|Year : 2018 | Volume
| Issue : 5 | Page : 262-266
The relationship between the consumption of yerba mate (Ilex paraguariensis) and the presence of micronuclei in the oral mucosa
Jéssica Pegoraro1, C M Marlon Dickemann1, S Iuri Martins1, Laura Kirsch1, L Ana Carolina Ecker1, Paula Wiethölter1, Carolina Barreto Mozzini2
1 Faculty of Dentistry, Faculty of Health of Rio Grande Do Sul, Fasurgs, Passo Fundo, RS, Brazil
2 Oncology Physical Therapy Clinic, Passo Fundo City Hospital, Passo Fundo, RS, Brazil
|Date of Web Publication||24-Oct-2018|
Prof. Paula Wiethölter
160 Angelica Otto Street, Passo Fundo, Postal code: 99025-270, RS
Source of Support: None, Conflict of Interest: None
Aims: The risk factors for cancer can be internal or external to the body, and the consumption of chimarrão has been recently cited as a potential influencing factor. This herbal infusion, ingested at high temperatures, and chemical carcinogenesis may contribute to cell mutation. Among the existing methods used to identify cellular alterations, the evaluation of the presence of micronuclei stands out. The purpose of this study was to identify the association between the consumption of chimarrão and the presence of micronuclei in the oral mucosa that evaluates the extent of damage that an aggressor agent may cause in the oral cavity. Material and Methods: Data collection included the collection of cells of the buccal mucosa and the application of a questionnaire on the consumption habits of 120 individuals, among them students, faculty, and employees of an academic institution. The collected cells were fixed, stained with Giemsa and analyzed by scanning optical microscopy. Results: The data collected were analyzed using descriptive statistics, Student's t-test, Spearman's correlation test, and Pearson's correlation test (Statistical Program BioEstat, 2007). The results showed that there are no statistically significant differences regarding the presence of micronuclei between the periodic consumption of chimarrão and no consumption at all, as well as between daily consumption and periodic consumption. Conclusions: Consuming chimarrão does not represent a risk factor for the significant increase in the number of micronuclei in the oral mucosa.
Keywords: Ilex paraguariensis, micronuclei, mouth neoplasia, oral neoplasms, yerba mate
|How to cite this article:|
Pegoraro J, Dickemann C M, Martins S I, Kirsch L, Ecker L A, Wiethölter P, Mozzini CB. The relationship between the consumption of yerba mate (Ilex paraguariensis) and the presence of micronuclei in the oral mucosa. J Int Oral Health 2018;10:262-6
|How to cite this URL:|
Pegoraro J, Dickemann C M, Martins S I, Kirsch L, Ecker L A, Wiethölter P, Mozzini CB. The relationship between the consumption of yerba mate (Ilex paraguariensis) and the presence of micronuclei in the oral mucosa. J Int Oral Health [serial online] 2018 [cited 2022 Jan 28];10:262-6. Available from: https://www.jioh.org/text.asp?2018/10/5/262/243853
| Introduction|| |
Cancer is characterized by a failure in the functional integrity of the cell cycle, giving the cells the ability to generate mutations, affecting the genes that control its proliferation and apoptosis, and making them aggressive and uncontrollable. Therefore, the cells tend to divide autonomously, accumulating more units, and this may trigger extensive genetic mutations, leading to the formation of the tumor.
The risk factors for its development may be internal or external to the body. The internal risk factor includes genetic inheritance and individual factors, such as age and gender; the external factors are related to environmental factors and habits and customs established in the environment, such as smoking, alcoholism, ultraviolet radiation, and diet. The interaction and time of exposure to these factors may predispose cellular changes. However, there are reports of other risk factors for this type of cancer, such as the consumption of chimarrão.,,
Yerba mate is a herbal infusion, popularly known as chimarrão, consumed at high temperatures by a large part of the population in South America, which can cause thermal injury and induce chemical carcinogenesis, thus contributing to cell mutations.,
Among the existing methods used to identify cellular alterations, the evaluation of the presence of micronuclei in buccal mucosa cells stands out. This evaluation can provide information about several events that may occur in the cells, such as cell proliferation, cell death, and cytotoxic evaluations.
A study conducted by Crasta et al. found that the micronuclei may subsist for several generations and that the mutations acquired in them may be introduced into the genome of a cancer cell that is in development.
It is still not possible to state that the consumption of chimarrão represents a risk factor for oral cancer. However, the following hypotheses have been proposed: the temperature of the water used in the consumption, as well as the chemical action of yerba mate associated with other risk factors, prominently the use of tobacco and alcohol, would increase the chances of triggering tumoral processes. On the other hand, chimarrão has been cited as being a potential protector of the circulatory system and having antioxidant effect.
Thus, this study aimed to identify the association between the consumption of chimarrão and the presence of micronuclei in the oral mucosa of an academic community.
| Materials and Methods|| |
In order to evaluate the effect of the consumption of yerba mate (Ilex paraguariensis) in the form of chimarrão on oral mucosa cells, students, professors, and employees of a private institution in the countryside of Rio Grande do Sul, selected by the researchers, were evaluated from February to August 2016. This study was approved by the Research Ethics Committee under the number 1.559.043, and all the participants signed the free and informed consent form.
The inclusion criteria were individuals who were part of the academic institution, were over 18 years old, of both genders, did or did not consume chimarrão, and signed the Free and Informed Consent Term. The exclusion criteria were those who did not agree to participate and those who gave up participating in the research after data collection.
Initially, the personal data and the data regarding the habits of each individual were collected, and then their oral mucosa cells were collected using the brush “cytobrush” through the internal scraping of the buccal mucosa.
The cells were placed on the slides, smeared and left at room temperature until they were completely dry. The samples were fixed with methanol and acetic acid (30 ml of methanol and 10 ml of acetic acid) and placed in an oven for 30 min.
Then, the slides were stained with Giemsa stain (10 ml of stain and 30 ml of distilled water) for 15 min and left at room temperature. The slides were washed with distilled water and left at room temperature to dry. After completely drying, they were stored in boxes at room temperature.
The slides were analyzed with an optical microscope of the Brand Bioval (BRAZIL), at ×400, and 1000 cells were analyzed per slide.
The micronuclei count and classification followed a protocol suggested by Tolbert et al. and was carried out as follows:
- Cell counting: the counting considered cells that had normal and intact nuclei, with a smooth and distinct nuclear perimeter, and with an intact cytoplasm, with the exception of those with small folds and little or no overlap with adjacent cells
- Micronuclei counting: the counting considered the micronuclei whose structure had a surrounding halo that suggested a membrane, less than one-third of the main nucleus diameter, with an intensity of Giemsa staining similar to that of the nucleus, on the same microscopic focal plane, and without overlaps or bridges with the nucleus. The total number of micronuclei was considered, as well as the number of cells affected by their appearance in the cytoplasm.
The data collected were statistically analyzed using the BioEstat 5.0 Statistics Software. The clinical data were analyzed using descriptive statistics. The data on the amount of micronuclei found, and the habit of consumption of chimarrão were analyzed using Student's t-test. Correlations were performed using the Spearman and Pearson correlation test, considering a P = 0.05.
| Results|| |
A total of 120 individuals were studied, of which 84 (70%) were female and 36 (30%) were male, with an average age of 25.2 (±8.3) years old (from 18 to 54 years old); 110 (91.7%) individuals were white, four (3.3%) were black, and six (5%) were from another race.
Regarding the family history of cancer, 53 (45.7%) individuals reported having a history of the disease, while 63 (54.3%) did not have. When questioned about the habit of consuming chimarrão, 53 (44.2%) individuals reported consuming it as a daily habit, 50 (41.7%) consumed chimarrão sometimes, 16 (13.3%) reported not consuming it, and one (0.8%) did not respond.
The data on the daily and periodic consumption of chimarrão are shown in [Table 1].
|Table 1: Characteristics of the subjects that consumed chimarrão daily and sporadically|
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The data related to the consumption of chimarrão are presented in [Table 2]. The study analyzed 120 slides in which 118 cells with one micronucleus and 51 cells with two or more micronuclei were identified; there were individuals who had cells with one micronucleus and with two or more. In two slides there were no cells with micronuclei. Thus, the general average of micronuclei found in all analyzed blades was of 7.2 with a standard deviation of ±7.2.
|Table 2: Analysis of the consumption or not of chimarrão and the presence of micronuclei|
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When the presence of micronuclei between the daily and sporadic consumption of chimarrão, we observed a statistically significant difference in cells that had two or more micronuclei, and those who consumed it sporadically presented more cells in these situations than those who consumed daily (P = 0.048) [Table 3].
|Table 3: Analysis of daily and periodic consumption and the presence of micronuclei|
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When we correlated the consumption of chimarrão and the presence of micronuclei in oral mucosa cells of those who had the daily consumption habit and those who never consumed the compound, the number of cells with 1 MN had an absent or very weak correlation (0.075) (P = 0.424); and when we correlated the number of cells with 2 or more micronuclei, the correlation was also weak (0.253) (P = 0.077).
When we correlated the consumption of chimarrão and the presence of micronuclei in cells of the oral mucosa of the individuals who had the daily consumption habit and those who consumed the compound sporadically, the number of cells with 1 micronucleus had absent or very weak correlation (0.043) (P = 0.672); and when we correlated the number of cells with 2 or more micronuclei, the correlation was weak (0.286) (P = 0.060) [Table 4].
|Table 4: Correlation between the use of chimarrão and the number of micronuclei|
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| Discussion|| |
Chimarrão, as it is popularly known, is a traditional compound in southern Brazil, especially in the state of Rio Grande do Sul. This compound is prepared with hot water placed within a container known as “cuia” (gourd) that contains the yerba mate. The mixture of yerba mate with water results in an infusion that is sorbed through a metal tube known as chimarrão pump.
Regarding the temperature of the beverages consumed by the population, it can be seen that temperatures above 65°C may influence the development of tumors in the oral cavity, pharynx, esophagus, and stomach. When consumed at high temperatures, they can potentiate the effect of carcinogenic substances on mucous membranes of the mouth, pharynx, and esophagus, especially when there is associated consumption of alcohol and tobacco.
However, in this study, we did not find any statistically significant evidence of the effect of the consumption of chimarrão on cellular stability.
De Stefani et al., Muñoz et al., and Castellsagué et al. investigated the relationship between the consumption of chimarrão and esophageal cancer, based on the temperature of the consumed water and the chemical composition of yerba mate. The results showed that the thermal action of the water in contact with the aerodigestive tract would increase the chances of developing neoplasia.
In a literature review made by Loria et al., which investigated a possible association between cancer and the consumption of yerba mate, the studies found hypothesized that the consumption of gourd at high temperatures could increase the risk of esophagus, larynx, and oral cavity neoplasia. It is also hypothesized that high water temperature tends to cause damage to the mucosa, leaving it susceptible to the penetration of carcinogenic substances present in tobacco and alcohol.
Kamangar et al., when analyzing the exposure to the polycyclic aromatic hydrocarbons (PAHs) present in the yerba mate, reported that these contaminants can be found in several kinds of food and beverage. In small quantities, they are not harmful to health, but in greater quantity, they tend to increase the risk of developing neoplasia in the long term. The analysis carried out with several commercial brands of yerba mate found high concentrations of hydrocarbons in hot and cold infusions, thus leading to the conclusion that the carcinogenic potential of this compound may be related to the presence of PAHs.
In the present study, it was difficult to evaluate the amount of damage that the yerba mate and the temperature of the water can cause in the oral cavity of the individuals, since there was no accurate measurement of the water temperature in which they consumed the compound, and the self-evaluation occurred from subjective answers such as “hot,” “very hot,” and “warm.”
Furthermore, there was a limitation in analyzing the size of the gourd, because the concept of size is subjective and varies from individual to individual, thus what may be a small gourd for some, may be big for others. Likewise, the amount of herb used by each person also varies, which consequently changes the amount of water added. All these factors may interfere with changes in oral mucosa and may have limited the results of this study. Moreover, the habit of drinking chimarrão is part of the traditions in Rio Grande do Sul, and it is most often consumed in a group of people who share the same gourd, making it even more difficult to discern the volume of liquid ingested by the individual, and this was one of the questions answered by the individuals in this study.
Although much has been discussed about the harmful effects of yerba mate, mainly in relation to cancer in the aerodigestive tract, other studies have evidenced the antioxidant action of this compound in tests in vitro and in vivo., In the phytochemical studies made with I. paraguariensis, several classes of chemical constituents were found, with emphasis to the amount of polyphenols and flavonoids. They seem to play an important role in the protection against various diseases, and can aid in the prevention of atherosclerosis due to the inhibition of oxidation of low-density lipoprotein (LDL), thus also reducing the risk of coronary arterial disease, and they also seem to contribute to reduce the risk of developing neurodegenerative diseases related to the oxidative process, such as the Alzheimer's Disease.
Another study performed sought to investigate the effects of the yerba mate in the process of alveolar cicatrization after dental extraction in mice, which received the compound during 28 days before the procedure and 28 days after it. The results showed an increase in osseous mass in the alveoli, higher levels of antioxidants and of biochemical markers of osseous formation, thus supposing that the ingestion of yerba mate during the postsurgical period of a dental extraction is beneficial for the osseous cicatrization.
This evidence may help to explain the results found in our study.
Similarly, in a study conducted by Gugliucci, who investigated the LDL oxidation of plasma from three healthy individuals, before and after the consumption of the extract of I. paraguariensis, it was possible to conclude that the extract inhibits the copper-induced LDL oxidation in vivo in human plasma, and that this effect is related to the polyphenols and flavonoids present in the extract.
In the study carried out by Rocha et al. to evaluate the effect of the yerba mate in different tissues of diabetic mice exposed during 30 days to the compound, it is possible to notice that the consumption of the yerba mate resulted in the reduction of the adipose tissue in animals, improvement in the levels of blood glycose, creatinine, urea, and total protein, as well as in the absorption of glucose by the muscles, thus demonstrating that the consumption of the yerba mate may be beneficial and assist in the metabolic disturbs of the diabetes.
The next steps will involve the study of the effect of the daily dose of chimarrão over the buccal mucosa, the brand of the yerba mate, and the active ingredient existing in the species, as an attempt to understand with more clarity the action of these compounds in the human organism.
| Conclusions|| |
Thus, we conclude in this study that the consumption of chimarrão was not related to the presence of MNs in the oral cavity. Among the main evidence, it is suggested that the phytochemicals constituents of I. paraguariensis may act to protect the oral mucosa, preventing the installation of a process of cellular disorder that may be stimulated by the use of hot water.
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Conflicts of interest
There are no conflicts of interest.
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[Table 1], [Table 2], [Table 3], [Table 4]