© Borgis - Nowa Stomatologia 4/2004, s. 176-179
Joële Luc1, Elżbieta Dybiżbańska2, Christine Roques3
Bactericidal and fungicidal activity of selected mouthrinses - an in vitro study
1Fonderephar, Toulouse, Francja
2z Zakładu Stomatologii Zachowawczej IS AM w Warszawie
Kierownik Zakładu: prof. dr hab. Maria Wierzbicka
3Laboratoire de Bacteriologie, Virologie et Microbiologie Industrielle, Toulouse, Francja
INTRODUCTION
Dental caries as well as periodontal diseases are disorders of bacterial origin. The microorganisms responsible for caries are bacteria of the Streptococcus mutans and Lactobacillus acidophilus species. Among the microorganisms considered pathogenic for the periodontium are Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Tannarella forsythensis, while Prevotella intermedia, Campylobacter rectus, Peptostreptococcus micros, Fusobacterium, Eubacterium and Treponema species, as well as yeast-like fungi, are classified as potentially pathogenic (1). The above mentioned bacteria are deposited on the tooth surface forming a biofilm, generally known as plaque. Plaque control plays a fundamental role in the prevention of dental caries and periodontitis. Despite of this, mechanical methods of plaque control are often insufficient, and the supplemental use of chemical agents is recommended. It is often the case that manual skills are deficient, in individuals with physical or mental handicap, as well as following oral surgical procedures (2, 3). The aim of chemical plaque control is to prevent the proliferation of microorganisms (bacteriostatic agents), or the eradication of the microorganisms already present in the biofilm, as well as the newcomers (bactericidal agents) (4). The most frequently used carrier of an antibacterial chemical agent is a mouthrinse. Other carriers used include toothpaste, sprays, irrigation liquids, chewing gum, lozenges and dental varnishes (5). Among the most frequently used substances in plaque control are chlorhexidine, belonging to the bis-biguanides, as well as cetylpiridinium chloride, a quaternary ammonium compound, phenolic compounds such as triclosan, oxidizing agents e.g. hydrogen peroxide, fluoride compounds e.g. a combination of fluoroamines and stannous fluoride, metal salts e.g. tin, zinc, and others e.g. hexetidine (5).
The effectiveness of the chemical agents used for plaque control has been assessed through in vitro and in vivo assays. For the in vitro studies bacteriological tests were used, which assess the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the tested agents (4).
The aim of this study was an in vitro assessment of the bactericidal and fungicidal action of eight mouthrinses used in home oral hygiene, with reference to selected species of bacteria and yeast most frequently observed in dental caries and periodontitis.
MATERIALS AND METHODS
The tested mouthrinses are presented in Table 1. The bactericidal and fungicidal action of the rinses was evaluated with respect to standard strains of the following microorganisms: Streptococcus mutans CIP 103220T, Lactobacillus acidophilus CIP 7316T, Fusobacterium nucleatum CIP 101130T, Prevotella intermedia CIP 103607 and Actinobacillus actinomycetemcomitans CIP 52109, as well as Candida albicans IP 4872, originating from the microorganism collection of l´Institut Pasteur (Paris).
Table 1. Mouthrinses tested.
Mouthrinse | Active ingredient | Concentration used |
Corsodyl | 0.20% chlorhexidine digluconate | pure |
Eludril | 0.10% chlorhexidine digluconate | diluted: 1:2, 1:3 |
Hextril/Oralden | 0.10% hexetidine | pure |
Meridol | fluoroamines + 0.025% stannous fluoride | pure |
Tantum verde | 0.15% benzydamine hydrochloride | diluted: 1:2 |
Alodont | 0.005% cetylpiridinium chloride | pure |
Lacalut | 0.20% chlorhexidine digluconate, aluminium lactate, betain | pure |
Plak out | 0,12% chlorhexidine digluconate | pure |
Bacterial suspensions were prepared directly prior to use in sterile, distilled water at a concentration of 108-109 bacteria/ml, whereas for C. albicans – at a concentration of 107/ml. Bactericidal and fungicidal action was determined using the „dilution-neutralization” method (European Standard). The test was conducted by adding 1 ml of the microorganism suspension to 9 ml of the tested solution. In order to stimulate conditions in vivo in which there is interaction with organic components 0.3% bovine albumin was added to the test tube. Contact time was 1 minute at a temperature of 32°C. Contact was interrupted by transferring 1 ml from the test tube into 9 ml of a neutralizing substrate (10% polysorbate, 2% saponin, 2% lecithin, 0,5% sodium thiosulfate, soy-tryptone broth). Following 10 minutes of neutralization, the remaining live microorganisms were counted by incubating 1 ml of the suspension in Schaedler´s agar substrate (in the case of S.mutans), agar MRS substrate (for L. acidophilus), or agar plus malt extract (for C. albicans). In the case of F. nucleatum, P. intermedia and A. actinomycetemcomitans, counts were performed by plating 100 ml on Columbia sheep blood agar. The cultures were incubated at a temperature of 37°C for 48-72 hours under anaerobic or aerobic conditions for the bacteria and at a temperature of 30°C for C.albicans. After that, the colonies were counted. All tests were done twice.
RESULTS
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Piśmiennictwo
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