MATRIX FORMULATED WITH PROBIOTICS TO PROVIDE STABILITY TO THE BENEFICIAL CHEMICAL-MICROBIAL PROPERTIES IN ORAL HEALTH

Abstract

The present invention relates to a matrix composition formulated with probiotics to provide stability to the beneficial chemical-microbial properties in oral health, allowing a decrease in the development of caries due to the inhibition of Streptococcus mutans and other pathogens that are the main causes of oral diseases, characterized by comprising 3-5 mg / 100 mL of pectin, 6-9 g / 100 mL of dextrose, 25-50 mg / 100 mL of citric acid, 2-4 mL / 100 mL of natural flavoring; and 4x10^12 CFU / mL of an active microorganism selected from Streptococcus salivarius and Lactobacillus plantarum or a mixture of both;The invention contemplates the production process of the formulated matrix to provide stability to the beneficial chemical-microbial properties in oral health and the use of the composition of said formulated matrix to improve oral health, comprising administering said composition to a patient to prevent tooth decay, prevent bad breath, remove tartar, eliminate yellowing of teeth and other oral conditions.

Description

MATRIX FORMULATED WITH PROBIOTICS TO PROVIDE STABILITY TO THE BENEFICIAL CHEMICAL-MICROBIAL PROPERTIES IN ORAL HEALTH FIELD OF INVENTION The present invention relates to biotechnology and microbiology in general, and more specifically to compositions for preventing and treating diseases associated with oral health. More specifically, it relates to a matrix formulated with probiotics to provide stability to the beneficial chemical-microbial properties for oral health. BACKGROUND OF THE INVENTION Tooth decay and periodontitis are caused by an imbalance in the bacterial populations of biofilms that form naturally and help maintain the normal state of the oral cavity. A healthy biofilm can consist of more than 700 bacteria, of which less than 1% are potentially pathogenic, such as Streptococcus mutans, which is the main cause of tooth decay in 95% of cases. The tooth surface is an essential natural habitat for this bacterium, and its tropism for the dental biofilm is reflected in its adaptation to synthesize glucans, bind compounds, and adjust its acidity. The glycosyltransferases (Gtfs) of Streptococcus mutans adsorb onto the enamel to produce glucans in situ, providing sites for avid colonization and adhering firmly via peptides they develop. This makes simple removal impossible, requiring a visit to a dentist. Currently, dentistry is the only health science that combats these diseases in critical stages through techniques such as extraction and restoration using endodontics or crowns. On the other hand, biotechnology applied to dentistry is an innovative and minimally invasive alternative. Species such as Streptococcus salivarius and Lactobacillus plantarum, considered Generally Recognized As Safe (GRAS) by the Food and Drug Administration (FDA), when used as probiotics, can locally inhibit bacterial growth and adhesion, modulating the immune response through the production of antiviral agents. The present invention relates to a matrix formulated to provide stability to the beneficial chemical-microbial properties in oral health. A search was conducted to determine the closest prior art, and the following document was found. The patent application WO201 0077795A3, filed by Roughead Zamzam Kabiry and Kaspar Kala Marie on December 14, 2009, was located. This document discloses compositions and methods for preventing and treating diseases associated with oral health. The compositions and methods can improve oral health by reducing the incidence of aspiration pneumonia, community-acquired pneumonia, and nosocomial pneumonia, resulting in less gingivitis and plaque, and promoting an improved tongue flora. By utilizing probiotics naturally present in the oral cavity or gastrointestinal tract, the compositions and methods can reduce anaerobic gram-negative bacilli and increase the presence of normal flora. The document discloses a composition for improving oral health comprising a therapeutically effective amount of beneficial bacteria, wherein said beneficial bacteria comprise at least one bacterium normally selected from the group consisting of: i) Streptococcus salivarius; ii) Lactobacillus reuteri; Lactobacillus platarum 299 or 299v; iv) Streptococcus salivarius K1 2; v) Lactobacillus reuteri ATCC55730; vi) Lactobacillus johnsonii L1; vii) Lactobacillus rhamnosus GG; viii) Streptococcus thermophilus NCC 1561; ix) Lactococcus lactis NCC2211 (Pelargon strain); yx) Lacteol. The composition is in a form such as liquid, solid, semi-solid, or a combination thereof. The composition may be a complete oral nutritional supplement. The composition may also be in a form such as tablets, lozenges, sachets, soluble films, or a combination thereof. The document also reveals that the bacteria are inactive. The oral microbiota of the gastrointestinal tract (GIT) is composed of more than 500 bacteria that assist in the digestion and detoxification of food, as well as stimulating the immune system. This flora is regulated by peristalsis, the secretion of gastric acid and bile salts; it also generates a greater bacterial concentration gradient in the colon, which can be affected by ingested food and external factors such as pollution, sunlight, etc. Therefore, the use of probiotics that reach the intestinal microflora is important, since the mucosa of this organ helps regulate the symbiotic interaction and strengthens the immune system, reducing diseases such as pneumonia or dysphagia; diseases whose reduction is claimed in claims 7, 14, and 17 of the patent application published as WO2010077795A3.However, for a probiotic to reach the gastrointestinal system safely and effectively, it must be in an inactive formulation, commonly known as "freeze-dried." The same patent, in claim 9, mentions the protection of various microorganisms of claim 7 in this "freeze-dried" and / or inactive state. This allows these inactivated bacteria to be activated by gastric juices and strengthen the gastrointestinal flora. While the oral microbiota is a completely different environment, due to its pH, oxygenation, and the presence of over 700 different bacteria, etc., and is primarily regulated by saliva and the proteins derived from it. Because of these different conditions, an inactivated bacterium cannot easily colonize the oral microbiota; it needs to be activated, and the conditions in the mouth make this impossible or limit its effect, both in the extent of the change and its duration. However, it is important to strengthen the oral microbiota, as it also harbors pathogenic bacteria that can develop a tropism in the microbial population and give rise to diseases such as gingivitis, cavities, bad breath, and many others. Claim 1 of the aforementioned document discloses the microorganisms Lactobacillus plantarum 299 or 299v and Streptococcus salivarius K12. Lactobacillus plantarum is a probiotic strain capable of residing in the human colonic mucosa because it adheres to the intestinal mucosa, where it modulates the composition of the intestinal microflora, helping to maintain adequate colonization of the gastrointestinal tract and improving digestion and metabolism. By colonizing the gastrointestinal tract, the probiotic can form a protective barrier, thus preventing the adhesion of pathogens, protecting against infections, and stimulating the immune system. Streptococcus salivarius K12, on the other hand, is a strain isolated from the throat of a New Zealand child capable of producing antibiotic bacteriocin that helps counteract the growth of S. pyogenes and inhibits the growth of pathogens such as Haemophilus influenzae and S.pneumoniae and Moraxella catarrhalis, all of which are involved in the etiopathogenesis of acute otitis media. Claims 10 through 12 of the aforementioned document disclose the use of lozenges, lozenges, sachets, soluble films, and topical applications. Our invention (viscous solution) will be the filling for chewing gum, gummies, toothpaste, or any confectionery product that may contain a filling. This is because the probiotic bacteria must adhere to the surface of the teeth for an extended period to achieve an inhibitory effect against caries and other diseases. This is because the probiotic bacteria will be persistent and will become part of the oral biofilm that constitutes the oral microbiota. The resulting biofilm keeps bacterial pathogens out of the oral tissues, filling the space that pathogens would invade in the absence of the biofilm, while simultaneously competing with cariogenic bacteria. Document WO201 0077795A3 reveals in claims 7 and 17 that its product helps reduce the incidence of aspiration pneumonia and dysphagia. Our invention, however, focuses on preventing cavities, reducing tartar buildup, halitosis, and yellowing of the teeth, etc. Dental caries is one of the main oral health problems, and its consequences range from inflammation or infection of the pulp tissue to tooth loss. Naturally, the surface of the tooth is covered by a biofilm composed of millions of bacterial cells, salivary polymers, and food debris, which over time and without proper cleaning begins to turn into plaque, providing an adhesion site for the colonization and growth of pathogenic bacterial species such as Streptococcus mutans. Streptococcus mutans plays a central role in the etiology of dental caries because it can adhere to the salivary film on the enamel and to other plaque bacteria. This is because it produces weak organic acids as metabolic byproducts of fermentable carbohydrates, which lower the local pH, causing demineralization of the tooth tissue. Typically, the presence of Streptococcus mutans in tooth cavities is followed by caries after 6–24 months. The adhesion of this microorganism to the tooth surface is due to the interaction between the PAc protein and other salivary proteins that are adsorbed by the tooth enamel. The number of Streptococcus mutans increases with the onset of dental caries and decreases when the caries is treated, thus establishing a correlation between this pathological process. Plaque control can be achieved through mechanical oral hygiene procedures, but in many cases this is insufficient. Therefore, the direct addition of active probiotic products according to our invention to the oral cavity helps reduce the overall rate of new plaque accumulation, reduce or eliminate existing plaque, and inhibit the growth of pathogenic species. Document WO201 0077795A3 has a different focus; it is primarily aimed at the gastrointestinal flora and reducing or preventing diseases such as dysphagia and pneumonia. On the other hand, our invention reduces and prevents various diseases derived from the oral microbiota to a better degree, due to the effectiveness of the inoculation and the state of our bacteria. Thanks to the components used, our microorganisms can be active, unlike the bacteria in the document cited here, which only discloses the use of other microorganisms in an inactive state. Document WO201 0077795A3 reveals that the compositions can have presentations in the form of tablets, iodine, lozenges, sachets, soluble films, whereas our invention can be in liquid, semi-viscous and / or viscous form that is used as a filling for any confectionery product. Given the need for an organoleptically friendly matrix for human consumption with natural components that give stability and viability to the active biomass of L. plantarum and S. salivaria integrated into the matrix, which upon contact with the mouth allows reactivation and incorporation into the oral flora, inhibiting the bacterial biofilm adhered to the teeth and reducing oral problems such as cavities, bad breath, tartar, among others; the present invention was developed. OBJECTIVES OF THE INVENTION The main objective of the present invention is to make available a matrix formulated with probiotics to provide stability to the beneficial chemical-microbial properties in oral health that allows a decrease in the development of caries due to the inhibition of Streptococcus mutans and other pathogens that are the main causes of oral diseases. Another objective of the invention is to provide said matrix formulated with probiotics to provide stability to the beneficial microbial chemical properties in oral health, which also offers viability and stability to the microorganisms Lactobacillus plantarum and Streptococcus salivariae that, upon contact with the mouth, manage to colonize and strengthen the oral flora. Another objective of the invention is to provide said matrix formulated with probiotics to provide stability to the beneficial chemomicrobial properties in oral health, which also allows to maintain viable and stable microorganisms in an active state that, upon contact with the mouth, manage to colonize the oral flora. Another objective of the invention is to provide said matrix formulated with probiotics to provide stability to the beneficial chemomicrobial properties in oral health, which also combats the formation of caries, prevents bad breath, eliminates tartar and other oral conditions. And all those qualities and objectives that will become apparent when making a general and detailed description of the present invention supported by the illustrated modalities. BRIEF DESCRIPTION OF THE INVENTION To develop a probiotic-formulated matrix that provides stability to the beneficial chemical-microbial properties for oral health, research and selection of the active component (microorganisms) began. A susceptibility test was performed using filter paper discs coated with different probiotic microorganisms or natural substances. These discs were then allowed to dry (2-4 min) and placed in Petri dishes inoculated with the caries consortium, including Streptococcus mutans. They were incubated for 24 h at 37°C and checked every 12 h to determine inhibition. After 24 h, a zone of inhibition was observed around the rings impregnated with Streptococcus salivarius and Lactobacillus plantarum, surprisingly demonstrating the antimicrobial effect of these probiotics against the caries consortium. Surprisingly, it was found that the microorganism Streptococcus salivarius manages to produce two classes of antibiotic bacteriocins: salivaricin A2 and salivaricin B, both strongly antagonistic to the growth of Streptococcus mutans, which in 95% of cases is the main cause of the formation of cavities. On the other hand, it was determined that Lactobacillus plantarum has greater hydrophobicity and adhesion affinity than Streptococcus mutans, since it contains the lac 4 and lac 5 genes that produce a greater amount of Beta-D-galactosidase, counteracting the cohesion of the pathogen. Testing began with the microorganisms Streptococcus salivarius and Lactobacillus plantarum, which were freeze-dried in a tablet; this tablet was tested to determine the viability of the strain and its effectiveness since they are a type of dental probiotic. To test its effect, the tablet was dissolved in BHI medium (calf brain and heart infusion, peptic hydrolysate of animal tissues, pancreatic casein hydrolysate, sodium chloride, glucose, disodium phosphate), left to incubate for 24 hours, and afterwards, it was examined under a microscope to determine the number of cells; however, no growth was obtained because in this state the microorganisms require an acidic medium to reactivate, so it was concluded that said tablet only has an effect at the gastrointestinal level; consequently, it was decided to make a medium that simulated the acids of the stomach (pH 4.5). The tablet was again dissolved in BHI medium (calf brain and heart infusion, animal tissue peptic hydrolysate, pancreatic casein hydrolysate, sodium chloride, glucose, disodium phosphate) to which a medium simulating stomach acid (pH 4.5) was added. It was incubated for 24 hours, after which growth was observed under a microscope. To validate viability, a Petri dish was plated, and colonies were obtained after 16 hours. These colonies had a chewy consistency, which may be due to the compounds in the tablets. For a probiotic microorganism to have an effect on the human body, it must be active and stable. Currently, the simplest method is freeze-drying, which allows the microorganisms to remain stable but inactive until activated by gastric juices. However, these microorganisms do not interact with the oral flora but rather with the intestinal flora once they manage to colonize part of it. Therefore, it was determined that the active component (microorganisms) must be placed in a matrix that allows for its stability in an active and stable state to be effective. To determine the composition of the matrix, several formulations were made, focusing on four formulations. The first consisted of a gelatinous paste based on fruit, which was inoculated with the microorganisms of interest, but there was no growth. The other three were made with a pectin base and only the flavoring was varied (cinnamon, cherry, and mint). From these, it was surprisingly found that the best composition of the formulated matrix to provide stability to the beneficial chemical-microbial properties in oral health consists of a viscous solution, which is composed of pectin (3-5 g / 100 mL), dextrose (6-9 g / 100 mL), citric acid (25-50 mg / 100 mL) and natural flavoring (2-4 mL / 100 mL) selected from mint and spearmint; and as an active ingredient, a mixture of microorganisms Streptococcus salivarius and Lactobacillus plantarum. These components, when combined, form a protective barrier that keeps microorganisms in stable conditions. Pectin acts as a natural thickener, and when combined with sugar and citric acid, it forms gels that keep the microorganisms stable and simultaneously develop anti-adhesive activity against pathogens. Dextrose provides the necessary nutrients to keep the microorganisms alive without stimulating their proliferation. Citric acid regulates the pH, increasing the gel's preservation and acting as an emulsifier. Finally, the flavoring provides the organoleptic properties. The matrix gives viability and stability to the microorganisms Streptococcus salivarius and Lactobacillus plantarum which, upon contact with the mouth, manage to colonize and strengthen the oral flora. The matrix of the present invention had an unexpected effect on the direct attack on the oral flora, since it reduces dental bacterial plaque, because the presentation of the probiotics within the matrix is ​​active and stable, which allows it to attack and inhibit Streptococcus mutans and other pathogens contained in the consortium, which are the main cause of diseases such as caries, that is, the greatest unexpected effect was the visible reduction of caries both in the halos in Petri dishes, as well as in the extracted teeth that presented a high degree of contamination. In the case of teeth, the probiotic matrix managed to remove bacterial plaque and cavities (black spots) non-invasively in just 5 days. The matrix formulated as described can be used in confectionery, medicinal, dental products, mouthwashes, gums, etc. The matrix provides stability and viability to the active ingredient (at least one microorganism in an active state selected Streptococcus salivarius and Lactobacillus plantarum or a mixture of both) which, upon contact with a patient's mouth, allows easy colonization by the oral flora, inhibiting bacterial biopesticide adhered to the teeth, reducing oral problems such as cavities, bad breath, tartar, among others; since these microorganisms stabilize the oral flora by counteracting the tropism generated by S. mutans. At least one active microorganism selected from Streptococcus salivarius and Lactobacillus plantarum, or a mixture of both in the formulated matrix, manages to inhibit S. mutans and the microbial consortium found in caries because the S. salivarius and L. plantarum strains produce antagonistic substances that inhibit the growth of S. mutans. Naturally, teeth form a classic or native biofilm, composed of bacteria embedded in an exopolysaccharide matrix. Colonization occurs through the growth of primitive bacteria (Actinomyces naeslundii and various streptococcal species) that are part of the oral microbiota, whose function is to promote oral health. When these primitive bacteria adhere to tooth surfaces, they produce adhesins that facilitate the attachment of secondary and tertiary bacteria. All these microorganisms form part of a microenvironment that, under stable conditions, contributes to oral health. However, when an imbalance occurs in the oral microbiota, the microorganisms form bacterial plaque, which resembles a classic biofilm but causes cavities. If left untreated, this can lead to tooth loss. Therefore, the matrix developed in accordance with the present invention simulates the same mechanism since by keeping the probiotic microorganisms viable, they manage to come into contact with the bacterial plaque and inhibit the growth of species such as S. mutans, which allows reducing the number of pathogens, and at the same time the probiotic strains adhere through receptors or anchoring proteins, which allows colonization with beneficial bacteria that help to rebalance the oral microbiota. In the case of in vitro cultures (Figures 6a-8b) the reduction of pathogens is perceived in the decrease of black spots, which correspond to the formation of caries, as well as in vivo. Species such as Streptococcus salivarius and Lactobacillus plantarum, generally recognized as safe (GRAS) by the Food and Drug Administration, commonly known by its acronym FDA; when used as a probiotic in the matrix formulated in accordance with the present invention, manage to locally inhibit bacterial growth and adhesion, modulating the immune response through the production of antiviral agents. The mechanism of action of probiotics is based on a nutritional competition with oral microorganisms that are considered pathogenic, thus creating an ideal oral environment in which the risk of suffering from cavities is reduced due to the production of antimicrobial substances such as bacteriocins and hydrogen peroxide. The invention also contemplates a production process for the formulated matrix to provide stability to the beneficial chemomicrobial properties in oral health, comprising the steps of: a) Pour distilled water at a temperature between 80°C-100°C into a beaker, and add the ingredients in the following order: dextrose powder [6-9 g / 100 mL], citric acid powder [25-50 mg / 100 mL], pectin powder [3-5 g / 100 mL]; b) Stir at a temperature of 80°C for 5-10 minutes at a speed of 250 to 500 rpm until dissolved; c) Add the flavoring [(1-2 mL / 100 mL] and mix for 2-5 minutes at a speed of 100 to 150 rpm; d) Pour the formulated matrix into a container forming a maximum layer of 1 cm thick and subject it to UV light sterilization for 15 min, then pour it into a previously sterilized container for storage; e) Prepare the inoculum by reactivating the strains of at least one selected microorganism of Streptococcus salivarius and Lactobacillus plantarum or a mixture of both, performing a count using a Neubauer chamber to determine the initial concentration, which will allow calculating the mL needed to obtain the final cell concentration of 4×10Û12 CFU / ml; f) Take 1 mL aliquots and place them in Eppendorf tubes (the number of samples depends on the initial concentration) to perform three washes with physiological solution (2500 rpm, 5 min), between each wash the microorganism tablet is resuspended; g) Finally, resuspend the tablet in sterile physiological saline solution (0.9% NaCl) and concentrate in the same diluent to obtain a final concentration of 4 x 10¹² CFU / mL. This will ensure that for every 0.5 mL there are 2 x 10¹⁹ CFU. h) Inoculate the matrix by pouring 1.5 - 2 mL of the matrix into sterile Eppendorf tubes and inoculate each tube with the corresponding probiotic microorganism, injecting the cell suspension, mixing gently and storing at room temperature (25°C) for 5 days; i) The entire process from inoculum preparation to matrix inoculation is carried out in a laminar flow hood under aseptic conditions and with sterile material. To better understand the characteristics of the invention, the following drawings, which are illustrative but not limiting, are included as an integral part of this description. BRIEF DESCRIPTION OF THE FIGURES Figures 1a and 1b show a top view of a Petri dish where a disc with four radial divisions is defined with BHI medium (calf brain and heart infusion, peptic hydrolysate of animal tissues, pancreatic hydrolysate of casein, sodium chloride, glucose, disodium phosphate) inoculated with the caries consortium and then inoculated with natural coccus and the microorganisms Saccharomyces Boulardii, Streptococcus salivarius and Lactobacillus plantarum. Figures 2a and 2b show a top view of a Petri dish where a disc is defined with BHI medium (calf brain and heart infusion, animal tissue peptic hydrolysate, pancreatic casein hydrolysate, sodium chloride, glucose, disodium phosphate) first inoculated with the caries consortium and then inoculated with Streptococcus salivarius and Lactobacillus plantarum, showing the growth inhibition halos of the caries consortium in the areas where they were subsequently inoculated with Streptococcus salivarius and Lactobacillus plantarum. Figure 3 shows a top view of a Petri dish where a disc with nine radial divisions is defined with BHI culture medium (calf brain and heart infusion, peptic hydrolysate of animal tissues, pancreatic hydrolysate of casein, sodium chloride, glucose, disodium phosphate), inoculated with Streptococcus salivarius with the determination of the viability of the strains contained in the matrices with cinnamon, mint and cherry flavorings, defining three quadrants (each division corresponds to 11% of the total of the dish). Figures 4a and 4b show a top view of two Petri dishes where a disc with six radial divisions is defined (each division corresponds to 16% of the total of the dish) with BHI culture medium (calf brain and heart infusion, peptic hydrolysate of animal tissues, pancreatic hydrolysate of casein, sodium chloride, glucose, disodium phosphate), inoculated one with Streptococcus salivaria and the other with Lactobacillus plantarum, with the determination of the viability of the strains contained in the matrices. Figure 4c shows a top view of four Petri dishes where a disc with six radial divisions (each division corresponds to 16% of the total of the dish) is defined in each one with BHI culture medium (calf brain and heart infusion, peptic hydrolysate of animal tissues, pancreatic hydrolysate of casein, sodium chloride, glucose, disodium phosphate), inoculated one with Streptococcus salivaria and another with Lactobacillus plantarum, with the determination of the viability of the strains contained in twelve matrices; where six of the twelve matrices prepared maintained the viability of the strains, which is reflected in the formation of colonies in figures 4a and 4b. Figure 5 shows a top view of five Petri dishes that were first seeded with the caries consortium and then the rings containing the probiotic matrix with the different flavorings were placed on them. For the two dishes on the left, the matrix 20 included the microorganism Streptococcus salivarius and for the two dishes on the right, the matrix included the microorganism Lactobacillus plantarum, while the central dish was a control dish of matrix with water. Figures 6a and 6b show the before and after of a damaged molar with caries on a support, which in an in vitro test has been treated with the mint flavor matrix of the present invention. Figures 7a and 7b show the before and after of another damaged molar with caries on a support, which in an in vitro test has been treated with the mint flavor matrix of the present invention. Figures 8a and 8b show the before and after of another damaged molar with caries on a support, which in an in vitro test has been treated with the spearmint flavor matrix of the present invention. Figures 9a and 9b show the before and after of another damaged molar with caries on a support, which in an in vitro control test has been given the matrix without prebiotic microorganisms. Figures 10a and 10b show a before-and-after photograph of the oral cavity of a patient exhibiting dentures that were treated with the matrix formulated according to the present invention incorporated into chewing gums that were chewed twice a day for one week. For a better understanding of the invention, a detailed description of some of its modalities will be given, shown in the drawings that are attached to this description for illustrative but not limiting purposes. DETAILED DESCRIPTION OF THE INVENTION The characteristic details of the matrix formulated to provide stability to the beneficial chemical-microbial properties in oral health, in accordance with the present invention, are clearly shown in the following detailed description and in the attached illustrative drawings, the same reference signs serving to indicate the same parts. According to Figures 1a and 1b, two Petri dishes were prepared with BHI medium (calf brain and heart infusion, animal tissue peptic hydrolysate, pancreatic casein hydrolysate, sodium chloride, glucose, disodium phosphate). These were first inoculated with the caries consortium. Subsequently, filter paper discs containing 50, 100, and 150 pL of natural substances such as natural cocci and the microorganisms Saccharomyces boulardii, Streptococcus salivarius, and Lactobacillus plantarum were inoculated. The discs were allowed to dry and then placed onto the surface of the Petri dishes using sterile forceps. The Petri dishes were incubated at 37°C for 24 hours. The objective of this experiment was to determine the inhibitory components and quantities of the caries consortium made up mainly of Streptococcus mutans. Table 1 - Components for the inhibition ring test Radial Division Inhibitory component % of division Al Coco Natural 25 A-2 S. boulardii 25 A-3 S. salivarius 25 A-4 L. plantarum 25 Bl Coco Natural 25 B-2 S. boulardii 25 B-3 S. salivarius 25 B-4 L. plantarum 25 The above was an experiment using the Kirby-Bauer technique, where the following results were obtained. The discs impregnated with Streptococcus salivarius and Lactobacillus plantarum (A-3, A-IV, B-111, and B-IV) showed high interference activity against the microorganisms of the caries consortium due to the generation of joint inhibition. The other components, however, showed no signs of inhibition. According to Figures 2a and 2b, derived from the experiment in accordance with Table 1 and Figures 1a and 1b, it was decided to perform the same experiment, but only with those two microorganisms (Streptococcus salivarius and Lactobacillus plantarum). As shown in Figures 2a and 2b, where inhibition halos of approximately 3±0.3 cm in diameter can be observed in each of the rings, bacterial growth of the strain from a caries does not occur in said halo. This indicates that the 100 pL of the solution with both probiotic strains had an inhibitory effect on the growth of the bacterial consortium. To determine the matrix composition, several formulations were made, and it was found that those that included pectin, dextrose and a natural flavoring (cinnamon, cherry and mint) offered the best results in stability for the probiotic microorganism Streptococcus salivarius and Lactobacillus plantarum, as shown in Table 2 and Figure 3. Table 2. Design of experiments to determine the concentration of the different reagents Radial Division Flavoring [ml / 20ml] Pectin [g / 20ml] Dextrose [g / 20ml] Citric Acid [mg / 20ml] Flavoring Active Microorganism % division 1.1 0.5 0.75 1.5 3 Mint S. salivarius 11% 1.2 1 1.25 2.25 5 Mint S. salivarius 11% 1.3 2 1.75 3 9 Mint S. salivarius 11% 2.1 0.5 0.75 1.5 3 Cherry S. salivarius 11% 2.2 1 1.25 2.25 5 Cherry S. salivarius 11% 2.3 2 1.75 3 9 Cherry S. salivarius 11% 3.1 0.5 0.75 1.5 3 Cinnamon S. salivarius 11% 3.2 1 1.25 2.25 5 Cinnamon S. salivarius 11% 3.3 2 1.75 3 9 Cinnamon 5. salivarius 11% According to Figure 3, a Petri dish containing BHI medium (calf brain and heart infusion, animal tissue peptic hydrolysate, pancreatic casein hydrolysate, sodium chloride, glucose, disodium phosphate) was radially divided, and each prepared matrix, as specified in Table 2, was inoculated into each compartment. This test was performed to determine the ideal concentration of each ingredient based on the growth of the probiotic microorganisms contained within them, as this would indicate their stability and the active presence of the strains. Several flavorings were also evaluated to select one that would not interfere with the growth and viability of the probiotic microorganisms. Figure 3 shows a top view of a Petri dish where a disc with nine radial divisions is defined (each division corresponds to 11% of the total of the dish) with BHI culture medium (calf brain and heart infusion, peptic hydrolysate of animal tissues, pancreatic hydrolysate of casein, sodium chloride, glucose, disodium phosphate), inoculated with Streptococcus salivarius with the determination of the viability of the strains contained in the matrices with cinnamon, mint and cherry flavorings, defining three quadrants 1.1, 1.2, 1.3 for mint; 2.1, 2.2, 2.3 for cherry and 3.1, 3.2, 3.3 for cinnamon. It can be shown that the quadrants that did not show inhibition were the formulations with the Mint flavoring (quadrants 1.1, 1.2, 1.3), concluding that the cherry and cinnamon flavorings contain antimicrobial components.The first contains various probiotic agents, while the second contains eugenol between 70-95%, components used as bactericides. Based on the results obtained in the first experimental design, a second design was carried out (Table 3) where the same concentrations of all reagents were tested again, but the two probiotic strains Streptococcus salivarius and Lactobacillus plantarum were involved, spearmint was added as a new flavoring, which has characteristics similar to mint, and three brands of both flavorings were evaluated to determine those that did not inhibit the growth of the microorganisms. Table 3. Design of experiments to determine the concentration of the different reagents. Radial Division Flavoring [ml / 20ml] Pectin [g / 20ml] Dextrose [g / 20ml] Citric Acid [mg / 20ml] Flavoring Active Microorganism % of division 1 1.5 1.75 3 7 Spearmint S.salivarius / L.plantarum 16% 2 1.5 1.75 3 7 Spearmint S.salivarius / L.plantarum 16% 3 1.5 1.75 3 7 Peppermint S.salivarius / L.plantarum 16% 4 1.5 1.75 3 7 Peppermint S.salivarius / L.plantarum 16% 5 0.5 0.75 1.5 3 Peppermint S.salivarius / L.plantarum 16% 6 0.5 0.75 1.5 3 Peppermint S.salivarius / L.plantarum 16% 7 0.5 0.75 1.5 3 Spearmint S.salivarius / L.plantarum 16% 8 0.5 0.75 1.5 3 Spearmint S.salivarius / 16% 1 L.plantarum 9 1 1.25 2.25 5 Peppermint S.salivarius / L.plantarum 16% 10 1 1.25 2.25 5 Mint S.salivarius / L.plantarum 16% 11 1 1.25 2.25 5 Mint S.salivarius / L.plantarum 16% 12 1 1.25 2.25 5 Peppermint S.salivarius / L.plantarum 16% As shown in Figure 4c, six of the twelve matrices prepared maintained the viability of the strains, which is reflected in the colony formation shown in Figures 4a and 4b. This result verified that the average concentration is adequate to keep the strains alive and viable within the matrix at room temperature, because the properties of each component work together to promote the growth of the microorganisms used. With the results obtained, it was possible to establish a range for each component of the matrix: a) Pectin (3-5 g / 100 mL), b) Dextrose (6-9 g / 100 mL), c) Citric acid (25-50 mg / 100 mL), and d) Natural flavoring-Mint / Spearmint (1-2 mL / 100 mL), e) Microorganisms S. salivarius and / or f) Microorganisms L. plantarum Table 4,- Properties of the matrix at 27±2° X. Physical Properties Chemical Properties Color: Semi-transparent pH: 4 ± 2 Odor: Mint or Spearmint Brix Degrees: 13° Flavor: Mint or Spearmint Viscosity: 1.56 ± 0.3 CPS To determine the appropriate flavorings, a susceptibility test was performed with each of the matrices, in which sterile filter paper rings were impregnated with 100 pL of each flavoring and left to incubate for 24 hours. According to Figure 5, which shows five Petri dishes, each Petri dish was first inoculated with the caries consortium, and then rings containing the probiotic matrix with different flavorings were placed on top. After the incubation period, the dishes with matrices 9-12 showed an inhibition zone of approximately 1-2 mm around each ring, indicating that the matrix, in conjunction with the probiotics, inhibits the growth of microorganisms originating from caries. This result was corroborated by observing that the rings in the control dish (water and flavored matrix without probiotics) did not show any inhibition zone. To determine the concentration of the matrix components, an experimental design was carried out (Tables 2 and 3) in which the stability of the matrix was evaluated by means of the growth (conservation) of the microorganisms, their viability (Figures 4a and 4b) and subsequently their inhibitory effect on the consortium of bacteria from caries (Figure 5). For the flavoring selection process, the viability of the strains contained in each of the matrices was determined. To do this, a 10 pL sample was taken, then a seed was made in BHI medium (calf brain and heart infusion, peptic hydrolysate of animal tissues, pancreatic hydrolysate of casein, sodium chloride, glucose, disodium phosphate) and it was left to incubate for 24 h. It was concluded that the matrix should be prepared with the aforementioned quantities, as the microorganisms have the necessary nutrients to maintain them in the stationary phase. Furthermore, with these quantities, the matrix can be stored unrefrigerated without any alteration. To verify the viability of the strains, a culture was taken after 30 days, which resulted in a stationary state with the desired minimum growth. In the case of microorganisms, these are removed from the BHI medium (calf brain and heart infusion, peptic hydrolysate of animal tissues, pancreatic casein hydrolysate, sodium chloride, glucose, disodium phosphate) upon reaching the stationary phase. They are then subjected to three washes with physiological saline solution to prevent cell lysis due to osmotic pressure. The pellet is then resuspended and concentrated in sterile physiological saline solution (0.9% NaCl) to achieve a final concentration of 4 x 10¹² CFU / mL. This ensures that 0.5 mL contains 2 x 10¹⁹ CFU. According to Figures 6a-9b, to determine the matrix's function, teeth affected by caries were selected and placed in BH I medium (calf brain and heart infusion, animal tissue peptic hydrolysate, pancreatic casein hydrolysate, sodium chloride, glucose, disodium phosphate). Using a sterile syringe, the teeth were coated with 100 pL of the corresponding matrix twice daily for 5 days. Between each application, the remnants of the previous matrix were removed with a sterile swab. After each application, the Petri dishes were stored in an incubator at 37°C to simulate the temperature of the mouth. The in vitro study determined that the matrix alone (without the probiotic microorganisms Streptococcus salivarius and Lactobacillus plantarum) did not produce a positive response, as the control tooth became contaminated after two days, leading to increased caries and almost complete darkening of the tooth. In cases where the matrix formulated according to the present invention was applied, it inhibited the bacterial biofilm adhering to the teeth, reducing the problem of caries and thus potentially eliminating bad breath, tartar, and other issues resulting from the tropism generated by S. mutans. The study was conducted qualitatively, and for this purpose, five people with some type of tooth decay were selected and given the product to consume twice a day for one week. A photograph was taken of one patient at the beginning of the trial and another at the end to visualize the reduction in tooth decay (see figures 10a and 10b). Participants were also asked if they had noticed any change while consuming the chewing gum with the probiotic matrix. All reported feeling a reduction in bacterial plaque upon first contact with the gum containing the matrix formula of the present invention. The invention has been sufficiently described to allow a person of average skill to reproduce it and obtain the results mentioned herein. However, any person skilled in the art to which this invention pertains may be able to make modifications not described herein. Nevertheless, if the application of these modifications to a particular structure or to the manufacturing process thereof requires the subject matter claimed in the following claims, such structures shall be considered within the scope of the invention.

Claims

1. - A matrix formulated with probiotics to provide stability to the beneficial chemical-microbial properties in oral health characterized by comprising pectin, dextrose, citric acid, natural flavoring and an active ingredient consisting of at least one microorganism selected from Streptococcus salivarius and Lactobacillus plantarum or the mixture of both.

2. The matrix formulated with probiotics to provide stability to the beneficial chemical-microbial properties in oral health according to claim 1, characterized by comprising 35 g / 100 mL of pectin, 6-9 g / 100 mL of dextrose, 25-50 mg / 100 mL of citric acid, 2-4 mL / 100 mL of natural flavoring; and 4x10'12 CFU / mL of a microorganism selected from Streptococcus salivarius and Lactobacillus plantarum or a mixture of both.

3. The matrix formulated with probiotics to provide stability to the beneficial chemical-microbial properties in oral health according to the preceding claims, characterized in that said natural flavoring is selected from mint and spearmint.

4. The matrix formulated with probiotics to provide stability to the beneficial chemical-microbial properties in oral health according to claim 1, characterized in that it is a viscous solution.

5. The matrix formulated with probiotics to provide stability to the beneficial chemical-microbial properties in oral health according to claim 1, characterized in that pectin acts as a natural thickener which, when combined with sugar and citric acid, allows the formation of gels, which keep the microorganisms stable and at the same time develop an anti-adhesive activity against pathogens; wherein dextrose provides necessary nutrients to the microorganisms to keep them alive but without stimulating their proliferation; wherein said citric acid regulates the pH, increasing the preservation of the gel and acts as an emulsifier; and wherein the flavoring provides the organoleptic properties.

6. The matrix formulated with probiotics to provide stability to the beneficial chemical-microbial properties in oral health according to the preceding claims, characterized in that said at least one microorganism selected from Streptococcus sal i vari us and Lactobacillus plantarum or mixture of both, are in an active state.

7. The matrix formulated with probiotics to provide stability to the beneficial chemical-microbial properties in oral health according to the preceding claims, characterized in that it is integrated into confectionery, medicinal, dental products, rinses and gums.

8. A process for producing a probiotic-formulated matrix to provide stability to the beneficial chemical-microbial properties in oral health, with a composition selected from any of claims 1 to 6, characterized by comprising the steps of: a) Pouring distilled water into a beaker at a temperature between 80°C and 100°C, and adding the ingredients in the following order: dextrose powder [6-9 g / 100 mL], citric acid powder [25-50 mg / 100 mL], pectin powder [3-5 g / 100 mL]; b) Stirring at a temperature of 80°C for 5-10 minutes at a speed of 250 to 500 rpm until dissolved; c) Add the flavoring [(1-2 mL / 100 mL] and mix for 2-5 minutes at a speed of 100 to 150 rpm; d) Pour the formulated matrix into a container forming a maximum layer of 1 cm thick and subject to UV light sterilization for 15 min, then pour into a previously sterilized container for storage;e) Prepare the inoculum by reactivating the strains of at least one selected microorganism from Streptococcus salivarius and Lactobacillus plantarum or a mixture of both, performing a count using a Neubauer chamber to determine the initial concentration, which will allow calculating the mL needed to obtain the final cell concentration of 4x10'12 CFU / mL; f) Take 1 mL aliquots and place them in Eppendorf tubes (the number of samples depends on the initial concentration) to perform three washes with physiological solution (2500 rpm, 5 min), between each wash the microorganism tablet is resuspended; g) Finally, resuspend the tablet in sterile physiological solution (0.9% NaCl) and concentrate in the same diluent to obtain a final concentration of 4x10'12 CFU / mL;h) Inoculate the matrix by pouring 1.5 - 2 mL of the matrix into sterile Eppendorf tubes and inoculate each tube with the corresponding probiotic microorganism, injecting the cell suspension, mixing gently and storing at room temperature (25°C) for 5 days; 9. The production process of the probiotic-formulated matrix according to claim 8, characterized in that steps e” to “h” are carried out in a laminar flow hood under aseptic conditions and with sterile material. 10.- The use of a probiotic-formulated matrix to improve oral health comprising administering to a patient a composition selected from any of claims 1 to 7. 11.- The use of a matrix formulated with probiotics according to claim 10, characterized in that it counteracts or prevents caries in teeth, prevents bad breath, eliminates tartar, eliminates yellowing of teeth and other oral conditions.