Elsevier

Archives of Oral Biology

Volume 59, Issue 9, September 2014, Pages 887-896
Archives of Oral Biology

Antiadherent activity of Schinus terebinthifolius and Croton urucurana extracts on in vitro biofilm formation of Candida albicans and Streptococcus mutans

https://doi.org/10.1016/j.archoralbio.2014.05.006Get rights and content

Abstract

Objective

To evaluate the antiadherent property of crude, methanol and acetate methanol extract fractions from Schinus terebinthifolius and Croton urucurana in hydroalcoholic (HA) and dimethylsulfoxide (DMSO) solvents on in vitro biofilms formed by Streptococcus mutans and Candida albicans strains.

Design

The minimal concentration of adherence (MICA) was determined to evaluate the antiadherent potential of extracts on the in vitro biofilm formation. The extracts of plants were subjected to thin layer chromatography (TLC) in order to detect what class of compounds was responsible for the antiadherent activity. Data were estimated by analysis of variance (ANOVA) complemented by Tukey test level of significance set at 5%.

Results

Both plants demonstrated inhibition of S. mutans and C. albicans on in vitro biofilm formation. The biofilms of C. albicans were more efficiently inhibited by the S. terebinthifolius fraction of acetate–methanol and methanol in hydroalcoholic solvents (p < 0.05). The S. mutans biofilms adherence was best inhibited by the S. terebinthifolius crude extract and its methanolic fraction, both in hydroalcoholic solvent (p < 0.05). TLC of crude extracts and fractions of S. terebinthifolius detected the presence of several active compounds, including phenolic compounds, anthraquinones, terpenoids, and alkaloids. C. urucurana extracts confirmed activity for both microorganisms (p < 0.05). However, higher concentrations were needed to achieve antiadherent activity, mainly to inhibit in vitro biofilm formation of C. albicans.

Conclusion

The antiadherent potential of both plants on in vitro biofilms formed by C. albicans and S. mutans were confirmed, suggesting the importance of studies about these extracts for therapeutic prevention of oral diseases associated with oral biofilms.

Introduction

In Brazil, the use of plants in folk medicine for the prevention and treatment of several health disorders is a common practice.1, 2, 3, 4 Among popular plants used for this purpose is Schinus terebinthifolius Raddi, from the Anacardiaceae family, popularly known as aroeira, aroeira-vermelha, aroeira-mansa, aroeira branca, aroeira-da-praia, aroeira-do-sertão, aroeira-do-paraná, araguaraiba, corneiba, fruto-de-sabiá and peppertree. This is a native species prominent in South and Central America, and also has been found in subtropical and tropical regions of United States and Africa. Compounds produced by S. terebinthifolius have been found to exhibit antimicrobial, anti-inflammatory and antiulcer properties.5, 6, 7

The Croton urucurana is a tree belonging to the Euphorbiaceae family and can grow up to 15 m tall, with an open canopy and bright stem. It is wide spread in wetlands and riparian areas and commonly found in southern Brazil, northern Argentina, Paraguay and Uruguay. It is popularly known by the name of dragon blood, blood water, capixingui, urucuana, lucurana, tapexingui and tapixingui. C. urucurana has anti-inflammatory, anti-diarrhoeal, anti-haemorrhagic and anti-fungal therapeutic properties. It is also used among other forms of treatments for ailments such as anaemia, bronchitis and asthma.8, 9, 10

The human oral cavity is a dynamic environment often exposed to variations such as changes of pH, nutrient sources, carbohydrate availability, oxygen tension, and is a reservoir of more than 350 different species of microorganisms which when balanced with the host immune defenses, help to prevent a wide variety of diseases. However, this frail balance can be disturbed by endogenous factors associated to the host such as immune response, salivary flow and buffer capacity, as well as factors related to the environment as the frequent presence of sucrose in the diet.11, 12

The development of the oral biofilm is a major factor responsible for the emergence of various diseases, including dental cavities, gingivitis and periodontitis.13, 14 Amongst various species of microorganisms present in the oral microbiota, S. mutans bacteria and the yeast C. albicans represent the major and most common microbial agents associated with oral biofilm.15, 16 Streptococcus mutans stands out for being the main etiological agent of dental caries.17, 18 Candida albicans, when present in the microbiota as a colonizer, seems to increase the risk of the caries disease.15, 16 Gregoire et al.18 revealed a novel cross-kingdom interaction that is mediated by bacterial GtfB, which readily attaches to the yeast cell surface. Surface-bound GtfB promotes the formation of a glucan-rich matrix in situ and may enhance the accumulation of S. mutans on the tooth enamel surface, thereby modulating the development of virulent biofilms. In addition, the overgrowth of C. albicans results in candidiasis, which is often observed in patients using dental prostheses,19, 20 demonstrating the action of these agents in the formation of biofilm.

Some herbal products have been investigated due to their potential to control biofilms, including extract of pea (Pisum sativum),21 lectin extracted from “pitombeira” (Telisia esculenta)22 and crude extract from mulberry leaves (Morus alba).23 After pharmaceutical evaluations, plant extracts were added to mouthwashes, toothpastes, and other oral hygiene products, in order to make them more efficient at preventing the formation of the oral biofilm.

The specific nature of the antiadherent properties of S. terebinthifolius and C. urucurana to control oral biofilm development is not certain and is still under observation.24, 25 In general, the study of these plants is mainly focused on their anti-inflammatory, antifungal, and antibacterial properties.6, 22, 26, 27 Nevertheless, the present study evaluated the antiadherent potential of S. terebinthifolius and C. urucurana extracts on in vitro biofilms formed by S. mutans and C. albicans.

Section snippets

Botanical specimens

S. terebinthifolius leaves were collected from a sample of latitude 26.827 S 25°, 49° 13.997 W longitude and altitude of 940 m. The botanical identification was performed in the Botany Department Herbarium of the Federal University of Paraná – Brazil (UPCB), a sample of specimen has been deposited at the Herbarium under registration UPCB-30848. The stem bark of C. urucurana was collected 606628.67mE, 749192925mS, in altitude of 960 m, the botanical identification was performed in the Herbarium,

Results

A total of 29.901 g of concentrated crude methanolic extract was obtained from 123 g of S. terebenthifolius, which means a yield of 24.31% from dried leaves. Whereas 57.284 g of C. urucurana extract were obtained from 400 g of the stem bark, showing a yield of 14.32%.

Minimum inhibitory adherence concentration of each extract was determined by reading the smallest rate of absorbance, which corresponded to the least developed biofilm in which cells remained viable by plating and cultivation on BHI

Discussion

The extensive application of plants in traditional medicine is among the most common practices for the treatment and prevention of diseases in Brazil due to the great biodiversity of herbal resources, popular culture, low cost and usage facilities.8, 37 This resource of oral disease prevention deserves particular attention, especially concerning control of oral biofilms, which is the precursor of most oral infectious diseases.2, 3, 38, 39, 40 The ability to impair bacterial adhesion represents

Funding

This work was supported by a Brazilian Government, (Grant number: 05 9533/2006 - 2862007) from Fundação Araucária, Coordenação de Pessoal de Nível Superior (CAPES) and Conselho Nacional de Pesquisa (CNPq), (Grant number: 30 7547/2012-4).

Competing interests

None declared.

Ethical approval

Not required.

Acknowledgements

The authors acknowledge Professor Dr. Juarez Gabardo, Department of Genetics, Federal University of Parana State, Paraná, Brazil for assistance with statistical analysis and the Electron Microscopy Center – Federal University of Parana State, Paraná, Brazil, for the use and technical support of the scanning electron microscopy, particularly to Prof. Dr. Lucelia Donatti. We also emphasize the collaboration of MSc. Patricia Fernanda Herkert and Dr. Renata Rodrigues Gomes in the experimental

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    Centro Politécnico, Universidade Federal do Paraná, Departamento de Química, Post Code: 81.531-980 Curitiba, PR, Brazil.

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