WO1992014475A1

WO1992014475A1 - Prophylactic ferments for combatting dental caries

Info

Publication number: WO1992014475A1
Application number: PCT/FR1992/000126
Authority: WO
Inventors: Isabelle Madinier
Original assignee: Isabelle Madinier
Priority date: 1991-02-13
Filing date: 1992-02-12
Publication date: 1992-09-03
Also published as: FR2672494A1

Abstract

Prophylactic ferments for combatting dental caries on account of their competitive ecological properties in relation to possible cariogenic strains and agro-food properties enabling them to be used in industry. The prophylactic ferments consist of different bacteria capable of entering into competition with other oral flora bacteria, such as avirulent strains of $i(Streptococcus), $i(Lactobacillus) and $i(Stomatococcus), or any other avirulent bacterial or mutant species corresponding to the oral health. Said prophylactic ferments can be used alone, or preferably in association, for example, as coferments in the manufacture of yoghurts, kefirs and other fermented milk-based products, or as lyophilizing products in the form of tablets. The prophylactic ferments according to the invention are for use especially in the agro-food, healthfoods and pharmaceutical industries.

Description

PROPHYLACTIC FERMENTS TO PROMOTE DENTAL CARIES PREVENTION

The present invention relates to prophylactic enzymes which promote the prevention of dental caries by virtue of bacteriological properties which are competitive vis-à-vis possible .cariogenic flora and with agro-food properties enabling their industrial use. These ferments are particularly suitable for the food and dietetic industries, for example associated with lactic ferments for the manufacture of yogurts and other products based on fermented milks, as well as for the pharmaceutical industry, for example in the form of lyophilized tablets.

Tooth decay is a widely spread infectious disease primarily responsible for Streptococcus mutans, a bacteria capable of destroying enamel and dentin by releasing acids in the oral environment during the breakdown of carbohydrates. According to the current nomenclature (Bergey's Manual ^8th edition, 1984 and 1986) the species S. mutans is actually made up of eight serotypes referred a to h: S. mutans itself (serotypes c, e, f), S rattus (serotype a), S. cricetus (serotype b) and S. sobrinus (serotypes d, g, h). The cariogenic species of this group (S. mutans c and f, S. sobrinus d and g) are characterized in particular by their ability to colonize soft tissues using specific adhesins, to colonize hard tissues with extracellular polysaccharides (dextran or glucans), to coaggregate with other oral bacteria, to quickly form acids from sugars and to survive at very low pH. Other oral bacteria can potentiate this cariogenic power in early or established carious lesions, in particular other species of the genus Streptococcus and certain species of the genera Actinomyces (A. naeslundii and A. odontolyticus) and Lactobacillus (casei and L. fermentum ). Caries leads to progressive and irreversible destruction of the affected tooth which, if left untreated, can cause localized or generalized infections. Current preventive techniques are based on correct oral hygiene, reasoned consumption of carbohydrates and the use of fluoride hygiene and dental care products. These methods, although effective, require active individual cooperation which is far from being systematically observed, especially in children, so that cavities are still a major public health problem. The device according to the invention is based on the principle of interbacterial competition and ecological balance within the oral flora: it makes it possible to promote the prevention of dental caries by a repeated exogenous contribution, at the level of the plaque. dental, small quantities of non-virulent prophylactic ferments capable of opposing the colonization of dental surfaces by a possible cariogenic flora dominated by pathogenic strains of S. mutans Indeed, the early plaque which recolonizes enamel in uninjured subjects of active caries is dominated by bacteria type

5 Gram positive cocci, immobile, aerotolerant and / or encapsulated, in particular S. sanguis (serotype 1: S. gordoπii, serotype 2: S. mitior or S. oralis), S. mitis (serotypes 1 and 2) and Stomatococcus mucilaginosus , a non-cariogenic encapsulated commensal of the oral flora. These different species are found in variable proportions depending on the individuals, the location and the age of the deposits. The species included in the constitution of

10 prophylactic enzymes are therefore chosen:

- among all non-virulent strains of bacteria compatible with the maintenance of oral integrity,

- among the early colonizers of the plate mentioned above in particular S. mitis (ATCC 15 909, ATCC 15 912), S. salivarius (ATCC 9758, ATCC 13 419, ATCC 27

15 945), and Stomatococcus mucilaginosus (ATCC 25 296, CCM 2417, NCTC 10 663),

- among the non-virulent strains of S. mutans, L casei (in particular L casei subsp. rhamnosus ATCC 7469, DSM 20 012, DSM 20 021, DSM 20 022, DSM 20 023, DSM 20 245, DSM 20 247), and L fermentum,

- among the non-virulent strains of L. acidophilus (ATCC 4356), L. crispatus (ATCC 33 20 820, VR 3199), L. gasseri (DSM 20 243) and L salivarius (L. salivarius salivarius

ATCC 1 1 741, L salivarius salicinus ATCC 1 1 742) which have the double advantage of presenting a tropism for the oral environment and of belonging to the group of thermophilic lactobacilli directly usable as lactic ferments.

- among all avirulent mutants derived from all the species mentioned above, 5 - among all avirulent mutants derived from other bacterial species and having new characteristics allowing them to enter into ecological competition with possible cariogenic oral flora, such as, for example, strains used in the dairy industry (such as Lactobacillus bulgaricus and Streptococcus thermophilus) and modified so as to exhibit specific adhesion and coaggregation phenomena 0 (pili, fimbriae, adhesins) and / or non-specific (glycocalyx, capsules, extra-cellular poiysaccharides of the glucan / dextran or fructan / levan type) in the oral environment.

This list is in no way limiting and is likely to evolve as a function of the progress made in the knowledge of the oral flora and of the taxonomic updating of the species in progress or to come. Prophylactic ferments are formed from one, or better, of several of the species mentioned above associated by

REPLACEMENT SHEET example in ratios of 1/1, and can be stored frozen in liquid nitrogen or freeze-dried depending on the industrial objectives sought. It should be noted that these prophylactic enzymes are also very different from the flora associated with periodontopathies, which does not exclude a possible additional prophylactic effect in the context of periodontal diseases.

Yogurts, khefirs and other products based on fermented milks constitute an excellent support for this type of prevention since the usual lactic ferments used for their manufacture belong precisely to the genera Lactobacillus and Streptococcus. These dairy products are therefore perfectly suited to the culture and survival of prophylactic enzymes by simultaneously allowing a supply of immediately competitive live germs at the level of dental plaque, and a supply of decalcium which promotes the remineralization of demineralized carious fronts. In this case, the prophylactic ferments can be used in addition to the usual lactic ferments at an optimal concentration taking into account the biological, technical and gastronomic imperatives sought, for example in a range of final concentrations going from 1/10 ** to 1 / 10 ^{1 0} compared to lactic ferments. They are used either as co-ferments if they support the manufacturing processes and in particular the thermal constraints (for example the association L. salivarius, L. acidophilus and L casei subsp. Rhamnosus in 1/1/1 proportion), either by simple addition as by stirring at the end of manufacture (for example the association L. salivarius, S. salivarius and S. mucilaginosus in a 1/1/1 proportion). These prophylactic ferments are also interesting lyophilized; and distributed for example by 1 to 2 million units in the form of pharmaceutical tablets to be sucked slowly with a suitable excipient such as a non-fermentable carbohydrate. In fact, the oral environment is a favorable environment for retention (bacterial coaggregation) and then for the growth of lyophilized bacteria (temperature, pH, humidity, presence of nutrients and growth factors, bacterial synergy, etc.). These examples of use of the prophylactic enzymes are not limiting, and they can be used in any other form whatsoever, in particular associated with all the chemical compounds reputed to have a preventive effect against dental caries, such as for example the compounds fluorinated or non-fermentable carbohydrates.

The device according to the invention is particularly intended for the dietetic, pharmaceutical and especially agrifood industries due to the widely / widespread consumption of yogurts and other products based on fermented milks, in particular in children for whom the prevention of dental caries is a major concern.

REPLACEMENT SHEET

Claims

1) Prophylactic ferments other than the genera Bifidobacterium, Lactobacillus, Streptococcus and the species Actinobacillus bovis characterized in that they promote the prevention of dental caries thanks to competitive bacteriological properties vis-à-vis possible cariogenic flora, and to agro-food properties allowing their industrial use in any form whatsoever.

2) ferments according to claim 1 characterized in that the bacterial species entering into their constitution are chosen, alone or in combination, from the non-virulent strains of bacteria compatible with the maintenance of oral integrity, in particular the species of type cocci, Gram positive, immobile, aerotolerant and / or encapsulated such as for example Stomatococcus mucilaginosus. These prophylactic enzymes can also promote the prevention of periodontal diseases thanks to their competitive bacteriological properties vis-à-vis possible pathogenic flora associated with periodontopathies. 3) Ferments according to claims 1 and 2 characterized in that the species entering into their constitution are chosen, alone or in combination, from non-virulent strains of bacteria known to be pathogenic such as certain members of the genus Actinomyces (for example A. naeslundii and A . odontolyticus).

4) Ferments according to claims 1, 2 and 3 characterized in that the bacterial species entering into their constitution are chosen, alone or in combination, from all possible avirulent mutants derived from the species mentioned above.

5) Ferments according to claims 1 and 2 characterized in that the species entering into their constitution are chosen, alone or in combination, from all possible avirulent mutants derived from other bacterial species, such as lactic ferments modified so as to present new characters allowing them to enter into ecological competition with possible pathogenic oral flora.

6) Device according to any one of the preceding claims, characterized in that the prophylactic ferments are formed from one, or preferably from several of the bacterial species mentioned above associated for example in ratios of 1/1 and preserved from preferably frozen in liquid nitrogen or lyophilized, this list not being in any way limiting and being likely to evolve according to the progress made in the knowledge of the oral flora and the taxonomic updating of

SUBSTITUTE SHEET current or future cash.

7) Prophylactic ferments according to any one of the preceding claims, characterized in that they are particularly intended for use in the manufacture of yogurts, khefirs and other products based on fermented milks in addition to the usual lactic ferments. The prophylactic ferments can be used either as co-ferments if they support the manufacturing processes, and in particular thermal stresses, or by simple addition by stirring at the end of manufacture. The optimal final concentration is for example in a range of 1/10 ⁶ to 1/10 ¹ ° compared to lactic ferments taking into account the biological, technical and gastronomic imperatives sought.

8) Prophylactic ferments according to any one of the preceding claims, characterized in that they can be used to promote the prevention of dental caries or periodontal diseases associated with all the chemical compounds known to have a preventive effect against these oral pathologies, such as, for example, fluorinated compounds or non-fermentable carbohydrates.

REPLACEMENT

SHEET OF MODIFIED REVEb-DICATIO S

[received by the International Bureau on 10 July 1992 (10.07.92); claims 1-3,7 and 8 amended; other claims unchanged (2 pages)]

1) Prophylactic ferments other than the genera Bifidobacterium, Lactobacillus, Streptococcus and the species Actinobacillus bovis characterized in that they promote the prevention of dental caries thanks to competitive bacteriological properties vis-à-vis possible cariogenic flora, and to agro-food properties. allowing their industrial use in any form whatsoever.

2) ferments according to claim 1 characterized in that the bacterial species entering into their constitution are chosen, alone or in combination, from the non-virulent strains of bacteria compatible with the maintenance of oral integrity, in particular the species of type cocci, Gram positive, immobile, aerotolerant and / or encapsulated such as for example Stomatococcus mucilaginosus. These prophylactic enzymes can also promote the prevention of periodontal diseases thanks to their competitive bacteriological properties vis-à-vis possible pathogenic flora associated with periodontopathies. 3) Ferments according to claims 1 and 2 characterized in that the species entering into their constitution are chosen, alone or in combination, from non-virulent strains of bacteria reputed to be pathogenic such as certain members of the genus Actnomyces (for example A. naeslundiietA. Odontolyticυs ).

6) Device according to any one of the preceding claims, characterized in that the prophylactic ferments are formed from one, or preferably from several of the bacterial species mentioned above associated for example in ratios of 11l and preferably stored frozen. in liquid nitrogen or lyophilised, this list being in no way limiting and being likely to evolve according to the progress made in the knowledge of the oral flora and the taxonomic updating of current or future cash.

7) Prophylactic ferments according to any one of the preceding claims, characterized in that they are particularly intended for use in the manufacture of yogurts, khefirs and other products based on fermented milks in addition to the usual lactic ferments. The prophylactic ferments can be used either as co-ferments if they support the manufacturing processes, and in particular thermal stresses, or by simple addition by stirring at the end of manufacture. The optimal final concentration is for example in a range of 1/10 ⁶ to 1/10 ¹⁰ compared to the lactic ferments taking into account the biological, technical and gastronomic imperatives sought.

8) Prophylactic ferments according to any one of the preceding claims, characterized in that they can be used to promote the prevention of dental caries or periodontal diseases associated with all the chemical compounds known to have a preventive effect against these oral pathologies, such as, for example, fluorinated compounds or non-fermented carbohydrates.