Micronutrient delivery to the gingival pocket

Abstract

The present invention includes compositions and methods for delivering one or more nutrients to a subject, the compositions and methods comprising the step of delivering an effective amount of one or more nutrients into the gingival sulcus, the amount being sufficient to supplement the subject's diet in terms of nutrients.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Patent Application No. 17 / 956,270, filed September 29, 2022, the entire contents of each of which are incorporated herein by reference.

[0002] The present invention relates generally to the field of nutrient targeting, and more particularly to targeting the delivery of nutrients and / or micronutrients to the junctional epithelium (JE) of the gingival sulcus.

[0003] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH none

[0004] INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ON A COMPACT DISC none [Background technology]

[0005] Without limiting the scope of the invention, its background is described in relation to the delivery of model proteins, antigens, allergens, and micronutrients and vitamins.

[0006] Micronutrient deficiencies in humans impair healthy development, impair health maintenance, and cause disease. Severe forms can lead to premature death. Eliminating these deficiencies promotes healthy development, health maintenance, prevents disease, and potentially prevents premature death. The general population worldwide will benefit from this simple intervention, and subpopulations at particular risk of micronutrient deficiencies, such as obese populations, and even obese populations undergoing surgery to treat obesity, will benefit dramatically.

[0007] Micronutrients, with the exception of vitamin D, are not produced in the body and must be consumed in the diet. It is estimated that more than half of children under the age of five worldwide suffer from vitamin and mineral deficiencies. Over two billion people worldwide suffer from micronutrient deficiencies. Iron deficiency is the leading cause of anemia, affecting 40% of children under the age of five and 30% of pregnant women worldwide. Vitamin D deficiency is 8% in the U.S. population but is likely much higher globally. In the U.S. population, micronutrient deficiencies vary by age, sex, or race / ethnicity, but are closer to 10% of the population overall, with certain groups much higher in vitamin D deficiency, such as non-Hispanic blacks (31%) and Mexican Americans (12%).

[0008] Iron, zinc, and iodine are essential for preventing anemia, boosting immune function, fighting diseases such as diarrhea, pneumonia, and malaria, and promoting healthy cognitive development in infants. Vitamin A is essential for healthy vision and proper immune function, preventing blindness and death from infectious diseases such as measles and diarrhea. Vitamin D and calcium are essential for healthy bone mineralization, preventing rickets in children and osteoporosis and osteomalacia in adults.

[0009] Micronutrient deficiencies have been established in obese populations due to factors such as fat severing, inflammation-associated malabsorption in the intestine, and hyperinsulinemia-associated excretion in the urine. This association worsens with ever-increasing obesity and is highest among premenopausal obese women. The issue of micronutrient deficiencies is also confounded by the increasing prevalence of morbid obesity and the subsequent increase in the frequency of the only known effective treatment for morbid obesity, bariatric surgery. In the United States, the percentage of the population with Class / Grade I obesity (BMI 30-34.9) is 20.6%, Class II (BMI 35-39.9) is 8.8%, and Class III (BMI > 40) is 6.9%. The number of bariatric procedures performed each year continues to increase; in 2019, 256,00 bariatric procedures were performed in the United States alone. While bariatric surgery is highly effective in treating morbid obesity and its associated metabolic comorbidities, increasing life expectancy, and reducing health-related costs due to obesity, a downside is that it can lead to micronutrient deficiencies, which can lead to serious health problems and even death if postoperative vitamin regimens are not adhered to. For example, vitamin B1 or thiamine deficiency can lead to Wernicke's encephalopathy and potentially permanent neurological damage and even death. Morbidly obese individuals who undergo bariatric surgery are at highest risk for developing micronutrient deficiencies. Fat-soluble vitamins, such as A, D, E, and K, and water-soluble vitamins, such as B1, B3, B6, B9, and B12, copper, zinc, calcium, and iron, have all been documented to decline in this population after surgery for a variety of complex reasons. Micronutrients obtained from diet alone after bariatric surgery are insufficient to prevent these micronutrient deficiencies. For this reason, it is essential that individuals undergo bariatric surgery supplement their diet with oral vitamins and minerals to prevent these deficiencies. These deficiencies are due to many complex reasons, firstly, that bariatric procedures alter the normal gastrointestinal anatomy by changing the shape of the stomach and / or bypassing certain portions of the intestinal tract to different degrees.These procedures can cause malabsorption, particularly of fat, to varying degrees, alter pH, reduce total caloric intake, food intake, and fluid intake, alter food preferences, and reduce the levels of several substances critical for vitamin absorption, such as "intrinsic factor." The micronutrients most commonly affected by these changes, with the greatest clinical relevance, are vitamins B1, B12, D, K, A, E, folic acid, iron, and calcium. For this reason, absorption of oral supplements is also affected, requiring specialized dosage forms. Current supplementation regimens can be difficult due to the number and frequency of intakes or the taste of these supplements, potentially leading to noncompliance. Regimens require education and counseling, which can be difficult and often result in noncompliance. There are also commercially available skin patches from PatchMD®. The company has not presented scientific evidence of efficacy, and only a few clinical trials conducted by third-party sponsors have shown that patches from PatchMD® are significantly inferior to oral supplementation and that the rate of micronutrient deficiencies is significantly higher when the patch is used as the sole supplement. Some vitamins, particularly important in post-bariatric surgery populations, such as B12 or B1, are often administered by subcutaneous or intramuscular injection weekly or monthly for maintenance, which can be costly and painful. Nasal sprays and sublingual applications of vitamin B12 are also effective, but are expensive and limited to this vitamin. The lack of adequate monitoring, diagnosis, and timely treatment of some of these micronutrient deficiencies, such as vitamin B1 or thiamine deficiency, has led to serious health events, such as Wernicke's encephalopathy, and even death, and has significantly increased medical malpractice lawsuits. Micronutrient deficiencies that are diagnosed as severe are often treated with parenteral delivery methods. This often occurs in hospitalized patients. Delivery methods include intravenous, subcutaneous, or intramuscular injection. These methods can be costly, painful, and reserved for patients who are already hospitalized and require long-term treatment. Summary of the Invention [Problem to be solved by the invention]

[0010] Simplification and practicality of micronutrient supplementation methods are needed to improve compliance and therefore reduce the risk of these deficiencies. Despite these advances, there remains a need for nutrient-targeting strategies that maximize the delivery of nutrients or micronutrients to subjects. [Means for solving the problem]

[0011] As embodied and broadly described herein, aspects of the present disclosure relate to a method for delivering one or more nutrients to a subject, the method comprising delivering an effective amount of one or more nutrients into the gingival sulcus, the amount being sufficient to supplement the subject's diet in terms of nutrients. In one aspect, the one or more nutrients are not targeted for delivery to the vestibular mucosa. In another aspect, the one or more nutrients are targeted to the gingival sulcus epithelium, the lining epithelium, the junctional epithelium, or a combination thereof. In another aspect, the one or more nutrients are encapsulated to maximize delivery of the one or more nutrients. In another aspect, the method further comprises adding one or more agents that increase the permeability of the one or more nutrients to the gingival sulcus epithelium. In another aspect, 0.001% to 100% of the one or more nutrients are present in a depot in the junctional epithelium (JE) of the gingival sulcus. In another aspect, the one or more nutrients are repeatedly provided to the junctional epithelium of the gingival sulcus. In another embodiment, delivery of the one or more nutrients to the junctional epithelium is after consuming food or drink or brushing teeth. In another embodiment, the one or more nutrients are applied one or more times daily, weekly, or monthly, for example, 1, 2, 3, 4, 5, or 6 times daily, or 1, 2, 3, 4, 5, 6, or 7 times weekly, or 1, 2, 3, or 4 times monthly. In another embodiment, the one or more nutrients provide 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the daily nutritional needs by flossing one, two, or three times daily. In another embodiment, delivery of one or more nutrients to the endothelium, junctional epithelium, or a combination thereof is 0 hours, 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or more before the subject eats food, drinks liquid, or brushes their teeth.In another embodiment, delivery of the one or more nutrients to the marginal epithelium, junctional epithelium, or a combination thereof is 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, or more hours after the subject eats food, drinks liquid, or brushes their teeth. In another embodiment, the amount of the one or more nutrients delivered to the marginal epithelium, junctional epithelium, or a combination thereof is in the picogram to milligram range. In another embodiment, the one or more nutrients are selected from iron, phosphorus, zinc, thiamine, riboflavin, niacin, vitamin B6, folate, vitamin B12, pantothenic acid, biotin, boron, choline, chromium, copper, manganese, selenium, molybdenum, iodine, chloride, coenzyme Q10 (CoQ10), vitamin A, calcium, potassium, magnesium, vitamin E, vitamin C, carotenoids, vitamin D, or vitamin K.

[0012] As embodied and broadly described herein, embodiments of the present disclosure relate to a nutrient delivery system comprising an effective amount of one or more nutrients in a delivery device that targets the junctional epithelium of the gingival sulcus, wherein the amount of the one or more nutrients is a nutritionally effective amount. In one embodiment, the one or more nutrients are not delivered to the vestibular mucosa. In another embodiment, the delivery device comprises a cross-sectional shape that maximizes delivery of the one or more nutrients to the gingival sulcus. In another embodiment, the nutrient delivery system further comprises one or more agents that increase the permeability of the one or more nutrients to the gingival sulcus epithelium, lining epithelium, junctional epithelium, or a combination thereof. In another embodiment, 0.001% to 100% of the one or more nutrients are present in a depot in the gingival sulcus epithelium, junctional epithelium, or a combination thereof. In another embodiment, the nutrient delivery system further comprises one or more pharmaceutically acceptable carriers, excipients, diluents, buffers, or salts. In another embodiment, the one or more nutrients are selected from iron, phosphorus, zinc, thiamine, riboflavin, niacin, vitamin B6, folate, vitamin B12, pantothenic acid, biotin, boron, choline, chromium, copper, manganese, selenium, molybdenum, iodine, chloride, coenzyme Q10 (CoQ10), vitamin A, calcium, potassium, magnesium, vitamin E, vitamin C, carotenoids, vitamin D, or vitamin K.

[0013] As embodied and broadly described herein, aspects of the present disclosure relate to a method of making a floss containing a predetermined amount of one or more nutrients, the method comprising the steps of providing a floss and depositing on the floss one or more deposits of one or more nutrients in a pharmacologically acceptable carrier, each deposit having a known predetermined amount of the one or more nutrients. In one aspect, each adjacent deposit contains the same one or more nutrients or a different one or more nutrients, or each adjacent deposit contains the same one or more nutrients at different concentrations; or each adjacent deposit contains different one or more nutrients at different concentrations; or each adjacent deposit is located on a different plane from its adjacent deposit; or each adjacent deposit is located on an opposite side of the floss from the deposit; or each adjacent deposit contains one or more nutrients that are different from the previous adjacent deposit. In another embodiment, each adjacent deposit comprises one or more nutrients having different solvent requirements, selected from one or more nutrients soluble in aqueous solvents and one or more other nutrients soluble in organic solvents. In another embodiment, two or more nutrients are deposited stacked on top of each other in the form of deposits at the same or different distances / lengths. In another embodiment, one or more nutrients having different solvent requirements are deposited on opposite sides at the same or different distances / lengths. In another embodiment, one or more nutrients having the same solvent requirement are deposited stacked on top of each other at the same or different distances / lengths. In another embodiment, one or more nutrients having different solvent requirements are deposited on opposite sides at the same or different distances / lengths. In another embodiment, the floss is solid, worn, comprises multiple strands, is treated to be adhesive, is treated to adhere to a pharmacologically acceptable carrier, or is treated to adhere to an active agent. In another embodiment, each adjacent deposit comprises a dye or indicia that identifies adjacent drops or patches. In another embodiment, the floss is not soaked in one or more nutrients, a pharmacologically acceptable carrier, or both.In another embodiment, the one or more nutrients are selected from iron, phosphorus, zinc, thiamine, riboflavin, niacin, vitamin B6, folic acid, vitamin B12, pantothenic acid, biotin, boron, choline, chromium, copper, manganese, selenium, molybdenum, iodine, chloride, coenzyme Q10 (CoQ10), vitamin A, calcium, potassium, magnesium, vitamin E, vitamin C, carotenoids, vitamin D, or vitamin K. In another embodiment, the floss has a thickness of less than 5 mm, preferably less than 3 mm, preferably less than 1 mm, or preferably 35-175 microns. In another embodiment, the floss comprises a natural or synthetic polymer, an organic material, a metal, an inorganic material, or a combination thereof. In another embodiment, the floss comprises a mucoadhesive layer, or a hydrophobic or hydrophilic layer, or a combination. In another embodiment, the floss comprises a microporous structure that allows diffusion of the one or more nutrients into the gingival sulcus. In another embodiment, the floss delivers one or more nutrients to the marginal epithelium, junctional epithelium, or a combination thereof 0 hours, 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or more before the subject eats food, drinks liquid, or brushes their teeth. In another embodiment, the floss delivers one or more nutrients to the endodontic epithelium, junctional epithelium, or a combination thereof 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, or more hours after the subject eats food, drinks liquid, or brushes their teeth. In another embodiment, the viscosity of the deposit is 0.01 centipoise (cp), 1 cp, 10 cp, 100 cp, 1000 cp, 10000 cp, 100,000 cp, 200,000 cp, 300,000 cp, 500,000 cp, 1,000,000, or 100,000,000 cp.

[0014] As embodied and broadly described herein, aspects of the present disclosure relate to a floss containing a predetermined amount of one or more nutrients, the floss comprising: the floss; and one or more depots of the one or more nutrients in a pharmacologically acceptable carrier, deposited on the floss, each droplet or patch having a known, predetermined amount of an active agent. In one aspect, each adjacent deposit contains the same one or more nutrients or a different one or more nutrients; or each adjacent deposit contains the same one or more nutrients at different concentrations; or each adjacent deposit contains a different one or more nutrients at different concentrations; or each adjacent deposit is located on a different plane from the adjacent deposit; or each adjacent deposit is located on the opposite side of the floss from the adjacent deposit; or each adjacent deposit contains one or more nutrients that are different from the previous adjacent deposit. In another embodiment, each adjacent deposit comprises one or more nutrients having different solvent requirements, selected from one or more nutrients soluble in aqueous solvents and one or more nutrients soluble in organic solvents. In another embodiment, the one or more nutrients are deposited in the form of drops or patches stacked one on top of the other at the same or different distances / lengths. In another embodiment, one or more nutrients having different solvent requirements are deposited on opposite sides at the same or different distances / lengths. In another embodiment, one or more nutrients having the same solvent requirements are deposited on opposite sides at the same or different distances / lengths. In another embodiment, the floss is solid, worn, comprises multiple strands, is treated to be adhesive, is treated to adhere to a pharmacologically acceptable carrier, or is treated to adhere to an active agent. In another embodiment, each adjacent drop or patch comprises a dye or indicia that distinguishes between adjacent deposits. In another embodiment, the floss is not soaked in one or more nutrients, a pharmacologically acceptable carrier, or both.In another embodiment, the one or more nutrients are selected from iron, phosphorus, zinc, thiamine, riboflavin, niacin, vitamin B6, folic acid, vitamin B12, pantothenic acid, biotin, boron, choline, chromium, copper, manganese, selenium, molybdenum, iodine, chloride, coenzyme Q10 (CoQ10), vitamin A, calcium, potassium, magnesium, vitamin E, vitamin C, carotenoids, vitamin D, or vitamin K. In another embodiment, the floss has a thickness of less than 5 mm, preferably less than 3 mm, preferably less than 1 mm, or preferably 35-175 microns. In another embodiment, the floss comprises a natural or synthetic polymer, an organic material, a metal, an inorganic material, or a combination thereof. In another embodiment, the floss comprises a mucoadhesive layer, or a hydrophobic or hydrophilic layer, or a combination thereof. In another embodiment, the floss comprises a microporous structure that allows diffusion of antigens into the gingival sulcus. In another embodiment, the one or more nutrients are delivered to the junctional epithelium in the picogram to milligram range. In another embodiment, the one or more nutrients provide 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the daily nutritional needs by flossing one, two, or three times per day. In another embodiment, delivery of the one or more nutrients to the endodontic epithelium, junctional epithelium, or a combination thereof is 0 hours, 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or more before the subject eats food, drinks liquid, or brushes their teeth. In another aspect, delivery of the one or more nutrients to the endothelium, junctional epithelium, or a combination thereof is 0 hours, 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or more after the subject eats food, drinks liquid, or brushes their teeth.In another embodiment, the viscosity of the deposit is 0.01 centipoise (cp), 1 cp, 10 cp, 100 cp, 1000 cp, 10,000 cp, 100,000 cp, 200,000 cp, 300,000 cp, 500,000 cp, 1,000,000, or 100,000,000 cp. [Brief explanation of the drawings]

[0015] For a more complete understanding of the features and advantages of the present invention, the following detailed description of the invention should be read in conjunction with the accompanying drawings, in which: [Figure 1-1] ~ [Figure 1-3] Figure 1A shows the gingival sulcus and junctional epithelium of the human mouth. Figure 1B shows the structure of the gingival sulcus and junctional epithelium (JE). Figure 1C shows the delivery of an active agent to the junctional epithelium of the gingival sulcus and its diffusion over time in the junctional epithelium and adjacent tissues. Figure 1D shows the delivery of the antigen molecule ovalbumin (Ova) coated on floss and the effect of flossing on mouse gingival tissue. Figure 1E shows the diffusion of rhodamine-conjugated Ova in gingival tissue (in vitro). Flossing was performed by placing the antigen-coated floss around each incisor tooth and flossing approximately 10-15 times to deposit the coated antigen into the gingival pocket. Figure 1F shows the delivery efficiency of floss coated with fluorescein isothiocyanate (FITC)-conjugated ovalbumin (Ova). [Figure 2] 1 shows deposition ability, deposition of peptides, nanoparticles, proteins, oligonucleotides, microparticles onto floss on only one side in different deposition patterns as a single deposition area of ​​short or longer length or as multiple discontinuous deposition areas. [Figure 3] FIG. 1 shows the deposition ability of water-soluble materials and water-insoluble materials, including pollen grain microparticles. [Figure 4]Figure 1 shows the deposition capacity and, as an example, the different deposition patterns using two different compounds: one formulation was ovalbumin (a protein) conjugated to NHS-rhodamine (a fluorescent reagent) in water, designated "A," and the other was M2e peptide conjugated to gold nanoparticles and CpG (single-stranded DNA) in water, designated "B." [Figure 5] FIG. 1 is a diagram illustrating the deposition capabilities of multiple materials, in this case four different food colors (blue, green, yellow, red) deposited as four separate portions. [Figure 6] FIG. 1 shows the coating ability of coating both sides of a floss with different formulations. [Figure 7] FIG. 1 shows an example of an automated coating station for coating floss. [Figure 8] 1A and 1B show two examples of flosser system designs. [Figure 9] FIG. 1 shows floss coated with vitamins B1, B12, and D. [Figure 10] FIG. 1 illustrates the passage of floss through teeth. [Figure 11] FIG. 1 shows floss coated with vitamin B12. [Figure 12] FIG. 1 shows the percent change in serum vitamin B12 levels in human subjects between the pill and floss groups. DETAILED DESCRIPTION OF THE INVENTION

[0016] Although the making and using of various embodiments of the invention are described in detail below, it should be understood that the present invention provides many applicable inventive concepts that can be embodied in a wide variety of specific contexts. The specific embodiments described herein are merely illustrative of specific ways to make and use the invention and do not limit the scope of the invention.

[0017] To facilitate understanding of the present invention, certain terms are defined below. Terms defined herein have meanings commonly understood by one of ordinary skill in the art in the areas relevant to the present invention. Terms such as "a," "an," and "the" do not refer to only a single entity, but include general classes for which specific examples may be used for purposes of illustration. While the terminology herein is used to describe specific embodiments of the present invention, their usage does not limit the present invention except as outlined in the claims.

[0018] The present invention seeks to address micronutrient deficiencies in the general population, which are highly prevalent and occur worldwide, stemming from complex socioeconomic issues. Micronutrient deficiencies in the bariatric surgery population are also highly prevalent, and the current mainstay of prevention is oral supplementation. Regimen requires education and counseling, can be difficult, and often leads to non-compliance. There are also commercially available skin patches from PATCHMD®. The company has not presented scientific evidence of efficacy, and only a few clinical trials conducted by third-party sponsors have shown that patches from PATCHMD® are significantly inferior to oral supplementation and that the rate of micronutrient deficiencies is significantly higher when the patch is used as the sole supplement (Saurabh, S., et al., Is Transdermal Multivitamin Patch Effective in Gastric Bypass Patients? Obes Surg, 2019. 29(12): pp. 3818-3823; McCormick, R., et al., The Effectiveness of Transdermal Iron Patches in Athletes With Suboptimal Iron Status (Part 1). Int J Sport Nutr Exerc Metab, 2020. 30(3): pp. 185-190). Some vitamins that are particularly important in post-bariatric surgery populations, such as B12 or B1, are often administered subcutaneously or intramuscularly weekly or monthly for maintenance, which can be costly and painful. Vitamin B12 nasal sprays and sublingual applications are also effective, but are expensive and limited to this vitamin.

[0019] Micronutrient deficiencies that are diagnosed as severe are often treated with parenteral delivery methods. This often occurs in hospitalized patients. Delivery methods are intravenous, subcutaneous, or intramuscular injections. These methods are costly, can be painful, and may be reserved for patients who are already hospitalized and require long-term treatment.

[0020] The present invention addresses the problem of nutrient / micronutrient deficiency using floss, particularly floss coated with nutrients and / or micronutrients. Because some individuals in the general population prefer not to take pills, using coated floss to deliver micronutrients can increase micronutrient supplementation. This is particularly effective for highly efficient and effective delivery of nutrients / micronutrients that are readily available using daily routines, as demonstrated herein.

[0021] In the bariatric surgery population, where adherence to a postoperative vitamin / mineral supplement regimen is critical in preventing serious illness and possible death, any method that facilitates compliance with such a regimen significantly reduces morbidity and mortality. Passing coated floss through the gum pockets several times daily is preferable for most individuals to swallowing multiple tablets or capsules multiple times daily.

[0022] To date, no one has described an effective method of coating floss with micronutrients, then delivering that coating to the gum pockets by flossing, and then demonstrating elevated blood levels of these micronutrients after delivery.

[0023] The rationale for using the gingival pocket for this purpose is the presence of a highly permeable tissue called the junctional epithelium, which is vascularized and highly permeable, thus providing immediate access to the systemic circulation (Garant, PR, Oral cells and tissues. 2003, Chicago: Quintessence Pub. Co. ix, 430 p.; Moutsopoulos, NM and JE Konkel, Tissue-Specific Immunity at the Oral Mucosal Barrier. Trends Immunol, 2018. 39(4): pp. 276-287). When teeth erupt through the gingiva, they create a break in the otherwise continuous and uninterrupted mucosal surface (Pollanen, M.T., J.I. Salonen, and V.-J. Uitto, Structure and function of the tooth-epithelial interface in health and disease. Periodontology 2000, 2003. 31(1): pp. 12-31; Schroeder, H.E. and M.A. Listgarten, The Junctional Epithelium: From Strength to Defense. Journal of Dental Research, 2003. 82(3): pp. 158-161). To seal this discontinuity, gingival tissue attaches to the tooth through the junctional epithelium (Nakamura, M., Histological and immunological characteristics of the junctional epithelium. Jpn Dent Sci Rev, 2018. 54(2): pp. 59-65).

[0024] Interestingly, the junctional epithelium is leaky and highly permeable (Nakamura, M., Histological and immunological characteristics of the junctional epithelium. Jpn Dent Sci Rev, 2018. 54(2): p. 59-65). The dentogingival junction is only three to four cell layers thick (often becoming a single cell thick towards its base), the cells of the junctional epithelium are loosely packed, and there are only a few adherens junctions, desmosomes, and gap junctions (Luke, D.A., The structure and functions of the dentogingival junction and periodontal ligament. British Dental Journal, 1992. 172(5): pp. 187-190; Bosshardt, D.D. and N.P. Lang, The Junctional Epithelium: from Health to Disease. Journal of Dental Research, 2005. 84(1): pp. 9-20; Stern, I.B., Current Concepts of the Dentogingival Junction: The Epithelial and Connective Tissue Attachments to the Tooth. Journal of Periodontology, 1981. 52(9): p. 465-476), all of which are essential for creating tight connections and establishing a barrier between adjacent cells in the epithelium. These structures are abundant in the gastrointestinal epithelium, and therefore the oral route has low permeability. The lack of these structures in the junctional epithelium makes it attractive for systemic delivery.

[0025] Beyond the junctional epithelium's junctional zone, when gingival tissue approaches the upper surface of the tooth, it remains unattached to the tooth, forming a gingival sulcus, also known as a gingival pocket. This is the same crevice that dentists probe to determine the depth of the pocket during an examination. This unusually high permeability of the junctional epithelium can be exploited for micronutrient delivery. The inventors hypothesized that micronutrients administered through this niche region could diffuse freely into the systemic circulation.

[0026] However, the junctional epithelium is only about 2 mm long, and the gingival sulcus is only 1-3 mm deep. Therefore, administration to the gingival sulcus, which is close to the junctional epithelium, is not a trivial matter. The inventors further hypothesized that floss intended to clean the space between the teeth and gingival pockets could be coated with vitamins and micronutrients that would be targeted to the gingival sulcus for uptake through the junctional epithelium.

[0027] Delivery of micronutrients to the gingival space allows for direct uptake of micronutrients into the circulatory system without the need to inject them into the skin or muscle or into a vein. Application is easy for the individual and there is no pain associated with injections. Cost is significantly lower as no visit to a healthcare provider or specialized equipment is required. By allowing direct uptake into the circulatory system, the inherent problems of intestinal uptake associated with obesity are alleviated. A much lower dose than the recommended oral daily allowance is required because the gingival pocket epithelium does not pose the severe bioavailability issues associated with uptake via the intestine and potentially eliminates the first-pass effect (loss of bioavailability due to first-pass hepatic administration).

[0028] The present invention is directed to administering micronutrients and vitamins through the junctional epithelium of the gingival sulcus to achieve systemic delivery. Because the junctional epithelium is only 2 mm long and the gingival sulcus is 1-2 mm deep, the inventors coated vitamins and micronutrients onto dental floss, targeting them to the gingival sulcus for uptake through the junctional epithelium. While the inventors used floss targeted to the junctional epithelium, other approaches that can target the junctional epithelium can be used. For example, a thin, flat surface similar in size to the gingival sulcus can be used. This surface can be coated with the desired material, or the material can be encapsulated in the surface. Using a mouse model, the inventors demonstrated that floss can be coated with Ova solution and flossed on mouse teeth, demonstrating that this method is an effective delivery method. The inventors further demonstrated that vitamin-coated floss can deliver the coated cargo in humans, and that vitamin B12-coated floss increased serum levels after 1 or 2 weeks of flossing.

[0029] The present invention can be used with dental flosses known in the art. For example, the dental floss can be produced as nylon dental floss. In nylon dental floss, nylon is polymerized into a polymer, and then formed, pumped, or extruded to form a monofilament or multiple filaments. The polymer is cured, and the monofilament or multiple filaments are combined to form a strand of dental floss. The dental floss can be produced from polytetrafluoroethylene (PTFE or TEFLON®), polypropylene, polyethylene, styrene butadiene copolymer, or a combination thereof. Once formed, the polymer can be melted and extruded into thin strands. See, for example, U.S. Patent No. 6,270,890, the relevant portions of which are incorporated herein by reference.

[0030] In one non-limiting example, nylon or PTFE is mixed with a basic amino acid (or its salt) and molded or extruded to form one or more filaments. In the case of multiple filaments, these are typically twisted together to form dental floss. Alternatively, a single ribbon floss, such as PTFE, can be formed. Often, dental floss has a denier of about 450 to about 1350; in other examples, the floss denier is about 100 to about 900.

[0031] The dental floss is then coated with the vitamins and / or micronutrients of the present invention as known to those skilled in the art. For example, the dental floss is treated in a bath containing the vitamins and / or micronutrients. The bath can include one or more waxes that adhere to the floss, thereby adhering the vitamins and / or micronutrients to the floss. As an example, dental floss containing nylon or PTFE fibers is coated with the vitamins and / or micronutrients. Waxes or polymers, such as polyvinyl alcohol, polyvinyl acetate, etc., can be used to coat the vitamins and / or micronutrients in or around the dental floss. See, for example, U.S. Pat. No. 6,289,904, the relevant portions of which are incorporated herein by reference.

[0032] In filament dental floss, vitamins and / or micronutrients can be embedded in thin filament bundles, e.g., nylon filament bundles, before, during, or even after the bundles are formed. The bundles may then be coated with wax or a polymer. The number of filaments can range from about 2 to about 500, e.g., about 2 to about 250, depending on the denier of the dental floss filament. Dental floss filaments are often twisted at about 1 to 5 twists per inch to form the floss. The twisting provides the dental floss with integrity when placed on a spool and / or during subsequent handling. Upon application, the dental floss filaments unfold and spread against the tooth surface at the gingival junctional epithelium, thereby delivering the vitamins and / or micronutrients. Floss can also be formed from interlocking fibers. The dental floss product preferably has a thickness that allows it to fit between teeth while also reaching the gingival junctional epithelium. If multiple filaments are used, the coating may be applied before and / or after twisting, generally after application of the vitamins and / or micronutrients. Other additives may be applied to the dental floss to preserve the vitamins and / or micronutrients or to aid in the coating process or to achieve controlled release of the vitamins and / or micronutrients.

[0033] Additionally, flavors can be applied to dental floss as a liquid or solid. Flavors can be spray-dried in liquid or solid form. When flavors are applied as a liquid, they are generally dried before winding the floss onto a spool. Drying can be air-dried or heat-dried, and the floss is then wound onto a spool.

[0034] As used herein, the terms "deposited," "depot," and "deposition" refer to placement on the floss in the form of one or more deposits of active agent separated from adjacent deposits by gaps.

[0035] The terms "nutrient," "micronutrient," and "vitamin" are used interchangeably herein and generally refer to any substance that provides nutrients essential for growth and the maintenance of life, including, but not limited to, one or more of: iron, phosphorus, zinc, thiamine, riboflavin, niacin, vitamin B6, folic acid, vitamin B12, pantothenic acid, biotin, boron, choline, chromium, copper, manganese, selenium, molybdenum, iodine, coenzyme Q10 (CoQ10), vitamin A, calcium, potassium, magnesium, vitamin E, vitamin C, carotenoids, vitamin D, or vitamin K. Other examples of nutrients include proteins, amino acids, lipids, carbohydrates, nucleic acids, or physical or chemical combinations thereof. These can be pure substances or mixtures or extracts from natural sources.

[0036] As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier that does not cause adverse effects in subjects to whom it is administered (e.g., humans and pets such as dogs, cats, cows, pigs, or other domesticated or even non-domesticated animals). Suitable pharmaceutically acceptable carriers include, for example, one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol, dimethyl sulfoxide, and the like, and combinations thereof. Additionally, if desired, the vitamins and / or micronutrients can contain minor amounts of auxiliary substances such as wetting or emulsifying agents, pH buffering agents, plasticizers, thickening agents, and / or surface tension adjusting agents.

[0037] As used herein, the term "subject" refers to a human, a pet (such as a dog, cat, cow, sheep, goat, horse, rabbit, or pig), or other domesticated animal, or a non-domesticated animal such as a deer, buffalo, or wild horse.

[0038] The junctional epithelium is located at the bottom of the gingival sulcus and is 1 to 2 mm deep in healthy gums. Furthermore, the apical tissue of the gingival cavity tightly holds the teeth, allowing only thin instruments less than 1 mm, preferably less than 500 μm, to enter the cavity. Therefore, administering materials to the gingival sulcus is not a trivial task. To overcome this challenge, the present invention uses vitamins and / or micronutrients coated on dental floss. Dental floss is used daily by millions of people to clean the gingival sulcus, and the present invention describes a method for coating dental floss with vitamins and / or micronutrients that are targeted to the gingival sulcus for uptake through the junctional epithelium. Dental floss offers the additional advantage of being non-invasive, painless, and self-administerable in the comfort of one's home. Dental floss should be considered a non-limiting example of a system whose ultimate goal is to deliver materials to the junctional epithelium. Other approaches based on the principle of allowing devices such as tapes, films, strips, strings, threads, sutures, gels, hydrogels, polymers, viscous materials, particles, or combinations thereof to enter the gingival sulcus and help target the junctional epithelium are included in the present invention. These systems and devices can be inserted into the gingival sulcus, but can also be placed on the apical side of the sulcus rather than within it to penetrate the tissue, allowing the desired molecules to then diffuse from the system and device into the sulcus and ultimately to the junctional epithelium. These systems placed on the apical side of the sulcus can be designed to maximize diffusion of molecules into the sulcus while minimizing loss outward and into the general oral cavity. In one such approach, the delivery system can be coated with an impermeable layer on the side facing the sulcus.

[0039] To coat the floss, we developed a simple manual coating method using a pipette to apply the material to the floss. After conducting a preliminary feasibility study, we selected Oral-B® Glide Pro-Health Original Floss from among five different flosses. Using this method, we were able to coat floss with different molecules, including proteins, small molecules (such as nutrients and / or micronutrients), peptides, nanoparticles, single-stranded DNA oligonucleotides, and influenza viruses. Next, we established the feasibility of flossing the teeth of mice. We chose to floss the lower anterior incisors because of their ease of access. Flossing was performed while the mice were under anesthesia. The figure shows the incisors before, during, and after flossing. Images under a fluorescent stereomicroscope confirmed that the coated fluorescent ovalbumin (Ova) penetrated the junctional epithelium and entered the gingival tissue in less than 30 minutes. We determined the percentage of Ova delivered to the gingival sulcus by quantifying the Ova coated on the floss (M1) and the Ova remaining on the floss after flossing (M2). For n = 4 mice, the delivery efficiency ([M1-M2] / M1 x 100) was approximately 75%.

[0040] We used a pipette tip to manually coat the floss with a solution containing antigens / allergens and / or vitamins and / or micronutrients. To increase coating reproducibility and delivery efficiency, an automated coating technique using a computer-controlled linear stage and fluid dispensing system can be used. Using a floss-coating machine, specific lengths of floss can be coated with any vitamin and / or micronutrient simply by switching vials for the coating liquid. Other coating options include dip coating, spray coating, inkjet printing, pipette-based coating, cartridge printing, or a combination thereof. Coatings may require excipients such as thickeners or surface tension-reducing agents to improve coating and delivery efficiency. Additionally, trehalose and other substances known to protect molecules from drying forces can be used to improve molecule stability. Delivery efficiency can be evaluated, and image processing can be used to characterize the coating.

[0041] Figures 1A-1F show the junctional epithelium of the oral cavity: (Figure 1A) Human mouth; (Figure 1B) Structure of the gingival sulcus and junctional epithelium (JE); (Figure 1C) Delivery of an active agent to the junctional epithelium of the gingival sulcus and its diffusion over time in the junctional epithelium and adjacent tissues; (Figure 1D) Delivery of antigen molecules coated on floss and the effect of flossing on mouse gingival tissue; and (Figure 1E) Diffusion of rhodamine-conjugated ovalbumin (Ova) in gingival tissue. Flossing was performed by placing antigen-coated floss around each incisor tooth and flossing 10 times to deposit the coated antigen at the gum line. (Figure 1F) Delivery efficiency of floss coated with fluorescein isothiocyanate (FITC)-conjugated ovalbumin (Ova). [Example]

[0042] How to attach to dental floss The majority of floss on the market is coated with a continuous coating of material (wax, flavorings, etc.). Currently, the entire floss length (hundreds of feet in a floss cartridge) is coated. These coatings are not properly characterized and cannot be used for medical applications where it is important to deliver a known amount of agent, such as in the case of vaccine delivery, or drug / therapeutic molecule delivery, or vitamin and / or micronutrient delivery, where deviations from the recommended dose can be harmful or cause side effects.

[0043] The compositions and methods of the present invention allow for the simple coating of any molecule by fluid dispensing. The methods can be used to coat one or more active agents / molecules along specific lengths of floss, on specific surfaces of the floss, and in distinct locations. Furthermore, the methods can be used to coat both sides of the floss, if desired.

[0044] The present invention provides a novel method for coating dental floss. Substances (e.g., synthetic molecules or polymers, amino acids or polymers thereof, nucleotides or polymers thereof, lipids, carbohydrates, natural materials, antigens / allergens / adjuvants / drugs / vitamins, and / or micronutrients, or combinations thereof) can be coated onto the surface of the floss for delivery to the gingival tissue. Delivery can have any desired application, such as modulating immune response, systemic effects, or local effects. The surface of the floss can be coated by depositing a substance over a shorter or longer distance / length of the floss (deposition methods include depositing a single continuous portion of the floss or two or more separate portions of the floss with the same or different spacing between each of the deposited areas) (deposition methods include placing drops on the floss, or dragging drops on the floss and spreading them over a distance / length of the floss, or spray coating, or inkjet printing, or pipette-based coating, or cartridge printing, including different printing techniques such as piezo or ultrasonic printing, or combinations thereof), and allowing the coating to dry. To demonstrate the invention, antigens / allergens / peptides / microparticles / nanoparticles / single-stranded deoxyribonucleic acid (DNA) / vitamins and / or micronutrients were used to show efficacy and proof of concept. Varying amounts of substances can be coated onto the surface of the floss. The coated material can be easily delivered to the gum tissue by simple flossing actions.

[0045] For medical applications using coated floss, it is important that it have the following properties: (a) the coating should be consistent over a short length of the coated floss to allow consistent delivery to the gum pocket by the user; (b) a known amount of formulation should be coated onto the floss; and / or (c) the coating should remain adhered to the surface until intended use.

[0046] Floss is often made of hydrophobic materials (such as TEFLON® or nylon), and these surfaces are difficult to wet using coating solutions. Because of the poor wetting, it is difficult to achieve a continuous, uniform coating on the floss. Many different solvents can be used to make the coating solution, but water is preferred for biological materials that must be coated onto the floss, and water-based coating solutions are much more difficult to coat onto the floss. However, non-aqueous solutions can also be used in the present invention, where an active agent that is not soluble (or partially soluble) in water is present in the solvent, the active agent is deposited on the floss, and the solvent is allowed to evaporate, leaving the active agent behind.

[0047] Instead of creating a continuous coating, discrete drops can be deposited on the floss surface. This reduces the need to spread the coating evenly across its length, allowing for reproducible coatings and patterns to be achieved. (1) A fluid dispensing system (manual or automatic, or a combination thereof) can be used to place drops on the floss. For proof-of-concept purposes, manual dispensing was performed. (2) The surface of the floss can be made hydrophilic (e.g., by coating with a hydrophilic polymer, or by oxygen plasma treatment or other common surface treatment techniques that can alter the surface energy of the floss surface to better allow for spreading of the coating liquid). (3) The coating liquid is placed on the floss surface. After a period of time, after enough solvent has evaporated, the liquid on the floss can be mechanically spread. After some solvent has evaporated, the viscosity of the coating liquid increases, improving its ability to be spread across the floss.

[0048] Advantages of using a dispensing system (manual, automated, or a combination thereof) to apply to the floss include: (1) reduced loss of applied formulation compared to spray / dip coating; (2) precise amount application; (3) application of multiple applied formulations; and / or (4) avoidance of surface modification of the floss, since even aqueous solutions can be applied as drops to create a uniform pattern.

[0049] Spray or dip coating can result in material wastage. In contrast, the use of deposition on the surface of the floss to separate deposits minimizes or eliminates material loss. Depositing floss and achieving precise control (e.g., when the goal is to deposit small spots on the floss less than approximately 1 mm in length / diameter) is difficult. However, with fluid dispensing, even nanoliter to picoliter volumes can be easily deposited on the floss at known, precise locations. With fluid dispensing, it is also straightforward to deposit different materials with small gaps between different deposition spots. This level of accuracy and precision is difficult with spray / dip coating. Using this approach, deposition devices can be developed and constructed and deployed in pharmacies, homes, or clinics. Furthermore, the present invention can be used to separately and independently deposit active agents with different solvent requirements for solubility (e.g., one active agent requires water as a solvent and another active agent requires an organic solvent) on the floss.

[0050] Figure 2 shows the deposition capability and deposition of peptides, nanoparticles, proteins, oligonucleotides, and microparticles onto floss in different deposition patterns, either as a single deposition area of ​​short or long length or as multiple discontinuous deposition areas, on only one side of the floss. Figure 3 shows the deposition capability for depositing water-soluble materials and water-insoluble materials, including pollen grain microparticles. Figure 4 shows the deposition capability and different deposition patterns using two different compounds as examples. One formulation was ovalbumin (a protein) conjugated to NHS-rhodamine (a fluorescent reagent) in water, designated "A," while the other was M2e peptide conjugated to gold nanoparticles and CpG (single-stranded DNA) in water, designated "B." Figure 5 shows the deposition capability for multiple materials, in this case, four different food colors (blue, green, yellow, and red) deposited as four separate portions. Figure 6 shows the coating capability for coating both sides of the floss with different formulations. Figure 7 shows an example of an automated coating station for coating floss. Automated coating station 10 includes a stand 12 including a controlled linear motion stage 14 (shown as a two-dimensional stage in this embodiment) that allows movement in one or two dimensions, with a syringe assembly 18 coupled to a back surface 16 that controls the delivery of droplets 20 to floss 22. The stage is controlled by a computer 24 that can be connected to controlled linear motion stage 14 and / or syringe assembly 18. Figure 8 shows two example flosser system designs. [Example]

[0051] Micronutrient delivery to the gingival pocket The purpose of this example was to provide a method for delivering micronutrients (such as vitamins and minerals) to a mammalian subject via their gingival pockets using floss coated with said micronutrients. The method includes the steps of (1) employing the micronutrient-coated floss, (2) inserting the floss into the gingival pocket, and (3) flossing.

[0052] In some embodiments, the floss can be coated with a single micronutrient, while in other embodiments, the floss can be coated with multiple micronutrients, either mixed together or individually on the same floss.

[0053] Some micronutrients are water-soluble, while some can be water-insoluble or have poor water solubility. The goal is to obtain uniformity of composition and reproducibility of dosage when more than one micronutrient is coated onto the floss as a mixture. To achieve this, any approach can be followed, including: (1) using a co-solvent system (such as a polar solvent like water + an organic solvent like ethanol) that allows solubilization of all of the micronutrients; and / or (2) forming a stable suspension, such that some micronutrients are fully soluble in the solvent system while others are suspended as particles to form a homogeneous suspension or emulsion. These suspended micronutrients can have partially solubilizing molecules that solubilize them in the solvent system.

[0054] To allow for the formation of reproducible coatings, the substrate of the floss can be modified chemically or physically, or by a combination thereof. For example, the floss material can be coated with a material containing a polymer that can alter surface properties, such as surface energy, so that the wetting tendency of the floss surface becomes more receptive to the coating. An example of this material can be poly(lactic-co-glycolic acid). The substrate can be exposed to oxygen, ozone, plasma, or other gas and radiation sources. Substrates can be selected in addition to those commonly used in floss (Teflon® and nylon are the most common, but silk and other natural fibers also exist).

[0055] Micronutrients can be formulated to modify their release kinetics, such as slow-release formulations. This can be achieved in a number of ways: (1) encapsulating the micronutrients individually or in combination into particles; and / or (2) formulating water-soluble and water-insoluble (slightly soluble) micronutrients together, whereby the water-insoluble (slightly soluble) micronutrients act as the slow-release component without the need for extra materials.

[0056] Highly water-soluble micronutrients can be formulated in a way that increases their residence time in the gum pocket, allowing for better bioavailability. The floss can have a solid material or a braided structure.

[0057] Figure 9 shows a floss (CVS brand) coated with vitamins B1, B12, and D. The coated floss is passed through the gum pocket, depositing most of the coating there. Figure 10 shows the floss being passed through the teeth. The majority of the coated vitamin mixture (B1, B12, and D) is deposited in approximately two passes of the floss. [Example]

[0058] Efficacy of vitamin B12-coated floss in increasing serum B12 levels in human subjects CVS brand floss was coated with 6 μg of vitamin B12 by manually pipetting an aqueous solution containing vitamin B12 onto the floss. Obese pre-bariatric surgery patients were recruited and divided into an experimental floss group and a micronutrient pill / tablet (Centrum™ Adult) group. The pill also contained 6 μg of vitamin B12. In the floss group, subjects self-applied the coated floss for 7 days (D), 14 days, or 21 days. In the pill group, subjects took a tablet orally once per day for 7 days or 30 days. Blood was collected before treatment began (baseline) and at the end of treatment. Clinical laboratory tests were performed to measure serum vitamin B12 levels, and the percent change in serum vitamin B12 was calculated. Figure 11 shows the vitamin B12-coated floss. Figure 12 shows the percent change in serum vitamin B12 levels for the pill and floss groups. The percent change in the floss group was found to be significantly higher compared to the pill group, demonstrating the implementation of the present invention of this patent.

[0059] As embodied and broadly described herein, aspects of the present disclosure relate to a method for delivering one or more nutrients to a subject, the method comprising, consisting essentially of, or consisting of delivering an effective amount of one or more nutrients into the gingival sulcus, the amount being sufficient to supplement the subject's diet in terms of nutrients. Each of the following aspects can be an individual aspect or can be combined with one or more of the following aspects. In one aspect, the one or more nutrients are not targeted for delivery to the vestibular mucosa. In another aspect, the one or more nutrients are targeted to the gingival sulcus epithelium, lining epithelium, junctional epithelium, or a combination thereof. In another aspect, the one or more nutrients are encapsulated to maximize delivery of the one or more nutrients. In another aspect, the method further comprises adding one or more agents that increase the permeability of the one or more nutrients to the gingival sulcus epithelium. In another aspect, 0.001% to 100% of the one or more nutrients are present in a depot in the junctional epithelium (JE) of the gingival sulcus. In another embodiment, the one or more nutrients are repeatedly provided to the junctional epithelium of the gingival sulcus. In another embodiment, delivery of the one or more nutrients to the junctional epithelium is after consuming food or drink, or brushing teeth. In another embodiment, the one or more nutrients are applied one or more times daily, weekly, or monthly, for example, 1, 2, 3, 4, 5, or 6 times daily, or 1, 2, 3, 4, 5, 6, or 7 times weekly, or 1, 2, 3, or 4 times monthly. In another embodiment, the one or more nutrients provide 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the daily nutritional needs by flossing one, two, or three times daily. In another embodiment, delivery of one or more nutrients to the endothelium, junctional epithelium, or a combination thereof is 0 hours, 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or more before the subject eats food, drinks liquid, or brushes their teeth.In another embodiment, delivery of the one or more nutrients to the marginal epithelium, junctional epithelium, or a combination thereof is 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, or more hours after the subject eats food, drinks liquid, or brushes their teeth. In another embodiment, the amount of the one or more nutrients delivered to the marginal epithelium, junctional epithelium, or a combination thereof is in the picogram to milligram range. In another embodiment, the one or more nutrients are selected from iron, phosphorus, zinc, thiamine, riboflavin, niacin, vitamin B6, folate, vitamin B12, pantothenic acid, biotin, boron, choline, chromium, copper, manganese, selenium, molybdenum, iodine, chloride, coenzyme Q10 (CoQ10), vitamin A, calcium, potassium, magnesium, vitamin E, vitamin C, carotenoids, vitamin D, or vitamin K.

[0060] As embodied and broadly described herein, embodiments of the present disclosure relate to a nutrient delivery system comprising, consisting essentially of, or consisting of an effective amount of one or more nutrients in a delivery device that targets the junctional epithelium of the gingival sulcus, wherein the amount of the one or more nutrients is a nutritionally effective amount. Each of the following embodiments can be an individual embodiment or can be combined with one or more of the following embodiments. In one embodiment, the one or more nutrients are not delivered to the vestibular mucosa. In another embodiment, the delivery device includes a cross-sectional shape that maximizes delivery of the one or more nutrients to the gingival sulcus. In another embodiment, the nutrient delivery system further includes one or more agents that increase the permeability of the one or more nutrients to the gingival sulcus epithelium, lining epithelium, junctional epithelium, or a combination thereof. In another embodiment, 0.001% to 100% of the one or more nutrients are present in a depot in the gingival sulcus lining epithelium, junctional epithelium, or a combination thereof. In another embodiment, the nutrient delivery system further comprises one or more pharmaceutically acceptable carriers, excipients, diluents, buffers, or salts. In another embodiment, the one or more nutrients are selected from iron, phosphorus, zinc, thiamine, riboflavin, niacin, vitamin B6, folic acid, vitamin B12, pantothenic acid, biotin, boron, choline, chromium, copper, manganese, selenium, molybdenum, iodine, chloride, coenzyme Q10 (CoQ10), vitamin A, calcium, potassium, magnesium, vitamin E, vitamin C, carotenoids, vitamin D, or vitamin K.

[0061] As embodied and broadly described herein, aspects of the present disclosure relate to a method of making a floss containing a predetermined amount of one or more nutrients, the method comprising the steps of providing a floss and depositing on the floss one or more deposits of one or more nutrients in a pharmacologically acceptable carrier, each deposit having a known predetermined amount of the one or more nutrients. Each of the following aspects can be an individual aspect or can be combined with one or more of the following aspects. In one aspect, each adjacent deposit contains the same one or more nutrients or different one or more nutrients, or each adjacent deposit contains the same one or more nutrients at different concentrations; or each adjacent deposit contains different one or more nutrients at different concentrations; or each adjacent deposit is located on a different plane from its adjacent deposit; or each adjacent deposit is located on an opposite side of the floss from the deposit; or each adjacent deposit contains one or more nutrients that are different from the previous adjacent deposit. In another embodiment, each adjacent deposit comprises one or more nutrients having different solvent requirements, selected from one or more nutrients soluble in aqueous solvents and one or more other nutrients soluble in organic solvents. In another embodiment, two or more nutrients are deposited stacked on top of each other in the form of deposits at the same or different distances / lengths. In another embodiment, one or more nutrients having different solvent requirements are deposited on opposite sides at the same or different distances / lengths. In another embodiment, one or more nutrients having the same solvent requirement are deposited stacked on top of each other at the same or different distances / lengths. In another embodiment, one or more nutrients having different solvent requirements are deposited on opposite sides at the same or different distances / lengths. In another embodiment, the floss is solid, worn, comprises multiple strands, is treated to be adhesive, is treated to adhere to a pharmacologically acceptable carrier, or is treated to adhere to an active agent. In another embodiment, each adjacent deposit comprises a dye or indicia that identifies adjacent drops or patches. In another embodiment, the floss is not soaked in one or more nutrients, a pharmacologically acceptable carrier, or both.In another embodiment, the one or more nutrients are selected from iron, phosphorus, zinc, thiamine, riboflavin, niacin, vitamin B6, folic acid, vitamin B12, pantothenic acid, biotin, boron, choline, chromium, copper, manganese, selenium, molybdenum, iodine, chloride, coenzyme Q10 (CoQ10), vitamin A, calcium, potassium, magnesium, vitamin E, vitamin C, carotenoids, vitamin D, or vitamin K. In another embodiment, the floss has a thickness of less than 5 mm, preferably less than 3 mm, preferably less than 1 mm, or preferably 35-175 microns. In another embodiment, the floss comprises a natural or synthetic polymer, an organic material, a metal, an inorganic material, or a combination thereof. In another embodiment, the floss comprises a mucoadhesive layer, or a hydrophobic or hydrophilic layer, or a combination. In another embodiment, the floss comprises a microporous structure that allows diffusion of the one or more nutrients into the gingival sulcus. In another embodiment, the floss delivers one or more nutrients to the marginal epithelium, junctional epithelium, or a combination thereof 0 hours, 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or more before the subject eats food, drinks liquid, or brushes their teeth. In another embodiment, the floss delivers one or more nutrients to the endodontic epithelium, junctional epithelium, or a combination thereof 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, or more hours after the subject eats food, drinks liquid, or brushes their teeth. In another embodiment, the viscosity of the deposit is 0.01 centipoise (cp), 1 cp, 10 cp, 100 cp, 1000 cp, 10000 cp, 100,000 cp, 200,000 cp, 300,000 cp, 500,000 cp, 1,000,000, or 100,000,000 cp.

[0062] As embodied and broadly described herein, embodiments of the present disclosure relate to a floss containing a predetermined amount of one or more nutrients, the floss comprising, consisting essentially of, or consisting of the floss; and deposits on the floss, one or more depots of the one or more nutrients in a pharmaceutically acceptable carrier, each droplet or patch having a known, predetermined amount of an active agent. Each of the following embodiments can be an individual embodiment or can be combined with one or more of the following embodiments. In one embodiment, each adjacent deposit contains the same one or more nutrients or different one or more nutrients, or each adjacent deposit contains the same one or more nutrients at different concentrations; or each adjacent deposit contains different one or more nutrients at different concentrations; or each adjacent deposit is located on a different plane from the adjacent deposit; or each adjacent deposit is located on the opposite side of the floss from the adjacent deposit; or each adjacent deposit contains one or more nutrients that are different from the previous adjacent deposit. In another embodiment, each adjacent deposit comprises one or more nutrients having different solvent requirements, selected from one or more nutrients soluble in aqueous solvents and one or more nutrients soluble in organic solvents. In another embodiment, the one or more nutrients are deposited in the form of drops or patches stacked one on top of the other at the same or different distances / lengths. In another embodiment, one or more nutrients having different solvent requirements are deposited on opposite sides at the same or different distances / lengths. In another embodiment, one or more nutrients having the same solvent requirements are deposited on opposite sides at the same or different distances / lengths. In another embodiment, the floss is solid, worn, comprises multiple strands, is treated to be adhesive, is treated to adhere to a pharmacologically acceptable carrier, or is treated to adhere to an active agent. In another embodiment, each adjacent drop or patch comprises a dye or indicia that distinguishes between adjacent deposits. In another embodiment, the floss is not soaked in one or more nutrients, a pharmacologically acceptable carrier, or both.In another embodiment, the one or more nutrients are selected from iron, phosphorus, zinc, thiamine, riboflavin, niacin, vitamin B6, folic acid, vitamin B12, pantothenic acid, biotin, boron, choline, chromium, copper, manganese, selenium, molybdenum, iodine, chloride, coenzyme Q10 (CoQ10), vitamin A, calcium, potassium, magnesium, vitamin E, vitamin C, carotenoids, vitamin D, or vitamin K. In another embodiment, the floss has a thickness of less than 5 mm, preferably less than 3 mm, preferably less than 1 mm, or preferably 35-175 microns. In another embodiment, the floss comprises a natural or synthetic polymer, an organic material, a metal, an inorganic material, or a combination thereof. In another embodiment, the floss comprises a mucoadhesive layer, or a hydrophobic or hydrophilic layer, or a combination thereof. In another embodiment, the floss comprises a microporous structure that allows diffusion of antigens into the gingival sulcus. In another embodiment, the one or more nutrients are delivered to the junctional epithelium in the picogram to milligram range. In another embodiment, the one or more nutrients provide 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the daily nutritional needs by flossing one, two, or three times per day. In another embodiment, delivery of the one or more nutrients to the endodontic epithelium, junctional epithelium, or a combination thereof is 0 hours, 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or more before the subject eats food, drinks liquid, or brushes their teeth. In another aspect, delivery of the one or more nutrients to the endothelium, junctional epithelium, or a combination thereof is 0 hours, 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or more after the subject eats food, drinks liquid, or brushes their teeth.In another embodiment, the viscosity of the deposit is 0.01 centipoise (cp), 1 cp, 10 cp, 100 cp, 1000 cp, 10,000 cp, 100,000 cp, 200,000 cp, 300,000 cp, 500,000 cp, 1,000,000, or 100,000,000 cp.

[0063] It is contemplated that any embodiment discussed herein can be implemented with respect to any method, kit, reagent, or composition of the invention, and vice versa. Furthermore, the compositions of the invention can be used to practice the methods of the invention.

[0064] It will be understood that the specific embodiments described herein are shown by way of illustration and not as limitations of the invention. The principal features of this invention can be employed in various embodiments without departing from the scope of the invention. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, numerous equivalents to the specific procedures described herein. Such equivalents are considered to be within the scope of this invention and are covered by the claims.

[0065] All publications and patent applications mentioned in this specification are indicative of the level of skill of those skilled in the art to which this invention pertains. All publications and patent applications are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

[0066] The use of the words "a" or "an" when used in connection with the word "comprising" in the claims and / or specification can mean "one," but is also consistent with the meanings of "one or more," "at least one," and "one or more." While this disclosure supports definitions that refer to alternatives only and "and / or," the term "or" in the claims will be used to mean "and / or" unless clearly indicated to refer to alternatives only or unless the alternatives are mutually exclusive. Throughout this application, the term "about" is used to indicate that a value includes the inherent variation of error for the device, method being employed to determine the value, or the variation that exists among study subjects.

[0067] As used in this specification and the claims, the words "comprising" (and all forms of compris- ing, e.g., "comprise" or "comprises"), "having" (and all forms of having, e.g., "have" or "has"), "including" (and all forms of including, e.g., "includes" or "include"), or "containing" (and all forms of containing, e.g., "contains" or "contain") are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. In any of the embodiments of the compositions and methods provided herein, "comprising" may be substituted with "consisting essentially of" or "consisting of." As used herein, the phrase "consisting essentially of" requires the specified integers or steps and those that do not materially affect the characteristics or function of the claimed invention. As used herein, the term "consisting of" is used to indicate the presence of only the stated integer (e.g., feature, element, characteristic, property, method / process step or limitation) or group of integers (e.g., feature, element, characteristic, property, method / process step or limitation).

[0068] As used herein, the term "or combinations thereof" refers to all permutations and combinations of the listed items preceding the term. For example, "A, B, C, or combinations thereof" is intended to include at least one of A, B, C, AB, AC, BC, or ABC, and further includes, where order is important in the particular context, BA, CA, CB, CBA, BCA, ACB, BAC, or CAB. Continuing with this example, combinations involving repeats of one or more items or terms are expressly included, such as BB, AAA, AB, BBC, AAABCCCC, CBBAAA, CABABB, etc. Those of skill in the art will understand that there is typically no limit to the number of items or terms in any combination, unless a different interpretation is apparent from the context.

[0069] As used herein, approximation words such as, without limitation, "about," "substantial," or "substantially" refer to conditions that, when so modified, are understood not to be necessarily absolute or perfect, but that would be considered close enough to one of ordinary skill in the art to justify specifying the condition as existing. The extent to which the description can vary depends on how large a change can be made, while still allowing one of ordinary skill in the art to recognize that the modified feature still possesses the required characteristics and capabilities of the unmodified feature. Generally, but in accordance with the immediately preceding discussion, numerical values ​​herein modified by approximation words such as "about" can vary from the stated value by at least ±1, 2, 3, 4, 5, 6, 7, 10, 12, or 15%.

[0070] Additionally, the section headings herein are provided for consistency with implications under 37 CFR 1.77 or to otherwise provide organizational markers. These headings should not limit or characterize the inventions described in any claims that may issue from this disclosure. Specifically and by way of example, although the heading refers to the "Technical Field," such claims should not be limited by language under this heading that describes the so-called technical field. Furthermore, the description of technology in the "Background Art" section should not be construed as an admission that that technology is prior art to any invention in this disclosure. The "Summary of the Invention" should not be considered a characterization of the inventions described in the issued claims. Furthermore, any reference to the singular "invention" in this disclosure should not be used to argue that there is only one novelty in this disclosure. Multiple inventions may be set forth according to the limitations of the multiple claims issuing from this disclosure, and such claims will accordingly define and protect the invention(s), and their equivalents. In all cases, the scope of such claims shall be measured on their own merits in light of this disclosure, but should not be constrained by the headings set forth herein.

[0071] All of the compositions and / or methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations may be applied to the compositions and / or methods described herein and in the steps or sequence of steps of the methods without departing from the concept, spirit, and scope of the invention. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope, and concept of the invention as defined by the appended claims.

[0072] In order to assist the United States Patent Office, and any reader of any patent issued on this application, in interpreting the claims attached hereto, Applicants wish to point out that, unless the words "means for" or "step for" are expressly used in a particular claim, Applicants do not intend to subject any of the appended claims to the application of 35 U.S.C. § 112, paragraph 6, 35 U.S.C. § 112(f), or the doctrine of equivalents, as they exist on the filing date of this application.

[0073] For each claim, each dependent claim may depend on any independent claim and each previous dependent claim, so long as the previous claim is a proper antecedent basis for a term or element in the claim.

[0074] References 1 Saurabh, S., et al., Is Transdermal Multivitamin Patch Effective in Gastric Bypass Patients? Obes Surg, 2019. 29(12): p. 3818-3823. 2 McCormick, R., et al., The Effectiveness of Transdermal Iron Patches in Athletes With Suboptimal Iron Status (Part 1). Int J Sport Nutr Exerc Metab, 2020. 30(3): p. 185-190. 3 Garant, PR, Oral cells and tissues. 2003, Chicago: Quintessence Pub. Co. ix, 430 p. 4 Moutsopoulos, N.M. and J.E. Konkel, Tissue-Specific Immunity at the Oral Mucosal Barrier. Trends Immunol, 2018. 39(4): p. 276-287. 5 Pollanen, M.T., J.I. Salonen, and V.-J. Uitto, Structure and function of the tooth-epithelial interface in health and disease. Periodontology 2000, 2003. 31(1): p. 12-31. 6 Schroeder, H.E. and M.A. Listgarten, The Junctional Epithelium: From Strength to Defense. Journal of Dental Research, 2003. 82(3): p. 158-161. 7 Nakamura, M., Histological and immunological characteristics of the junctional epithelium. Jpn Dent Sci Rev, 2018. 54(2): p. 59-65. 8 Luke, D.A., The structure and functions of the dentogingival junction and periodontal ligament. British Dental Journal, 1992. 172(5): p. 187-190. 9 Bosshardt, D.D. and N.P. Lang, The Junctional Epithelium: from Health to Disease. Journal of Dental Research, 2005. 84(1): p. 9-20. 10 Stern, I.B., Current Concepts of the Dentogingival Junction: The Epithelial and Connective Tissue Attachments to the Tooth. Journal of Periodontology, 1981. 52(9): p. 465-476.

Claims

1. 1. A method for delivering one or more nutrients to a subject, comprising: The method comprising the step of delivering an effective amount of one or more nutrients into the gingival sulcus, said amount being sufficient to supplement the subject's diet with respect to said nutrients.

2. 10. The method of claim 1, wherein the one or more nutrients are not targeted for delivery to the vestibular mucosa.

3. The method of claim 1 , wherein the one or more nutrients target the sulcular epithelium, the lining epithelium, the junctional epithelium, or a combination thereof.

4. 10. The method of claim 1, wherein the one or more nutrients are encapsulated to maximize delivery of the one or more nutrients.

5. 10. The method of claim 1, further comprising adding one or more agents that increase the permeability of one or more nutrients to the epithelium of the gingival crevicular region.

6. 2. The method of claim 1, wherein 0.001% to 100% of the one or more nutrients are present in a depot in the junctional epithelium (JE) of the gingival sulcus.

7. The method of claim 1, wherein one or more nutrients are repeatedly provided to the junctional epithelium of the gingival sulcus.

8. 10. The method of claim 1, wherein the delivery of one or more nutrients to the junctional epithelium occurs after ingesting food or drink, or brushing teeth.

9. 10. The method of claim 1, wherein the one or more nutrients are applied one or more times on a daily or weekly or monthly basis, for example 1, 2, 3, 4, 5, or 6 times daily, or 1, 2, 3, 4, 5, 6, or 7 times weekly, or 1, 2, 3, or 4 times monthly.

10. 10. The method of claim 1, wherein the one or more nutrients provide 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the daily nutritional need by flossing one, two, or three times per day.

11. 10. The method of claim 1, wherein the delivery of one or more nutrients to the marginal epithelium, junctional epithelium, or a combination thereof is 0 hours, 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or more before the subject eats food, drinks liquid, or brushes their teeth.

12. 10. The method of claim 1, wherein the delivery of the one or more nutrients to the endothelium, junctional epithelium, or a combination thereof is 0 hours, 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or more after the subject eats food, drinks liquid, or brushes their teeth.

13. 10. The method of claim 1, wherein the amount of the one or more nutrients delivered to the marginal epithelium, the junctional epithelium, or a combination thereof is in the picogram to milligram range.

14. 2. The method of claim 1, wherein the one or more nutrients are selected from iron, phosphorus, zinc, thiamine, riboflavin, niacin, vitamin B6, folic acid, vitamin B12, pantothenic acid, biotin, boron, choline, chromium, copper, manganese, selenium, molybdenum, iodine, chloride, coenzyme Q10 (CoQ10), vitamin A, calcium, potassium, magnesium, vitamin E, vitamin C, carotenoids, vitamin D, or vitamin K.

15. A nutrient delivery system comprising an effective amount of one or more nutrients in a delivery device that targets the junctional epithelium of the gingival sulcus, wherein the amount of the one or more nutrients is a nutritionally effective amount.

16. 16. The nutrient delivery system of claim 15, wherein one or more nutrients are not delivered to the vestibular mucosa.

17. 16. The nutrient delivery system of claim 15, wherein the delivery device comprises a cross-sectional shape that maximizes delivery of one or more nutrients to the gingival sulcus.

18. 16. The nutrient delivery system of claim 15, further comprising one or more agents that increase the permeability of one or more nutrients to the gingival sulcus epithelium, lining epithelium, junctional epithelium, or a combination thereof.

19. 16. The nutrient delivery system of claim 15, wherein 0.001% to 100% of the one or more nutrients are present in a depot in the gingival sulcus lining epithelium, the junctional epithelium, or a combination thereof.

20. 16. The nutrient delivery system of claim 15, further comprising one or more pharmaceutically acceptable carriers, excipients, diluents, buffers, or salts.

21. 16. The nutrient delivery system of claim 15, wherein the one or more nutrients are selected from iron, phosphorus, zinc, thiamine, riboflavin, niacin, vitamin B6, folic acid, vitamin B12, pantothenic acid, biotin, boron, choline, chromium, copper, manganese, selenium, molybdenum, iodine, chloride, coenzyme Q10 (CoQ10), vitamin A, calcium, potassium, magnesium, vitamin E, vitamin C, carotenoids, vitamin D, or vitamin K.

22. 1. A method of making a floss containing a predetermined amount of one or more nutrients, comprising: providing a floss; and depositing on said floss one or more deposits of said one or more nutrients in a pharmacologically acceptable carrier. wherein each deposit has a known, predetermined amount of said one or more nutrients.

23. 23. The method of claim 22, wherein each adjacent deposit comprises the same one or more nutrients or a different one or more nutrients, or each adjacent deposit comprises the same one or more nutrients at different concentrations; or each adjacent deposit comprises a different one or more nutrients at different concentrations; or each adjacent deposit is located on a different plane than the adjacent deposit; or each adjacent deposit is located on an opposite side of the floss deposit; or each adjacent deposit comprises one or more nutrients that are different from the previous adjacent deposit.

24. 24. The method of claim 23, wherein each adjacent deposit comprises one or more different nutrients having different solvent requirements selected from one or more nutrients having solubility in aqueous solvents and one or more other nutrients having solubility in organic solvents.

25. 24. The method of claim 23, wherein two or more nutrients are deposited in the form of deposits stacked one on top of the other at the same or different distances / lengths.

26. 26. The method of claim 25, wherein one or more nutrients having different solvent requirements are deposited on opposite sides over the same or different distances / lengths.

27. 26. The method of claim 25, wherein one or more nutrients having the same solvent requirement are deposited stacked one on top of the other at the same or different distances / lengths.

28. 24. The method of claim 23, wherein one or more nutrients having different solvent requirements are deposited on opposite sides at the same or different distances / lengths.

29. The method of claim 22, wherein the floss is solid, attrited, comprises multiple strands, is treated to be adhesive, is treated to adhere to a pharmacologically acceptable carrier, or is treated to adhere to an active agent.

30. 23. The method of claim 22, wherein each adjacent deposit includes a dye or indicia that identifies adjacent drops or patches.

31. 23. The method of claim 22, wherein the floss is not soaked in one or more nutrients, a pharmacologically acceptable carrier, or both.

32. 23. The method of claim 22, wherein the one or more nutrients are selected from iron, phosphorus, zinc, thiamine, riboflavin, niacin, vitamin B6, folic acid, vitamin B12, pantothenic acid, biotin, boron, choline, chromium, copper, manganese, selenium, molybdenum, iodine, chloride, coenzyme Q10 (CoQ10), vitamin A, calcium, potassium, magnesium, vitamin E, vitamin C, carotenoids, vitamin D, or vitamin K.

33. 23. The method of claim 22, wherein the floss has a thickness of less than 5 mm, preferably less than 3 mm, preferably less than 1 mm, or preferably between 35 and 175 microns.

34. 23. The method of claim 22, wherein the floss comprises a natural or synthetic polymer, an organic material, a metal, an inorganic material, or a combination thereof.

35. 23. The method of claim 22, wherein the floss comprises a mucoadhesive layer, or a hydrophobic or hydrophilic layer, or a combination.

36. 23. The method of claim 22, wherein the floss comprises a microporous structure that allows diffusion of one or more nutrients into the gingival sulcus.

37. 23. The method of claim 22, wherein the floss delivers one or more nutrients to the marginal epithelium, junctional epithelium, or a combination thereof 0 hours, 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or more before the subject eats food, drinks liquid, or brushes their teeth.

38. 23. The method of claim 22, wherein the floss delivers one or more nutrients to the marginal epithelium, junctional epithelium, or a combination thereof at 0 hours, 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or more after the subject eats food, drinks liquid, or brushes their teeth.

39. 23. The method of claim 22, wherein the viscosity of the adherend is 0.01 centipoise (cp), 1 cp, 10 cp, 100 cp, 1000 cp, 10,000 cp, 100,000 cp, 200,000 cp, 300,000 cp, 500,000 cp, 1,000,000, or 100,000,000 cp.

40. A floss containing a predetermined amount of one or more nutrients, floss; and The floss is coated with one or more depots of said one or more nutrients in a pharmacologically acceptable carrier. wherein each droplet or patch has a known, predetermined amount of active agent.

41. 41. The floss of claim 40, wherein each adjacent deposit comprises the same one or more nutrients or a different one or more nutrients; or each adjacent deposit comprises the same one or more nutrients at different concentrations; or each adjacent deposit comprises a different one or more nutrients at different concentrations; or each adjacent deposit is located on a different plane than the adjacent deposit; or each adjacent deposit is located on the opposite side of the floss from the adjacent deposit; or each adjacent deposit comprises one or more nutrients that are different from the previous adjacent deposit.

42. 42. The floss of claim 41, wherein each adjacent deposit comprises one or more nutrients having different solvent requirements selected from one or more nutrients soluble in aqueous solvents and one or more nutrients soluble in organic solvents.

43. 42. The floss of claim 41, wherein one or more nutrients are deposited in the form of drops or patches piled on top of one another at the same or different distances / lengths.

44. 43. The floss of claim 42, wherein one or more nutrients having different solvent requirements are deposited on opposite sides over the same or different distances / lengths.

45. 45. The floss of claim 44, wherein one or more nutrients having the same solvent requirements are deposited on top of each other at the same or different distances / lengths.

46. 42. The floss of claim 41, wherein one or more nutrients having different solvent requirements are deposited on opposite sides at the same or different distances / lengths.

47. The floss of claim 41, wherein the floss is solid, attrited, comprises multiple strands, is treated to be adhesive, is treated to adhere to a pharmacologically acceptable carrier, or is treated to adhere to an active agent.

48. 42. The floss of claim 41, wherein each adjacent drop or patch contains a dye or indicia that distinguishes between adjacent deposits.

49. 42. The floss of claim 41, wherein the floss is not soaked in one or more nutrients, a pharmacologically acceptable carrier, or both.

50. 42. The floss of claim 41, wherein the one or more nutrients are selected from iron, phosphorus, zinc, thiamine, riboflavin, niacin, vitamin B6, folic acid, vitamin B12, pantothenic acid, biotin, boron, choline, chromium, copper, manganese, selenium, molybdenum, iodine, chloride, coenzyme Q10 (CoQ10), vitamin A, calcium, potassium, magnesium, vitamin E, vitamin C, carotenoids, vitamin D, or vitamin K.

51. 42. The floss of claim 41, wherein the floss has a thickness of less than 5 mm, preferably less than 3 mm, preferably less than 1 mm, or preferably between 35 and 175 microns.

52. 42. The floss of claim 41, wherein the floss comprises a natural or synthetic polymer, an organic material, a metal, an inorganic material, or a combination thereof.

53. 42. The floss of claim 41, wherein the floss comprises a mucoadhesive layer, or a hydrophobic or hydrophilic layer, or a combination.

54. 42. The floss of claim 41, wherein the floss comprises a microporous structure that allows diffusion of antigens into the gingival sulcus.

55. 42. The floss of claim 41, wherein the one or more nutrients are delivered to the junctional epithelium in the picogram to milligram range.

56. 42. The floss of claim 41, wherein the one or more nutrients provide 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the daily nutritional needs by flossing one, two, or three times per day.

57. 42. The floss of claim 41, wherein the one or more nutrients are delivered to the endodontic epithelium, junctional epithelium, or a combination thereof 0 hours, 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or more before the subject eats food, drinks liquid, or brushes their teeth.

58. 42. The floss of claim 41, wherein the delivery of one or more nutrients to the endodontic epithelium, junctional epithelium, or a combination thereof is 0 hours, 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or more after the subject eats food, drinks liquid, or brushes their teeth.

59. 42. The floss of claim 41, wherein the viscosity of the deposit is 0.01 centipoise (cp), 1 cp, 10 cp, 100 cp, 1000 cp, 10,000 cp, 100,000 cp, 200,000 cp, 300,000 cp, 500,000 cp, 1,000,000, or 100,000,000 cp.

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