Journal of Dental Science and Medicine
Open Access

Biofilms; Waterways for dental units; Disinfection; Microorganisms; Nanoemulsion

Introduction

Due to the rising number of elderly and immunocompromised patients seeking dental care, the contamination of dental unit waterlines (DUWLs) is a growing concern in dentistry [1]. It is conceivable that microscopic organisms got from spit and plaque from the mouth of one patient can vaccinate different patients by means of dental unit water needles and hand pieces. Nanoemulsions are one-of-a-kind disinfectants that have a uniform population of high-energy droplets ranging in diameter. Nanoemulsions have broad biocidal efficacy against bacteria, enveloped viruses, and fungi by disrupting their outer membranes. There are a wide variety of commercial waterline cleaning products and systems available, some of which can be retrofitted to existing DUWLs [2]. The use of nanoemulsions to control adhesion and the formation of biofilm is a logical approach to the issues presented because we have demonstrated that nanoemulsions are effective against biofilms. Additionally, cetylpyridinium chloride has biocidal activity through mechanisms that are distinct from those of nanoemulsions.

Yogurt is the most well-known type of probiotic food, and dairy products seem to be the most natural way to get probiotic bacteria. Another benefit is that milk products contain essential nutrients for a growing child. Due to their natural content of casein, calcium, and phosphorous, they are also considered safe for the teeth [3]. However, the fermented milks that are available on the market have an acidic pH and contain sucrose. According to, these characteristics facilitate the formation of biofilms as well as Lactobacillus adhesion to the dental surface and decay development [4]. The dental biofilm that forms in the presence of sucrose contains a high concentration of alkali-soluble carbohydrates in addition to a low concentration of fluoride, calcium, and phosphorous. As a result, biofilm research is now crucial for a variety of therapeutic approaches.

Hence, taking into account both the presence of sucrose in the matured milks containing probiotic and its significance in the inorganic arrangement of the dental biofilm [5], the point of this study was to in situ assess: the pH of the dental biofilm before and after the application of the tested products, the concentration of fluoride (F), calcium (Ca), phosphate (P), and insoluble extracellular polysaccharides (EPS) in the dental biofilm, and the superficial microhardness test that was performed on the bovine dental enamel following the application of probiotic-fermented milk.

Materials and Procedures

The local Human Ethical Committee had previously approved the experimental design of this study [6]. The informed consent was signed by ten healthy volunteers between the ages of 21 and 34. The volunteers used dentifrice that did not contain fluoride one week prior to the beginning of the experiment and throughout the entirety of it. The study was conducted in three 14-day phases using an in situ blind crossover design. In order to allow for the accumulation of dental biofilm and to shield it from disturbance, each volunteer wore acrylic palatal devices with four enamel bovine blocks. Additionally, there was a gap of 1 mm between the enamel and the plastic mesh.

Volunteers dripped the following treatment solutions onto the enamel bovine blocks eight times per day throughout each experimental phase: matured milk A, or matured milk B or 20% sucrose arrangement (control). The oral replacement for the device took place five minutes after the application [7]. Between each phase of the experiment, a washout period of seven days was established. The volunteers’ diets were not restricted, but they were instructed to remove the appliance during meals and for oral hygiene. During the experiment, they were not permitted to utilize any fluoride or antimicrobial products.

Lacquer blocks planning

Lacquer blocks estimating 4 mm × 4 mm were acquired from cowlike incisor teeth recently put away in 2% formaldehyde arrangement (pH 7.0) for 1 month. They had their surface sequentially cleaned and the chose blocks were separated in concurrence with the normal of hardness of the absolute of blocks and the trust stretch. According to, cross-sectional enamel hardness (CSH) and surface hardness (SH) measurements were carried out. Evaluation of dental biofilm acidogenicity pH estimations were finished utilizing a palladium microelectrode. After overnight fasting to ensure that no bacterial carbohydrate was stored and five minutes after dripping either fermented milk, fermented milk, or a 20% sucrose solution, the pH of the dental biofilm was determined in situ on day 13 of each phase. Extra-orally, the solution was dripped onto four blocks [8]. The appliance was replaced in the mouth after one minute, and a palladium microelectrode was used to measure the in situ pH. A salt bridge of 3 M KCl was constructed between the volunteer’s finger and the reference electrode. The pH difference (pHd) was then determined.

Dental biofilm analysis

The dental biofilm that had formed on the enamel blocks was weighed in preweighed microcentrifuge cap tubes on the 14th day of each phase was added to each tube in the ratio of 0.5 mL to 10 mg plaque wet weight. The same volume was added to the tube as a buffer after three hours of extraction at room temperature with constant agitation. The acid-soluble concentrations of F, Ca, and P were determined after the sample was centrifuged for one minute was added to the precipitate. On the vortex, the samples were homogenized for one minute before being kept at room temperature for three hours under agitation. The concentration of the insoluble extracellular polysaccharides (EPS) was determined following centrifugation [9]. Utilizing an ion-specific electrode, the Orion a potentiometer, the Orion and under the same conditions as the samples, fluoride was analyzed. Phosphorus was measured with both a colorimetric method23 and a spectrophotometer. The calcium was analyzed with a potentiometer Orion 720-A and a calcium ion specific electrode that was calibrated with standards. The phenol-sulfuric acid method was used to analyze carbohydrates. All samples were tested twice, and the results were expressed in g/mg of biofilm.

Analysis of enamel blocks

Following the experimental phase, the surface hardness (SH2) and cross-sectional hardness were tested in accordance. Statistical analysis In every analysis, each volunteer was counted as “n.” The tested null hypothesis was that the fermented milk would not cause dental enamel to become demineralized. Means from biofilm analysis (fluoride, calcium, phosphorus, pH, and concentration of insoluble extracellular polysaccharides) and enamel analysis (%SH and KHN) did not exhibit a normal or homogeneous distribution according to the Kolmogorov– Smirnov test [10]. After that, they were subjected to the Kruskal–Wallis test, the Miller test, and the Kruskal–Wallis test with a significance level of 5% using the software.

Results and Discussions

Because it is fermented to acids and serves as a substrate for the production of extracellular polysaccharides (EPS) in dental biofilm, sucrose is regarded as the most carcinogenic carbohydrate. In this in situ study, the pH of the dental biofilm decreased for all treatments after the enamel was exposed to fermented glasses of milk and sucrose. These EPS make it easier for new bacteria to stick to the biofilm that is growing. On the other hand, when comparing the products that were used in this study, it was discovered that fermented milk contained a high amount of total carbohydrates. The difference between the reducing and total carbohydrates in this product could be explained by a high content of reducing carbohydrate caused by the hydrolysis from non-reducing carbohydrates. However, there was a significantly higher biofilm pH fall after fermented milk than fermented milk and sucrose, which could be justified by the lowest In this in-situ study, the fermented milk produced less EPS content on dental biofilm than the other treatments, which may partially explain its probiotic action despite the fact that both of the products’ carbohydrates can be metabolized by cariogenic bacteria [11]. Another point that could contribute is that the treatment that created lower EPS content on the dental biofilm likewise advanced lower rate change of surface hardness (%SH) and lower incorporated loss of subsurface hardness.

Different information that could affirm these outcomes could be the inorganic structure of dental biofilm, which diminished when shaped in presence of sucrose. The current discoveries showed that the convergence of the particles was lower for the benchmark group and matured milk than matured milk. The treatment with matured milk B brought about an essentially higher convergence of all particles in dental biofilm when contrasted with different gatherings, who demonstrated that biofilm formed in response to sucrose contained a high carbohydrate content and a low inorganic concentration of F, Ca, and P, reported the current outcome. Even though the dental biofilm contained a higher concentration of Ca and P when fermented milk was used than when fermented milk was used, the calcium content of the two fermented milks was comparable. Additionally, the percentage change in surface hardness that fermented milk produced was comparable to that of sucrose, which contains little calcium.

In situ studies have shown that high concentrations and frequency of sucrose exposure increased EPS concentration in the biofilm matrix, slowed fasting pH values, and enhanced enamel demineralisation when compared to the biofilm formed in the absence of sucrose. In relation to the demineralisation evaluated in both the enamel surface and in the cross-section microhardness, fermented milk produced less enamel demineralisation in comparison to fermented milk and sucrose [12]. This is in contrast Despite the fact that these results show a correlation with the concentration of EPS, they are not conclusive. In any case, the most elevated particles fixation couldn’t guarantee a higher microhardness, since certain examinations showed that even with a fluoride fuse in the treatment there is no change in the mineral loss.

The principal focal point of the examinations on oral probiotic potential has been the caries counteraction, essentially the collaboration among probiotic and dental biofilm microbiota, and the chance of lessening the quantity of Streptococcus mutans. In this manner, the creation of natural acids from dietary sugars has been a vital consider the caries cycle, because of the low pH produced by acids challenges that contributed for caries sores improvement [13]. By altering the metabolism of Streptococcus mutans, the probiotic action on the bacterial activity of fermented milk B may have reduced the EPS production in the biofilm. However, more research is needed to verify this assertion.

Conclusion

It was inferred that all treatment diminished the pH of dental biofilm and advanced demineralisation of the polish, in spite of the fact that matured milk B introduced the least EPS content and rate change and coordinated loss of surface hardness. After the literature has shown that probiotics are a promissory agent for reducing caries, more research should be done to evaluate how probiotics affect bacterial activity and how they affect demineralization.

Acknowledgement

None

Conflict of Interest

None

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