION: The SF therapy enhanced enamel remineralization with no biocompatibility concerns. CLINICAL SIGNIFICANCE: With the enhanced enamel remineralization potential discerned in the present study, the SF therapy can be used as a promising caries-preventive agent targeted for high caries-risk individuals.

Physicochemical Properties and Inductive Effect of Calcium Strontium Silicate on the Differentiation of Human Dental Pulp Stem Cells for Vital Pulp Therapies: An In Vitro Study.

The development of biomaterials that exhibit profound bioactivity and stimulate stem cell differentiation is imperative for the success and prognosis of vital pulp therapies. The objectives were to (1) synthesize calcium strontium silicate (CSR) ceramic through the sol−gel process (2) investigate its physicochemical properties, bioactivity, cytocompatibility, and its stimulatory effect on the differentiation of human dental pulp stem cells (HDPSC). Calcium silicate (CS) and calcium strontium silicate (CSR) were synthesized by the sol−gel method and characterized by x-ray diffraction (XRD). Setting time, compressive strength, and pH were measured. The in vitro apatite formation was evaluated by SEM-EDX and FTIR. The NIH/3T3 cell viability was assessed using an MTT assay. The differentiation of HDPSC was evaluated using alkaline phosphatase activity (ALP), and Alizarin red staining (ARS). Ion release of Ca, Sr, and Si was measured using inductive coupled plasma optical emission spectroscopy (ICP-OES). XRD showed the synthesis of (CaSrSiO4). The initial and final setting times were significantly shorter in CSR (5 ± 0.75 min, 29 ± 1.9 min) than in CS (8 ± 0.77 min, 31 ± 1.39 min), respectively (p < 0.05). No significant difference in compressive strength was found between CS and CSR (p > 0.05). CSR demonstrated higher apatite formation and cell viability than CS. The ALP activity was significantly higher in CSR 1.16 ± 0.12 than CS 0.92 ± 0.15 after 14 d of culture (p < 0.05). ARS showed higher mineralization in CSR than CS after 14 and 21 d culture times. CSR revealed enhanced differentiation of HDPSC, physicochemical properties, and bioactivity compared to CS.

Qualitative and Quantitative Micro-CT Analysis of Natal and Neonatal Teeth.

The objective of this study was to characterize natal and neonatal teeth using micro-computed tomography. A total of 4 natal and 11 neonatal teeth were used for the analysis. The reconstructed scans were assessed for the maximum enamel/dentin thickness and mineral density (MD). The scanned specimens were 3D reconstructed to qualitatively determine the surface topography. The dentin thickness was two-fold greater than enamel thickness for both natal and neonatal teeth (p < 0.05). The cervical third enamel MD remained undetermined in natal and neonatal teeth. The dentin MD at the cervical third for neonatal teeth was significantly lower than the incisal and middle third dentin (p < 0.05). Similarly, the dentin MD at the cervical third of neonatal teeth was significantly lower than the cervical third dentin MD of natal teeth (p < 0.05). Our qualitative analysis suggests that the cervical thirds of both natal and neonatal teeth are peculiar of an anomalous structure, with neonatal teeth showing an irregular outline. Under the conditions of the present study, it can be concluded that the neonatal teeth studied exhibited a distinguishable aberrant structure compared to the natal teeth. Therefore, the natal teeth unfold as a more organized, three-dimensional structure compared to the neonatal teeth.

Biofilm modulatory response of arginine-fluoride varnish on multi-species biofilm.

OBJECTIVES: To examine the biofilm modulatory effect of arginine (Arg)-fluoride (F) varnish on multi-species biofilms. METHODS: Experimental varnishes were prepared by incorporating L-Arg (1, 2, and 4%) in 5% NaF varnish, which served as the control. Multi-species biofilms comprising Streptococcus mutans, Streptococcus sanguinis and Streptococcus gordonii were grown on hydroxyapatite (HA) discs and treated with the Arg and F released from the experimental and control groups. The HA discs with treated biofilms were examined for biofilm composition. The biofilm thickness and live/dead counts were investigated using confocal microscopic imaging, while biofilm polysaccharides, proteins, and extracellular DNA (eDNA) were assessed spectrophotometrically. Bacterial composition in biofilms was analysed using viability real-time quantitative polymerase chain reaction (qPCR). The relative gene expression (RGE) was determined for gtfB, SMU.150, nlmD, arcA, and sagP. RESULTS: Both the 2 and 4% Arg-NaF groups reduced biofilm thickness, with the 4% Arg-NaF group showing a significantly greater proportion of dead bacteria, followed by 1 and 2% Arg-NaF (p < 0.001). All Arg-NaF groups significantly reduced the carbohydrate content of the biofilm, while the 4% Arg-NaF-treated biofilms demonstrated higher concentration of eDNA and proteins compared to the control NaF (p < 0.001). Expression of gtfB, SMU.150, and nlmD were significantly downregulated in 4% Arg-NaF-treated biofilms; while 2% Arg-NaF enhanced the expression of arcA. Both 2% Arg-NaF and 4% Arg-NaF significantly increased the expression of sagP. CONCLUSION: The incorporation of L-arginine (2%/4%) enhances the biofilm modulatory effect of 5% NaF varnish through released Arg and F. CLINICAL SIGNIFICANCE: The study results indicate that Arg-F varnish (at 2%/4% w/v. Arg) has the potential to modulate cariogenic biofilms in high-risk individuals.

The acid-resistance potential of arginine-fluoride varnish treated enamel.

The study objective was to examine the acid-resistance potential of enamel carious lesions treated with arginine (Arg)-sodium fluoride (NaF) varnishes using nano-mechanical testing and chemical mapping. L-arginine (at 1%, 2%, & 4%) was incorporated in 5% NaF varnish. The experimental/control groups were: 1% Arg-NaF, 2% Arg-NaF, 4% Arg-NaF, NaF, and no treatment. Enamel specimen blocks were subjected to incipient carious lesion formation. After treatment, the specimens underwent chemical pH-cycling for 8-days and acid challenge for 2 h. The specimens were characterised for surface nano-hardness (SNH) and calcium/phosphate content of the treated lesions to determine enamel solubility reduction (ESR). Post-acid challenge, X-ray diffraction crystallography (XRD), and energy dispersive X-ray spectrophotometry (EDX) were performed. The SNH for 2%/4% Arg-NaF demonstrated a higher resistance to acid challenge with significantly higher SNH recovery than NaF varnish (p<0.05). The ESR potential of 2%/4% Arg-NaF varnish was significantly higher than NaF varnish (p<0.05). The XRD crystalline phases demonstrated that 2%/4% Arg-NaF had intense hydroxyapatite peaks discerning its increased potential to resist demineralization than NaF varnish. The EDX results showed that 2%/4% Arg-NaF demonstrated Ca/P ratio closer to hydroxyapatite (~1.67) post-acid challenge. Incorporating 2%/4% L-arginine in a 5% NaF varnish enhances the acid-resistance potential of NaF varnish.

Concentration-Dependent Multi-Potentiality of L-Arginine: Antimicrobial Effect, Hydroxyapatite Stability, and MMPs Inhibition.

This study's objective was to examine L-arginine (L-arg) supplementation's effect on mono-species biofilm (Streptococcus mutans/Streptococcus sanguinis) growth and underlying enamel substrates. The experimental groups were 1%, 2%, and 4% arg, and 0.9% NaCl was used as the vehicle control. Sterilised enamel blocks were subjected to 7-day treatment with test solutions and S. mutans/S. sanguinis inoculum in BHI. Post-treatment, the treated biofilms stained for live/dead bacterial cells were analysed using confocal microscopy. The enamel specimens were analysed using X-ray diffraction crystallography (XRD), Raman spectroscopy (RS), and transmission electron microscopy (TEM). The molecular interactions between arg and MMP-2/MMP-9 were determined by computational molecular docking and MMP assays. With increasing arg concentrations, bacterial survival significantly decreased (p < 0.05). The XRD peak intensity with 1%/2% arg was significantly higher than with 4% arg and the control (p < 0.05). The bands associated with the mineral phase by RS were significantly accentuated in the 1%/2% arg specimens compared to in other groups (p < 0.05). The TEM analysis revealed that 4% arg exhibited an ill-defined shape of enamel crystals. Docking of arg molecules to MMPs appears feasible, with arg inhibiting MMP-2/MMP-9 (p < 0.05). L-arginine supplementation has an antimicrobial effect on mono-species biofilm. L-arginine treatment at lower (1%/2%) concentrations exhibits enamel hydroxyapatite stability, while the molecule has the potential to inhibit MMP-2/MMP-9.

The effect of arginine-fluoride varnish on biochemical composition of multi-species biofilm.

OBJECTIVE: To examine the biochemical components of multi-species biofilm on the arginine (Arg)-sodium fluoride (NaF) varnish-treated enamel following bacterial pH-cycling. METHODS: l-arginine (at 1%, 2%, & 4% w/v.) was incorporated in a 5% NaF varnish. The experimental and control groups were: 1%, 2%, 4% Arg-NaF, NaF, and no treatment. Enamel blocks were prepared, acid-etched, varnish-treated and then subjected to 72 h bacterial pH-cycling in an oral biofilm reactor. The organic (carbohydrates, proteins, amyloids, and eDNA) and inorganic components (calcium, inorganic phosphate, F) were assayed for the obtained biofilm suspensions. The biofilms were stained for exopolysaccharides (EPS)/bacteria and the respective proportions of live/dead bacteria was determined using confocal imaging. RESULTS: The total carbohydrate content of the biofilm was the lowest for the 2% and 4% Arg-NaF (p < 0.05). Except for 2% Arg-NaF, the biofilm proteins for 4% Arg-NaF were significantly higher than the other groups (p < 0.05). The amyloids for Arg-NaF groups were significantly lower than the controls (p < 0.05). The eDNA for 4% Arg-NaF was significantly higher than the controls (p < 0.05). The 2% and 4% Arg-NaF-treated enamel had increased biofilm Pi and F compared to the NaF-treated enamel (p < 0.05). The proportion of biofilm EPS matrix to bacteria was significantly reduced in Arg-NaF groups compared to the controls (p < 0.05). The dead bacterial proportions of 4% Arg-NaF were significantly higher than the controls (p < 0.05). CONCLUSION: Higher concentrations (i.e. 2%/4%) of Arg in 5% NaF varnish have the potential to modulate the biochemical composition of the biofilm growing on the treated enamel.

Reinforced Universal Adhesive by Ribose Crosslinker: A Novel Strategy in Adhesive Dentistry.

Enzymatic biodegradation of demineralized collagen fibrils could lead to the reduction of resin-dentin bond strength. Therefore, methods that provide protection to collagen fibrils appear to be a pragmatic solution to improve bond strength. Thus, the study's aim was to investigate the effect of ribose (RB) on demineralized resin-dentin specimens in a modified universal adhesive. Dentin specimens were obtained, standardized and then bonded in vitro with a commercial multi-mode adhesive modified with 0, 0.5%, 1%, and 2% RB, restored with resin composite, and tested for micro-tensile bond strength (µTBS) after storage for 24 h in artificial saliva. Scanning electron microscopy (SEM) was performed to analyze resin-dentin interface. Contact angles were analyzed using a contact angle analyzer. Depth of penetration of adhesives and nanoleakage were assessed using micro-Raman spectroscopy and silver tracing. Molecular docking studies were carried out using Schrodinger small-molecule drug discovery suite 2019-4. Matrix metalloproteinases-2 (MMP-2) and cathepsin-K activities in RB-treated specimens were quantified using enzyme-linked immunosorbent assay (ELISA). The significance level was set at α = 0.05 for all statistical analyses. Incorporation of RB at 1% or 2% is of significant potential (p < 0.05) as it can be associated with improved wettability on dentin surfaces (0.5% had the lowest contact angle) as well as appreciable hybrid layer quality, and higher resin penetration. Improvement of the adhesive bond strength was shown when adding RB at 1% concentration to universal adhesive (p < 0.05). Modified adhesive increased the resistance of collagen degradation by inhibiting MMP-2 and cathepsin-K. A higher RB concentration was associated with improved results (p < 0.01). D-ribose showed favorable negative binding to collagen. In conclusion, universal adhesive using 1% or 2% RB helped in maintaining dentin collagen scaffold and proved to be successful in improving wettability, protease inhibition, and stability of demineralized dentin substrates. A more favorable substrate is created which, in turn, leads to a more stable dentin-adhesive bond. This could lead to more advantageous outcomes in a clinical scenario where a stable bond may result in longevity of the dental restoration.

Combined effect of arginine and fluoride on the growth of Lactobacillus rhamnosus GG.

The objectives of the in vitro study were: (1) to investigate the effect of combining L-arginine (Arg) and NaF on the growth of Lactobacillus rhamnosus GG (LRG); and (2) to identify an optimum synergistic concentration for the synbiotic (Arg + LRG)-fluoride (SF) therapy. 1% Arg + 2000-ppm NaF (A-SF) and 2% Arg + 2000-ppm NaF (B-SF) demonstrated antagonism against LRG (FIC > 4.0). Both XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) and WST-8 (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt) assays showed that A-SF and B-SF enhanced the growth of LRG when compared to 2000-ppm NaF and LRG control. Colony forming units, bacterial weight, and biofilm thickness of A-SF and B-SF were significantly higher than 2000-ppm NaF and LRG control. Biofilm imaging depicted that 2000-ppm NaF inhibited biofilm formation; while 1%/2% Arg, A-SF, and B-SF increased biofilm growth of LRG. Lactic acid formation was the lowest for 2000-ppm NaF, followed by A-SF and then B-SF. The SF buffer potential after 24 h was the highest for B-SF, and then A-SF. Biofilm pH for B-SF was closest to neutral. Fluoride, Arg and LRG bioavailability remained unaffected in B-SF. The relative gene expression for arcA, argG, and argH was significantly higher for B-SF than the respective controls. In conclusion, combining 2% Arg, 2000-ppm NaF, and LRG provides an optimum synbiotic-fluoride synergism.

Enamel remineralization potential of arginine-fluoride varnish in a multi-species bacterial pH-cycling model.

OBJECTIVES: To examine the enamel fluoride uptake and remineralization potential of arginine-fluoride (Arg-NaF) varnishes in a simulated clinical condition using a multi-species bacterial pH-cycling model. METHODS: L-Arginine (at 1 %, 2 %, and 4 % by wt.) was incorporated in a 5 % NaF varnish. Experimental and control groups were: 1 % Arg-NaF; 2 % Arg-NaF; 4 % Arg-NaF; NaF and no treatment. Artificial incipient caries-like lesions were formed on 30 enamel specimen blocks (n = 6). The specimens underwent multi-species bacterial pH-cycling in an artificial mouth system using oral biofilm reactor for 72 h after treatment. The specimens were evaluated for mineral density using micro-CT, Ca/P ratio with SEM-EDX, enamel fluoride uptake (EFU) and plaque fluoride uptake (PFU). RESULTS: Increasing concentrations of Arg in NaF varnish significantly increased the EFU of incipient caries-like lesions (p < 0.001). The PFU for 1 % Arg-NaF was significantly higher than 4 % Arg-NaF and the control NaF (p < 0.05). Post pH-cycling, Ca/P ratio with 1 %/2 % Arg-NaF was closest to hydroxyapatite (1.67). Mineral gain and % remineralization of 1 %/2 % Arg-NaF was significantly higher than the control NaF varnish (p < 0.05). CONCLUSION: The prebiotic L-arginine (at 1 %/2 % by wt.) in a 5 % NaF varnish enhanced the enamel fluoride uptake and remineralization potential of the conventional 5 % NaF varnish. CLINICAL SIGNIFICANCE: The Arg-NaF varnish addresses the limitations of fluorides on cariogenic biofilms. The Arg-NaF varnish appears a promising caries-preventive regimen that counters the pathogenic biofilms by Arg and promotes remineralization with fluorides. In high caries-risk patients, professional application of Arg-NaF varnish might aid to alleviate the global burden of dental caries.

Physicochemical Characteristics of Arginine Enriched NaF Varnish: An In Vitro Study.

The in vitro study objectives were to investigate the effect of arginine (Arg) incorporation in a 5% sodium fluoride (NaF) varnish on its physical and chemical properties including F/Arg release. Six experimental formulations were prepared with L-arginine (L-Arg) and L-arginine monohydrochloride at 2%, 4%, and 8% w/v in a 5% NaF varnish, which served as a control. The varnishes were subjected to assessments for adhesion, viscosity, and NaF extraction. Molecular dynamics were simulated to identify post-dynamics total energy for NaF=Arg/Arg>NaF/ArgArg concentration denotes the stabilized environment compared to NaF

Inter-method reliability for determining total and soluble fluorides in child low-fluoride formula dentifrices.

The study aimed to compare three methods for determining total (TF) and total soluble fluorides (TSF) in 5 child formula dentifrices (CFD) using Inter-method reliability (IMR) statistical approach. The methods were direct acid-hydrolysis (DM), the least-time-consuming method; Modified direct acid-hydrolysis with standard-addition method (MDM), ISO 19448:2018 method; and modified Taves acid-HMDS diffusion analysis (TAD), the claimed gold standard method. A significant difference in the mean difference was observed for all methods at all levels (p < 0.001), except DM and TAD for TF (p = 0.622). A proportional bias was discerned in the agreement distribution between DM and TAD for TF (p < 0.001). The ICC analysis identified significant reliability between all measurements, irrespective of the model, measure, and fluoride type (p < 0.001). For TF and TSF, the IMR between DM and TAD was lower than MDM and TAD for consistency/absolute agreement reliability at single/average measures. The reliability measure for DM and MDM was higher than MDM and TAD for TSF, but was lower than MDM and TAD for TF. The ICC measure for DM-TAD was significantly lower than DM-MDM and MDM-TAD (p < 0.05). The ISO 19448:2018 MDM is a reliable test that can be used as an alternative to TAD/DM for determining TF/TSF in CFD.

Enhanced Fluoride Bioavailability with Incorporation of Arginine in Child Dentifrices.

OBJECTIVE(S): To: 1) examine the fluoride concentrations in commercial child formula dentifrices (CFD)s; and 2) investigate the effect of arginine incorporation in CFDs on fluoride bioavailability. STUDY DESIGN: Five commercial CFDs were examined for fluoride concentrations. Total, total soluble, and insoluble fluorides in CFDs were determined by the modified Taves acid-diffusion method (TAD). Ionic F and MFP were estimated by modified direct method with standard addition technique. L-arginine (L-Arg)/L-arginine monohydrochloride (L-Arg.HCl) were incorporated at 2% w/w in the commercial CFDs. The pH of the toothpaste slurries, buffer capacity of the added Arg, potentially available fluorides (PAF) and 1-min PAF by TAD were determined. RESULTS: The CFDs had 4 to 32% of insoluble fluorides. Addition of L-Arg/L-Arg.HCl significantly improved the fluoride bioavailability in CFDs (p<0.05). Incorporation of L-Arg significantly increased the pH of toothpaste slurries (p<0.05); while L-Arg.HCl decreased the pH. Principal component analysis showed that L-Arg.HCl decreased the pH of toothpaste slurries due to the presence of Cl in the form of HCl; whereas the inherent elements/molecules (Na/P/Pi/F) remain distinct with unidentified influence on the variables. CONCLUSION(S): The CFDs containing NaF only have higher concentrations of bioavailable fluoride. Incorporating arginine (L-arginine or L-arginine monohydrochloride) at 2% w/w improves fluoride bioavailability of the child formula dentifrices.

A Scoping Review on Arginine in Caries Prevention.

OBJECTIVES: Emerging science on arginine or arginine formulations has driven the need to examine the research in the field. The scoping study objectives were (1) to identify the extent, range, and type of evidence on the role of arginine or arginine formulations in caries prevention and (2) to explore the future scope of research on arginine-containing caries-preventive agents. METHODS: A systematic search was performed in PubMed, Scopus, and Web of Science. In vitro studies, clinical trials, narrative reviews, systematic reviews and or meta-analysis, and umbrella reviews or meta-evaluation examining arginine or arginine formulations for caries prevention were included. The data-charting process involved extracting variables followed by evidence synthesis. Arginine variants investigated up to date were discussed to explore future scope of research. RESULTS: Thirty-nine articles were included for review from 105 identified citations comprising of in vitro studies, clinical trials, and reviews. Most articles studied 1.5% arginine-fluoride toothpaste. Most studies were from Asia, followed by North America, with fewest studies from Europe and South America. Arginine or arginine formulations demonstrated a superior caries-preventive effect compared with their matched controls (including fluorides); however, the evidence is with high risk of bias. Until now, three arginine variants have been investigated with l-arginine monohydrochloride as the least explored variant. CONCLUSIONS: The evidence on the caries-preventive effect of arginine or arginine formulations has a high risk of bias. High-quality clinical trials are needed to assess the caries-preventive potential of arginine in commercial formulations. The role of l-arginine monohydrochloride in caries prevention can further be explored by incorporating in self-applied and professionally applied caries-preventive agents.