JP6047105B2 - Disinfectant - Google Patents
Disinfectant Download PDFInfo
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- JP6047105B2 JP6047105B2 JP2013557541A JP2013557541A JP6047105B2 JP 6047105 B2 JP6047105 B2 JP 6047105B2 JP 2013557541 A JP2013557541 A JP 2013557541A JP 2013557541 A JP2013557541 A JP 2013557541A JP 6047105 B2 JP6047105 B2 JP 6047105B2
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- honey
- porphyromonas gingivalis
- royal jelly
- disinfectant
- present
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- 239000000645 desinfectant Substances 0.000 title description 16
- 235000012907 honey Nutrition 0.000 claims description 29
- 241000605862 Porphyromonas gingivalis Species 0.000 claims description 24
- 229940109850 royal jelly Drugs 0.000 claims description 24
- 239000002324 mouth wash Substances 0.000 claims description 6
- 229940051866 mouthwash Drugs 0.000 claims description 4
- -1 oral ointment Substances 0.000 claims description 4
- 235000013305 food Nutrition 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 2
- 239000007937 lozenge Substances 0.000 claims 1
- 229940042125 oral ointment Drugs 0.000 claims 1
- 229940034610 toothpaste Drugs 0.000 claims 1
- 239000000606 toothpaste Substances 0.000 claims 1
- 208000028169 periodontal disease Diseases 0.000 description 11
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 3
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 210000000214 mouth Anatomy 0.000 description 3
- 239000002953 phosphate buffered saline Substances 0.000 description 3
- 230000003449 preventive effect Effects 0.000 description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 210000004556 brain Anatomy 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 238000001802 infusion Methods 0.000 description 2
- DYKFCLLONBREIL-KVUCHLLUSA-N minocycline Chemical compound C([C@H]1C2)C3=C(N(C)C)C=CC(O)=C3C(=O)C1=C(O)[C@@]1(O)[C@@H]2[C@H](N(C)C)C(O)=C(C(N)=O)C1=O DYKFCLLONBREIL-KVUCHLLUSA-N 0.000 description 2
- 229960004023 minocycline Drugs 0.000 description 2
- 239000002674 ointment Substances 0.000 description 2
- 230000003239 periodontal effect Effects 0.000 description 2
- 201000001245 periodontitis Diseases 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 238000013207 serial dilution Methods 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 235000005979 Citrus limon Nutrition 0.000 description 1
- 244000248349 Citrus limon Species 0.000 description 1
- 241000104430 Elsholtzia rugulosa Species 0.000 description 1
- 208000005888 Periodontal Pocket Diseases 0.000 description 1
- 241000605894 Porphyromonas Species 0.000 description 1
- 208000014151 Stomatognathic disease Diseases 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 235000008429 bread Nutrition 0.000 description 1
- 210000004534 cecum Anatomy 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000002079 cooperative effect Effects 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 239000000551 dentifrice Substances 0.000 description 1
- 230000000249 desinfective effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 235000013325 dietary fiber Nutrition 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 230000008029 eradication Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 235000015203 fruit juice Nutrition 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000005445 natural material Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 235000020185 raw untreated milk Nutrition 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 230000017423 tissue regeneration Effects 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/63—Arthropods
- A61K35/64—Insects, e.g. bees, wasps or fleas
- A61K35/644—Beeswax; Propolis; Royal jelly; Honey
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
- A61K8/987—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of species other than mammals or birds
- A61K8/988—Honey; Royal jelly, Propolis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/005—Antimicrobial preparations
Description
本発明は、歯周疾患の原因菌の除菌剤及び歯周疾患の予防又は改善剤に関する。 The present invention relates to a disinfectant for causative bacteria of periodontal disease and a preventive or ameliorating agent for periodontal disease.
歯槽膿漏などの歯周疾患に罹患した歯の一般的な治療は、歯周外科、歯周組織再生療法、歯石除去及び歯肉マッサージ等であり、失われた機能を回復することはほぼ不可能であることから、疾患の進行を止めることに主眼が置かれている。 Common treatments for teeth suffering from periodontal diseases such as alveolar pyorrhea are periodontal surgery, periodontal tissue regeneration therapy, calculus removal and gingival massage, and it is almost impossible to restore lost function Therefore, the main focus is on stopping the progression of the disease.
歯周疾患の主な原因菌は、ポルフィロモナス・ジンジバリス（Porphyromonas gingivalis）であり、この菌を抑制することによって歯周疾患の進行を抑制することができると考えられている。 The main causative bacterium of periodontal disease is Porphyromonas gingivalis, and it is considered that the progression of periodontal disease can be suppressed by suppressing this bacterium.
従来行われている方法には、抗菌薬のミノサイクリンの軟膏を盲嚢内に貼付する方法、消毒薬を含有する洗口剤による洗口等があるが、ミノサイクリンの盲嚢内適用は歯科医師が行わなければならないという制約があり、週に一回歯科医院に通院しなければならないという問題があった。また、消毒剤によるものには、決定的な効果が見られにくいという欠点があった。 Conventional methods include the application of an antibacterial minocycline ointment in the cecum and the mouthwash with a disinfectant-containing mouthwash, but dentist must apply minocycline in the sac. There was a problem of having to go to the dental clinic once a week. Moreover, the thing by a disinfectant had the fault that a decisive effect was hard to be seen.
歯科疾患実態調査によると、日本においては歯周疾患の目安となる歯周ポケットが４ｍｍ以上存在している割合が、５０代の人で約半数に達しており、また、高齢者の歯周疾患患者が増加していることが示されている。 According to a survey on the actual conditions of dental diseases, the proportion of periodontal pockets of 4 mm or more, which is a standard for periodontal diseases, in Japan is about half of those in their 50s. It has been shown that patients are increasing.
したがって、このような欠点を有さないポルフィロモナス・ジンジバリスの除菌剤の開発が待たれていた。 Therefore, development of a disinfectant for Porphyromonas gingivalis that does not have such drawbacks has been awaited.
本発明は、歯周疾患の原因菌の除菌剤及び歯周疾患の予防又は改善剤を提供する。 The present invention provides a disinfectant for periodontal diseases and a preventive or ameliorating agent for periodontal diseases.
上記課題を解決するために、本発明者らは、ヤハシハチミツ及び／又はローヤルゼリーを用いた抗ポルフィロモナス・ジンジバリス活性の検討を行った。 In order to solve the above-mentioned problems, the present inventors have examined anti-porphyromonas gingivalis activity using yah honey and / or royal jelly.
そして、驚くべきことに、ヤハシハチミツ及び／又はローヤルゼリーがポルフィロモナス・ジンジバリスを効果的に除菌することができることを見出し、本発明を完成させた。 Surprisingly, the present inventors have found that Yahashi honey and / or royal jelly can effectively sterilize Porphyromonas gingivalis and completed the present invention.
すなわち、本発明は、下記：
１．ヤハシハチミツ及び／又はローヤルゼリーを含む、ポルフィロモナス・ジンジバリス除菌剤、
２．ヤハシハチミツ及び／又はローヤルゼリーを含む、歯周疾患の予防又は改善剤、
に関する。That is, the present invention provides the following:
1. Porphyromonas gingivalis disinfectant containing Yahashi honey and / or royal jelly,
2. A preventive or ameliorating agent for periodontal disease, including yah honey and / or royal jelly,
About.
本発明のポルフィロモナス・ジンジバリス除菌剤は、ポルフィロモナス・ジンジバリスに対して優れた除菌作用を有し、かつ天然物質であることから安全性が高く、耐性菌の出現等の問題がなくなると同時に、歯科医の手を借りずに適用することができる。したがって、ポルフィロモナス・ジンジバリスの除菌、ポルフィロモナス・ジンジバリスに関連する疾患の予防または改善に非常に有用である。 The Porphyromonas gingivalis disinfectant of the present invention has an excellent disinfecting action against Porphyromonas gingivalis and is a natural substance, so it is highly safe and has problems such as the appearance of resistant bacteria. At the same time, it can be applied without the help of a dentist. Therefore, it is very useful for eradication of Porphyromonas gingivalis and prevention or improvement of diseases related to Porphyromonas gingivalis.
本発明において、ポルフィロモナス・ジンジバリス除菌剤とは、ポルフィロモナス・ジンジバリスの増殖を抑制することができる組成物を意味する。本発明の除菌剤は、ポルフィロモナス・ジンジバリスに関連する疾患の予防または改善のために用いることができる。 In the present invention, the Porphyromonas gingivalis disinfectant means a composition that can suppress the growth of Porphyromonas gingivalis. The disinfectant of the present invention can be used for prevention or amelioration of diseases associated with Porphyromonas gingivalis.
ヤハシハチミツは、中国雲南省の比較的高地に自生するヤハシ（Elsholtzia rugulosa）の花から採蜜される独特のハチミツであり、バターのように硬化することから硬蜜とも言われる，ヤハシハチミツは、最近、わが国でも販売されている（ジャパンローヤルゼリー社、皇蜜（登録商標））。 Yahashi honey is a unique honey harvested from Elsholtzia rugulosa flowers that grow naturally in relatively high altitudes in Yunnan, China. It is also sold in Japan (Japan Royal Jelly Co., Ltd., Honey (registered trademark)).
ローヤルゼリーとは、若い働き蜂であるミツバチの、上顎と下顎の咽頭腺から分泌されるそれぞれ異なった成分が反応する事により生成される分泌物及びそれを原料とする健康食品である。 Royal jelly is a secreted product produced by the reaction of different components secreted from the pharyngeal glands of the upper and lower jaws of a young worker bee, and a health food made from it.
本発明の除菌剤は、任意の食品素材、例えば、牛乳、オレンジやレモン等の果汁、ヨーグルト等の乳製品、パン、その他の食品に添加して用いることもできる。 The disinfectant of the present invention can be added to any food material, for example, milk, fruit juices such as orange and lemon, dairy products such as yogurt, bread, and other foods.
また、本発明の除菌剤においては、ヤハシハチミツの成人１日当たりの摂取量は、通常０.０１〜１０.０ｇである。通常、除菌剤中のヤハシハチミツの濃度は、その形態により異なるが、錠剤（タブレット）、トローチ剤、顆粒、ガム等の固形の形態の場合は、総質量に対して１０〜１００重量％の範囲である。また、ペースト、液状の場合には総重量に対して１０〜８０重量％の範囲である。好ましくは、唾液中の最終濃度が５〜１０ｍｇ/ｍｌになるように調製する。 Moreover, in the disinfectant of the present invention, the daily intake amount of yah honey per adult is usually 0.01 to 10.0 g. Usually, the concentration of honey honey in the disinfectant varies depending on the form, but in the case of solid forms such as tablets (tablets), troches, granules, and gums, it is in the range of 10 to 100% by weight based on the total mass. It is. In the case of paste or liquid, it is in the range of 10 to 80% by weight based on the total weight. Preferably, the final concentration in saliva is adjusted to 5 to 10 mg / ml.
本発明の除菌剤の形態としては、歯磨剤、口腔軟膏剤、洗口剤、含嗽剤、トローチ剤、錠剤（タブレット・口中錠）、顆粒を挙げることができる。 Examples of the disinfectant of the present invention include dentifrices, oral ointments, mouth washes, mouthwashes, troches, tablets (tablets and tablets in mouth), and granules.
さらに本発明の除菌剤には種々の添加物、例えば各種栄養素、種々のビタミン、ミネラル、食物繊維、多価不飽和脂肪酸、分散剤および乳化剤等の安定剤、甘味料、呈味成分、フレーバー等を配合することができる。また、液状の場合には、最初から液状として調製してもよいが、粉末またはペースト状で調製し、所定量の水性液体に溶解することができる。 Further, the disinfectant of the present invention includes various additives such as various nutrients, various vitamins, minerals, dietary fiber, polyunsaturated fatty acids, stabilizers such as dispersants and emulsifiers, sweeteners, taste ingredients, flavors. Etc. can be blended. In the case of a liquid, it may be prepared from the beginning as a liquid, but it can be prepared in a powder or paste form and dissolved in a predetermined amount of an aqueous liquid.
本発明の除菌剤は、口腔内のポルフィロモナス・ジンジバリスを除去することができることから、ポルフィロモナス・ジンジバリスに関連する疾患を予防または改善するために用いることができる。ポルフィロモナス・ジンジバリスに関連する疾患としては、口腔内の代表的なものとして歯周病（歯槽膿漏）がある。また、歯周病と強く関連すると考えられている糖尿病、循環器疾患等がある。 Since the disinfectant of the present invention can remove Porphyromonas gingivalis in the oral cavity, it can be used to prevent or ameliorate diseases related to Porphyromonas gingivalis. As a disease related to Porphyromonas gingivalis, there is periodontal disease (alveolar pyorrhea) as a typical one in the oral cavity. In addition, there are diabetes, cardiovascular disease and the like which are considered to be strongly associated with periodontal disease.
本発明を下記の実施例により詳細に説明するが、本発明はこれに限定されるわけではない。 The present invention will be described in detail by the following examples, but the present invention is not limited thereto.
使用薬物
１）ローヤルゼリー
ＲＪ原乳２００５０４０７はジャパンローヤルゼリー株式会社（東京）から供与された。
２）皇蜜
ヤハシ純粋蜂蜜（４ ５８２２６０ ８９０９９８）はジャパンローヤルゼリー株式会社（東京）から供与された。Drugs Used 1) Royal Jelly RJ raw milk 20050407 was provided by Japan Royal Jelly Co., Ltd. (Tokyo).
2) Imperial honey Yahashi pure honey (4 582260 890998) was provided by Japan Royal Jelly Co., Ltd. (Tokyo).
方法
１） 皇蜜およびローヤルゼリー溶液の調整
皇蜜溶液は２０％ｗ／ｖ（２００ｍｇ／ｍｌ）となるようにリン酸緩衝生理食塩液（ＰＢＳ）、またはポルフィロモナス・ジンジバリス用ブレインハートインフージョン（ＢＨＩ）培地に溶解した後、０．２２μｍフィルターにて濾過滅菌し調製した。ローヤルゼリー溶液は５０％ｗ／ｖになるようにＰＢＳ中に懸濁し、４℃、１２，０００ｒｐｍで１０分間遠心分離し、上清を０．２２μｍフィルターにて濾過滅菌し調製した。
２) ポルフィロモナス・ジンジバリスに対する皇蜜およびローヤルゼリーの最少発育阻止濃度（minimum inhibitory concentration；MIC)測定
ポルフィロモナス・ジンジバリスに対する皇蜜およびローヤルゼリーのＭＩＣを９６ウェルマイクロプレートを用い、連続希釈法により測定した。
３）、皇蜜とローヤルゼリー（ＲＪ）の協力作用
ローヤルゼリ（ＲＪ）０、１、２、３、および５％と皇蜜０、２．５、５、１０、２０％を含有するポルフィロモナス・ジンジバリス用ブレインハートインフージョンを作成し、９６ウェルマイクロプレートを用い測定した。Method 1) Preparation of honey and royal jelly solution The honey solution is 20% w / v (200 mg / ml) in phosphate buffered saline (PBS) or Brain Heart Infusion for Porphyromonas gingivalis ( (BHI) After being dissolved in the medium, the solution was sterilized by filtration through a 0.22 μm filter. The royal jelly solution was suspended in PBS to 50% w / v, centrifuged at 12,000 rpm at 4 ° C. for 10 minutes, and the supernatant was sterilized by filtration through a 0.22 μm filter.
2) Minimum inhibitory concentration (MIC) measurement of honey and royal jelly against Porphyromonas gingivalis MIC of honey and royal jelly against Porphyromonas gingivalis was measured by a serial dilution method using a 96-well microplate. did.
3) Co-operation of honey and royal jelly (RJ) Porphyromonas containing 0, 1, 2, 3, and 5% royal jelly (RJ) and 0, 2.5, 5, 10, 20% honey A brain heart infusion for Gingivalis was prepared and measured using a 96-well microplate.
結果
連続希釈法にてポルフィロモナス・ジンジバリスに対するＭＩＣを測定した。その結果陽性対照のＡＢＰＣのＭＩＣは０．２５μｇ／ｍｌ、皇蜜のＭＩＣは５０ｍｇ／ｍｌであった（図１）。 なお、この皇蜜溶液のｐＨは６．７４であった。
次に、皇蜜とローヤルゼリー（ＲＪ）の協力作用を検討した。その結果ローヤルゼリーが１％以上存在することにより皇蜜５％で完全にポルフィロモナス・ジンジバリスを死滅させることができた。ローヤルゼリーが５％存在する場合には、皇蜜２．５％で完全にポルフィロモナス・ジンジバリスを死滅させることができた。なお、ローヤルゼリーが存在しない場合には、皇蜜１０％で完全にポルフィロモナス・ジンジバリスを死滅させることができた。Results MIC against Porphyromonas gingivalis was measured by a serial dilution method. As a result, the MIC of the positive control ABPC was 0.25 μg / ml, and the MIC of honey was 50 mg / ml (FIG. 1). The pH of this honey solution was 6.74.
Next, we examined the cooperative action of honey and royal jelly (RJ). As a result, it was possible to completely kill Porphyromonas gingivalis with 5% of honey by the presence of 1% or more of royal jelly. When 5% of royal jelly was present, Porphyromonas gingivalis could be completely killed with 2.5% of honey. In the absence of royal jelly, Porphyromonas gingivalis could be completely killed with 10% of honey.
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