In Vitro Model to Evaluate the Development of Discolorations on Human Enamel Caused by Treatment with Mouth Rinses and Black Tea Considering Brushing.

OBJECTIVES: The study aimed to develop and test an in vitro model to investigate the staining potential of mouth rinses on human enamel, considering alternating intake of black tea and tooth brushing, thus mimicking the situation in the oral cavity more realistically. MATERIALS AND METHODS: Eight mouth rinses with six different active ingredients (benzydamine hydrochloride [BNZ], polyhexamethylene biguanide hydrochloride [PHMB], chlorhexidine digluconate [CHX], hexetidine gluconate [HEX], octenidine dihydrochloride [OCT] and octenidine dihydrochloride + 2-phenoxyethanol [OCTP]) and concentrations were tested. Sets of six halved human molar crowns were initially pretreated by soaking in artificial saliva (30 min). Afterward, the cyclic treatment was started by soaking in artificial saliva (2 min), staining with black tea (1 min), brushing with toothpaste (5 s), and soaking in the mouth rinse (30 s). Samples were rinsed with distilled water after each treatment step. The cyclic treatment was repeated 30 times, mimicking the consumer behavior after 15 days. Photographic images were taken after 0, 10, 20, and 30 cycles. Color measurements were conducted after each staining and brushing step using a VITA-Easyshade spectrophotometer to determine the difference in lightness ∆L and the total color difference ∆E. STATISTICAL ANALYSIS: Analysis of variance and post-hoc Tukey test (α = 0.05) were applied. RESULTS: The new testing model with included brushing sequences allowed to assess the staining behavior on human teeth and provided a clear differentiation between the different investigated products. In detail, up to cycle 10, ΔE values increased for all mouth rinses with each additional cycle number. However, while ΔE values continued to increase for 0.15% BNZ, 0.1% PHMB, and 0.2% CHX between treatment cycle 10 and 30, ΔE values only slightly increased after treatment with 0.08% OCTP, 0.1% OCTP, 0.1% OCT, and 0.1% HEX. After 20 and 30 cycles, significantly less staining was found for 0.08% OCTP, 0.1% OCT, 0.1% HEX as compared to 0.2% CHX, 0.15% BNZ, and 0.1% PHMB (p < 0.05). ΔE-values were significantly lower after treatment with 0.1% OCTP as compared to 0.2% CHX1 and 0.2% CHX2 (p < 0.05). CONCLUSION: The proposed new methodology was found to be appropriate for assessing the staining progression of mouth rinses over a simulated application period of 15 days. The model allows differentiation of products with different active ingredients and concentrations.

Influence of the Amount of Toothpaste on Cleaning Efficacy: An In Vitro Study.

OBJECTIVES: The aim of this in vitro study was to test the influence of the amount of toothpaste on enamel cleaning efficacy. MATERIALS AND METHODS: The hydrated silica-based test toothpaste (radioactive dentin abrasion: 60.19 ± 1.35) contained all ingredients of a regular fluoride toothpaste. The cleaning efficacy of four different toothpaste amounts (1.00 g, 0.50 g [both "full length of brush"], 0.25 g ["pea-size"], and 0.125 g ["grain of rice-size"]) diluted in 1.00 mL water were each tested for different brushing times (10, 30, 60, 120, 180, and 300 seconds) using a standardized staining model on human molars with a brushing machine. Photographic documentation and colorimetric measurements were conducted, respectively, initially, after staining and after each brushing step. Colorimetric measurements were used to calculate the stain removal (in %). STATISTICAL ANALYSIS: Results were analyzed by one-way analysis of variance with post hoc Tukey test and Levene's test for analysis of homogeneity of variance. The level of significance α was set at ≤ 0.05. RESULTS: The cleaning efficacy decreased significantly when using smaller toothpaste amounts. Stain removal after 120 seconds brushing time was: 77.4 ± 5.0% (1.00 g toothpaste), 75.7 ± 3.4% (0.50 g toothpaste), 54.1 ± 6.7% (0.25 g toothpaste), and 48.2 ± 7.1% (0.125 g toothpaste), respectively. CONCLUSION: In this in vitro study the cleaning efficacy of a medium-abrasive, hydrated silica-based toothpaste was analyzed. Note that 1.00 g toothpaste showed for all brushing times a significantly higher cleaning efficacy than 0.25 g toothpaste and 0.125 g toothpaste.

Robot and mechanical testing of a specialist manual toothbrush for cleaning efficacy and improved force control.

BACKGROUND: Toothbrushes require flexibility to access all dental surfaces and remove plaque effectively, but they should also aim to prevent or limit overbrushing and consequent damage to teeth and gums. In two studies, the physical properties and cleaning performance of specialist test toothbrushes with flexible necks were compared to a reference rigid-necked toothbrush. METHODS: In Study 1, a universal testing machine (Instron E 10,000) with a specially designed setup was used to test the deflection behaviour of toothbrush head and neck. Untufted toothbrushes were fixed in a custom holder and force was applied to the head while the deflection was measured. In Study 2, one control and five test toothbrushes were assessed using a robot system to simulate the cleaning of artificial plaque from defined surfaces of artificial replicated human teeth in a model oral cavity (typodonts). RESULTS: Study 1 showed that the flexible-neck toothbrush deflected 2 to 2.5 times more than the rigid-neck reference toothbrush when same force was applied to the toothbrush head. Study 2 revealed that all five test toothbrushes showed statistically superior simulated plaque removal to the reference toothbrush. This superiority was observed for all test toothbrushes employing horizontal and rotating brushing action (all p = 0.001) but only three of the five toothbrushes when vertical brushing was employed (all p = 0.001). Cleaning efficacy of the test toothbrushes was demonstrated both interdentally and at the gumline locations. The Complete Protection toothbrush showed the most effective cleaning performance followed by the Repair and Protect and Rapid Relief toothbrushes. CONCLUSION: The addition of a flexible-neck component to the toothbrush designs helped to reduce stiffness and may allow more effective cleaning compared to rigid designs with controlled force distribution on the teeth and gums. This may help to provide plaque control at all potential risk areas in an in vitro robot model and could support good oral hygiene in-use.

The Impact on Dental Staining Caused by Beverages in Combination with Chlorhexidine Digluconate.

OBJECTIVES: There are several hypotheses regarding how chlorhexidine (CHX) digluconate causes staining with the role of beverages, specifically the precipitation of anionic dietary chromogens onto adsorbed cations, the most probable cause. The aim of this study was to investigate and compare the staining potential of common beverages using an in vitro staining and brushing model to better understand the interactions between chromogens from different beverage categories and the teeth. MATERIALS AND METHODS: Human enamel samples were exposed to a cyclic treatment of artificial saliva and 0.2% CHX mouthwash combined with a range of beverages, with and without brushing, simulating a period equivalent to 2 weeks. Eleven beverages were tested: diet coke, diet lemonade, white wine, red wine, lager beer, black tea, coffee, black tea with milk, coffee with milk, ginger and lemon infusion, and water. Toothbrushing was performed in a brushing simulator with toothpaste and also with water. Colorimetric differences were determined by ΔE using a VITA Easyshade dental spectrophotometer. Statistical analyses were performed by one-way analysis of variance with post hoc Tukey's honestly significant difference test and Levene's test. RESULTS: Black tea and red wine produced highest staining, which agrees with the literature. Significant staining was also observed for a ginger and lemon infusion, coffee, coffee with milk, tea with milk, and lager beer compared with water (p < 0.05). The staining potential of diet coke in combination with brushing appeared to be connected to its low pH. Both white wine and diet lemonade produced stain comparable to the water control. After treatment with high staining beverages, scanning electron microscope evaluation confirmed the formation of a surface layer. The mechanical resistance of the stain differed depending on the beverage, black tea stain was the most resistant. The addition of milk to tea and coffee considerably modified the stain layer and the adhesion to the tooth surface. CONCLUSION: The data may help demonstrate that appropriate user guidance can avoid stain and in turn help improve user compliance during short-term use of this gold standard antimicrobial treatment.

In Vitro Whitening Effect of a Hydroxyapatite-Based Oral Care Gel.

OBJECTIVE: Oral care formulations aim to prevent oral diseases such as dental caries and gingivitis. Additionally, desire for white teeth still exists across all age groups. It is known that most whitening toothpastes are highly abrasive and can be harmful to teeth and gingiva. Therefore, a gel formulation with biomimetic hydroxyapatite (HAP; Ca5[PO4]3[OH]) as active ingredient was developed. This formulation was tested with respect to its tooth whitening properties in an in vitro study. MATERIALS AND METHODS: Enamel samples were allocated to either group (a) HAP gel, (b) whitening mouth rinse with phosphates, or (c) negative control (distilled water). Test products were applied by finger (a) or were rinsed (b, c) for 1, 3, and 9 (b and c only) cycles, respectively. RESULTS: Color changes (ΔE) were measured spectrophotometrically. Group (a) showed a significant increase in color changes with respect to whitening after one cycle (mean ΔE = 5.4 [±2.66], p ≤ 0.006) and three cycles (mean ΔE = 11.2 [±3.11], p < 0.0001) compared to groups (b) and (c). For group (b), a significant increase in color change was measured after three (mean ΔE = 2.77 [±1.01], p = 0.02) and nine cycles (mean ΔE = 3.27 [±1.61], p = 0.006) compared to (c). Group (c) showed only minor and statistically insignificant color changes. CONCLUSION: This in vitro study demonstrated a significantly higher ad hoc whitening effect of the HAP gel compared to the mouth rinse and water after short-time application.

Controlled release minocycline-lipid-complex extrudates for the therapy of periodontitis with enhanced flexibility.

We describe the development of flexible minocycline-lipid-complex extrudates with optimized mechanical and drug release properties. These extrudates contain a minocycline - magnesium stearate chelate complex with a higher stability in aqueous media, which has now been incorporated in a PEG-PLGA (polyethylene glycol - poly(lactic-co-glycolic acid)) matrix. PEG 1500 has been utilized in different concentrations to serve as plasticizer. The novel formulations have been characterized by texture analysis, X-Ray powder diffraction (XRPD) and differential scanning calorimetry (DSC). Extrudates with a reduced diameter of 300 µm (previously 600 µm) were introduced, and a more sensitive quantification method with a tandem-mass spectrometry detector was developed. From all tested formulations, the extrudates consisting of Expansorb DLG 50 - 6P (PEG-PLGA, molar weight 30-60 kDa) paired with 10% PEG 1500 emerged as best formulation. These extrudates feature a drug content of 11.5% and a controlled release over at least 42 days. The release profile is without a lag time and shows initially a slightly higher release rate, which is desired. Compared to previous developments, the extrudates now offer a high flexibility combined with a large mechanical resilience, which will ease the handling and administration.

In Vitro Evaluation of Antimicrobial Activity of Minocycline Formulations for Topical Application in Periodontal Therapy.

Periodontal therapy using antimicrobials that are topically applied requires slow or controlled release devices. The in vitro antimicrobial activity of biodegradable polymer formulations that contain a new minocycline lipid complex (P-MLC) was evaluated. The new P-MLC formulations that contained 11.5% minocycline were compared with pure minocycline or an existing commercial formulation, which included determination of minimal inhibitory concentration (MIC) values against two oral bacteria and activity on six-species periodontal biofilm. Moreover, the flow of gingival crevicular fluid (GCF) was modeled up to 42 d and the obtained eluates were tested both for MIC values and inhibiting biofilm formation. In general, MICs of the P-MLC formulations were slightly increased as compared with pure minocycline. Biofilm formation was clearly inhibited by all tested formulations containing minocycline with no clear difference between them. In 3.5 d old biofilms, all formulations with 250 µg/mL minocycline decreased bacterial counts by 3 log10 and metabolic activity with no difference to pure antimicrobials. Eluates of experimental formulations showed superiority in antimicrobial activity. Eluates of one experimental formulation (P503-MLC) still inhibited biofilm formation at 28 d, with a reduction by 1.87 log10 colony forming units (CFU) vs. the untreated control. The new experimental formulations can easily be instilled in periodontal pockets and represent alternatives in local antimicrobials, and thus warrant further testing.

Extrudates of lipophilic tetracycline complexes: A new option for periodontitis therapy.

The objective of this study was to develop an improved drug delivery system for the local antimicrobial treatment of periodontitis, that offers enhanced drug stability, easy application and controlled release over several weeks. Chelate complexes consisting of a tetracycline antibiotic and a fatty acid salt were developed. Minocycline and doxycycline were paired with magnesium- and calcium stearate in different molar ratios. These chelate complexes stabilize the active pharmaceutical ingredient and enable the incorporation into a PLGA (poly(lactic-co-glycolic acid)) polymer matrix via hot melt extrusion. The chelate complexes were characterized via UV/Vis- and IR-spectroscopy. A high antibiotic activity of the complex was observed in a disc diffusion test. The drug complex was mixed with different PLGA-polymers and cryomilled in advance of the extrusion. The hot melt extrusion yielded homogeneous extrudates with a diameter from 600 to 900 µm. They contain 11.5% of minocycline, are adjustable in length and are easy to handle. In vitro release studies revealed a controlled release of the drug over 42 days. In conclusion, the developed extrudates are promising systems to improve the treatment of periodontitis.

Material compatibility and antimicrobial activity of consumer products commonly used to clean dentures.

STATEMENT OF PROBLEM: Regular denture cleaning is essential to good oral health, but only limited evidence is available regarding the effects of common cleaning routines. PURPOSE: The purpose of this in vitro study was to determine the compatibility of denture materials with and the antimicrobial effects of typical cleaning regimens. MATERIAL AND METHODS: The evaluated treatments were derived from a study of dental professional recommendations and consumer habits, including denture cleanser tablets, toothpaste, mouthwash, isopropyl alcohol (IPA), household bleach, soap, and vinegar. The material integrity of denture materials, including polymethyl methacrylate (PMMA) and metals, was evaluated by scanning electron microscopy and profilometry after treatment with laboratory regimens simulating 2 years of typical consumer use. Treatments were also evaluated in a microbial kill time assay against a range of oral microorganisms with typical treatment regimens. RESULTS: Alcohol-based mouthwash and IPA damaged the surface of PMMA, and brushing with toothpaste caused scratching and surface material loss. Bleach caused limited damage to PMMA, but corroded CoCr alloy (pitting) and solder (layer formation). Denture tablets caused little damage to any materials apart from the layer formation on silver solder. Vinegar and soap were compatible with all materials. In antimicrobial assays, bleach gave excellent results, and IPA and mouthwash required concentrated dilutions to be effective. Cleanser tablets were effective at 5 minutes treatment time against all organisms. Toothpaste was effective against bacteria but not Candida albicans. Vinegar, soaps, salt, and sodium bicarbonate were microbially ineffective. CONCLUSIONS: Bleach was highly antimicrobial but incompatible with metal dental prosthesis components. IPA and mouthwash were antimicrobial but damaged PMMA. Specialist denture cleanser tablets gave a good combination of microbial efficacy and reasonable material compatibility.