JP6464370B2 - 細胞修復剤および殺菌消毒システム - Google Patents
細胞修復剤および殺菌消毒システム Download PDFInfo
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- JP6464370B2 JP6464370B2 JP2016542471A JP2016542471A JP6464370B2 JP 6464370 B2 JP6464370 B2 JP 6464370B2 JP 2016542471 A JP2016542471 A JP 2016542471A JP 2016542471 A JP2016542471 A JP 2016542471A JP 6464370 B2 JP6464370 B2 JP 6464370B2
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- electrolyzed water
- acidic electrolyzed
- cells
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- sterilization
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
- A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/32—Ingredients for reducing the noxious effect of the active substances to organisms other than pests, e.g. toxicity reducing compositions, self-destructing compositions
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/36—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids
- A01N37/38—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids having at least one oxygen or sulfur atom attached to an aromatic ring system
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/36—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids
- A01N37/38—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids having at least one oxygen or sulfur atom attached to an aromatic ring system
- A01N37/40—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids having at least one oxygen or sulfur atom attached to an aromatic ring system having at least one carboxylic group or a thio analogue, or a derivative thereof, and one oxygen or sulfur atom attached to the same aromatic ring system
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
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Description
以下に、様々な試験の結果に基づいて、本発明の実施の形態の細胞修復剤について説明する。なお、本発明は、これらの試験例(実施例)になんら限定されるものではない。
増殖期および定常期のマウス線維芽細胞に対する酸性電解水の影響を調べる試験を行った。試験に使用する酸性電解水(AEW)は、0.06〜0.08%塩化ナトリウム水溶液を電解質溶液として、電気分解装置(株式会社アルテック社製「ALTRON-MINI AL-700」)を用いて、交流電圧100Vおよび定格電流0.6Aで15分間電気分解することにより得た。得られた酸性電解水のpH、酸化還元電位(ORP)および残留塩素濃度は、pH/ORPメーター(Mettler-Toled社製「SG2」)および残留塩素メーター(HANNA Instruments Japan社製「HI96771C」)を用いて測定した。その結果、酸性電解水は、pHが2.2〜2.7、ORPが1100mV以上、残留塩素濃度が30〜60mg/lであることを確認した。
定常期のマウス線維芽細胞に酸性電解水を接触させたときに発生する、細胞内の活性酸素種(ROS)の量を調べる試験を行った。まず、試験1と同様に調製・培養した定常期のマウス線維芽細胞をリン酸緩衝化生理食塩水で洗浄後、純水にて希釈した4倍希釈系列の酸性電解水を100μlずつ各wellに加えた。30秒後に2倍希釈系列の酸性電解水を除去し、PBSで洗浄した後、直ちに細胞内ROS量を測定キット(Cell Biolabs社製「OxiselectTM Intracellular ROS Assay Kit」)を用いて測定した。
試験2と同様に、定常期のマウス線維芽細胞を、4倍希釈の酸性電解水および酸性電解水原液で30秒間処理した。PBSで洗浄後、ヒドロキシルラジカルのスカベンジャーとして、140mM DMSO(和光純薬工業社製)あるいは1mg/mlのプロアントシアニジン(Indeda社製「Leucoselect(登録商標)」)を含む1mM DCFH−DAを、各wellに100μl加え、37℃、5%CO2条件下で1時間培養した。培養後、試験2と同様に、細胞内細胞内ROS量を定量した。なお、各群間の有意差は、Tukey-Krammerの多重比較解析法で検定した。
試験1と同様に、増殖期のマウス線維芽細胞を、4倍希釈の酸性電解水および酸性電解水原液で30秒間処理した。リン酸緩衝化生理食塩水で洗浄後、血清無添加DMEMで調製した140mM DMSOあるいは1mg/mlのプロアントシアニジンを、各wellに100μl加え、37℃、5%CO2条件下で30分間培養した。PBSで洗浄後、新鮮培地を加え、さらに37℃、5%CO2条件下で24時間培養後の細胞増殖の程度をMTT法で測定した。なお、各群間の有意差は、Tukey-Krammerの多重比較解析法で検定した。
増殖期のヒト歯肉線維芽細胞について、試験4と同様の試験を行った。まず、25cm2フラスコで、37℃、5%CO2条件下で培養したヒト歯肉線維芽細胞(株式会社プライマリーセルより購入)を、マウス線維芽細胞の場合と同様に、細胞密度が2×104cells/mlになるよう培地にて細胞懸濁液を調製した。この細胞懸濁液を、100μlずつ96−wellマイクロプレートの各wellに播種した。37℃、5%CO2条件下で24時間培養後、試験4と同様に、増殖期の細胞を4倍希釈の酸性電解水および酸性電解水原液で30秒間処理した。PBSで洗浄後、血清無添加DMEMで調製した140mM DMSOあるいは1mg/mlのプロアントシアニジンを、各wellに100μl加え、37℃、5%CO2条件下で30分間培養した。PBSで洗浄後、新鮮培地を加え、さらに37℃、5%CO2条件下で24時間培養後の細胞増殖の程度をMTT法で測定した。なお、各群間の有意差は、Tukey-Krammerの多重比較解析法で検定した。
試験4と同様に、増殖期のマウス線維芽細胞を、4倍希釈の酸性電解水および酸性電解水原液で30秒間処理した。PBSで洗浄後、血清無添加DMEMで調製した1mg/mlの(+)−カテキン(東京化成工業社製)を各wellに100μl加え、37℃、5%CO2条件下で30分間培養した。PBSで洗浄後、新鮮培地を加え、さらに37℃、5%CO2条件下で24時間培養後の細胞増殖の程度をMTT法で測定した。なお、各群間の有意差は、Tukey-Krammerの多重比較解析法で検定した。
試験4と同様に、増殖期のマウス線維芽細胞を、酸性電解水原液で30秒間処理した。PBSで洗浄後、血清無添加DMEMで調製した1mg/mlのクロロゲン酸(Sigma-Aldrich社製)、カフェイン酸(東京化成工業社製)および没食子酸(東京化成工業社製)を、各wellに100μl加え、37℃、5%CO2条件下で5分あるいは30分間培養した。PBSで洗浄後、新鮮培地を加え、さらに37℃、5%CO2条件下で24時間培養後の細胞増殖の程度をMTT法で測定した。なお、各群間の有意差は、Tukey-Krammerの多重比較解析法で検定した。
試験4と同様に、増殖期のマウス線維芽細胞を、酸性電解水原液で30秒間処理した。PBSで洗浄後、血清無添加DMEMで調製した1mg/mlのプロアントシアニジンを各wellに100μl加え、それぞれ室温で1分間および5分間静置した。PBSで洗浄後、新鮮培地を加え、さらに37℃、5%CO2条件下で24時間培養後の細胞増殖の程度をMTT法で測定した。なお、各群間の有意差は、Tukey-Krammerの多重比較解析法で検定した。
試験4と同様に、増殖期のマウス線維芽細胞を、クロルヘキシジン原液(CHL;Xttrium Laboratories社製)あるいは、PBSで15倍希釈したイソジンガーグル液7%(ISO;明治製菓社製)で30秒間処理した。PBSで洗浄後、血清無添加DMEMで調製した1mg/mlのプロアントシアニジンを各wellに100μl加え、室温で5分間静置した。PBSで洗浄後、新鮮培地を加え、さらに37℃、5%CO2条件下で24時間培養後の細胞増殖の程度をMTT法で測定した。なお、各群間の有意差は、Tukey-Krammerの多重比較解析法で検定した。
試験5と同様にして得られた増殖期のヒト歯肉線維芽細胞を、PBSで洗浄後、500mM過酸化水素水を100μl加え、405nmのレーザー光を出力100mW(放射照度は310mW/cm2)で30秒間照射した。レーザー光照射は、インジウムガリウム窒化物をレーザーダイオードとして装着したレーザー装置(リコー光学株式会社製「RV−1000」)を用いて行った。
10 殺菌消毒システム
11 噴射器本体
11a 噴射口
12 殺菌消毒手段
13 後処理手段
Claims (2)
- プロアントシアニジン、(+)−カテキン、クロロゲン酸、カフェイン酸および没食子酸からなる群から選ばれるポリフェノールのうちの少なくとも1種を含むことを特徴とする細胞修復剤。
- 生体に対して殺菌処理および/または消毒処理を行う殺菌消毒手段と、
前記殺菌消毒手段による前記生体の被処理部に対して、請求項1記載の細胞修復剤を接触可能に設けられた後処理手段とを、
有することを特徴とする殺菌消毒システム。
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