The aim of this study was to evaluate the antimicrobial effect and sensorial analysis of the gel developed with the essential oil of Melaleuca alternifolia. Thirty-four volunteers, divided into 2 groups, were monitored for 4 weeks. Initially, clinical biofilm (plaque index) and saliva samples (bacteria count) were collected, from which the standard values for each patient were obtained. For 7 days, group 1 used the melaleuca gel (Petite Marie/All Chemistry, São Paulo, Brazil), and group 2 used Colgate Total (S.B. Campo, São Paulo, Brazil). After 7 days, the plaque index was performed again, as well as the bacteria count and the sensorial analysis (appearance, color, odor, brightness, viscosity, and first taste sensation). The volunteers were instructed to return to their usual dental hygiene habits for 15 days. After this, group 1 started using Colgate Total, and group 2 started using the melaleuca gel, with the same evaluation procedures as the first week. The data were analyzed statistically with a significance level of 5%. In the bacteria count and clinical disclosure, the melaleuca gel was more effective in decreasing the dental biofilm and the numbers of bacteria colonies. According to the data from the sensory evaluation, Colgate Total (the control) showed better results regarding flavor and first sensation ( P <0.05). We concluded that melaleuca gel is efficient in bacteria control but needs improvement in taste and first sensation.
The prevention of caries and periodontal disease is a primary goal in dentistry. During orthodontic therapy, great changes occur in oral biofilm, resulting from the increase in retention niches of the orthodontic appliances. The oral cavity, colonized by supragingival biofilm and bacteria that normally inhabit the gingival sulcus, might predispose the patient to clinical and inflammatory alterations in the periodontal tissues.
Control of dental microbial biofilm can be achieved by interruption, reduction, or elimination of the microorganisms. Dentifrices help in the removal of dental microbial biofilm and are excellent vehicles for the release of therapeutic substances, such as antimicrobial agents.
Mechanical dental microbial biofilm removal by tooth brushing with dentifrices is the most common way of controlling caries. However, patients have difficulty in maintaining an acceptable level of oral hygiene, which is difficult to achieve by the mechanical method. Therefore, antimicrobial agents are being incorporated into mouthwash solutions and dentifrices as supplements to traditional cleaning methods.
Orthodontic treatments should not only have malocclusion correction and esthetic improvements as goals. During orthodontic treatment, professionals can teach preventive dentistry and control of caries, and encourage their patients to form new habits.
The use of phytotherapeutic products for the prevention, treatment, or cure of diseases is one of humanity’s most ancient forms of medical practice. Many medicinal plants inhibit the growth or development of bacteria that form part of dental microbial biofilm.
Therefore, the antimicrobial activity of vegetable oils and extracts has been recognized. Nevertheless, few investigations have compared a large number of oils and extracts using directly comparable methods.
The therapeutic formulations of dentifrices can contain components to reduce microbial biofilm, gingivitis, caries, and stomatitis, in addition to active antimicrobial ingredients such as Melaleuca alternifolia .
Various diseases, including inflammation, infections by microorganisms of the oral cavity, some viruses (including the herpes simplex virus), and fungi, have been treated with melaleuca oil because of its proven antimicrobial action. The oil has also demonstrated an anti-inflammatory response when applied topically. Among the diverse uses of the oil, the most interesting is the elimination of bacteria causing infection.
The action of melaleuca in microbial control has been reported in several studies. Nevertheless, there are no reports in the literature about this phytotherapeutic agent in patients with fixed orthodontic appliances. Therefore, the aims of this study were to verify the antibacterial effect with gel developed with the essential oil of Melaleuca alternifolia (Petite Marie/All Chemistry, São Paulo, Brazil) through clinical biofilm disclosure, to quantify Streptococcus mutans in saliva, and to perform comparative sensorial analyses in orthodontic patients.
Material and methods
Thirty-four volunteers were included, divided into 2 groups (groups 1 and 2, each containing 17 volunteers) of both sexes, between the ages of 11 and 29 years. The patients had a fixed orthodontic appliance and no caries, periodontal disease, or infiltrated restorations. The volunteers signed a term of free and informed consent, approved by the research ethics committee of the University of Araras in Brazil.
Initially, clinical biofilm (plaque index) and saliva samples (bacteria count) were collected, from which the standard value for each patient was obtained. For 7 days, group 1 used the melaleuca gel, and group 2 used the Colgate Total gel (S.B. Campo, São Paulo, Brazil). After 7 days, the plaque index was performed again, as well as bacteria counts and sensorial analyses (score analysis of appearance, color, odor, brightness, viscosity, first taste sensation). The volunteers were instructed to return to their usual dental hygiene habits for 15 days. After this period, group 1 started using Colgate Total, and group 2 started using melaleuca gel (switching the gels), with the same evaluation procedures as the first week. A summary of the research design is shown in the Figure 1 .
The participants were instructed to brush their teeth in the habitual manner, 3 times a day, using the gel and brush provided by the researchers, taking care to use a sufficient quantity of the get to cover all the soft toothbrush bristles in the longitudinal direction (Reach; Eco Johnson & Johnson, New Brunswick, NJ). Thus, approximately 1.5 g of dentifrice would be used in each brushing session. The participants were given tubes of melaleuca dental gel and Colgate Total 12 Fresh gel, each containing 30 g.
The melaleuca gel was composed of 2% carboxymethylcellulose, distilled water, 0.5% essential oil of Melaleuca alternifolia , 5% silica, 4.0% laurel sodium sulphate, 0.2% essence, 0.5% green coloring agent, 5.0% polyethylenoglycol 400, 1.0% menthol, 5.0% glycerin, 15.0% propylenoglycol, 0.15% saccharine, 0.15% cyclamate, and 0.25% sodium benzoate.
For dental microbial biofilm disclosure under artificial light, a wooden spatula to withdraw the mucosa was used and, when necessary, an exploratory probe, in case of doubt. The upper and lower first molars were evaluated.
The patients were followed for 4 weeks. On the first day of the first week, clinical biofilm disclosure was performed in accordance with the plaque index of Quigley and Hein and Ciancio et al, with scores to determine the clinical disclosure of biofilm. The plaque index is shown in Table I , as modified by Al-Anezi and Harradine. For each patient, the mean was calculated as the sum of the index value of each tooth divided by the number of teeth evaluated.
|1||Discontinuous plaque at the gingival margin or plaque only on the bracket or ligatures|
|2||Continuous line of plaque at the gingival margin less than 1 mm wide, or plaque only around the bracket|
|3||Gingival third of the surface is covered with plaque|
|4||Two thirds of the surface is covered with plaque|
|5||More than two thirds of the surface is covered with plaque|
The data were entered on clinical record charts. The clinician involved in the research (K.D.P.) was given no information about the products supplied to the volunteers because they were visually similar and identified as gels A and B.
Saliva was collected for counting Streptococcus mutans . The volunteers were given 2 g of solid paraffin to chew for 2 minutes to stimulate saliva formation. This sample of saliva was discarded after 1 mL of saliva from each volunteer was placed in a sterilized tube.
Petri dishes (5 × 2 cm) were used, as well as 20 mL of Mitis salivarius agar, which is the selective medium used for microorganisms of the mutans group. Rough surface colonies identified as Streptococcus mutans were counted for each Petri dish’s Mitis salivarius agar using a manual colony counter. The results were expressed in colony forming units.
The plaque index and the microbiologic evaluation data were analyzed for differences between the control and treatment groups by means with the Fisher exact test.
Sensorial analysis was performed after the volunteers used the gels during each week (7 days). Questionnaires were not used, to prevent induced answers. The parameters—general appearance, color, odor, brightness, viscosity, first taste sensation, and overall quality—were presented, which each subject had to classify on a scale from 1 to 9 to be recorded.
The scores were evaluated by means of a scale of points, in which the scores represented the following: 1 to 3, “I disliked it very much”; 4 to 6, “I neither liked nor disliked it”; and 7 to 9, “I liked it very much.”
Sensorial analysis was done with the nonparametric Wilcoxon test for paired data. A software program (version 9; SAS, Cary, NC) was used.
Data analysis by difference of means in dental microbial biofilm index per gel, as shown in Table II , showed statistically significant differences. There was a greater decrease in biofilm index, shown as a negative result, when the subject used the melaleuca gel compared with the reduction from the Colgate gel. Brushing the teeth with the melaleuca dentifrice promoted a significant reduction in dental microbial biofilm.
|Gel||n||Mean||SD||Minimum||Quartile 1||Median||Quartile 3||Maximum|
Bacterial counts, in Table III , showed statistically significant differences between the Colgate and the melaleuca gels, with a lower mean count of bacterial colonies. The melaleuca gel effectively promoted a greater reduction in the number of bacterial colonies.
|Gel||n||Mean||SD||Minimum||Quartile 1||Median||Quartile 3||Maximum|
In the results of the sensorial data analysis, Colgate Total gel (control) gave a better result regarding flavor and first taste sensation ( P <0.01). For the other criteria, there was no statistically significant difference between the 2 dentifrices ( Table IV ).
|Gel||n||Variable||Mean||SD||Minimum||Quartile 1||Median||Quartile 3||Maximum|
|First sensation ∗||8.35||0.98||5||8||9||9||9|
|First sensation ∗||7.03||2.18||1||6||7,5||9||9|