CN107929816A - It is a kind of that there is hemostasis, antibacterial, the adherence preventing material and preparation method thereof of promoting healing - Google Patents
It is a kind of that there is hemostasis, antibacterial, the adherence preventing material and preparation method thereof of promoting healing Download PDFInfo
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- CN107929816A CN107929816A CN201711023353.3A CN201711023353A CN107929816A CN 107929816 A CN107929816 A CN 107929816A CN 201711023353 A CN201711023353 A CN 201711023353A CN 107929816 A CN107929816 A CN 107929816A
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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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/148—Materials at least partially resorbable by the body
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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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/04—Macromolecular materials
- A61L31/042—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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/141—Plasticizers
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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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/16—Biologically active materials, e.g. therapeutic substances
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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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/41—Anti-inflammatory agents, e.g. NSAIDs
-
- 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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/418—Agents promoting blood coagulation, blood-clotting agents, embolising agents
-
- 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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
- A61L2300/602—Type of release, e.g. controlled, sustained, slow
- A61L2300/604—Biodegradation
Abstract
The present invention provides a kind of new material for preventing tissue adhesion and preparation method thereof.The raw material sources of use avoid the use of small molecule crosslinking agent, have excellent biological safety in natural polysaccharide and modified natural polysaccharide.The present invention can be used for preventing adhesion for pelvic cavity, abdominal cavity, heel string, nerve etc. by natural polysaccharide and the compound prepared by the chemical reaction of modified natural polysaccharide, while have the function of hemostasis, antibacterial, promoting healing.Adherence preventing material provided by the invention, can be prepared into the sponge that prevents adhesion, Antiadhesive film and anti-adhesion gel by different technology conditions, and surgical procedure convenience is strong, have good potential applicability in clinical practice.
Description
Technical field
The present invention relates to a kind of technical field of biomedical materials, particularly one kind directly acts on people, mammal etc.
There is the blood surface of a wound, there is the degradable adherence preventing material and preparation method thereof of hemostasis, antibacterial, promotion wound healing.
Background technology
After post-operation adhesion refers to operation wound, in tissue healing process with tissue, the abnormal fibrous between organ connects
Connect.The adhesion of surgical site infections is medically one of still unsolved problem for a long time, almost all of postoperative theoretically
Recovery process can all be related to the adhesion problems of tissue.Adhesion is organized as complication common in surgical operation, it is caused
Consequence but can not look down upon, and tissue adhesion can cause to include small intestine obstruction, more including infertility and chronic abdominal cavity, pelvic pain etc.
Kind of complication, and organize adhesion also to increase the difficulty and risk of operation, cause operating time to extend, intraoperative hemorrhage increase with
And surrounding tissue organ damage, cause great body and mind pain and financial burden to patient.So far, using adherence preventing material
The method that most effective preventing adhesions have been turned out to be by clinic.
At present, common tissue sticking resistant connects material and plays an important role during clinic prevents adhesion, but also deposits
It can not be ignored at some, have the problem of to be solved.It is anti-sticking that Chinese patent 201410580360.3 discloses a kind of medical injectable
Even gel and preparation method thereof, this anti-adhesion gel being formed in situ by duplex injection device, have internal retention it is long,
The advantages that easy to use, but using gel degradation prepared by patent when can bring internal small molecule crosslinking agent residue problem,
Security risk is caused, is unfavorable for clinical promotion and application.Chinese patent 201410748936.2 discloses a kind of starch/hyalomitome
Acid/gelatin Antiadhesive film and preparation method thereof, the Antiadhesive film which prepares overcome single transparent matter acid constituents to prevent adhesion effect
The shortcomings that fruit is poor, while starch and the preferable gelatin of biocompatibility with certain water imbibition are added, make prepared anti-
Adhesion membrane has hemostasis, promoting healing effect concurrently.But there are two shortcomings for the Antiadhesive film：On the one hand the Antiadhesive film is to have
The Geniposide for having extremely strong crosslinking ability is crosslinking agent, and Antiadhesive film may be made to be difficult to degrade, and increases Clinical practice risk；It is another
Aspect, bacterium infection are also one of the main reason for causing tissue adhesion, especially for the humidity for being present in a large amount of tissue fluid
There is the blood surface of a wound in environment.And the Antiadhesive film does not possess antibacterial functions, Clinical practice is limited.Chinese patent
201610870752.2 disclose a kind of adherence preventing material with antibacterial promoting healing and preparation method thereof, although this tool
The gel rubber material for having temperature sensitive performance shows good preventing adhesiving effect in the secondary lysis of adhesion, but fine caused by bleeding
Fibrillarin deposition is one of the main reason for causing adhesion to occur, and due to lacking anthemorrhagic performance, above-mentioned patented product is not suitable for
The more adhesion of the bleeding surface of a wound occurred frequently.
In conclusion lack on Vehicles Collected from Market it is a kind of have concurrently hemostasis, antibacterial, promote wound healing function biological safety
High adherence preventing material.This patent using natural polysaccharide and modified natural polysaccharide as raw material, using the schiff base reaction of the two and/
Or acylation reaction improves the stability of natural polysaccharide, by the good liquid absorption capacity of prepared adherence preventing material, realize
Concentration to clotting factor, so that effectively activation coagulation pathway reaches quick-acting haemostatic powder effect, and utilizes the antibacterial of modification of polysaccharides
Property and promoting healing performance, successfully developing a kind of has the function of hemostasis, antibacterial, the adherence preventing material of promoting healing.
The content of the invention
It is an object of the invention to overcome the shortcomings of current anti-sticking by-product, there is provided one kind has hemostasis, antibacterial, promotes wound
The material for preventing tissue adhesion of mouthful healing function, which can be done directly on people, animal body has a blood surface of a wound, including to body
Interior and body cavity inner tissue, organ, are somebody's turn to do when the bleeding surface of a wound under surgical operation, trauma care, laryngoscope, endoscope and hysteroscope uses
During material, using the teaching of the invention it is possible to provide hemostasis, antibacterial, promote wound healing and prevent tissue adhesion function.
Present invention solution above-mentioned technical problem, which adopts the technical scheme that, utilizes natural polysaccharide and modified natural polysaccharide warp
Chemical reaction formed it is a kind of have the function of hemostasis, antibacterial, promotion wound healing degradable adherence preventing material, its mechanism bag
Include：The adherence preventing material can absorb the moisture in blood, concentrate clotting factor, activate coagulation pathway, shorten the clotting time, together
When can block blood outlet, reach hemostasis purpose；The adherence preventing material of the present invention uses the modified natural with antibacterial functions more
Sugar is raw material components, can isolate the surface of a wound, bacterium infection is prevented, so as to reach antibacterial purpose；The adherence preventing material energy of the present invention
The opposite moistening of the surface of a wound is enough kept, meets wound " wet union is theoretical ", promotes wound healing purpose so as to reach.
It is described that there is hemostasis, antibacterial, promote the degradable adherence preventing material of wound healing to include one or more days
The solution of right polysaccharide and the solution of one or more modified natural polysaccharide form compound through schiff base reaction and/or acylation reaction
Afterwards, it is prepared through vacuum freeze drying, wherein the mass volume ratio of natural polysaccharide solution is 0.05% ~ 50%, and modified natural is more
The mass volume ratio of sugar juice is 0.05% ~ 50%, and the volume ratio of natural polysaccharide solution and modified natural polysaccharide solution is 100:1~
1:100.The volume ratio of the natural polysaccharide solution and modified natural polysaccharide solution is 20:1~1:20.
Specifically, vacuum freeze drying is that wet stock or solution are condensed into solid-state at -10 DEG C ~ -50 DEG C, then true
Reciprocal of duty cycle is that moisture by solid state sublimation is gaseous state under the conditions of 1 ~ 10 Pascal, so that material dewatering.
Specifically, natural polysaccharide includes chitin derivativ, derivatives of hyaluronic acids, sodium alginate derivative, starch derivatives
Biology, glucan derivative, pulullan polysaccharide derivative, bletilla polysaccharide derivative, cellulose derivative, in lignin derivative
One or more.
Modified natural polysaccharide includes amination derivatives of hyaluronic acids, aminated alginic acid sodio-derivative, amination starch
Derivative, amination glucan derivative, amination pulullan polysaccharide derivative, amination bletilla polysaccharide derivative, amination
Cellulose derivative, amination lignin derivative, oxidated chitin derivative, oxidized hyaluronic acid derivative, aoxidizes seaweed
Sour sodio-derivative, oxidized starch derivative, oxidized dextran derivative, pullulan oxide derivative, aoxidizes bletilla polysaccharide
Derivative, oxycellulose derivative, the one or more in derivatives oxi-lignin.
In order to increase the antibiotic effect for the sponge that prevents adhesion, the sponge that prevents adhesion further includes antibacterials, the antimicrobial
Thing includes penicillins, cephalosporins, beta-lactamase inhibitor, aminoglycoside, macrolides, Tetracyclines, woods
One kind that can be in mycin class, polypeptide, quinolones, sulfamido or its combination, wherein antibacterials account for the sponge quality that prevents adhesion
0.05% ~ 10%, after the antibacterials are mixed with adherence preventing material, compound anti-adhesion sponge is prepared through vacuum freeze drying.
There is hemostasis, antibacterial, the degradable adherence preventing material for promoting wound healing the invention also discloses described
Preparation method and technological forming approach, using natural polysaccharide and modified natural polysaccharide through chemically reacting formed compound system
Standby adherence preventing material, then takes the technologies such as freeze-drying, casting film-forming and gel in-situ to be prepared into sponge, film, gel respectively
Etc. the product of form, it can be adapted for different operation antiblocking needs, surgical procedure facility, product is adapted to industrialized production.
The preparation method of the sponge that prevents adhesion, its feature, which comprises the following steps, to be made：
（a）With deionized water dissolving or swelling natural polysaccharide and modified natural polysaccharide, respective solution is prepared.Wherein natural polysaccharide is molten
The mass volume ratio of liquid is 0.05% ~ 50%, and the mass volume ratio of modified natural polysaccharide solution is 0.05% ~ 50%.
（b）Natural polysaccharide solution is mixed with modified natural polysaccharide solution, wherein natural polysaccharide solution and modified natural is more
The volume ratio of sugar juice is 100:1~1:100, preferably 20:1~1:20.
（c）Solid-state is condensed at -10 DEG C ~ -50 DEG C of the mixed solution warp, then under the conditions of vacuum is 1 ~ 10 Pascal
Moisture is gaseous state and vacuum freeze drying by solid state sublimation, and preparation prevents adhesion sponge.
In order to increase the pliability for the sponge that prevents adhesion, the present invention by the solution of plasticizer and one or more natural polysaccharides with
And after the solution mixing of one or more modified natural polysaccharide, prepare flexibility through vacuum freeze drying and prevent adhesion sponge.
The plasticizer selects glycerine, sorbierite, ethanol, one kind in poly- ethanedioic acid or its combination, wherein plasticizer
Account for and prevent adhesion the 0.05% ~ 10% of sponge quality.
The preparation method of the Antiadhesive film, it is characterised in that comprise the following steps：
（a）With deionized water dissolving or swelling natural polysaccharide and modified natural polysaccharide, respective solution is prepared.Wherein natural polysaccharide
The mass volume ratio of solution is 0.05% ~ 50%, and the mass volume ratio of modified natural polysaccharide solution is 0.05% ~ 50%.
（b）Natural polysaccharide solution is mixed with modified natural polysaccharide solution, wherein natural polysaccharide solution and modified natural is more
The volume ratio of sugar juice is 100:1~1:100, preferably 20:1~1:20.
（c）By the mixed solution by 300 mesh screens, vacuum deaerator, casting film-forming after room temperature is stood.
The volume ratio of wherein natural polysaccharide solution and modified natural polysaccharide solution is 100:1~1:100, preferably 20:1~1:
20。
In order to increase the antibiotic effect of Antiadhesive film, set as further, the present invention is using clinical widely used anti-
After bacterium medicine is mixed with the solution of one or more natural polysaccharides and the solution of one or more modified natural polysaccharide, this is mixed
Solution is closed by 300 mesh screens, vacuum deaerator, casting film-forming after room temperature is stood, is drying to obtain Antiadhesive film.
In order to increase the pliability of Antiadhesive film, the present invention by the solution of plasticizer and one or more natural polysaccharides and
It is after the solution mixing of one or more modified natural polysaccharide, the mixed solution is quiet by 300 mesh screens, vacuum deaerator, room temperature
Casting film-forming is postponed, is drying to obtain Antiadhesive film.The plasticizer selects one in glycerine, sorbierite, ethanol, poly- ethanedioic acid
Kind or its combination, wherein plasticizer accounts for the 0.05% ~ 10% of Antiadhesive film quality.
The preparation method of the anti-adhesion gel, it is characterised in that comprise the following steps：
（a）With deionized water dissolving or swelling natural polysaccharide and modified natural polysaccharide, respective solution is prepared.Wherein natural polysaccharide is molten
The mass volume ratio of liquid is 0.05% ~ 50%, and the mass volume ratio of modified natural polysaccharide solution is 0.05% ~ 50%.
（b）Natural polysaccharide solution is mixed with modified natural polysaccharide solution, wherein natural polysaccharide solution and modified natural is more
The volume ratio of sugar juice is 100:1~1:100, preferably 20:1~1:20.
（c）By 4 ~ 60 DEG C of the mixed solution stand 10 minutes ~ 48 it is small when, prepare anti-adhesion gel.
In order to increase the antibiotic effect of anti-adhesion gel, set as further, the present invention is using clinical widely used
After antibacterials are mixed with the solution of one or more natural polysaccharides and the solution of one or more modified natural polysaccharide, by this
When 4 ~ 60 DEG C of standings 10 minutes ~ 48 of mixed solution are small, anti-adhesion gel is prepared.
The antibacterials select penicillins, cephalosporins, beta-lactamase inhibitor, aminoglycoside, big
Cyclic lactone class, Tetracyclines, lincomycin class, polypeptide, quinolones, one kind in sulfamido or its combination, wherein antibacterial
Medicine accounts for the 0.02% ~ 5% of anti-sticking gel quality.
In order to increase the intensity of anti-adhesion gel, the present invention by the solution of plasticizer and one or more natural polysaccharides and
After the solution mixing of one or more modified natural polysaccharide, by 4 ~ 60 DEG C of the mixed solution stand 10 minutes ~ 48 it is small when, prepare anti-
Adhesion gel.
The plasticizer selects glycerine, sorbierite, ethanol, one kind in poly- ethanedioic acid or its combination, wherein plasticizer
Account for the 0.02% ~ 5% of anti-adhesion gel quality.
, can be to the material that prevents adhesion of the present invention in order to ensure adherence preventing material is having the security directly used at the blood surface of a wound
Sterilize after material packaging, sterilization method includes but not limited to ethylene oxide sterilizing, Co-60 radiation sterilization, γ radiation sterilization, smelly
Oxygen sterilizes.
Advantages of the present invention：
Compared with prior art, the present invention it the advantage is that：(1) change that the present invention passes through natural polysaccharide and modified natural polysaccharide
Reaction is learned, has obtained having concurrently the adherence preventing material of two kinds of polysaccharide excellent properties, has solved single polysaccharide in the prior art and degraded
It hurry up, the problem of preventing adhesiving effect is bad（Fig. 7）.(2) adherence preventing material prepared by the present invention can be provided simultaneously with stopping blooding（Figure
2）, antibacterial（Fig. 5）And wound healing function（Fig. 6）.(3) the adherence preventing material shaping prepared by the present invention is easy, energy
The product of the forms such as sponge, film, gel is enough prepared into, is widely used in different operation antiblocking needs, is adapted to industry metaplasia
Production, has good potential applicability in clinical practice.
Brief description of the drawings
Fig. 1 is the sponge electronic scanner microscope figure that prevents adhesion prepared by the embodiment of the present invention 1.
Fig. 2 is the sponge whole blood coagulation results that prevent adhesion prepared by the embodiment of the present invention 2, wherein with anticoagulated whole blood in deionization
Control group is dissolved as in water.
Fig. 3 is Antiadhesive film cytotoxicity result prepared by the embodiment of the present invention 3, wherein thin with the logarithmic phase of normal growth
It is negative control that born of the same parents cultivate in the fresh sterile RAPI1640 culture mediums containing 10% hyclone, with the logarithmic phase of normal growth
It is positive control that cell is cultivated in 6% phenol solution, with the logarithmic phase cell of normal growth in Antiadhesive film leaching liquor
Cultivate as experimental group.
Fig. 4 be the embodiment of the present invention 4 prepare anti-adhesion gel at 37 DEG C storage modulus (G ') and loss modulus
The time sweep rheology of (G ") is as a result, as seen from the figure, the gelation time of the anti-adhesion gel is less than 6 seconds.
Fig. 5 be different anti-adhesion gels to the fungistatic effects of Escherichia coli, wherein a is the commercially available gel of Shandong company, b
The anti-adhesion gel prepared for the embodiment of the present invention 5, c are the commercially available gel of Hangzhou company, and d is the bacterium of normal growth.
Fig. 6 is the effect of the postoperative 7 days wound healings of SD rat skin wound models.A is SD rat skin surface of a wound moulds
The foundation of type, B are growing state of the surface of a wound after 7 days, and wherein control group is the blank group for being not added with any material, and experimental group is this hair
Anti-adhesion gel prepared by bright embodiment 6.
Fig. 7 crosses the postoperative 7 days preventing adhesiving effects of wiping model for SD rat cecals.Wherein control group is not added with any material that prevents adhesion
Material, experimental group are anti-adhesion gel prepared by the embodiment of the present invention 7, and commercial product is solidifying for preventing adhesion for Shandong company production
Glue.
Embodiment
Embodiment 1：
（1）At 4 DEG C that molecular weight is soluble in water for 1,000,000 derivatives of hyaluronic acids, obtaining 10mL mass volume ratios is
0.05% derivatives of hyaluronic acids solution；
（2）At 4 DEG C that molecular weight is soluble in water for 200,000 amination glucan derivative, obtaining 10mL mass volume ratios is
50% amination glucan derivative solution；
（3）By step at 4 DEG C（1）The derivatives of hyaluronic acids solution and step of gained（2）The amination glucan of gained spreads out
Biological solution by volume 20:Above-mentioned mixed solution, is then stood 48h, then in temperature by the lower mixing of 1 stirring at -20 DEG C
For -45 DEG C, vacuum is vacuum freeze drying 96h, so that material dewatering obtains the sponge that prevents adhesion under the conditions of 5 Pascals.
Embodiment 2：
（1）It is at 4 DEG C that molecular weight is soluble in water for 800,000 sodium alginate derivative, 10mL mass volume ratios are obtained as 50%
Sodium alginate derivative solution；
（2）At 4 DEG C that molecular weight is soluble in water for 300,000 oxidized hyaluronic acid derivative, obtaining 10mL mass volume ratios is
0.05% oxidized hyaluronic acid derivative solution；
（3）By step at 4 DEG C（1）The sodium alginate derivative solution and step of gained（2）The oxidized hyaluronic acid of gained is spread out
Biological solution by volume 2:The lower mixing of 8 stirrings, then the sorbitol solution that 0.2 mL mass fractions are 0.5% is slowly added dropwise
Into mixed solution, 30 minutes are stood at 4 DEG C.The mixed solution of above-mentioned addition sorbierite is then stood into 48h at -20 DEG C,
Then it is -45 DEG C in temperature, vacuum is under the conditions of 5 Pascals, and vacuum freeze drying 96h, finally obtains the anti-of plasticizer-containing
Adhesion sponge.
Embodiment 3：
（1）It is at 4 DEG C that molecular weight is soluble in water for 600,000 cellulose derivative, obtain the fibre that 10mL mass volume ratios are 3%
The plain derivative solution of dimension；
（2）It is at 4 DEG C that molecular weight is soluble in water for 300,000 amination bletilla polysaccharide derivative, obtain 10mL mass volume ratios
For 6% amination bletilla polysaccharide derivative solution；
（3）By step at 4 DEG C（1）The solution of cellulose derivative and step of gained（2）The amination bletilla polysaccharide of gained spreads out
Biological solution by volume 5:The lower mixing of 5 stirrings, is then stood the mixed solution by 300 mesh screens, vacuum deaerator, room temperature
Casting film-forming afterwards, so as to obtain Antiadhesive film.
Embodiment 4
（1）It is at 4 DEG C that molecular weight is soluble in water for 600,000 pulullan polysaccharide derivative, 10mL mass volume ratios are obtained as 8%
Pulullan polysaccharide derivative solution；
（2）It is at 4 DEG C that molecular weight is soluble in water for 300,000 oxidated chitin derivative, 10mL mass volume ratios are obtained as 8%
Oxidated chitin derivative solution；
（3）By step at 4 DEG C（1）The pulullan polysaccharide derivative solution and step of gained（2）The oxidated chitin of gained spreads out
Biological solution by volume 1:The lower mixing of 9 stirrings, then prevents adhesion the mixed solution when 25 DEG C of standings 24 are small so as to prepare
Gel.
Embodiment 5
（1）At 4 DEG C that molecular weight is soluble in water for 1,000,000 chitin derivativ, it is 10% to obtain 10mL mass volume ratios
Chitin derivativ solution；
（2）It is at 4 DEG C that molecular weight is soluble in water for 300,000 derivatives oxi-lignin, 10mL mass volume ratios are obtained as 8%
Derivatives oxi-lignin solution；
（3）By step at 4 DEG C（1）The chitin derivativ solution and step of gained（2）The derivatives oxi-lignin of gained
Solution by volume 2:The lower mixing of 8 stirrings, then by the mixed solution in 25 DEG C stand 24 it is small when so that prepare prevent adhesion it is solidifying
Glue.
Embodiment 6
（1）It is at 4 DEG C that molecular weight is soluble in water for 500,000 bletilla polysaccharide derivative, 10mL mass volume ratios are obtained as 15%
Bletilla polysaccharide derivative solution；
（2）It is at 4 DEG C that molecular weight is soluble in water for 300,000 oxidated chitin derivative, obtain the oxygen that mass volume ratio is 5%
Change chitin derivativ solution；
（3）By step at 4 DEG C（1）The bletilla polysaccharide derivative solution and step of gained（2）The oxidated chitin of gained derives
Thing solution by volume 2:The lower mixing of 8 stirrings, then by the mixed solution in 4 DEG C stand 48 it is small when so that prepare prevent adhesion it is solidifying
Glue.
Embodiment 7
（1）At 4 DEG C that molecular weight is soluble in water for 600,000 derivatives of hyaluronic acids, it is 5% to obtain 10mL mass volume ratios
Derivatives of hyaluronic acids solution；
（2）It is at 4 DEG C that molecular weight is soluble in water for 300,000 amination starch derivatives, 10mL mass volume ratios are obtained as 5%
Amination starch derivative solution；
（3）By step at 4 DEG C（1）The derivatives of hyaluronic acids solution and step of gained（2）The amination starch of gained derives
Thing solution by volume 1:The lower mixing of 9 stirrings, then prevents adhesion the mixed solution liquid when 50 DEG C of standings 12 are small so as to prepare
Gel.
Embodiment 8
（1）The hyaluronic acid that sodium alginate derivative and molecular weight that molecular weight is 800,000 are 600,000 is derived respectively at 4 DEG C
Thing is soluble in water, obtain sodium alginate derivative solution that 5mL mass volume ratios are 8% and 5mL mass volume ratios be 2% it is transparent
Matter acid derivative solution, respectively takes 5mL sodium alginates derivative solution and derivatives of hyaluronic acids solution to mix, is stirred under 300 rpm
Mix 3 it is small when obtain the mixed solution of homogeneous sodium alginate derivative and derivatives of hyaluronic acids.
（2）It is at 4 DEG C that molecular weight is soluble in water for 200,000 pullulan oxide derivative, obtain 10mL mass bodies
Product is than the pullulan oxide derivative solution for 3%；
（3）By step at 4 DEG C（1）The sodium alginate derivative of gained and the mixed solution and step of derivatives of hyaluronic acids
（2）The pullulan oxide derivative solution of gained by volume 3:The lower mixing of 7 stirrings, then by above-mentioned mixed solution in 20
DEG C stand 48 it is small when, so as to prepare anti-adhesion gel.
Embodiment 9
（1）It is at 4 DEG C that molecular weight is soluble in water for 300,000 chitin derivativ, obtain the first that 10mL mass volume ratios are 1%
Shell element derivative solution；
（2）It is respectively that the oxidation bletilla polysaccharide derivative that molecular weight is 200,000 and the oxidation that molecular weight is 100,000 is wooden at 4 DEG C
Plain derivative is soluble in water, obtains the oxidation bletilla polysaccharide derivative solution and 5mL mass volume ratios that 5mL mass volume ratios are 2%
For 4% derivatives oxi-lignin solution, respectively take 5mL oxidation bletilla polysaccharide derivative solutions and derivatives oxi-lignin molten
Liquid mixes, and the mixing of homogeneous oxidation bletilla polysaccharide derivative and derivatives oxi-lignin is obtained when stirring 3 is small under 300 rpm
Solution.
（3）By step at 4 DEG C（1）The chitin derivativ solution and step of gained（2）The oxidation bletilla polysaccharide of gained
The mixed solution of derivative and derivatives oxi-lignin is 5 by volume:The lower mixing of 5 stirrings, will then mix under above-mentioned stirring
Solution is closed when 37 DEG C of standings 18 are small, so as to prepare anti-adhesion gel.
Embodiment 10
（1）It is at 4 DEG C that molecular weight is soluble in water for 600,000 lignin derivative, obtain the wood that 10mL mass volume ratios are 1%
Quality derivative solution；
（2）It is at 4 DEG C that molecular weight is soluble in water for 200,000 aminated alginic acid sodio-derivative, obtain 10mL mass volume ratios
For 5% aminated alginic acid sodio-derivative solution；
（3）By step at 4 DEG C（1）The lignin derivative solution and step of gained（2）The aminated alginic acid sodium of gained spreads out
Biological solution by volume 1:The lower mixing of 9 stirrings, then by ethanol solution and 0.3 mL mass of the 0.5 mL mass fractions for 2%
The glycerite that fraction is 5% is slowly dropped in above-mentioned mixed solution, and 30 minutes are stood at 4 DEG C.Then by above-mentioned addition ethanol
48h is stood at -20 DEG C with the mixed solution of glycerine, is then -45 DEG C in temperature, vacuum is vacuum under the conditions of 5 Pascals
96h is freeze-dried, finally obtains the sponge that prevents adhesion of plasticizer-containing.
Embodiment 11
（1）It is at 4 DEG C that molecular weight is soluble in water for 500,000 sodium alginate derivative, 10mL mass volume ratios are obtained as 0.8%
Sodium alginate derivative solution；
（2）It is at 4 DEG C that molecular weight is soluble in water for 200,000 oxidized starch derivative, 10mL mass volume ratios are obtained as 10%
Oxidized starch derivative solution；
（3）By step at 4 DEG C（1）The sodium alginate derivative solution and step of gained（2）The oxidized starch derivative of gained
Solution by volume 8:0.1g tetracyclines, are then added in above-mentioned mixed solution, 30 points are stood at 4 DEG C by the lower mixing of 2 stirrings
Clock.The mixed solution of above-mentioned addition tetracycline is then stood into 48h at -20 DEG C, is then -45 DEG C in temperature, vacuum 5
Under the conditions of Pascal, vacuum freeze drying 96h, finally obtains the sponge that prevents adhesion containing antiseptic.
Embodiment 12
（1）It is at 4 DEG C that molecular weight is soluble in water for 1,000,000 starch derivatives, obtain the shallow lake that 10mL mass volume ratios are 8%
Powder derivative solution；
（2）At 4 DEG C that molecular weight is soluble in water for 200,000 oxidized sodium alginate derivative, obtaining 10mL mass volume ratios is
10% oxidized sodium alginate derivative solution；
（3）By step at 4 DEG C（1）The starch derivative solution and step of gained（2）The oxidized sodium alginate of gained presses volume
Than for 4:0.2g penicillin and 0.1g lincomycins, are then added in above-mentioned composite solution by the lower mixing of 6 stirrings, quiet at 4 DEG C
Put 30 minutes., then the mixed solution of above-mentioned addition penicillin and lincomycin is contained when 24 DEG C of standings 36 are small so as to prepare
The anti-adhesion gel of antiseptic.
Experimental example 1：Prevent adhesion sponge pattern observation experiment
Sample holder is got out, two-sided conductive copper glue is posted on stent, by the sponge that prevents adhesion as prepared by embodiment 1 on a small quantity
Bonded naturally with conductive copper glue, surface gold-plating processing 30 carries out sample using sputter (Hitachi, E-1045, Japan)
S, surface topography observation is carried out using scanning electron microscope (SEM, Hitachi S-4800, Japan).Experimental result table
Bright, the sponge that prevents adhesion prepared by embodiment 1 has good pore structure.
Experimental example 2：Whole blood clotting assay
Weigh prevent adhesion sponges of the 10mg as prepared by embodiment 2 to be positioned over inside surface plate, cultivated at 37 DEG C in 5min, then
200ul anticoagulations are slowly dropped to sample surfaces, are subsequently added into 20ul, 0.2mol/L calcium chloride solutions, continue 37 DEG C of cultures
5min.Later, 10ml distilled water is carefully added into surface plate and the shake culture 10min under 30rpm rotating speeds in shaking table, together
When repeated the above steps using being not added with adherence preventing material as control group, observation prevents adhesion suction-operated of the sponge to red blood cell.Experiment
The results show that there is the sponge that prevents adhesion as prepared by embodiment 1 good hemadsorption to act on, there is hemostasia effect.
Experimental example 3：Cytotoxicity experiment
Experimental cell uses the eugonic L929 cell line cells of h of 48 h ~ 72.Culture medium is 10% (V/V) tire ox of addition
The RAPI1640 of serum.Negative control（Fresh sterile RAPI1640 culture mediums containing 10% hyclone）, experimental group（Take 0.5 g
The fresh sterile RAPI1640 cultures of Antiadhesive film and 10 mL containing 10% hyclone described in embodiment 3 are based on 37 DEG C of extractions
24h).Positive controls（Mass fraction is 5% phenol solution）, test and carried out on 96 orifice plates.The cell of initial incubation liquid is close
Degree is about 50,000/mL, and 200 μ L cell culture fluids are added per hole, make number of cells about 10000 in every hole.Three are repeated per hole
Secondary, 24 h of culture, then discard former culture medium in the constant incubator of 37 DEG C of 5% (V/V) carbon dioxide/air.It is negative right
According to the fresh sterile RAPI1640 culture mediums for adding 200 μ L to contain 10% hyclone in group, experimental group adds 20 μ L, 40 μ L, 60 respectively
μ L, 80 μ L, 100 μ L leaching liquors, adding negative controls makes liquor capacity in every hole be 200 μ L.Positive controls add respectively
20 μ L, 40 μ L, 60 μ L, 80 μ L, the phenol solution that 100 μ L mass fractions are 5%, adding negative controls makes solution in every hole
Volume is 200 μ L.Then, which is put into after being further cultured for 48 h in 37 DEG C of constant incubators and takes out culture plate, discard training
Solution in plate is supported, 20 μ L MTT solution are added per hole, continues to cultivate 4h, then sucks stoste, add 150 μ L dimethyl sulfoxides,
Concussion ten minutes, absorbance (ABS) is measured in immune microplate reader at 570 nm wavelength.
Versus cell activity (%)=ABS570 experimental groups/ ABS570 negative controls×100%
According to GB/T16886.5-2003 vitro cytotoxicity judgment criteria, Antiadhesive film leaching liquor described in embodiment 35 ~
25 μ g/mL have 0 grade of cytotoxicity, show good biocompatibility.
Experimental example 4：Gel time determination experiment
Using rotary cone and plate rheometer（TA, DHR-1, USA）Detect the elasticity modulus according to the gel prepared by embodiment 4
G' and loss modulus G " change with time, and wherein temperature is set to 37 °C, 40 mm of aluminium sheet diameter, 25 μm of sample gap, frequency
Rate is 0.1-100 rad/s, detection time 600s.Test result indicates that the gelation time according to the gel prepared by embodiment 4
Less than 6 seconds.
Experimental example 5：Bacteriostatic experiment
Test bacterium and select Escherichia coli.Add gels of the 200 μ L as prepared by embodiment 5 per hole in 48 orifice plates, treat completely into
After glue, add 10 μ L Escherichia coli bacteria liquids (10 per hole6CFU/mL) in gel surface, 37 DEG C of 2 h of incubation, then add 1 per hole
Bacterium is resuspended in mL PBS, with 10 μ L bacterium solutions (106CFU/mL it is control group) to be dissolved in 1 mL PBS, is then incubated 24 h, with
Afterwards, 40 μ L Escherichia coli bacteria liquids are taken out per hole after 37 DEG C of 24 h of incubation of agar plate, observe agar plate surface bacterial growth
Situation.Wherein a is the commercially available gel of Shandong company, and b is anti-adhesion gel prepared by the embodiment of the present invention 5, and c is public for Hangzhou
The commercially available gel of department, d are the bacterium of normal growth.
Test result indicates that the anti-adhesion gel as prepared by embodiment 5 has good suppression Escherichia coli Growth work(
Effect.
Experimental example 6：Wound healing is tested
The surface of a wound that diameter is about 4 cm will be taken after experimental animal SD rat anesthesias, disinfection respectively in backbone both sides, experimental group is will
Adherence preventing material prepared by embodiment 6 is injected in the surface of a wound, then with aseptic dressing flap coverage；The surface of a wound of control group is then only with disappearing
Malicious dressing flap coverage.Postoperative 7 days the results show that using 6 adherence preventing material of embodiment SD rats, back wound healing is more
It hurry up, have the function that to promote healing.
Experimental example 7：Prevent adhesion experiment
Abdomen will be opened after experimental animal SD rat anesthesias, disinfection, be about 2cm circular wounds in stomach wall peel diameter, form bleeding table
Face；Then there is obvious bleeding in caecum surface operation brush friction to the caecum placenta percreta being in contact with the stomach wall surface of a wound, then by 1
ML anti-adhesion gels are applied to the stomach wall surface of a wound and the caecum surface of a wound respectively, and after gel solidification, suturing them, after 7 days, it is blind to open abdomen observation
Intestinal adhesion situation.Wherein control group is not added with any adherence preventing material, and experimental group is solidifying for preventing adhesion for the preparation of the embodiment of the present invention 7
Glue, commercial product are the anti-adhesion gel of Shandong company production.
Claims (10)
1. a kind of have the function of hemostasis, antibacterial, the degradable adherence preventing material for promoting wound healing, it is characterised in that：Described
Adherence preventing material includes the solution of one or more natural polysaccharides and the solution of one or more modified natural polysaccharide through schiff bases
After reaction and/or acylation reaction form compound, it is prepared through vacuum freeze drying, wherein the mass body of natural polysaccharide solution
For product than being 0.05% ~ 50%, the mass volume ratio of modified natural polysaccharide solution is 0.05% ~ 50%, natural polysaccharide solution and modified day
The volume ratio of right polysaccharide solution is 100:1~1:100.
2. having the function of hemostasis, antibacterial, the degradable adherence preventing material for promoting wound healing according to claim 1, it is special
Sign is：Wherein natural polysaccharide is chitin derivativ, derivatives of hyaluronic acids, sodium alginate derivative, starch derivatives, Portugal
Polysaccharid derivative, pulullan polysaccharide derivative, bletilla polysaccharide derivative, cellulose derivative, one kind in lignin derivative
It is or a variety of.
3. having the function of hemostasis, antibacterial, the degradable adherence preventing material for promoting wound healing according to claim 1, it is special
Sign is：Wherein modified natural polysaccharide includes amination derivatives of hyaluronic acids, aminated alginic acid sodio-derivative, and amination is formed sediment
Powder derivative, amination glucan derivative, amination pulullan polysaccharide derivative, amination bletilla polysaccharide derivative, amino
Cellulose derivative, amination lignin derivative, oxidated chitin derivative, oxidized hyaluronic acid derivative, oxidation sea
Alginic acid sodio-derivative, oxidized starch derivative, oxidized dextran derivative, pullulan oxide derivative, the oxidation bletilla striata are more
Sugar derivatives, oxycellulose derivative, the one or more in derivatives oxi-lignin.
4. having the function of hemostasis, antibacterial, the degradable adherence preventing material for promoting wound healing according to claim 1, it is special
Sign is：The sponge that prevents adhesion further includes antibacterials, and the antibacterials include penicillins, cephalosporins, β-interior
Lactamase inhibitor, aminoglycoside, macrolides, Tetracyclines, lincomycin class, polypeptide, quinolones, sulfamido
In one kind or its combination, the antibacterials mixed with adherence preventing material after preparation the adherence preventing material containing antibacterials.
5. a kind of preparation method using the sponge that prevents adhesion made of claim 1-4 any one of them adherence preventing materials, its
Feature, which comprises the following steps, to be made：
（a）With deionized water dissolving or swelling natural polysaccharide and modified natural polysaccharide, respective solution is prepared；Wherein natural polysaccharide is molten
The mass volume ratio of liquid is 0.05% ~ 50%, and the mass volume ratio of modified natural polysaccharide solution is 0.05% ~ 50%；
（b）Natural polysaccharide solution is mixed with modified natural polysaccharide solution, wherein natural polysaccharide solution and modified natural polysaccharide is molten
The volume ratio of liquid is 100:1~1:100, preferably 20:1~1:20；
（c）Solid-state is condensed at -10 DEG C ~ -50 DEG C of the mixed solution warp, then the moisture under the conditions of vacuum is 1 ~ 10 Pascal
It is gaseous state and vacuum freeze drying by solid state sublimation, preparation prevents adhesion sponge.
6. prevent adhesion sponge according to claim 5, it is characterised in that：Further include plasticizer, the step（c）It is middle to mix
After solution is mixed with plasticizer, through condensing into solid-state at -10 DEG C ~ -50 DEG C, then vacuum be 1 ~ 10 Pascal under the conditions of water
Point it is gaseous state and vacuum freeze drying by solid state sublimation, preparation prevents adhesion sponge, and the plasticizer accounts for the sponge quality that prevents adhesion
0.05% ~ 10%, the plasticizer selects glycerine, sorbierite, ethanol, one kind in poly- ethanedioic acid or its combination.
7. a kind of preparation method using Antiadhesive film made of claim 1-4 any one of them adherence preventing materials, it is special
Sign is to comprise the following steps：
（a）With deionized water dissolving or swelling natural polysaccharide and modified natural polysaccharide, respective solution is prepared；Wherein natural polysaccharide
The mass volume ratio of solution is 0.05% ~ 50%, and the mass volume ratio of modified natural polysaccharide solution is 0.05% ~ 50%；
（b）Natural polysaccharide solution is mixed with modified natural polysaccharide solution, wherein natural polysaccharide solution and modified natural polysaccharide is molten
The volume ratio of liquid is 100:1~1:100, preferably 20:1~1:20；
（c）By the mixed solution by 300 mesh screens, vacuum deaerator, casting film-forming after room temperature is stood.
8. Antiadhesive film according to claim 7, it is characterised in that：Further include antibacterials, the step（c）It is middle to mix
After solution is mixed with antibacterials, by 300 mesh screens, vacuum deaerator, casting film-forming after room temperature is stood, is drying to obtain and prevents adhesion
Film.
9. Antiadhesive film according to claim 7, it is characterised in that：Further include plasticizer, the step（c）In will mix it is molten
After liquid is mixed with plasticizer, by 300 mesh screens, vacuum deaerator, casting film-forming after room temperature is stood, is drying to obtain Antiadhesive film,
The plasticizer selects glycerine, sorbierite, ethanol, one kind in poly- ethanedioic acid or its combination, and wherein plasticizer, which accounts for, prevents adhesion
The 0.05% ~ 10% of film quality.
10. a kind of preparation method using anti-adhesion gel made of claim 1-4 any one of them adherence preventing materials, its
It is characterized in that comprising the following steps：
（a）With deionized water dissolving or swelling natural polysaccharide and modified natural polysaccharide, respective solution is prepared；Wherein natural polysaccharide is molten
The mass volume ratio of liquid is 0.05% ~ 50%, and the mass volume ratio of modified natural polysaccharide solution is 0.05% ~ 50%；
（b）Natural polysaccharide solution is mixed with modified natural polysaccharide solution, wherein natural polysaccharide solution and modified natural polysaccharide is molten
The volume ratio of liquid is 100:1~1:100, preferably 20:1~1:20；
（c）By 4 ~ 60 DEG C of the mixed solution stand 10 minutes ~ 48 it is small when, prepare anti-adhesion gel.
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Address after: 325001 No.1, Jinlian Road, Longwan District, Wenzhou City, Zhejiang Province Patentee after: Wenzhou Research Institute of Guoke (Wenzhou Institute of biomaterials and Engineering) Address before: 325001 enterprise development headquarters building, science and Technology City, 156 Wenchang Road, high tech Zone, Wenzhou City, Zhejiang Province Patentee before: Wenzhou Research Institute of Chinese Academy of Sciences (Wenzhou Institute of Biomaterials and Engineering) Address after: 325001 enterprise development headquarters building, science and Technology City, 156 Wenchang Road, high tech Zone, Wenzhou City, Zhejiang Province Patentee after: Wenzhou Research Institute of Chinese Academy of Sciences (Wenzhou Institute of Biomaterials and Engineering) Address before: 325000 enterprise development headquarters building, science and Technology City, No.156 Wenchang Road, high tech Zone, Longwan District, Wenzhou City, Zhejiang Province Patentee before: WENZHOU INSTITUTE OF BIOMATERIALS & ENGINEERING, CNITECH, CAS |
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| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20221221 Address after: 246000 No. 2, Shengli Road, east of Wenyuan Road and south of Longgang Road, Daqiao Street, Yixiu District, Anqing City, Anhui Province Patentee after: Anhui Zhongke Maide Medical Technology Co.,Ltd. Address before: 325001 No.1, Jinlian Road, Longwan District, Wenzhou City, Zhejiang Province Patentee before: Wenzhou Research Institute of Guoke (Wenzhou Institute of biomaterials and Engineering) |