CN110464871A - A kind of injection aquagel and preparation method thereof - Google Patents
A kind of injection aquagel and preparation method thereof Download PDFInfo
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- CN110464871A CN110464871A CN201910763325.8A CN201910763325A CN110464871A CN 110464871 A CN110464871 A CN 110464871A CN 201910763325 A CN201910763325 A CN 201910763325A CN 110464871 A CN110464871 A CN 110464871A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
- A61L26/0023—Polysaccharides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
- A61L26/0028—Polypeptides; Proteins; Degradation products thereof
- A61L26/0047—Specific proteins or polypeptides not covered by groups A61L26/0033 - A61L26/0042
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/008—Hydrogels or hydrocolloids
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
- C08J3/03—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
- C08J3/075—Macromolecular gels
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/06—Flowable or injectable implant compositions
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
- C08J2305/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2389/00—Characterised by the use of proteins; Derivatives thereof
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2405/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2401/00 or C08J2403/00
- C08J2405/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2489/00—Characterised by the use of proteins; Derivatives thereof
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Abstract
The invention discloses a kind of injection aquagels and preparation method thereof, it is characterized in that the injection aquagel is the injection aquagel being cross-linked to form at HRP and hydrogen peroxide oxidation catalytic condition using para hydroxybenzene propionic acid modification of chitosan and fibroin albumen.Preparation method includes the preparation of para hydroxybenzene propionic acid modification of chitosan, fibroin albumen extraction and solution allocation and the responsiveness of hydrogel synthesizes under the redox of horseradish peroxidase (HRP) and hydrogen peroxide.The hydrogel material has good biocompatibility and mechanical property, also has the characteristics that preparation condition mildly and without chemical residue.It can be used for filling irregular skin trauma cavity with in-situ injection, promote skin repair and regeneration.
Description
Technical field
The present invention relates to a kind of hydrogel biological materials, and specifically modified chitosan and fibroin albumen use horseradish mistake
The injection aquagel and preparation method thereof that oxide enzyme (HRP) is cross-linked to form.Belong to biomaterial and biomedical work
Journey field.
Background technique
Skin is played as the maximum organ of human body in prevention infection and the loss of Water-Electrolyte and in terms of adjusting body temperature
Vital effect.The healing of large area full thickness dermal is an important clinical problem.Although traditional dressing is being hurt
Mouth healing aspect has certain advantage, but due to lacking the adaptability to environment, is not suitable for large area, irregular wound.It can
Solid-like object is formed in situ by minimally invasive injection in defect point in injection hydrogel, and can fill complicated cavity in vivo.Water-setting
Glue is alternatively arranged as tissue engineering bracket and is conducive to cell migration, promotes regeneration.Therefore, research has the novel wound of syringeability
Hurt dressing to be of great significance.
Chitosan and fibroin albumen are the excellent natural materials for designing porous matrix, have good biocompatibility and life
Biodegradable can also form a variety of different structures by chemical modification, thus have required a variety of different performances.
There is chitosan more amino to exist, and is easily modified by sulphation, is hopeful to graft on hydroxy phenylpropionic acid on chitosan, can pass through
HRP enzyme crosslinking forms hydrogel.Fibroin albumen has good mechanical property, and the mechanical property of composite material can be improved, and
The tyrosine contained can form network structure by HRP enzyme crosslinking.
Summary of the invention
The present invention there are problems that being difficult to heal traditional dressing in the irregular wound of widespread skin, prepare HRP enzyme
The modification of chitosan of crosslinking/fibroin albumen injection aquagel can fill complicated wound cavity in vivo, promote skin
Skin wound healing.
The modification of chitosan of HRP enzyme crosslinking of the present invention/fibroin albumen injection aquagel is to utilize para hydroxybenzene third
The injection aquagel that sour modification of chitosan and fibroin albumen are cross-linked to form at HRP and hydrogen peroxide oxidation catalytic condition.Its
Specific preparation method the following steps are included:
1) 0.5-1.5%g/mL chitosan and 0.4%g/mL para hydroxybenzene propionic acid the preparation of modification of chitosan solution: are prepared
Mixed solution stirs 6h, and 0.726%g/mLEDC and 0.346%g/mLNHS is added, dialyses 3 days after 48h is stirred at room temperature, and freezes
0.4-2%g/mL para hydroxybenzene propionic acid modification of chitosan aqueous solution is configured to after dry 48h；
2) prepared by silk fibroin protein solution: weighing silk and is dissolved in the silkworm for being made into the lithium-bromide solution of 9.3M and preparing 20%g/mL
Silk solution, is placed in water-bath after 60 DEG C of heating 4h, using 3500D bag filter dialysis 48h；Then, the PEG of 20%g/mL is replaced
Solution continues dialysis 6-10h and obtains concentration silk fibroin protein solution；Using 10000rpm, 4 DEG C, centrifugation removes impurity after twenty minutes,
And concentration fibroin albumen concentration is determined by weight method, appropriate distilled water is added and is configured to the fibroin that concentration is 0.4-2%g/mL
Protein solution；
3) preparation of hydrogel: taking the modification of chitosan aqueous solution obtained in step 1), and addition volume ratio is 0.3-1.2%
Concentration be 10mg/mL HRP enzyme aqueous solution, be put into after mixing in double syringe A pipe；Then the silk obtained in step 2) is taken
Fibroin solution, the aqueous hydrogen peroxide solution that the concentration that addition volume ratio is 1-4% is 0.5%g/mL, is put into two-tube after mixing
In syringe B pipe；Solution in syringe A, B pipe is squeezed out in equal volume, is prepared into hydrogel within 1-5 seconds after mixing.
The present invention has the prominent advantages that compared with prior art:
1) in material selection, biodegradable natural fibroin albumen and chitosan possess preferable biocompatibility.Shell
Glycan has more chemical reaction site, is conducive to material modification.Fibroin albumen has good mechanical property, can enhance shell
The mechanical strength of glycan.
2) in preparation process, hydrogel preparation is carried out using the method for enzyme crosslinking, has preparation condition mildly and without chemistry
The characteristics of substance remains.
3) product functionally, the hydrogel of preparation can in-situ injection, can be used for filling irregular skin trauma cavity, promote
Skin repair and regeneration.
Specific embodiment:
Embodiment 1:
1) 1%g/mL chitosan and 0.4%g/mL para hydroxybenzene propionic acid mixed solution are prepared, 6h is stirred, is added 0.726%
G/mL EDC and 0.346%g/mL NHS dialyses 3 days after 48h is stirred at room temperature, and is configured to 1%g/mL to hydroxyl after being freeze-dried 48h
Base benzenpropanoic acid modification of chitosan aqueous solution；
2) it weighs 5g silk and is dissolved in the silk solution for being made into the lithium-bromide solution of 9.3M and preparing 20%g/mL, be placed in water-bath
In pot after 60 DEG C of heating 4h, using 3500D bag filter dialysis 48h；Then, the PEG solution for replacing 20%g/mL continues the 10h that dialyses
Obtain concentration silk fibroin protein solution；Using 10000rpm, 4 DEG C, centrifugation removes impurity after twenty minutes, and is determined by weight method dense
Contracting fibroin albumen concentration is added appropriate distilled water and is configured to the silk fibroin water solution that concentration is 1%g/mL；
3) the 5mL modification of chitosan aqueous solution obtained in step 1) is taken, the concentration that 30 μ L are added is the HRP enzyme of 10mg/mL
Aqueous solution is put into double syringe A pipe after mixing；Then the silk fibroin protein solution obtained in 5mL step 2) is taken, 100 μ are added
The concentration of L is the aqueous hydrogen peroxide solution of 0.5%g/mL, is put into double syringe B pipe after mixing；It will be in syringe A, B pipe
Solution squeezes out in equal volume, and 1-5s is prepared into hydrogel after mixing.
Embodiment 2:
1) 1%g/mL chitosan and 0.4%g/mL para hydroxybenzene propionic acid mixed solution are prepared, 6h is stirred, is added 0.726%
G/mL EDC and 0.346%g/mL NHS dialyses 3 days after 48h is stirred at room temperature, and is configured to 1.2%g/mL pairs after being freeze-dried 48h
Hydroxy phenylpropionic acid modification of chitosan aqueous solution；
2) it weighs 5g silk and is dissolved in the silk solution for being made into the lithium-bromide solution of 9.3M and preparing 20%g/mL, be placed in water-bath
In pot after 60 DEG C of heating 4h, using 3500D bag filter dialysis 48h；Then, the PEG solution for replacing 20%g/mL continues the 10h that dialyses
Obtain concentration silk fibroin protein solution；Using 10000rpm, 4 DEG C, centrifugation removes impurity after twenty minutes, and is determined by weight method dense
Contracting fibroin albumen concentration is added appropriate distilled water and is configured to the silk fibroin water solution that concentration is 0.4%g/mL；
3) the 5mL modification of chitosan aqueous solution obtained in step 1) is taken, the concentration that 30 μ L are added is the HRP enzyme of 10mg/mL
Aqueous solution is put into double syringe A pipe after mixing；Then the silk fibroin protein solution obtained in 5mL step 2) is taken, 100 μ are added
The concentration of L is the aqueous hydrogen peroxide solution of 0.5%g/mL, is put into double syringe B pipe after mixing；It will be in syringe A, B pipe
Solution squeezes out in equal volume, and 1-5s is prepared into hydrogel after mixing.
Embodiment 3:
1) 1%g/mL chitosan and 0.4%g/mL para hydroxybenzene propionic acid mixed solution are prepared, 6h is stirred, is added 0.726%
G/mL EDC and 0.346%g/mL NHS dialyses 3 days after 48h is stirred at room temperature, and is configured to 0.4%g/mL pairs after being freeze-dried 48h
Hydroxy phenylpropionic acid modification of chitosan aqueous solution；
2) it weighs 5g silk and is dissolved in the silk solution for being made into the lithium-bromide solution of 9.3M and preparing 20%g/mL, be placed in water-bath
In pot after 60 DEG C of heating 4h, using 3500D bag filter dialysis 48h；Then, the PEG solution for replacing 20%g/mL continues the 10h that dialyses
Obtain concentration silk fibroin protein solution；Using 10000rpm, 4 DEG C, centrifugation removes impurity after twenty minutes, and is determined by weight method dense
Contracting fibroin albumen concentration is added appropriate distilled water and is configured to the silk fibroin water solution that concentration is 1.2%g/mL；
3) the 5mL modification of chitosan aqueous solution obtained in step 1) is taken, the concentration that 30 μ L are added is the HRP enzyme of 10mg/mL
Aqueous solution is put into double syringe A pipe after mixing；Then the silk fibroin protein solution obtained in 5mL step 2) is taken, 100 μ are added
The concentration of L is the aqueous hydrogen peroxide solution of 0.5%g/mL, is put into double syringe B pipe after mixing；It will be in syringe A, B pipe
Solution squeezes out in equal volume, and 1-5s is prepared into hydrogel after mixing.
Claims (2)
1. injection aquagel is to be urged using para hydroxybenzene propionic acid modification of chitosan and fibroin albumen in HRP and hydrogen peroxide oxidation
The injection aquagel being cross-linked to form under the conditions of change.
2. the preparation method of injection aquagel, comprising the following steps:
1) 0.5-1.5%g/mL chitosan and the mixing of 0.4%g/mL para hydroxybenzene propionic acid the preparation of modification of chitosan solution: are prepared
Solution stirs 6h, and 0.726%g/mL EDC and 0.346%g/mL NHS is added, dialyses 3 days after 48h is stirred at room temperature, and freezing is dry
0.4-2%g/mL para hydroxybenzene propionic acid modification of chitosan aqueous solution is configured to after dry 48h；
2) silk fibroin protein solution prepare: weigh silk be dissolved in be made into the lithium-bromide solution of 9.3M prepare 20%g/mL silk it is molten
Liquid is placed in water-bath after 60 DEG C of heating 4h, using 3500D bag filter dialysis 48h；Then, the PEG solution of 20%g/mL is replaced
Continue dialysis 6-10h and obtains concentration silk fibroin protein solution；Using 10000rpm, 4 DEG C, centrifugation removes impurity after twenty minutes, and leads to
It crosses weight method and determines concentration fibroin albumen concentration, appropriate distilled water is added and is configured to the fibroin albumen that concentration is 0.4-2%g/mL
Aqueous solution；
3) preparation of hydrogel: taking the modification of chitosan aqueous solution obtained in step 1), and it is the dense of 0.3-1.2% that volume ratio, which is added,
Degree is the HRP enzyme aqueous solution of 10mg/mL, is put into double syringe A pipe after mixing；Then the fibroin egg obtained in step 2) is taken
White solution, the aqueous hydrogen peroxide solution that the concentration that addition volume ratio is 1-4% is 0.5%g/mL, is put into two-tube injection after mixing
In device B pipe；Solution in syringe A, B pipe is squeezed out in equal volume, is prepared into hydrogel within 1-5 seconds after mixing.
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| CN201910763325.8A CN110464871A (en) | 2019-08-21 | 2019-08-21 | A kind of injection aquagel and preparation method thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112159532A (en) * | 2020-07-03 | 2021-01-01 | 中国科学院大学温州研究院(温州生物材料与工程研究所) | Oxygen-containing hydrogel dressing and preparation and application thereof |
| CN112316211A (en) * | 2020-10-30 | 2021-02-05 | 南开大学 | Cartilage extracellular matrix bionic injectable hydrogel and preparation method thereof |
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| CN112159532A (en) * | 2020-07-03 | 2021-01-01 | 中国科学院大学温州研究院(温州生物材料与工程研究所) | Oxygen-containing hydrogel dressing and preparation and application thereof |
| CN112316211A (en) * | 2020-10-30 | 2021-02-05 | 南开大学 | Cartilage extracellular matrix bionic injectable hydrogel and preparation method thereof |
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