Material for demineralizing dentin collagen fibril, preparation method and application thereof

Abstract

a The invention relates to a dentin collagen fibril demineralizing material, and a preparation method and application thereof. The invention relates to a conjugate of water-soluble chitosan and an aminocarboxylic acid metal chelating agent used for dental demineralization, preferably dentin demineralization. The invention also discloses a dentin collagen fibril demineralizing material and a preparation method and application thereof, wherein the material is prepared from 0.5-1 part by mass of water-soluble chitosan, 10-30 parts by mass of EDTA, and 0.8-1.5 parts by mass of cross-linking agent. The invention relates to a dental demineralization composition comprising a conjugate and a material of the invention. The conjugate, material and composition can be used for dentin demineralization treatment before dental bonding and restoration. When used as a dentin collagen demineralizing agent, they have excellent selective extrafibrillar demineralization performance, good biological compatibility, effective anti-bacterial activity, endogenous matrix protease-inhibiting activity and extremely low cytotoxicity.

1 . A conjugate of a water-soluble chitosan and an aminocarboxylic acid metal chelating agent, for use in dental demineralization, preferably dentin demineralization. 
     
     
         2 . The conjugate according to  claim 1 , wherein the aminocarboxylic acid metal chelating agent is selected from the group consisting of ethylenediaminetetraacetic acid, hydroxyethyl ethylenediaminetriacetic acid, diethylenetriaminepentaacetic acid, aminotriacetic acid, dihydroxyethyl glycine, ethylene glycol tetraacetic acid, ethylenediamine diacetic acid, triethylenetetraaminehexaacetic acid, cyclohexanediamine tetraacetic acid, 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, 1,4,8,11-tetraazacyclododecane-1,4,8,11-tetraacetic acid or a mixture thereof, and preferably ethylenediaminetetraacetic acid. 
     
     
         3 . A material for demineralizing dentin collagen fibril, wherein the material is composed of 0.5-1 part by mass of water-soluble chitosan, 10-30 parts by mass of ethylenediaminetetraacetic acid, and 0.8-1.5 parts by mass of a crosslinking agent as starting materials. 
     
     
         4 . The conjugate or material according to  claim 1  or  3 , wherein the pH value of the conjugate or the material is neutral or alkaline. 
     
     
         5 . The conjugate or material according to  claim 1  or  3 , wherein the water-soluble chitosan is selected from the group consisting of glycol chitosan, carboxymethyl chitosan, chitosan hydrochloride, chitosan quaternary ammonium salt, chitosan sulfate ester, chitosan oligosaccharide or hyaluronic acid-like chitosan or a mixture thereof, preferably glycol chitosan. 
     
     
         6 . The conjugate or material according to  claim 1  or  3 , wherein the content of free amino group of the conjugate or the material is less than 15%, preferably 1% to 10%, more preferably 3% to 5%, as compared to the water-soluble chitosan prior to the conjugation reaction. 
     
     
         7 . The conjugate or material according to  claim 1  or  3 , wherein the Mn of the conjugate or the material is greater than 40 kDa, preferably 60 kDa to 300 kDa, more preferably 100 kDa to 200 kDa. 
     
     
         8 . The conjugate or material according to  claim 1  or  3 , wherein the Mw of the conjugate or the material is greater than 100 kDa, preferably 200 kDa to 800 kDa, more preferably 400 kDa to 600 kDa. 
     
     
         9 . The material of  claim 3 , wherein the cross-linking agent is selected from the group consisting of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide, N-hydroxysuccinimide, glutaraldehyde, bisimidate ester, or maleimide, or a mixture thereof. 
     
     
         10 . The conjugate or material according to  claim 1  or  3 , wherein the conjugate or the material comprises a dental demineralizing composition and further includes an orally acceptable excipient or vehicle. 
     
     
         11 . The conjugate or material according to  claim 1  or  3 , wherein the conjugate or the material comprises a dental bonding and restorative kit and further includes a dental bonding agent and/or restorative agent. 
     
     
         12 . The conjugate or material according to  claim 1  or  3 , wherein the conjugate or the material is applied to a tooth surface in need of restoration followed by applying a dental bonding agent and/or restorative agent to the tooth surface in need of restoration. 
     
     
         13 . A method for preparing a material according to  claim 3 , wherein the method comprises:
 mixing a water-soluble chitosan with an aminocarboxylic acid metal chelating agent in the presence of a cross-linking agent to provide a conjugate of the water-soluble chitosan and an aminocarboxylic acid metal chelating agent.   
     
     
         14 . The method according to  claim 13 , wherein the method comprises:
 mixing an aqueous water-soluble chitosan solution with an aqueous EDTA solution, and dissolving a cross-linking agent in the mixed solution for reaction at room temperature, wherein the water-soluble chitosan is 0.5-1 part by mass, EDTA is 10-30 parts by mass, and the cross-linking agent is 0.8-1.5 parts by mass.   
     
     
         15 . The method according to  claim 14 , wherein the method comprises:
 mixing an aqueous water-soluble chitosan solution at a concentration of 5 to 10 mg/ml with an aqueous EDTA solution at a concentration of 100 to 300 mg/ml in an equal volume,   adjusting the pH of the mixed solution to pH 6.0,   adding the cross-linking agent at a concentration of 8 to 15 mg/ml,   reacting for 12-16 hours at room temperature,   removing the unreacted residue via dialysis, and   freeze-drying to provide the material for demineralizing dentin collagen fibril.