WO2019031459A1 - Promoter of removal of dental and oral deposits - Google Patents

Abstract

Provided is nanobubble water for removing a smear layer, and for removing a dental plaque or biofilm and for removing a tongue plaque as a treatment immediately before a teeth filling/restoring treatment or a prosthesis, or as the washing of a root canal for a pulpectomy or a treatment of an infected root canal, or as a pretreatment for the application of a medical patch, or as the enlargement of an obliterated root canal, or a pretreatment for the filling of a root canal. An oral deposit removal promoter for removing a smear layer while keeping the strength of dentin. The nanobubble water is nanobubbles to be injected into a root canal. The nanobubbles have a zeta-potential of -10 mV or less.

Description

Removal promoter of teeth and intraoral adhesions

The present invention relates to endodontic and external teeth, periodontal tissue, tongue used for dental caries treatment, dental pulp and infection root canal treatment, periodontal disease treatment, hypersensitivity treatment, repair and prosthetic treatment, peri-implant inflammation treatment etc. The present invention relates to cleaning enhancers and kits for palates, restorations and restorations in the oral cavity, dentures and the like.

1. Removal of smear layer Generally, when a tooth is cut, the cut pieces clog the dentinal tubules of dentin, and a smear layer is formed on the dentin surface. The presence of the smear layer reduces the adhesion of the resin, cement and the like, and further, bacteria may be present in the smear layer itself. The smear layer adheres to the dentin surface and can not be removed even by strong washing with a three-way syringe or foaming washing with a 3% hydrogen peroxide solution or the like. Therefore, the smear layer is treated with phosphoric acid, a primer (such as citric acid), and the like before the repair treatment or the attachment of the prosthesis. In addition, a smear layer is also formed at the time of enlargement of the root canal in pulpectomy, infected root canal treatment, or root canal treatment. This smear layer also reduces the adhesion of the root canal filler (material), which prevents close root canal filling and causes microleakage. Furthermore, in particular in infected root canal teeth, it is necessary to completely remove bacteria deeply invading from the root canal wall dentinal tubule. At this time, generally, after mechanically enlarging the root canal, the smear layer clogged in the root canal wall dentin tubule is removed and cleaned with the EDTA preparation, and an antibiotic, calcium hydroxide or the like is applied in the root canal. Do. However, although the EDTA preparation is effective for removing the smear layer, the demineralization of the root canal wall dentin may reduce mechanical strength and may cause fracture. The washing of other citric acid-containing formulations (MTAD) suffers from the same mechanical strength problem and also has the disadvantage that the adhesive strength of the adhesive at the time of repair is weakened.

Patent Document 1 describes a method for removing a smear layer using sorbitan ester as a detergent and citric acid as an organic acid. However, this method has a problem that demineralization of dentine by citric acid occurs.
2. Removal of plaque and biofilm Recently, periodontal disease has been found to be closely related to diabetes. In addition, when periodontitis-causing bacteria such as P gingivalis enter blood vessels, thrombi are easily formed in the heart, aorta, veins, etc., and the risk of heart disease and cerebral infarction increases. Oral care is not only maintenance of dental health by prevention of dental caries and periodontal disease, but also various infections spread over the whole body including prevention of aspiration pneumonia, dementia, myocardial infarction, cerebral infarction etc by oral bacteria and toxins It is considered important for the prevention of On the other hand, oral cleaning instruction mainly focuses on physical plaque removal with a toothbrush. However, in practice it is difficult for elderly people and carers to physically brush for a long time. With age, the periodontal pocket deepens, the gap between the teeth becomes large, and tooth decay occurs in the cervix, resulting in accumulation of dental plaque / biofilm that can not be removed by ordinary brushing or gargle. Bacteria and toxins are excreted to the whole body from the collection of bacteria, which can cause chronic inflammation in various tissues. As it is difficult for the elderly to receive frequent professional care, it is considered essential to develop new simple methods that promote voluntary self-care and do not deposit plaque or biofilm.

Patent Document 2 describes a method for removing dental plaque and biofilm in the oral cavity comprising an α-olefin sulfonate and alanine and / or lysine. However, in this method, the removal effect of dental plaque and biofilm is not sufficient because the temporal stability of α-olefin sulfonate is insufficient.
3. Drug removal When performing root canal treatment, it is possible to eliminate bacteria with a high concentration of 10 mg / mL antibiotics, etc., but at this concentration, it can not be washed by conventional methods, and the drug remaining on dentin sidewall has cytotoxicity is there. For example, when blood clots or Platelet-rich Plasma (PRP) are injected into the root canal to revascularize the unrooted tooth, the remaining drug on the dentin side wall may affect cell adhesion, proliferation, and differentiation. It will be. For example, Non-Patent Document 1 describes that cephem antibiotics may increase the risk of hypocarnitinemia.
4. Tongue removal There are hundreds of billions to one trillion bacteria in the oral cavity of adults. When the secretion of saliva is reduced due to aging, the self-cleaning action of saliva is no longer effective and the bacteria are more likely to be established. Among the bacteria, Candida, Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, Haemophilus influenzae, and the like cause bacteria of systemic diseases, and further increase when the immunity declines with age. In particular, the “tongue” where the keratin on the surface of the tongue has spread and becomes hard and bacteria and dirt have accumulated in the gaps is inadequate mouth cleaning, mouth breathing, reduction of saliva, tongue movement function decline, systemic diseases (diabetes, It is caused by Sjögren's syndrome, autonomic imbalance, duodenal ulcer, etc. Therefore, "volatile sulfur compounds" which are easily produced in the elderly and are produced cause halitosis and affect bright and healthy social life. It can also cause taste abnormalities and aspiration pneumonia. However, it is generally said that tongue care is difficult and it is not easy to clean. Tongue care should be brief, painless and uncomfortable, easy and easy to continue. It is a common practice to apply a moisturizing gel to the tongue and scrape it with a tongue brush or sponge brush, but when the brush is put in the back of the tongue, vomiting reflex often occurs. In addition, it is auxiliary in mouthwash and it is difficult to remove physically attached bacteria and dirt by itself.

Protease is known as an enzyme having the resolution of tongue-lid, and foods such as candies and tablets containing an enzyme such as cysteine protease have been reported (Patent Document 3). However, cysteine proteases are heat labile and have poor stability over time in hot and humid conditions such as summer.
5. Enlargement of the constricted root canal Aging or external stimulation to the dental pulp (stimulation of bacteria such as dental caries, mechanical stimulation at the time of tooth cutting, pharmaceutic chemical stimulation at the treatment of pulp capping, physical stimulation by trauma occlusion, etc.) are long-term In the latter case, dentin is added to the dentin side wall of the root canal of the tooth as a defense reaction of the living body, or calcification occurs in the pulp in the root canal. As a result, the entrance of the root canal is tightly closed, and the presence and position of the root canal can not be known, or the device for treatment (reamer, file) is tightly closed halfway through the root canal to the tip (apex) of the root canal It may not reach. If there is an apical lesion under the root and bacterial infection extends under the root, it is necessary to allow the instrument to reach the apex and to enlarge and clean the root canal (Non-patent Document 2). In particular, micro root canals such as side branches and accessory root canals, which can not be reached by instruments, exist intricately on the apical side 1/3. If the main root canal can not be opened to the apical end, the micro root canal and apical periodontal tissue may not be able to penetrate the drug applied to the root canal for eradication. In addition, in the case of a curved root canal, if it is attempted to expand forcibly, it may be drilled in a direction other than the original root canal. In addition, when the root canal can not be expanded to the apex, surgical treatment or tooth extraction is performed to remove the apical lesion from the bone under the root as a surgical operation.

Japanese Patent Publication No. 2005-516032 Unexamined-Japanese-Patent No. 2017-100964 International Publication No. 2007/026755

Naoko Miura, Basic research on the effect of warmth method on extravasation of cephem antibiotics, 24th Annual Meeting of the Japanese Society of Nursing Science Page 256 (2004.12) Clinical decision making in root canal treatment, Norihiro Sawada, Journal of the Japanese Prosthodontic Society Ann Jpn Prosthodont Soc 6: 362-367, 2014

The present invention has been made in view of the above problems, and the present invention relates to a filling and restoration treatment of a tooth, a treatment immediately before fitting to a prosthesis, a root canal lavage for pulpectomy and infected root canal treatment, a patch preparation, an enlarged root canal with stenosis It is an object of the present invention to provide a cleaning agent with nanobubble water or gel for smear layer removal, plaque / biofilm removal, tongue coating removal as tube filling pretreatment.

The cleaning agent for cleaning endodontic / external, periodontal tissue, tongue, restoration / prosthesis and implant according to the present invention is characterized by having nano bubbles. The surface is negatively charged, causing a crushing phenomenon, and is characterized by being a nano-sized (for example, a diameter of 10 nm to 1000 nm) water bubble of 10 ⁶ to 10 ⁹ cells / mL. Interfacial tension generates pressure between the interfaces of the gas phase and the liquid phase, and the liquid phase and the liquid phase, and when the bubble diameter becomes smaller, the internal pressure due to surface tension increases. By the self-crushing action, water, nitrogen and the like are decomposed to generate free radicals. The growth of bacteria is suppressed by the action of free radicals generated at the time of crushing. Further, the electric potential of the "slip surface" in the electric double layer formed around the particles in the solution, where liquid flow starts to occur, is referred to as the zeta potential, and is considered to be related to the fluidity, cohesion, storage, etc. of the particles.

According to the present invention, nanobubbles alone can effectively remove intra- and extra-lateral smear layers, remove plaques / biofilms, and remove tongue plaques without using drugs. When EDTA or citric acid and nanobubbles are used in combination, the demineralization effect as a root canal enlargement aid can be enhanced. For example, when benzalkonium chloride and nanobubbles are used in combination, plaque removal and biofilm removal effects as a gargle can be enhanced.

It is a photograph which shows the scanning electron microscope image of the smear layer removal in the root canal in the recommendation time, A is 5 minutes with distilled water, B is nano bubble water 5 minutes, C is 3% EDTA (Smear clean) 2 minutes, The image was washed for 5 minutes with D: 17% EDTA (17% EDTA liquid): 1 minute, E: 20% citric acid (Ultradent): 3 minutes, and F: 4.25% citric acid (MTAD): 5 minutes. It is the figure which analyzed the lumen area of the dentinal tubule after smear layer removal in a root canal. ^** P <0.01, ^* P <0.05, versus distilled water. ^## P <0.01, versus nano bubble water. It is a figure which shows the Vickers hardness in 100 micrometers from the root canal wall after smear layer removal in the root canal at recommendation time. ^* P <0.05, versus distilled water. ^# P <0.05, versus nano bubble water. It is a figure which shows the change of the smear layer removal effect by nano bubble density | concentration. A is a scanning electron microscope image, a is a concentration of 1.0 × 10 ⁸ cells / mL, b is 0.5 × 10 ⁸ cells / mL, c is 0.1 × 10 ⁸ cells / mL, d is 0.05 × 10 ⁸ cells / mL, e is 0.01 × 10 ⁸ cells / mL, f 2 is 0.005 × 10 ⁸ cells / mL, and g is distilled water. B is the figure which analyzed the lumen area per one of a dentinal tubule statistically. ^* P <0.05, versus distilled water. ^## P <0.01, versus 0.5 x 10 ⁸ cells / mL. It is a figure which shows the change of the smear layer removal effect by the difference in the zeta-potential of nano bubble water (7 * 10 < ⁷ > piece / mL). A is a scanning electron microscope image, a is zeta potential -21.7mV, b is -15.4mV, c is -11.2mV, d is -9.4mV, e is -8.6mV, f distilled water, g is untreated is there. B is the figure which analyzed the area per 1 of a dentinal tubule, and C statistically analyzed the density (piece / mm < ² >) of a dentinal tubule. D is a scanning electron microscope image examining the smear layer removing effect by pH change, a is nano bubble water pH 6.65, zeta potential-21.3 mV, b is nano bubble water pH 4.16, -16.0 mV, c is nano bubble water pH is 8.97, -8.6 mV, d is distilled water of pH 6.09, e is distilled water of pH 4.07, f is distilled water of pH 8.98, and g is untreated. E is a statistical analysis of the smear layer removing effect by the change of pH. It is a figure which shows the change of the smear layer removal effect by gas of nano bubble water. A is a photograph of a scanning electron microscope, a is air, b is carbon dioxide, c is nitrogen, d is oxygen, e is distilled water, and f is untreated. B is a statistical analysis diagram. It is the figure which compared the smear layer removal by nano bubble water or nano bubble gel. A shows a scanning electron microscope image, a is distilled water, b is nano bubble water, c is nano bubble gel (Alcox E-240), d is gel (Alcox E-240), e is untreated is there. B is a diagram statistically analyzing the area per dentinal tubule. ^** P <0.01, versus distilled water. ^## P <0.01, versus nano bubble gel. It is a figure which shows the scanning electron microscope image which removed the plaque formed on the hydroxyapatite disc for 30 minutes by culture | cultivating Streptococcus Mutans (ATCC 25175) for 48 hours under aerobic conditions for 30 minutes, A is distilled water, B is nano bubble water. , C is a nano bubble gel, D is untreated. FIG. 17A is a photographic image showing a scanning electron microscopic image of biofilms formed by culturing Enterococcus faecalis (ATCC 19433) for 14 days under anaerobic conditions and removing biofilms formed on hydroxyapatite disks with nanobubble water or nanobubble gel for 30 minutes. Is distilled water, B is nano bubble water, and C is nano bubble gel. It is a figure which shows the scanning electron microscope image which culture-removing Streptococcus Mutans (ATCC 25175) for 48 hours on aerobic conditions, and having removed the biofilm formed on the hydroxyapatite disc for 2 minutes, A is distilled water, B is nano bubble water , C: benzalkonium chloride, D: benzalkonium chloride + nanobubble water (50%), E: monoammonium glycyrrhizinate, F: monoammonium glycyrrhizinate + nanobubble water (50%), G: neostegreen (0.2% Benzethonium chloride), H is a concour (chlorhexidine gluconate, ammonium glycyrrhizinate, green tea extract). It is a fluorescence stereomicrograph which performed washing once for 5 minutes to medicine (tetracycline) which penetrated from the root canal wall (RC) deep inside pig tooth root dentinal tubule, A is physiological saline, B is only nano bubble, C: 1.5% EDTA, D: 1.5% EDTA + nanobubbles (50%), E: 8.5% EDTA, F: 8.5% EDTA + nanobubbles (50%). For the drug (tetracycline) that penetrated through the root canal wall (RC) deep in the pig tooth root dentinal tubule, apply a removal accelerator for 1 minute to the root canal, and then suction the removal accelerator to make a new removal accelerator again Is a fluorescent stereomicrograph that has been repeatedly removed several times, A is physiological saline, B is nanobubble water 1 minute application, C is nanobubble water 1 minute application, D is nanobubble water 1 minute Applied 3 times. Photographs showing removal of tongue coating, A is before surgery, B is gargle with 0.2% benzethonium chloride-containing mouthrinse (Neoste green) for 1 minute, C is after gargle with nanobubble water for 1 minute. It is a figure which shows Vickers hardness of 100, 300, 500 micrometers from the root canal wall 2 minutes after root canal enlargement adjuvant application. Distilled water is used as a control, using nanobubble water, 8.5% EDTA + nanobubbles (50%), 8.5% EDTA, 17% EDTA as a root canal enlargement aid. ^* P <0.05, ^** P <0.01, versus distilled water. ^# P <0.05, ^## P <0.01, versus nano bubble water. ^{† P <0.05, †† P <} 0.01, versus 17% EDTA. ^‡ P <0.05, versus 8.5% EDTA. It is a photograph by scanning electron microscope which shows the change of the plaque removal effect by the viscosity of nano bubble gel, a is distilled water, b is nano bubble water, c is nano bubble gel (1) (Alcox E-240 1.06 W%) viscosity 117 mPa · s, d is untreated, e is nano bubble gel PEG 20 k 5% (polyethylene glycol PEG 20 k 5%) 6 mPa · s, f is nano bubble gel A (Alcox E-240) 52 mPa · s, g is nano bubble gel B (cellogen BSH-12) 199 mPa · s, h: nano bubble gel HPC-M 2.5% (hydroxypropyl cellulose HPC-M 2.5%) 447 mPa · s, i: PEG 20 K, j: gel A, k: gel B, l: HPC-M It is. It is a figure which shows the change of the smear layer removal effect by the difference in the internal pressure of a nano bubble. A is a photograph by a scanning electron microscope image, a is air pressure 0.07 water pressure 0.18 MPa, b is air pressure 0.09 water pressure 0.24 MPa, c is air pressure 0.13 water pressure 0.20 MPa, d is air pressure 0.17 water pressure 0.28 MPa, e is The nano bubble water produced by 0.34 MPa of air pressure 0.34 water pressure, and f wash | cleaned with distilled water for 5 minutes. B is a diagram statistically analyzing the area per dentinal tubule. ^* P <0.05, ^** P <0.01, versus distilled water. ^# P <0.05, ^## P <0.01, versus air pressure 0.17 water pressure 0.28 Mpa. ^{† P <0.05, †† P <} 0.01, versus 0.13 water pressure 0.20MPa. C is the figure which analyzed the particle size distribution of each nano bubble water. It is a figure which shows measurement of the remaining Porphyromonas gingivalis number by PrestoBlue. It is a figure which shows the observation result of the remaining Porphyromonas gingivalis number by a scanning electron microscope, Among them, A is distilled water, B is nano bubble water, C is 0.025% benzalkonium chloride, D is 0.025% benzalkonium chloride (nano bubble dilution) is there. It is a figure which shows the observation result of the remaining Porphyromonas gingivalis by Live Dead dyeing | staining. It is a figure which shows the statistical analysis result of the effect of the nano bubble (gel) in a canine periodontal disease model.

The invention according to this embodiment is a removal promoter for intraoral adhesions. An intra-oral deposit is, for example, a smear layer, plaque, biofilm or tongue.

In the intraoral adhesion removal promoter for removing the smear layer, it is necessary not only to remove the smear layer but also to maintain the dentin strength. A part of the smear layer penetrates into the dentinal tubule, and the smear layer which penetrates into the dentinal tubule may be called a smear plug or a smear plug.

In the removal promoter of intra-oral deposit for removing plaque, biofilm or tongue, a nano bubble gel which is a gel containing nano bubbles is applied. The viscosity of the nano bubble gel is not particularly limited, but is, for example, 450 mPa · s or less. If the viscosity of the nano bubble gel is too high, the flowability is lowered, and the convenience may be lowered.

The nanobubbles according to the present embodiment have a liquid or gel-like form containing ultrafine bubbles. A gas (air, carbon dioxide, nitrogen, oxygen, ozone, etc.) is introduced into the nano-sized bubble diameter, a high internal pressure is given by surface tension, and it is negatively charged. That is, they are nanobubbles having high internal pressure and charge. Due to the effective removal promoting action based on the surface characteristics possessed by such nanobubbles and the movement characteristics such as Brownian motion, there is a feature that the removal can be effectively promoted without destroying air bubbles using an ultrasonic device etc. doing. In the present invention, two or more gases can be used as the gas contained in the nanobubbles. When two or more gases are used, for example, a mixture of nanobubbles consisting only of gas A and nanobubbles consisting only of gas B may be used, or nanobubbles containing a mixture of gas A and gas B may be used There is also.

When the intraoral deposit removal promoter according to the present invention is a liquid preparation, it is preferable that the solution constituting this is an aqueous solution. In the present invention, an aqueous solution containing a gas in the nanobubble state is referred to as nanobubble water.

In nano bubble water, the pH of the aqueous solution is not particularly limited, but may be, for example, weakly acidic of 5.00 to 6.00. The zeta potential of the nanobubbles is negative when the pH is 5.00 or more, and its absolute value does not change significantly over time. Further, in the nano bubble water, the hardness is not particularly limited, but for example, the hardness can be 20 to 30.

The size of the nano bubble is generally in the range of 10 to 1,000 nm, preferably in the range of 10 to 500 nm, preferably 10 to 300 nm, more preferably 10 to 200 nm, and still more preferably Is in the range of 50 to 200 nm, more preferably 50 to 150 nm. By setting the diameter (bubble diameter) of the nanobubbles in the above range, an effective removal promoting action can be obtained. In addition, if the size of the nano bubble becomes too large, the removal promoting action will be reduced. The smaller the size of the nanobubbles, the better the stability of long-term storage. The size of the nanobubbles can be adjusted to a desired size using a reverse osmosis membrane, air pressure or the like.

The nano bubble has a nano-sized bubble diameter, and its surface tension causes the internal pressure to be high, and is negatively charged. Here, the internal pressure of the nanobubbles is generally determined by the Young-Laplace equation corresponding to the diameter of the nanobubbles, and the nanobubbles used in the present invention have an internal pressure of about 3 atm to about 300 atm. In addition, the zeta potential of the nanobubbles in water is -30 to 0 mV, and the nanobubbles are considered to be negatively charged. The zeta potential of the nanobubbles is generally in the range of -30 to 0 mV, preferably in the range of -30 to -10 mV, preferably -30 to -10 mV, and more preferably -30. It is in the range of -10 mV. By setting the zeta potential of the nanobubbles in the above range, an effective removal promoting action can be obtained. In addition, when the zeta potential of the nanobubbles becomes too large, the removal promoting action is reduced. It is presumed that the removal can be promoted effectively based on the charging characteristics.

The concentration of nanobubbles is generally indicated as the number of bubbles in a specified volume, and in the present invention, preferably 1 × 10 ⁶ to 2 × 10 ⁹ cells / mL, preferably 5 × 10 ⁶ to 1 × 10 ⁹ / m L, more preferably distributed in the ratio of ^{1 × 10 7 ~ 5 × 10} 8 cells / mL, and contains. Here, if the amount of such nanobubbles is too small, it is not possible to advantageously exhibit an effective removal promoting effect by negatively charged nanobubbles having a high internal pressure. In addition, if the concentration of nanobubbles is too high, its removal promoting action tends to saturate, resulting in poor benefits from an economic point of view.

The size of the nano bubble as described above and the concentration thereof can be measured using a commercially available nanoparticle measuring device. For example, a nanoparticle distribution measuring apparatus (SALD-7100) manufactured by Shimadzu Corporation, a nanoparticle analyzing apparatus (Nanosite LM-20) which can be obtained from Nippon Quantum Design Co., Ltd., and the like can be mentioned. . In addition, the zeta potential can be obtained from Maruballoon Division Spectris Co., Ltd. Zeta Sizer Nano Z, Otsuka Electronics Co., Ltd. Zeta Potential Measurement System (ELSZ-2000Z), Kyowa Interface Science Co., Ltd. Zeta Potential Measurement Apparatus (ZC-3000) etc. can be mentioned.

According to the present invention, the removal enhancer containing nanobubbles can be used as a mixture by directly using the target nanobubbles, or directly introducing it into a predetermined drug (the drug itself or a pre-composition containing the drug). It is. Particularly advantageously, it is formed in the form of a mixture obtained by mixing a liquid or gel solid of water containing nanobubbles with a predetermined drug or liquid or gel-like pre-composition containing it Thus, the target nanobubbles can be easily and advantageously introduced into the drug composition. That is, it is easy to prepare various target drug compositions by preparing in advance a liquid or gel containing the target nanobubble, and uniformly mixing the predetermined drug or the preliminary composition thereto. You can get it. Alternatively, various liquids of the target pharmaceutical composition may be prepared by preparing a liquid containing the target nanobubble in advance, dissolving the gel therein, and uniformly mixing the liquid with a predetermined drug or the preliminary composition thereof. You can get things easily. In addition, the removal promoter of the intraoral adhesion thing containing the nano bubble concerning this embodiment is not limited to what is used in combination with a chemical | medical agent, For example, a cell extract, a cell culture supernatant, a microbial fermentation product, a plant extraction It can be used in combination with various compositions such as products and purified proteins.

The drug is not particularly limited, and is, for example, an antibiotic such as benzalkonium chloride or a tetracycline antibiotic.

In addition, a root canal enlargement aid for enlarging a narrowing root canal of middle-aged and elderly people according to the present invention includes nano-bubble water or nano-bubble gel and a root canal enlargement cleaning agent. The root canal enlargement cleaning agent is not particularly limited, and examples thereof include EDTA, citric acid, chlorhexidine, MTAD and the like. Preferably the root canal cleaning agent is EDAT.

By the way, in the present invention, nano bubbles having a nano-sized bubble diameter can be formed using various known nano bubble generating devices. In particular, a device in which nano bubbles are formed by releasing a predetermined gas through a breathable film formed by generating crazes on a polymer resin film and thereby controlling the amount of gas permeation, for example, a patent An apparatus as disclosed in Japanese Patent No. 3806008 and Japanese Patent No. 5390212 can be advantageously used. Among them, in the present invention, a gas-permeable film formed by generating crazes on a polymer resin film is disposed on a gas-permeable surface provided on a cylindrical outer peripheral surface, and A cylindrical gas permeable portion configured to release a predetermined pressurized gas under control; and an air supplying means for supplying the gas in a pressurized state into a cylinder of the cylindrical gas permeable portion; A cylindrical casing having an inner peripheral diameter larger than the outer peripheral diameter of the cylindrical gas permeable portion, and a gap formed by accommodating and arranging the cylindrical gas permeable portion in the cylindrical casing, An ultra-fine bubble generation device configured to include a flow means for flowing a predetermined fluid through the fluid flow path given by the above is preferably used. Then, there, the air bubbles formed by the gas released from the gas permeable surface of the gas permeable part by the fluid flowing through the fluid flow path are sheared and refined in the initial stage of their formation. An ultra-fine bubble (nano bubble / ultra fine bubble) having a nano-sized bubble diameter, to which a high internal pressure and a negative charge are imparted, can be effectively formed.

Although the manufacturing method of nano bubble water or nano bubble gel is not specifically limited, For example, it is the following. That is, at least a tubular gas permeation apparatus in which a breathable film formed by generating crazes on a polymer resin film capable of limiting the gas permeation amount is disposed in a gas permeation part, and a tank storing water or gel-like fluid And a pump for delivering water or gel-like fluid contained in the storage tank to the cylindrical circulation path, and the cylindrical gas permeation device is installed in the cylindrical circulation path. A fluid pressure is adjusted using a pump to introduce water or a gel-like fluid into a gap formed by the difference between the outer peripheral diameter of the cylindrical gas permeable portion and the inner peripheral diameter of the cylindrical circulation passage. By controlling the pressurized state and supplying the gas to the gas permeable portion of the gas permeable device, fine bubbles of nano size are mixed in the water or gel-like fluid.

Example 1
(Pine root canal dentin smear layer removal)
The root canal of pig fresh removed premolars is enlarged to # 60 with K file (Manny), and after alternate washing with 2 mL of 5% sodium hypochlorite and 2 mL of 3% hydrogen peroxide water, 5 mL physiological salt It was further washed with water and dried. Nanobubble water 5 minutes, 3% EDTA preparation (pH 9.5, smear clean, Nippon Dental Pharmaceutical) 2 minutes, 17% EDTA preparation (pH 7.3, 17% EDTA liquid, benton Japan) 1 minute, 20% citric acid preparation (PH 1.4, Ultradent citric acid 20%, Ultradent Japan) 3 minutes, 4.25% citric acid preparation (containing 3% doxycycline and 0.5% Tween 80, pH 2.15, BioPure MTAD, Dentsu ply Sirona) 5 minutes, and distilled The smear layer was washed for 5 minutes with water and 2 mL of 6 smear layer removing agents for a recommended time. The nanobubble water was nanobubble water manufactured using air with a dental nanobubble generator (FOAMEST 8 (registered trademark), Nac Corp.). After washing with physiological saline, it is divided in half with toothpicks, fixed with 2% glutaraldehyde for 12 hours, dehydrated with 30, 50, 70, 90, 100% ethanol and evaporated with platinum 10 kV ( Conductive film deposition (sputter coating) MSP-20-UM, vacuum device). Thereafter, each specimen was observed with a scanning electron microscope (VE9800, KEYENCE) from the apex of the root canal at 3 mm, 4.5 mm, and 6.0 mm.

The smear layer could not be removed at all by distilled water (FIG. 1A), but the smear layer could be completely removed by washing only with nanobubble water for 5 minutes (FIG. 1B). On the other hand, the smear layer remained considerably in the commonly used 3% EDTA 2 minutes (FIG. 1C), 17% EDTA 1 minute (FIG. 1D) and 4.25% citric acid (FIG. 1F). In addition, in 20% citric acid 3 minutes (Fig. 1E), the smear layer could be almost removed, but the effect was slightly inferior to that of nanobubble water.

A total of three images were taken at a magnification of 1,000 times, 3, 4.5 and 6.0 mm from the apex, and the lumen area of the dentinal tubule was measured with Image J and statistically analyzed. Nanobubble water, 17% EDTA, and 20% citric acid significantly showed a smear layer removing effect as compared to distilled water (P <0.01, P <0.01, P <0.05). Nano bubble water was found to have a smear layer removing effect significantly compared to other smear layer removing agents (P <0.01) (FIG. 2).
(Demineralization of dentin wall after removal of porcine root canal dentin smear layer)
The root canal of pig fresh removed premolars was enlarged to # 60 in the same manner as described above, washed with sodium hypochlorite only, washed with 5 ml of physiological saline, and stored wet. The smear layer removing agent similar to the above was applied to the root canal for a recommended time, washed, and further washed with physiological saline. The thickness was adjusted to 3 mm with a Zege microtome (Leica), polished to # 2000 with sandpaper, and stored in saline until measurement. The Vickers hardness at a point of 100 μm from the root canal wall of the prepared sample was measured with a micro Vickers hardness tester (Akashi Mfg. MVK-E) at a weight of 50 g for 15 seconds.

The nanobubbles alone did not show any change in Vickers hardness compared to distilled water. On the other hand, Vickers hardness at 100 μm from the root canal wall was significantly reduced with 17% EDTA 1 min, 20% citric acid 3 min, 4.25% citric acid 5 min compared to distilled water and nano bubbles (P <0.05) (FIG. 3). Although 3% EDTA did not change the mechanical strength, the smear layer removing effect was also weak.

Example 2
(Change of smear layer removal effect by concentration of nano bubble water)
Root canals of porcine fresh removed premolars were treated and dried as described above. Nanobubbles water nanobubble generating device (FOAMEST (registered trademark), Nac Corp.) after production using air, diluted with distilled water, concentration of 1 × 10 ⁸ cells /ML,0.5×10 ⁸ pieces /mL,0.1 × 10 ⁸ cells /mL,0.05×10 prepare ^eight /ML,0.01×10 ⁸ /ML,0.005×10 ⁸ / mL, 5 minutes into the root canal applications, after washing, the smear layer removal effect Compared. In addition, the particle size peak of this nano bubble was 109 nm, zeta potential-21.7 mV, and pH 6.38. A scanning electron microscope specimen is prepared by a common method, and four spots of 1.5, 3, 4.5 and 6.0 mm from the apex at 1,000 × magnification are taken, and the lumen area of the dentinal tubule is measured with Image J. And analyzed statistically.

At the concentration of 0.5 × 10 ⁸ cells / mL and 1.0 × 10 ⁸ cells / mL, the smear layer could be almost removed, but at 0.1 × 10 ⁸ cells / mL, considerable smear layer remaining was observed, and 0.05 × 10 ⁸ cells / mL. The removal effect was hardly seen below / mL (FIG. 4A). Doing statistical analysis, the concentration of 0.5 × 10 ⁸ cells / mL, 0.1 × 10 ⁸ pieces /ML,0.05×10 ⁸ pieces /ML,0.01×10 ⁸ pieces /ML,0.005×10 ⁸ cells / mL and Compared with distilled water, it was revealed that the area per dentinal tubule was significantly higher and the smear layer removing effect was higher. However, no significant difference was found between 0.5 × 10 ⁸ cells / mL and 1.0 × 10 ⁸ cells / mL (FIG. 4B).

(Change of smear layer removal effect by zeta potential of nano bubble water)
Root canals of porcine fresh removed premolars were treated and dried as described above. The concentration of nano bubble water (particle size about 100 nm) manufactured using air was adjusted to the same concentration (0.7 × 10 ⁸ cells / mL), and the zeta potential -21.7 mV, -15.4 mV, -11.2 mV, -9.4 mV and- The change in smear layer removal effect due to the difference of 8.6 mV was compared for 5 minutes after application and washing in the root canal. Distilled water application and no treatment were used as controls. A scanning electron microscope specimen is prepared by a common method, and a total of four photographs of 1.5, 3, 4.5 and 6.0 mm from the apex at 1,000 × magnification are taken, and the lumen area and density of the dentinal tubule are measured with Image J. (The number of capillaries per square millimeter) was measured and analyzed statistically.

The smear layer was removed at zeta potentials of -21.7 mV, -15.4 mV and -11.2 mV, but at -9.4 mV and -8.6 mV, the smear layer was not partially removed (Fig. 5A).

Statistical analysis shows that the zeta potential is -21.7mV, -15.4mV and -11.2mV, which is significantly higher than that of no treatment (P <0.05, P <0.01, P <0.05). It became clear that the smear layer removal effect was high. Also, at -21.7mV, -15.4mV and -11.2mV, the area per dentin tubule was significantly higher than -9.4mV and -8.6mV (P <0.05, P <0.01, P <0.05) (P <0.05) Figure 5B). In addition, statistical analysis of the density of dentinal tubules revealed that the zeta potential -15.4 mV was significantly higher than the other potentials (Fig. 5C).

(Change of smear layer removal effect by pH of nano bubble water)
Root canals of porcine fresh removed premolars were treated and dried as described above. The pH was adjusted to 4, 6, 9 for nanobubble water (particle size 88 nm) produced using air. In addition, distilled water of pH corresponding to it was prepared. That is, nanobubble water pH 4.16, -16.0mV, nanobubble water pH 6.65, zeta potential-21.3mV, nanobubble water pH 8.97-8.58mV, distilled water pH 4.07, pH 6.09 and pH 8.98 were produced. The smear layer removing effect due to the change in pH was applied for 5 minutes in the root canal and compared after washing. Untreated was used as a control. A scanning electron microscope specimen is prepared by a common method, and four spots of 1.5, 3, 4.5 and 6.0 mm from the apex at 1,000 × magnification are taken, and the lumen area of the dentinal tubule is measured with Image J. And analyzed statistically.

Nanobubble water pH 4.16 zeta potential-16.0mV, pH 6.65 zeta potential-21.3mV the smear layer was almost removed, but nanobubble water pH 8.97-8.6mV There is a portion where the smear layer is not removed only a part did. In distilled water, no smearing effect was observed at pH 4 and pH 6 (FIG. 5E). Therefore, it was revealed that the smear layer removing effect was not at the pH but at the zeta potential.

(Change of smear layer removal effect by gas during production of nano bubble water)
The gas at the time of nano bubble water production was changed from air to carbon dioxide, nitrogen and oxygen. As a result, it had the following properties. The smear layer removal effects of nanobubble water of each gas were compared.

As a result, in the scanning electron microscope image, the nanobubble water with carbon dioxide was somewhat inhomogeneous in smear layer removal, but in the other cases, the smear layer could be removed almost similarly (FIG. 6A). Statistical analysis of the area of the dentinal tubule per piece showed no significant difference depending on the type of gas (FIG. 6B).

(Change of smear layer removal effect by nano bubble gel)
In the nano bubble gel, powdery alcox E-240, serogen BSH-12, hydroxypropyl cellulose HPC-M 2.5%, or polyethylene glycol PEG 20 k 5% was completely uniformly mixed in nano bubble water.

The Alcox E-240 nanobubble gel was able to remove the smear layer more effectively than the nanobubble water (FIG. 7A). The smear layer could not be removed with distilled water or gel alone.

Statistical analysis revealed that the nanobubble gel had a significantly higher dentinal tubule area per piece (P <0.05) than the nanobubble water, and the smear layer removal effect was high (FIG. 7B).

[Example 3]
(Plate and biofilm removal of Streptococcus mutans by nano bubbles)
After adding 1 mL of BHI Brot (Kanto Chemical) + 5% sucrose to each well of a 48-well cell culture plate, 50 μl of Streptococcus mutans (ATCC 25175) culture solution was added to the same well. The hydroxyapatite (HA) disc (HA 48-3, Funakosi) was immersed and cultured for 48 hours to form plaque and a biofilm. In order to examine the cleaning effect of nano bubbles, Otsuka distilled water, nano bubble water, nano bubble gel (Alcox E-240) is allowed to act on this HA disc for 5 minutes, and the untreated ones are scanned according to the conventional method. Microscopic specimens were prepared and compared.

As shown in FIG. 8, plaque remained in distilled water even after 5 minutes. In both nanobubble water and nanobubble gel, most of the plaque decreased after 2 minutes, but nanobubble gel had a higher removal effect than nanobubble water.

(Plate and biofilm removal of Enterococcus faecalis by nano bubbles)
After adding 1 mL of BHI Broth + 5% sucrose to each well of a 48-well cell culture plate, 50 μl of Enterococcus faecalis (ATCC 19433) culture solution was added to the same well. The hydroxyapatite (HA) disc (HA 48-3, Funakosi) was immersed and cultured for 14 days to form plaque and biofilm. In order to examine the cleaning effect of nano bubbles, Otsuka distilled water, nano bubble water and nano bubble gel (Alcox E-240) were allowed to act on this HA disc for 30 minutes and compared with untreated ones by scanning electron microscope images did.

As shown in FIG. 9, the biofilm remained in the distilled water after 30 minutes, but in the nanobubble water, the considerable biofilm disappeared but slightly remained after 30 minutes, and almost all the nanobubble gel disappeared Was.

(Effect of drugs on plaque and biofilm removal of nanobubble Streptococcus mutans)
After adding 1 mL of BHI Broth + 5% sucrose to each well of a 48-well cell culture plate, 50 μl of Streptococcus mutans (ATCC 25175) culture solution was added to the same well. The hydroxyapatite (HA) disc (HA 48-3, Funakosi) was immersed and cultured for 48 hours to form plaque and a biofilm. Otsuka distilled water, nano bubble water, 0.02% benzalkonium chloride (Kanto Chemical), 0.02% benzalkonium chloride + nano bubble water (50%) against the HA disc to examine the influence of the drug on the cleaning effect of the nano bubble %) 0.5% monoammonium glycyrrhizinate (Kanto Chemical Co., Ltd.), 0.5% monoammonium glycyrrhizinate + nanobubble water (50%), neoste green (0.2% benzethonium chloride, Nippon Dental Pharmaceutical), contest (0.05% chlorhexidine gluconate, Reaction was carried out for 10 minutes with 0.5% ammonium glycyrrhizinate, and it was observed with a scanning electron microscope.

In distilled water (FIG. 10A), 0.02% benzalkonium chloride (FIG. 10C) and neoste green (FIG. 10G), most plaques / biofilms remained, and a considerable amount remained in the contest (FIG. 10H). In the nanobubbles, plaque and biofilm almost disappeared (FIG. 10B). On the other hand, the plaque / biofilm almost disappeared by the combined use of 0.02% benzalkonium chloride and nanobubbles (FIG. 10D). With 0.5% monoammonium glycyrrhizinate, the presence of nanobubbles remained unchanged, and almost no plaque / biofilm disappeared (Fig. 10E, F).

Example 4
(Drug removal in root canal dentin with nano bubbles)
Root canals of pig fresh removed premolars were enlarged to # 60, and the apexes were closed with unifast. After alternate washing with 2 mL of 5% sodium hypochlorite and 2 mL of 3% hydrogen peroxide solution, it is further washed with 5 mL saline, and smearin (3% EDTA) is further applied to the root canal for 2 minutes, and 5 mL saline It was further washed with water and stored in saline at 4 ° C. After drying in the root canal with a broach cotton plug, 5 mg / ml (final concentration) of tetracycline containing nanobubble water (50%) was applied for 5 minutes to allow the drug to penetrate deep into the root canal dentinal tubule (up to about 1 mm) . After washing with physiological saline, the inside of the root canal was dried. Next, in order to remove this deeply penetrated antibiotic, physiological saline, nano bubbles only, 1.5% EDTA, 1.5% EDTA + nanobubbles (containing 50%), 8.5% EDTA, 8.5% EDTA + nanobubbles (containing 50%) The root canal was washed once for 5 minutes with 2 mL of 6 ml of The teeth were fixed to a metal base with a dental wax, Unifast III so that the dental pulp cavity was parallel. After Unifast III had hardened, a section sample of about 300 μl in thickness was prepared with a Zege microtome and observed with a stereo fluorescent microscope.

The deeply penetrated antibiotic (tetracycline) could not be removed at all by distilled water (FIG. 11A), but could be removed considerably by nanobubbles, but remained on the surface of the root canal (FIG. 11B). Removal was not complete with 1.5% EDTA (FIG. 11C) and 8.5% EDTA (FIG. 11E), although removal proceeded by inclusion of nanobubbles, but removal was not complete and remained on the surface of the root canal (FIG. 11D) , F).

After impregnating with tetracycline in the same manner as described above, the root canal was washed with nanobubbles only, 2 mL, 1 minute, 2 times, 3 times.

The deeply infiltrated tetracycline could not be removed at all once in one minute of distilled water (FIG. 12A), and the removal progressed as the cycles were repeated by nanobubbles, and could be completely removed in three times (FIG. 12B-D).

[Example 5]
(Removal of tongue with nano bubble)
Tongue was rinsed with benzethonium-containing mouthwash for 1 minute or with nanobubbles for 1 minute.

FIG. 13A shows the nanobubble water before application. Nanobubble water (FIG. 13C) was able to remove more tongue brow compared to benzethonium-containing mouthwash (FIG. 13B). In addition, when applied to a tongue using a nano bubble gel prepared by uniformly mixing powder Alcox E-240 with nano bubble water, the tongue ring could be removed.

[Example 6]
(Promotion of decalcification by nanobubble-containing root canal cleaning agent)
The root canal of pig fresh removed premolars is enlarged to # 60 with K file (Manny), and after alternate washing with 2 mL of 5% sodium hypochlorite and 2 mL of 3% hydrogen peroxide water, 5 mL physiological salt It was further washed with water and dried. The root canal was treated with nanotubular water, 8.5% EDTA preparation + 50% nanobubble water, 8.5% EDTA preparation, and 17% EDTA preparation four root canal enlargement cleaning agents. Distilled water was used as a negative control. In addition, nano bubble water is nano bubble water manufactured using air with a nano bubble generator (FOAMEST (registered trademark), Nac Corp.), concentration 2 × 10 ⁸ particles / mL, particle size peak 100 nm, zeta potential −22.9 mV, It was pH 6.25.

The thickness was adjusted to 3 mm with a Zege microtome (Leica), polished to # 2000 with sandpaper, and stored in saline until measurement. The Vickers hardness at 100, 300 and 500 μm points from the root canal wall of the prepared sample was measured with a micro Vickers hardness tester (Akashi Mfg. MVK-E) at a weight of 50 g for 15 seconds.

As shown in FIG. 14, the Vickers hardness hardly changed with the nano bubbles alone as in the case of the distilled water. On the other hand, Vickers hardness decreased significantly only at the 100 μm point with 17% EDTA and 8.5% EDTA compared to nano bubbles and distilled water. Furthermore, at 8.5% EDTA + 50% nanobubble water, Vickers hardness decreases significantly at 100, 300 and 500 μm compared to 17% EDTA, and at 300 and 500 μm significantly compared to 8.5% EDTA. A decrease in Vickers hardness was observed. From the results, it was shown that the addition of nanobubbles to the root canal cleaning agent can deash to a significantly deeper point.

[Example 7]
(Change of plaque and biofilm removal of Streptococcus mutans by viscosity of nano bubble gel)
After adding 1 mL of BHI Brot (Kanto Chemical) + 5% sucrose to each well of a 48-well cell culture plate, 50 μl of Streptococcus mutans (ATCC 25175) culture solution was added to the same well. The hydroxyapatite (HA) disc (HA 48-3, Funakosi) was immersed and cultured for 48 hours to form plaque and a biofilm. In order to examine the washing effect of the viscosity of nano bubble gel, let the gel prepared with various viscosity act on this HA disc for 30 minutes, pass through Otsuka distilled water, nano bubble water, each gel only and untreated one, According to the method, scanning electron microscope specimens were prepared and compared. The viscosity was measured using a tuning fork viscometer. The gel and viscosity used are as follows. Gel nano bubble gel (1) (Alcox E-240 1.06 W%) viscosity 117 mPa · s, nano bubble gel PEG 20 k 5% (polyethylene glycol PEG 20 k 5%) 6 mPa, nano bubble gel A (Alcox E-240) 52 mPa · s, nano bubble Gel B (cellogen BSH-12): 199 mPa · s, nano bubble gel HPC-M 2.5% (hydroxypropyl cellulose HPC-M 2.5%) 447 mPa · s.

As shown in FIG. 15, plaque remained in the distilled water even after 30 minutes. In the nano bubble water, most of the plaque decreased after 30 minutes, but it remained a little. The nano bubble gel (1) was the most effective and almost completely removed plaque. The other nano bubble gels had a higher removal effect than nano bubble water, but a slight residual was observed, and the effect was not different due to the difference in viscosity.

[Example 8]
(Change of smear layer removal effect by difference of internal pressure of nano bubble water)
The following nano bubble characteristics were obtained by changing the air pressure and water pressure used at the time of nano bubble production.

In addition, air pressure and water pressure here are pressures at the time of creating nano bubble water according to the following manufacturing method. That is, a tubular gas permeation apparatus in which a gas permeable film is formed by generating a craze on a polymer resin film capable of limiting the gas permeation amount in a gas permeation part, a tank for storing water, and the storage tank. The pump comprises a pump for delivering the water to the tubular circulation path, and the tubular gas permeation device is installed in the tubular circulation path, whereby the outer diameter of the tubular gas permeation portion and the tubular shape are formed. A hydraulic pressure (this pressure is the water pressure) is adjusted using a pump to introduce water into the gap formed by the difference from the inner circumferential diameter of the circulation path, and water is introduced to the gas permeable portion of the gas permeable device, By adjusting the pressurized state and supplying a gas (the pressure of the gas is air pressure), nano-sized fine bubbles are mixed with water to produce nano-bubble water.

By these nanobubbles, the smear layer was completely removed and the pores were extended to the peritubular dentin at an air pressure of 0.17 water pressure 0.28 MPa (d) and an air pressure of 0.07 water pressure 0.18 MPa (a). In addition, the dentine wall surface became uneven at an air pressure of 0.34 MPa and a water pressure of 0.38 MPa (e) (FIG. 16A). That is, the smear layer removing effect was d> a> b> c> e.

Statistical analysis revealed that the air pressure 0.17 water pressure 0.28 MPa (d) has a significantly higher dentinal tubule area per piece and a higher smear layer removal effect than the other internal pressures ( Figure 16B).

Nano bubble (d) air pressure 0.17 water pressure 0.28 MPa, nano bubble (a) air pressure 0.07 water pressure 0.18 MPa, nano bubble (b) air pressure 0.09 water pressure 0.24 MPa, nano bubble (c) air pressure 0.13 water pressure 0.20 MPa, and nano bubble (e) The particle size distribution was measured for an air pressure of 0.34 and a water pressure of 0.38 MPa. The particle size distribution was measured using Malvern's Nanosite NS300 apparatus. As a result, as shown in FIG. 16C, d was 169 nm, a was 143 nm, b was 77 nm, c was 111 nm, and e was 151 nm.

[Example 9]
Porphyromonas gingivalis (ATCC 33277), which is one of the causative bacteria of marginal periodontitis, was cultured in a modified GAM broth medium to prepare a bacterial solution at 1 × 10 ⁸ CFU / ml.

(Biofilm removal by Nanobubbles of Porphyromonas gingivalis in an in vitro hydroxyapatite model)
After adding 1 ml of modified GAM broth + 5% sucrose to each well of a 48-well cell culture plate, 50 μl of Porphyromonas gingivalis culture solution was added to the same well. The hydroxyapatite (HA) disc (HA 48-3, Funakosi) was immersed and cultured for 6 days to form a biofilm. In order to examine the cleaning effect of the nanobubbles, nanobubbles, 0.025% benzalkonium chloride, and 0.025% benzalkonium chloride containing nanobubbles were allowed to act on the HA disc for 5 minutes. After the action, the cells were cultured at 37 ° C. for 90 minutes in a modified GAM broth containing 10% PrestoBlue® Cell Viability Reagent, and observed with a stereomicroscope (Leica: M205FA). Also, the culture supernatant was recovered, and the number of bacteria was counted using Molecular Devices: SpectraMax M5. In addition, after dehydrating hydroxyapatite with 30, 50, 70, 90, 100% ethanol, vapor deposition with platinum 10 kV (conductive film deposition (sputter coating) MSP-20-UM, vacuum device) and scanning electron microscope (VE9800) , KEYENCE).

When the number of bacteria remaining in hydroxyapatite after action is measured with PrestoBlue (registered trademark), the number of bacteria adapted to nanobubbles, 0.025% benzalkonium chloride and nanobubble containing 0.025% benzalkonium chloride tends to decrease. Was seen (Figure 17).

On scanning electron microscopy, the biofilm of Porphyromonas gingivalis could be significantly removed in 5 minutes of nanobubbles (FIG. 18B). Also, 0.025% benzalkonium chloride was able to remove Porphyromonas gingivalis (FIG. 18C), but a mixture of nanobubbles and 0.025% benzalkonium chloride was able to remove most of Porphyromonas gingivalis (FIG. 18D). ).

(Study on biofilm removal effect of Porphyromonas gingivalis nanobubbles by in vitro Live / Dead staining)
After adding 1 ml of modified GAM broth + 5% sucrose to each well of a 48-well cell culture plate, 50 μl of Porphyromonas gingivalis culture solution was added to the same well. The cells were cultured for 6 days to form a biofilm. In order to investigate the cleaning effect of nano bubbles, nano bubbles, 0.025% benzalkonium chloride, and 0.025% benzalkonium chloride (nano bubble dilution) were allowed to act for 5 minutes. After the action, the cells were stained with Live / Dead BacLight Bacterial Viability Kits component (Invitrogen) and observed with a fluorescence microscope (Keyence: VE7000).

Fluorescent coloration showing dead cells was particularly strong when treated with 0.025% benzalkonium chloride (FIG. 19). On the other hand, those containing nanobubbles were less likely to be seen in both live and dead bacteria. Containing nanobubbles 0.025% benzalkonium chloride has the same bactericidal action as 0.025% benzalkonium chloride, and since it is disinfected and washed away by the effect of nanobubbles, neither viable bacteria nor dead bacteria are found It can be considered.

(Removal of bacteria in periodontal pocket by nano bubbles in vivo)
All the treatment of beagle dogs (central science material) was performed after general anesthesia. Specifically, domitol (0.8 mg / 10 kg), dolmikam (1 mg / 10 kg), and vetofal (2 mg / 10 kg) were administered intramuscularly. After the sedation, the mesial, central, and centrifugal 3 points of the buccal periodontal pocket were measured with a probe, and then three plaques deep in the dog pocket were collected at the paper point, and the number of bacteria was measured. First, sterile paper point # 40 was inserted into the periodontal pocket, and the inside of the pocket was wiped five times in the near-distal direction. This paper point was set to a bacteria counter (Panasonic: DU-AA01NP-H) and measured. Hydroxypropyl Cellulose (HPC) gel with 0.025% benzalkonium chloride, 0.025% benzalkonium chloride HPC gel (nano bubble dilution) was injected into the periodontal pocket, and the number of bacteria was measured after 24 hours.

After measuring the number of bacteria in the periodontal pocket of the beagle dog, a benzalkonium chloride diluted to 0.025% with HPC gel or HPC gel containing nanobubbles was injected into the periodontal pocket. After 24 hours, the number of bacteria was significantly reduced for both benzalkonium chloride (gel) and benzalkonium chloride-containing nanobubbles (gel) as measured again (FIG. 20). Furthermore, when the benzalkonium chloride (gel) and the benzalkonium chloride-containing nanobubbles (gel) were compared, the drug-containing nanobubbles showed a significant reduction in the number of bacteria.

It can be used for caries treatment, pulpectomy / infective root canal treatment, periodontal disease treatment and the like.

Claims (12)

An intraoral adhesion removal promoter for removing smear layer while maintaining dentin strength,
An agent for promoting the removal of intra-oral deposits, comprising nanobubbles injected into a root canal. The intraoral deposit removal promoter according to claim 1, wherein the nanobubble is a nanobubble having a zeta potential of -10 mV or less. The agent for promoting removal of intra-oral deposit according to claim 1 or 2, characterized by having a nano-bubble gel which is a gel containing nano-bubbles. The agent for promoting removal of intra-oral adhesions according to claim 1 or 2, wherein the nanobubbles injected into the root canal are nanobubble water which is water containing nanobubbles. The nanobubble concentration in the nanobubble water is 1 × 10 ⁷ cells / mL to 2 × 10 ⁹ cells / mL, the nanobubble particle size is 10 nm to 200 nm, and the zeta potential is −10 mV or less. The intraoral adhesion removal promoter according to The agent for promoting removal of intra-oral deposit according to any one of claims 1 to 5, wherein the nano bubble contains any one of air, oxygen, carbon dioxide, nitrogen, or ozone. It is a removal promoter for intraoral adhesions which is continuously adhered to and removed from intraoral adhesions which are plaques, biofilms or tongues,
An agent for promoting removal of intra-oral deposit, characterized in that it has nano-bubble gel which is gel containing nano-bubbles. The removal promoter of the intraoral deposit according to claim 7, wherein the viscosity of the nano bubble gel is 450 mPa · s or less. The intraoral adhesion removal agent according to claim 7 or 8, wherein the nanobubbles are nanobubbles having a zeta potential of -10 mV or less. The removal promoter for intraoral adhesion according to any one of claims 1 to 9,
With drugs
An intraoral deposit cleaning accelerator characterized by including. 11. The agent for promoting the cleaning of the intra-oral deposit according to claim 10, wherein the agent is benzalkonium chloride or an antibiotic. Nano bubble water or nano bubble gel in which nano bubbles exist in water,
Root canal enlargement cleaning agent,
A root canal enlargement aid for enlarging a narrowing root canal of middle-aged and elderly people, characterized in that it comprises:

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