Oral hygiene composition comprising black cumin seed oil as active ingredient for prevention or alleviation of halitosis and tooth sensitivity

Abstract

The present invention relates to an oral hygiene composition comprising black cumin seed oil as an active ingredient for the prevention or alleviation of halitosis or tooth sensitivity. The oral hygiene composition according to the present invention comprises as an active ingredient black cumin seed oil containing thymoquinone, p-cymene, and carvacrol, thereby stably controlling bacteria in the oral cavity, inhibiting the production of organic acids in the oral cavity to prevent demineralization, and inhibiting the production of tartar or plaque, with the consequent prevention or alleviation of halitosis or tooth sensitivity.

Description

Oral hygiene composition for preventing or improving bad breath and tooth sensitivity, containing black cumin seed oil as an active ingredient

[0001] The present invention relates to an oral hygiene composition for preventing or improving bad breath and sensitive teeth symptoms, comprising black cumin seed oil as an active ingredient. Specifically, the present invention relates to an oral hygiene composition for preventing bad breath and sensitive teeth symptoms, comprising black cumin seed oil as an active ingredient, which suppresses the production of organic acids in the oral cavity, thereby suppressing oral acidification, inhibiting the production of dental plaque, and inhibiting the production of dental calculus / plaque.

[0002] Dental caries begins when the tooth surface is demineralized by organic acids (lactic acid being a prime example) produced by bacteria present in dental plaque through carbohydrate metabolism. S. mutans is known as a causative agent of dental caries. However, recent metagenomic studies have reported that S. mutans is not detected in dental caries lesions. However, this does not mean that the association between S. mutans and dental caries is completely negligible.

[0003] Dental plaque is a sticky, soft deposit that adheres to teeth and surrounding tissues. Tartar is formed when dental plaque is deposited with saliva and inorganic substances such as calcium and phosphorus within the gingival sulcus. Its composition is 75% inorganic matter, 6-15% organic matter, and the remainder water. Based on the formation of dental plaque, the plaque matures and undergoes mineralization (calcium, phosphorus, magnesium, and fluoride) before crystals are formed. The deposition mechanism of dental plaque is as follows.

[0004] In the first stage, a sterile, acquired film is formed, and in the second stage, microorganisms attach to this film, mature, and microorganisms, gingival exfoliating cells, leukocytes, and food debris attach to the film and mature. After 23 hours, aerobic cocci and bacilli account for 80-90%, and anaerobic bacteria gradually increase compared to aerobic cocci. Finally, inflammation occurs in the gingival tissue. In addition, dental plaque is formed through a process in which calcification begins 4-8 hours after the formation of dental plaque, 50% is calcified after 24 hours, and 60-90% is calcified after 12 days.

[0005] Under normal circumstances, the pH of the oral cavity is maintained at around 7.0–7.5 by saliva. However, when we eat, bacteria in the mouth convert the sugars in the food into acid, lowering the pH of the mouth below 7.0 and becoming acidic. This is because bacteria in plaque (mainly S. mutans) present on the surface of teeth break down sugars in the mouth, producing organic acids as a byproduct. These organic acids dissolve tooth enamel, forming cavities and lowering the surface acidity of the tooth to below pH 5.5, activating the activity of cavity-causing bacteria. When the acid produced by cariogenic bacteria attacks the tooth and damages the surface, caries occurs. Because the higher the oral acidity, the more active the bacteria become, maintaining an appropriate pH of the teeth is crucial for preventing caries. A pH of 5.2–5.6 is known to be the critical pH for tooth demineralization.

[0006] Therefore, controlling the production of organic acids and dental plaque and tartar, which cause dental caries and gingivitis, the initial stage of periodontal disease, at an early stage is an important factor in maintaining and improving oral health.

[0007] Antibiotics and mouthwashes are used to suppress pathogenic bacteria in the oral cavity. However, antibiotics can cause systemic side effects, the emergence of resistant bacteria in the oral cavity, and superinfection, making long-term use unacceptable. Furthermore, antibacterial agents such as Listerine, peroxide, and chlorhexidine are commonly used in mouthwashes. However, long-term use is not recommended due to concerns about tissue toxicity, the emergence of resistant bacteria, and carcinogenicity.

[0008] Tartar, which forms due to poor oral hygiene, cannot be removed through brushing alone. It can only be removed through scaling. Failure to remove tartar can easily lead to gingivitis, which can inflame not only the gums but also the ligaments and alveolar bone. However, improper scaling can cause gum damage, such as swelling of the gums or sensitivity to cold or hot foods. Therefore, there is a need to develop a composition that can be used safely and consistently for oral hygiene management and oral health promotion without adverse side effects on the teeth or gums.

[0009] Meanwhile, there are attempts to develop compositions for promoting oral health or maintaining oral hygiene using natural materials.

[0010] Korean Patent Publication No. 2023-0013235 discloses a lecithin enzyme reactant and an oral health material and product containing the same as an active ingredient.

[0011] Korean Publication No. 10-2023-0018335 proposes the combined use of a composition comprising an extract of cinnamon tree and superoxide dismutase and / or Bacillus strain spores for improving oral health.

[0012] Korean Patent Publication No. 2021-0121163 discloses a composition comprising thymoquinone and an additional biologically active compound, which composition is effective in treating or preventing an inflammatory condition.

[0013] Accordingly, the inventors of the present invention confirmed that black cumin seed oil, a natural antibacterial material with high antibacterial activity against Streptococcus mutans, a dominant species in the early stages of dental calculus and dental plaque formation, but is safe and has low toxicity, has an effect of inhibiting the formation of organic acids, dental calculus, and dental plaque, thereby completing the present invention.

[0014] The present invention provides an oral hygiene composition for preventing or improving bad breath or tooth sensitivity, comprising black cumin seed oil containing thymoquinone, p-cymene, and carvacrol. In particular, the present invention provides an oral hygiene composition that has the effect of reducing dental plaque and suppressing oral acidification, and inhibiting the formation of organic acids, dental plaque, and dental plaque.

[0015] In addition, the present invention aims to provide a health functional food, chewable soft capsule, toothpaste, oral cleanser, dietary film, oral spray, oral ointment, oral varnish, oral rinse, and gum massage cream containing the composition, which have an effect of preventing or improving bad breath or tooth sensitivity.

[0016] The above-mentioned task is achieved by an oral hygiene composition for preventing or improving bad breath or tooth sensitivity, which contains black cumin seed oil as an active ingredient.

[0017] Preferably, the black cumin seed oil is characterized by containing thymoquinone, p-cymene and carvacrol.

[0018] Also preferably, the oral hygiene composition has an effect of inhibiting the formation of dental calculus or dental plaque and an effect of inhibiting the formation of organic acids in the oral cavity.

[0019] Also preferably, the black cumin seed oil may be included in an amount of 0.001 to 10 wt% relative to the total weight of the oral hygiene composition.

[0020] Also preferably, the oral hygiene composition may have a selected formulation comprising a chewable soft capsule, toothpaste, mouthwash, dietary film, mouth spray, oral ointment, oral varnish, mouthwash, and gum massage cream.

[0021] The oral hygiene composition containing black cumin seed oil according to the present invention contains linoleic acid, thymoquinone, p-cymene, and carvacrol, which have high antibacterial activity against harmful microorganisms in the oral cavity, and thus can reduce dental caries, periodontal disease, and bad breath that affect oral health.

[0022] Figure 1 shows the results of an experiment confirming the inhibition of dental plaque acidification by black cumin seed oil according to the present invention.

[0023] All technical terms used in this invention, unless otherwise defined, have the following definitions and are consistent with the meanings commonly understood by those skilled in the art in the relevant fields of the present invention. Furthermore, while preferred methods and samples are described herein, similar or equivalent methods are also included within the scope of the present invention.

[0024] The term “about” means an amount, level, value, number, frequency percentage, dimension, size, quantity, weight, or length that varies by 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1% of the reference amount, level, value, number, frequency percentage, dimension, size, quantity, weight, or length.

[0025] Throughout this specification, unless the context otherwise requires, the words “comprise” and “comprising” are to be understood to imply the inclusion of a stated step or component, or group of steps or components, but not the exclusion of any other step or component, or group of steps or components.

[0026]

[0027] The present invention relates to an oral hygiene composition comprising black cumin seed oil containing thymoquinone, p-cymene, and carvacrol as an active ingredient. Preferably, the oral hygiene composition according to the present invention has an effect of preventing or improving bad breath or tooth sensitivity.

[0028] Nigella sativa (Nigella sativa) is a small flowering plant native to Southwest Asia, the Middle East, and Southern Europe. The small, black seeds of this shrub, Nigella sativa, are known by many names, including black cumin, black caraway, nigella, fennel, and Roman coriander. Black cumin seed oil, extracted from Nigella sativa seeds, has been used in traditional medicine for over 2,000 years due to its numerous therapeutic benefits.

[0029] In traditional medicine, black cumin seed oil has been used to treat a variety of health conditions, earning it the nickname "panacea" or "universal healer." While not all medicinal uses have been proven effective, studies have shown that black cumin seed oil, due to its high antioxidant content, may reduce inflammation and protect against conditions like heart disease and Alzheimer's disease. Further research is needed on its potential for asthma treatment. Studies have also shown benefits, including weight loss, reduced blood sugar levels, lower blood pressure, and lower cholesterol. However, not all black cumin seed oils are created equal when it comes to oral use.

[0030] The present inventors confirmed that black cumin seed oil contains tinoquinone, p-cymene, and carvacrol as effective ingredients. Black cumin seed oil has antibacterial effects against the oral pathogenic bacteria S. mutans, inhibiting dental plaque cell adhesion and dental plaque acidification, while commercially available Nigella sativa oil (from Turkey, manufactured by cold-pressing method) did not show effective results with regard to oral health when the same experiment was conducted.

[0031] The present invention provides an oral hygiene composition comprising black cumin seed oil containing thymoquinone, p-cymene, and carvacrol as an active ingredient, which has the effects of inhibiting dental plaque bacterial adhesion, inhibiting dental plaque acidification, and inhibiting demineralization, thereby having an excellent effect of preventing or improving bad breath or sensitive teeth symptoms. The black cumin seed oil can be prepared by a method such as oil extraction or solvent extraction, and is preferably prepared by oil extraction.

[0032] The black cumin seed oil according to the present invention contains C18:2 linoleic acid, thymoquinone, p-cymene, and carvacrol as active ingredients. Preferably, the black cumin seed oil according to the present invention contains 51 to 66% (v / v) of linoleic acid, 1 to 5% (v / v) of thymoquinone, 0.1 to 1.5% (v / v) of p-cymene, and 0.01 to 0.1% (v / v) of carvacrol.

[0033]

[0034] Hereinafter, the present invention will be described in detail with reference to examples and experimental examples, but the scope of the present invention is not limited by these examples.

[0035]

[0036] Manufacturing Example 1: Manufacturing of black cumin seed oil

[0037] Black cumin seeds (1,000 kg) were dried in a fluidized bed at room temperature and 0.05 bar for 20-40 minutes, yielding approximately 960 kg. After removing foreign substances, approximately 950 kg of seeds less than 4 mm in size were obtained, and oil was obtained by pressing at 20-40°C with a thymoquinone (TQ) content of 1.5-3.5% and a free fatty acid (FFA) content of less than 1.5%. 280 kg of oil was obtained by filtration at room temperature and 3 bar using a 10 μm filter. Sunflower lecithin and vitamin E were mixed with the above oil so that the thymoquinone (TQ) content was 1.5 to 3.5% (v / v), and the mixture was stirred at 400 to 500 RPM for 0.5 to 1 hour at room temperature to obtain black cumin seed oil having a weight ratio of free fatty acid (FFA) to thymoquinone (TQ) of approximately 2.33 to 0.57: 1.

[0038]

[0039] The experiment below was conducted through the 2023 Open Equipment Utilization Test Analysis Support Project of the Korea Food Industry Cluster Promotion Agency, and experiments on dental plaque cell adhesion, antibacterial activity, and dental plaque acidification were conducted according to the biomarker measurement method of the Ministry of Food and Drug Safety's Health Functional Food Functionality Evaluation Guide "Can Help Dental Health."

[0040]

[0041] Experimental Example 1: Strain Cultivation and Inoculation

[0042] Streptococcus mutans (S. mutans), a dental caries-causing bacteria used in the experiment on the adhesion of dental plaque to black cumin seed oil, was obtained as Streptococcus mutans KCTC 3065 strain from the Korea Center for Biological Resources (KCTC). Cryopreserved S. mutans (KCTC3065) was inoculated into Brain Heart Infusion liquid medium (hereinafter referred to as BHI liquid medium) and cultured under aerobic conditions at 130 rpm, 37°C, for 24 hours. Then, 1 x 10 5 Adjust the bacterial solution to the CFU / mL concentration.

[0043]

[0044] Experimental Example 2: Dental plaque bacterial cell adhesion (plaque formation) experiment

[0045] Add 10 mL of BHI liquid medium containing 1% (v / v) sucrose to a 50 mL glass tube, and add 50 μL of each concentration of black cumin seed oil (HT_01: 10% (v / v), 4% (v / v), 2% (v / v), prepared by dissolving in DMSO) prepared in Manufacturing Example 1. As a positive control, use 10% (v / v) of xylitol (a raw material for dental health notified by the Ministry of Food and Drug Safety), 10% (v / v) of Listerine, and as a negative control, use 100% (v / v) of DMSO, and 10% (v / v) of commercially available Turkish cold-pressed black cumin seed oil (NSO).

[0046] S. mutans concentration 1 x 10 5 After growing to CFU / mL, 100 μL was inoculated into each tube, and the glass tubes were tilted 30° and cultured in a 37°C incubator for 24 hours. After culture, 5 mL of 0.5 M NaOH solution was added to suspend the cells attached to the wall. 3 mL of the suspension was taken, and the absorbance was measured at 550 nm using a spectrophotometer (S-3100, SCINCO). The results are shown in Table 1.

[0047] Classification DMSO Turkish NSO 10% Xylitol 10% * Listerine 10% HT_012% HT_014% HT_0110% **** Absorbance (1st) 0.808 1.138 0.93 10.88 7 0.93 6 0.89 8 0.349 Absorbance (2nd) 0.73 5 1.02 2 0.88 5 0.75 10.88 4 0.81 6 0.392 Absorbance (3rd) 0.80 7 1.03 0 0.91 0 0.73 4 0.88 6 0.80 2 0.445 Average (00, 550 nm)0.7831.0630.9090.7910.9020.8390.395Standard deviation0.0340.0530.0190.0680.0240.0420.039

[0048] As shown in Table 1, it was confirmed that the absorbance decreased in a concentration-dependent manner in the black cumin seed oil (HT_01) manufactured in Manufacturing Example 1, and the turbidity of the culture medium was lowered (HT_01 10%, ****p<0.0001). The turbidity of xylitol, which was used as a positive control, increased significantly (*p<0.05), and the turbidity of Turkish NSO (cold-pressed extraction, negative control) also increased significantly (****p<0.0001).

[0049] Experimental Example 3-1: Antibacterial activity measurement experiment

[0050] 1 x 10 pre-cultured cells were cultured on Brain Heart Infusion solid medium (hereinafter, BHI solid medium) containing 1.5% (v / v) agar. 5 CFU / mLS. mutans were mixed in a volume ratio of 1% on 0.8% agar and layered. The test group (Preparation Example 1) and the control group were dissolved in DMSO and prepared at each concentration. 50 μL was absorbed into a filter disc, placed on a layered medium, and aerobically cultured at 37°C for 48 hours. The diameter (mm) of the Clear Zone around the disc was compared. The results are shown in Table 2.

[0051] - Test group (Preparation Example 1): 100% (v / v), 10% (v / v), 4% (v / v), 2% (v / v), 1% (v / v), 0.9% (v / v), 0.8%, 0.7% 0.6%

[0052] - Control group: Listerine 100% (v / v), xylitol (dental health notification raw material) 100% (v / v), Turkish cold-pressed NSO 100% (v / v), DMSO 100% (v / v)

[0053] Classification DMSO Turkish NSO 10% Xylitol 10% Listerine 10% HT_010.7% HT_012% HT_014% HT_0110% HT_01100% Clear Zone 1---131212244680 Clear Zone 2---18--325490 Clear Zone 3---16--266090 Mean (00)---15.67 121227.3353.3386.67 Standard deviation---2.05003.405.744.71

[0054] As shown in Table 2, the Clear Zone (mm) increased in a concentration-dependent manner in the black cumin seed oil (HT_01) manufactured in Manufacturing Example 1, confirming an increase in antibacterial activity. It was confirmed that Turkish NSO and xylitol had no antibacterial activity against S. mutans, and Listerine Gumcare 100% and black cumin seed oil (HT_01) 0.7%, 2%, 4%, 10%, and 100% had antibacterial activity against S. mutans. Compared to the control group Listerine Gumcare, black cumin seed oil (HT_01) 10% and 100% showed effectiveness (p<0.0001).

[0055] Experimental Example 3-2: Antibacterial activity measurement experiment

[0056] 1 x 10 of pre-cultured S. mutans (KCTC3065) 9 Adjusted to a concentration of CFU / mL, 200 μL was mixed with 3 mL top agar, and poured onto Brain Heart Infusion solid medium. The test group (Manufacturing Example 1) and the control group were prepared by dissolving them in DMSO, and 50 μL was absorbed into a filter disc, placed on a double-layer medium, and aerobically cultured at 37°C for 48 hours. The diameter (cm) of the Clear Zone around the disc was compared. The results are shown in Table 3. The control group used the Gumgard® product with a fluoride content of 1,000 ppm as is.

[0057] - Test group (Manufacturing Example 1): 5% (v / v)

[0058] - Control group: Geomgard® 100% (v / v) (original solution)

[0059] Category HT_01 5%Gumguard® Clear zone (cm) 3±0.071.25±0.05

[0060] Looking at Table 3, it can be confirmed that the antibacterial activity of black cumin seed oil (HT_01) manufactured in Manufacturing Example 1 against S. mutans is more than twice that of gum guard.

[0061] Experimental Example 4: Experiment to confirm inhibition of dental plaque acidification

[0062] Add 50 μL of each concentration of black cumin seed oil (HT_01: 10% (v / v), 4% (v / v), 2% (v / v), prepared by dissolving in DMSO) prepared in Manufacturing Example 1 to 10 mL of BHI liquid medium. The control group used 10% xylitol, 10% (v / v) Listerine, 100% (v / v) DMSO, and 10% (v / v) NSO. 1 x 10 5 Inoculate 100 μL of CFU / mL S. mutans. Incubate in a 37°C incubator for 48 hours, and then measure the pH of the medium. The results are shown in Table 4 and Figure 1.

[0063] Blank DMSO, Turkish NSO 10%, Xylitol 10%, Listerine 10%, HT_012%, HT_014%, HT_0110%, pH 7.345.255.155.25.165.185.187.25

[0064] Looking at Table 4 and Fig. 1, it can be confirmed that the pH increased in a concentration-dependent manner in the black cumin seed oil (HT_01) manufactured in Manufacturing Example 1, and the culture medium was neutralized. DMSO, which did not show antibacterial activity against S. mutans, showed an acidity of 5.25, Turkish NSO 5.15, and xylitol 5.2, while Listerine Gumcare, which showed antibacterial activity, showed an acidity of 5.16, and the 2% and 4% black cumin seed oil (HT_01) test groups showed an acidity of 5.18, and the 10% black cumin seed oil (HT_01) test group showed an acidity of 7.25. Compared to the control group, xylitol showed a decrease in acidity at the level of p<0.05, Turkish NSO showed a decrease in acidity at the level of p<0.0001, and the test substance NSO 10% showed a decrease in acidity at the level of p<0.0001, resulting in inhibition of acidification. The correlation between antibacterial activity and the degree of dental plaque adhesion and acidification inhibition is shown in Table 5 below.

[0065] Classification Blank DMSO Turkish NSO 10% Xylitol 10% Listerine 10% HT_012% HT_014% HT_0110% Antibacterial power-XXXOOOO Average absorbance 0.0000.7831.0630.9090.7910.9020.8390.395 pH 7.345.255.155.25.165.185.187.25

[0066] As shown in Table 5, the results of the experiment showed that the degree of bacterial attachment to dental plaque and the degree of oral acidification (organic acid production, inhibition of demineralization) in the negative control group, positive control group, and samples treated with black cumin seed oil (HT_01) manufactured in Manufacturing Example 1 were correlated, but the correlation with the antibacterial activity against S. mutans was low.

Claims

1. An oral hygiene composition containing black cumin seed oil as an active ingredient for preventing or improving bad breath or tooth sensitivity.

2. An oral hygiene composition according to claim 1, characterized in that the black cumin seed oil contains thymoquinone, p-cymene, and carvacrol.

3. An oral hygiene composition according to claim 1, characterized in that the oral hygiene composition has an effect of inhibiting the formation of dental calculus or dental plaque and an effect of inhibiting the formation of organic acids in the oral cavity.

4. An oral hygiene composition according to claim 1, characterized in that the black cumin seed oil is contained in an amount of 0.001 to 10 wt% relative to the total weight of the oral hygiene composition.

5. In the first paragraph, the oral hygiene composition is characterized in that it has a selected formulation consisting of a chewable soft capsule, toothpaste, mouthwash, dietary film, oral spray, oral ointment, oral varnish, oral rinse, and gum massage cream.

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