CN106983905A - A kind of injectable type self-healing hemostatic material and its preparation method and application - Google Patents
A kind of injectable type self-healing hemostatic material and its preparation method and application Download PDFInfo
- Publication number
- CN106983905A CN106983905A CN201710335184.0A CN201710335184A CN106983905A CN 106983905 A CN106983905 A CN 106983905A CN 201710335184 A CN201710335184 A CN 201710335184A CN 106983905 A CN106983905 A CN 106983905A
- Authority
- CN
- China
- Prior art keywords
- gelatin
- gel particles
- micro
- aqueous solution
- hemostatic material
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- 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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/10—Polypeptides; Proteins
- A61L24/104—Gelatin
-
- 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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/001—Use of materials characterised by their function or physical properties
-
- 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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/001—Use of materials characterised by their function or physical properties
- A61L24/0015—Medicaments; Biocides
-
- 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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/001—Use of materials characterised by their function or physical properties
- A61L24/0031—Hydrogels or hydrocolloids
-
- 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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/046—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
-
- 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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/08—Polysaccharides
-
- 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
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
- A61L2300/414—Growth factors
-
- 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
-
- 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
- A61L2400/04—Materials for stopping bleeding
Landscapes
- Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Surgery (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Dispersion Chemistry (AREA)
- Materials For Medical Uses (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a kind of injectable type self-healing hemostatic material and its preparation method and application.The present invention obtains injection-type self-healing hemostatic material using the polymer-network method particle of belt surface electric charge by the electrostatic interaction self assembly between particle or between particle and macromolecule.Because the electrostatic interaction between micro-gel particles or between particle and macromolecule is physical crosslinking and has invertibity, the hemostatic material that the inventive method is prepared has good syringeability, self-healing capability and preferable mechanical strength, and need not introduce chemical crosslink reaction in injection and solidification process, different from chemical macromolecule, also small molecule crosslinking agent is not introduced, good biocompatibility, it is degradable to absorb, have no toxic side effect, security is good, makes it possible its extensive use in biomedical sector.The hemostatic material of the present invention can also realize quick-acting haemostatic powder for the bleeding tissue that amount of bleeding is big, vascular pressure is high.
Description
Technical field
The invention belongs to biology medical material technical field, it is related to a kind of hemostatic material and preparation method thereof, specifically,
People and the other food in one's mouths that the reasons such as surgical operation (containing Minimally Invasive Surgery) and wound are caused can be directly applied to the present invention relates to one kind
The histoorgan surface of a wound of newborn animal, for stop blooding, close wound, reduce tissue ooze out, promotion organization reparation and protection the surface of a wound
The gelatin-based hemostasis pulvis and the preparation method of hemostasis gel material of the purposes such as tissue.
Background technology
Need that the wound that different reasons are caused is stopped blooding and closed in surgical procedures, therefore hemostatic material right and wrong
An often important class bio-medical material, with important research and development and application value.Hemostatic material clinical conventional at present is from work
It can be divided into three classes with mechanism：First kind hemostatic material is wound location is accelerated blood coagulation by the physically or chemically effect of material
(such as hemostatic gauze, high molecular polysaccharide, inorganic zeolite)；Equations of The Second Kind be directly or indirectly provide external hemostatic composition,
Using excite itself clotting mechanism speed up to blood coagulation (such as fibrin class hemostatic material, containing fibrin ferment and blood clotting factors
Hemostatic material)；3rd class is to organizing the very strong direct wound closure of adhesion strength, so as to realize hemostasis (such as α cyano group using material
Acrylate materials).But all there is many bottlenecks in different types of hemostatic material.
For realized using hemostatic mechanism blood clotting blood product for, its have the disadvantage they only it is less to flow velocity go out
Blood effectively, therefore must coordinate hemostasis clamp when using, this allows for above-mentioned hemostatic material usually needs surgery to cure when in use
Raw look-ahead amount of bleeding or repeated multiple times use haemostatic clamp.In addition, being loaded with the hemostatic article of thrombin class coagulant makes
During, thrombin class protein molecular is easily outer to be spilt in normal blood vessels, therefore there is induction normal blood flow generation blood coagulation generation
The risk of thrombus, thus independent of body clotting mechanism and can rapid wound closure hemostatic article in surgical operation more by green grass or young crops
Look at.
And the hemostatic article of hemostasis is realized by physical action, such as hemostatic gauze, sponge need to pass through in hemostasis
Physics compressing is carried out to the bleeding surface of a wound, while material by absorbing the water in blood, promotes blood coagulation, anthemorrhagic speed is slow, effect compared with
Difference.Also, this kind of material is only effective to the less bleeding of flow velocity, therefore must coordinate hemostasis clamp when using, and this is allowed for
State hemostatic material usually needs surgeon's look-ahead amount of bleeding or repeated multiple times use haemostatic clamp when in use.Meanwhile,
Traditional hemostatic material (such as cotton yarn, bandage) imitates for the hemostasis of irregular shape, the live common wound of deep, narrow, arteriorrhexis
Fruit is very undesirable.In addition, also there are other shortcomings：1) postoperative suture is influenceed, increases new surface of a wound bleeding；2) it can not drop naturally
Solution, forms residual in vivo；3) hemostasis needs prolonged pressed by external force to realize that physics stops blooding, for fragile nerve or
Brain tissue is not applied to.Another kind of inorganic matter hemostatic material, including zeolite, kaolin etc., this kind of material use natural silicon aluminium acid
The characteristics such as salt material loose structure, high-specific surface area, by absorbing the moisture in blood, concentrate the local hemostatic composition of the surface of a wound,
So as to realize quick-acting haemostatic powder.And the mesoporous silicon material manually prepared equally has the characteristics such as loose structure, high-specific surface area, and
It is same that effectively hemostasis can be achieved compared to the natural aluminosilicate silicate material advantage such as have composition, particle diameter, aperture controllable.However, this kind of
Material is helpless for the big bleeding of blood flow, also limited for large area surface of a wound effect；Although and this kind of material Jie is local
Tissue reaction is benign, but the mesoporous silicon grain for entering vascular and histoorgan can cause serious general toxicity.Therefore,
Exploitation turns into medical science and biomaterial science for scene and pre hospital care, quick, safe and effective new hemostatic material
Important topic in field.
The gel-like hemostatic article of injectable enters clinic gradually in recent years because of its clinical ease for operation, suitable for Minimally Invasive Surgery
Hemostat application.At present the hemostasis gel class product that clinically uses of the country relies primarily on import, wherein Johnson Co.Occupy the main market share.The material is made up of gelatin, water-soluble rear quick formation foam-like jello, production
Product have good syringeability.But there are problems in hemostasis.First, gel mechanics intensity difference, rheometer is surveyed
Obtain modulus of elasticity<1kPa, hemostasis can not be realized for the bleeding tissue that amount of bleeding is big, vascular pressure is high；Secondly, the material and group
It is less sticky between knitting, for the slightly larger surface of a wound of clinical amount of bleeding, it is difficult to realize quick-acting haemostatic powder.Therefore, exploitation has good injectable
Property, the clinical manipulation performance such as plasticity, available for CBF is big, high blood pressure histoorgan surface of a wound hemostasis new type gel material
With important Practical significance.
The content of the invention
In view of the problem of hemostatic material described above exists in the prior art, the present invention provides a kind of injectable type certainly
Heal hemostatic material and preparation method thereof.The hemostatic material for preparing of method of the present invention have good syringeability, from
Healing properties and preferable mechanical strength, quick-acting haemostatic powder can also be realized for the bleeding tissue that amount of bleeding is big, vascular pressure is high.
Technical scheme is as follows：
A kind of injectable type self-healing hemostatic material, is prepared by the following method：
(1) using gelatin as raw material, it is made to dissolve by heating in deionized water, configuration concentration is 1~10w/v% gelatin water
Solution, tune pH value is 1-6 or 8-14, is added into solution>The polar organic solvent of 2 times of liquor capacities, generates gelatin microgel
Particle dispersion, adds cross-linking agents and reacts 1~12h, centrifugation, cleaning obtain gelatin micro-gel particles；
Wherein, the zeta potentials of the gelatin micro-gel particles be -30~+30mV, the gelatin micro-gel particles it is straight
Footpath is 20nm~2 μm；
(2) the surface zeta potential potential for preparing step (1)>+ 10mV gelatin micro-gel particles, are dispersed in pH<5
Acidic aqueous solution or pH>In 9 alkaline aqueous solution, the dispersion liquid of positively charged gelatin micro-gel particles must be dispersed with, then with
Negatively charged polymeric particles dispersion liquid compares 1 according to granule number:10~10:1 blending, or and isoelectric point<6 it is negatively charged
The hydrophilic macromolecule aqueous solution according to gelatin micro-gel particles and hydrophilic macromolecule mass ratio 1:10~10:1 blending, is used
PH adjusting agent adjusts pH to 7.0, and freeze-drying obtains gelatin micro-gel particles freeze-dried powder I；
(3) the surface zeta potential potential for preparing step (1)<- 10mV gelatin micro-gel particles, are dispersed in pH<5
Acidic aqueous solution or pH>In 9 alkaline aqueous solution, obtain the dispersion liquid of gelatin micro-gel particles, then with positively charged macromolecule
Particle dispersion compares 1 according to granule number:10~10:1 mixing, or and isoelectric point>8 positively charged hydrophilic macromolecule water
Solution according to gelatin micro-gel particles and hydrophilic macromolecule mass ratio 1:10~10:1 blending, pH is adjusted extremely with pH adjusting agent
7.0, freeze-drying obtains gelatin micro-gel particles freeze-dried powder II；
(4) the surface zeta potential potential for preparing step (1) is dispersed in for -10~+10mV gelatin micro-gel particles
In neutral aqueous solution, then with another surface zeta potential potential -10~+10mV polymeric particles dispersion liquid according to granule number ratio
1:10~10:1 blending, or with the hydrophilic macromolecule aqueous solution of isoelectric point 6~8 according to gelatin micro-gel particles and hydrophily
High molecular mass ratio 1:10~10:1 blending, freeze-drying obtains gelatin micro-gel particles freeze-dried powder III；
(5) by gelatin micro-gel particles freeze-dried powder I, gelatin micro-gel particles freeze-dried powder II or gelatin micro-gel particles
Freeze-dried powder III is blended with aqueous solution respectively, obtains injectable type self-healing hemostatic material；
Wherein, the surface charge of described positively charged polymeric particles is+5~+60mV, negatively charged macromolecule
The surface charge of particle is -5~-60mV, and a diameter of 100nm~20 μm of the polymeric particles are obtained in step (5)
Colloidal solid percentage of the total volume is 50vol%~150vol% in injectable type self-healing hemostatic material；In step
(3), the molecular weight of the hydrophilic macromolecule described in (4) and (5) is 1k~500kDa.
In the preparation method of the above-mentioned injectable type self-healing hemostatic material of the present invention, the gelatin that step (1) is prepared
Micro-gel particles, are freeze-dried obtained gelatin micro-gel particles freeze-dried powder and are blended with aqueous solution, can also obtain this hair
Bright described injectable type self-healing hemostatic material, but its performance is worse than the injectable type that other method in the present invention is prepared
Self-healing colloidal gel.
In the preparation method of the above-mentioned injectable type self-healing hemostatic material of the present invention, in step (1), according to adjustment gelatin
The concentration of the aqueous solution, the addition of polar organic solvent, cross-linking reaction time etc., can be prepared with different zeta potentials
With the gelatin micro-gel particles of diameter.In above-mentioned technical proposal of the present invention, aqueous gelatin solution concentration is preferably 1~10w/
V%, more preferably 2.5~5w/v%；The addition of polar organic solvent is preferably aqueous gelatin solution>2 times, more preferably 3
~6 times, gelatin micro-gel particles diameter is preferably 20nm~5 μm, more preferably 100nm~2000nm.
In the preparation method of the above-mentioned injectable type self-healing hemostatic material of the present invention, in step (2), (3) and (4), gelatin
Colloidal gel prepared by the scale effect of the dispersion liquid of micro-gel particles and the granule number in each organic polymer particle dispersion
Modulus of elasticity and self-repair efficiency, the ratio of granule number is preferably 1 in two kinds of dispersion liquids in the present invention:10~10:1, it is more excellent
Elect 1 as:5~5:1, the two kinds of particles diameter ratio of blending is 1:5~5:When 1, it can obtain that there is more high elastic modulus and selfreparing
The injectable type self-healing hemostatic material of efficiency, if the difference of diameter is excessive, resulting colloidal gel modulus of elasticity declines,
Self-repair efficiency is reduced.
It is in the above-mentioned technical proposal of the present invention, the gelatin micro-gel particles freeze-dried powder I, the gelatin that prepare is micro- solidifying
Glue particle freeze-dried powder II or gelatin micro-gel particles freeze-dried powder III are applied directly as hemostatic material, can also with it is appropriate
Aqueous solution is blended, and prepares the injectable type self-healing hemostatic material of colloidal gel shape, its modulus of elasticity be 1Pa~
100kPa, preferably 20kPa~100kPa.
Further, in the above-mentioned technical solutions, described positively charged polymeric particles dispersion liquid is with chitosan, A
One or more in type gelatin, polyacrylamide, NIPA, polyethyleneimine are prepared into as raw material
Arrive, described negatively charged polymeric particles dispersion liquid is with hyaluronic acid, alginic acid, type A gelatin, type B gelatin or polypropylene
One or more in acid are prepared as raw material, and described surface zeta potential potential divides in -10~+10mV polymeric particles
Dispersion liquid is prepared using the one or more in collagen, albumin, gelatin as raw material.The positively charged macromolecule
The polymeric particles of particle dispersion or negatively charged polymeric particles dispersion liquid or surface zeta potential potential in -10~+10mV
Dispersion liquid, those skilled in the art can prepare according to the conventional formulation techniques of polymeric particles, in this application not
Statement in detail again.
Described isoelectric point>8 positively charged hydrophilic macromolecule is chitosan, type A gelatin, polyacrylamide, poly-
One or more in (NIPA), polyethyleneimine, the isoelectric point<6 negatively charged hydrophily is high
Molecule is the one or more in hyaluronic acid, alginic acid, type A gelatin, type B gelatin or polyacrylic acid, described isoelectric point 6~
8 macromolecule is the one or more in collagen, albumin, gelatin, polyvinyl alcohol, polyethylene glycol.
Further, in the above-mentioned technical solutions, the polar organic solvent described in step (1) be methanol, it is ethanol, different
One or more of combinations in propyl alcohol, butanol, acetone, acetonitrile, tetrahydrofuran；Described crosslinking agent be glutaraldehyde, glyceraldehyde,
One kind or several in formaldehyde, carbodiimide, saturated dihalide, isocyanates, diisocyanate, glutamine transaminage, Geniposide
Kind.
Further, in the above-mentioned technical solutions, crosslinking agent in the reaction system of the cross-linking reaction described in step (1)
Mol ratio with amino group in gelatin is 0.1~10.What the mol ratio influence of crosslinking agent and amino group in gelatin was formed
The degree of cross linking of gelatin micro-gel particles, the too high gelatin miniflow intensity of the degree of cross linking is higher, surface charge is more likely to negatively charged, crosslinking
Spend in the isoelectric point that low gelatine microsphere low intensity, surface charge depend on gelatin material, currently preferred technical scheme and hand over
Connection degree it is relatively low when more preferably, it is preferred that the mol ratio of amino group will be controlled 0.5 in heretofore described crosslinking agent and gelatin
~5.
Further, in the above-mentioned technical solutions, the acidic aqueous solution and alkalescence described in step (2) and step (3)
Contained ion concentration is respectively less than 200mM in the aqueous solution.In described acidic aqueous solution and alkaline aqueous solution it is contained from
The species of son is not particularly limited, can using this area it is conventional be used for adjust acid or alkalescence reagent, such as hydrochloric acid, sulfuric acid,
Acetic acid, calcium hydroxide, potassium hydroxide, ammoniacal liquor, sodium carbonate etc..
Further, in the above-mentioned technical solutions, the pH adjusting agent described in step (2) and step (3) includes acidity
Material and alkaline matter, the acidic materials are glucolactone, HCl, HNO3、H2SO4In one or more, the alkali
Property material be urea and the combination of urase or sodium hydroxide, calcium hydroxide, potassium hydroxide, ammoniacal liquor in one or more.
Further, in the above-mentioned technical solutions, the aqueous solution described in step (5) be ion concentration 120~
200mM, pH value is in 6~8 any aqueous solution, the aqueous solution of hydrophilic macromolecule, not water-soluble nanoparticulate dispersion
One or more of combinations.
Further, in the above-mentioned technical solutions, bioactive substance and/or excipient are contained in the aqueous solution.
Wherein described excipient is solvent, decentralized medium, coating agent, surfactant, antioxidant, preservative, isotonic agent, bonding
Agent, lubricant, pigment, and combinations thereof or the like；The bioactive substance is coagulant, anti-infectives, disappeared
At least one of scorching medicine, biological activity protein medicine.The coagulant is selected from collagen, gelatin, oxycellulose, carboxylic first
Base cellulose, chitosan, hyaluronic acid, sodium alginate, kaolin, fibrin ferment, fibrin, calcium agent, nucleoprotamine, polypeptide,
One kind or its combination in peptide, amino acid；The anti-infectives are selected from antibiotic, antibiotic preparation, anti-virus formulation, anti-
Fungi-medicine, anti-ulcer medicament, one kind of Chinese medicine preparation or its combination；The anti-inflammation drugs are selected from non-steroid, steroid
Antiphlogistic, anti-ulcer medicament, one kind of Chinese medicine preparation or its combination.
The present invention also provides injectable type self-healing hemostatic material described above and prepared in body surface organization, body cavity
Histoorgan have the stopping blooding of the blood surface of a wound, it is anti-be adhered, anti-infection, in the hemostatic article of promotion organization healing and/or closing wound
Application.Wherein described hemostatic article can using hemostasis pulvis, hemostasis granules, hemostatic ball, hemostasis aerosol, styptic sponge,
The modes such as hemostasis gel, haemostatic membrane, tampon or hemostatic adhesive bandage.
Beneficial effects of the present invention：
1. and difference and the advantage of traditional hemostatic material；
Need that the wound that different reasons are caused is stopped blooding and closed in surgical procedures, therefore hemostatic material right and wrong
An often important class bio-medical material, with important research and development and application value.Hemostatic material clinical conventional at present is from work
It can be divided into three classes with mechanism：First kind hemostatic material is wound location is accelerated blood coagulation by the physically or chemically effect of material
(such as hemostatic gauze, high molecular polysaccharide, inorganic zeolite)；Equations of The Second Kind be directly or indirectly provide external hemostatic composition,
Using excite itself clotting mechanism speed up to blood coagulation (such as fibrin class hemostatic material, containing fibrin ferment and blood clotting factors
Hemostatic material)；3rd class is to organizing the very strong direct wound closure of adhesion strength, so as to realize hemostasis (such as α cyano group using material
Acrylate materials).But all there is many bottlenecks in different types of hemostatic material.
For the hemostatic article using hemostatic mechanism to realize blood clotting, its have the disadvantage they only it is less to flow velocity go out
Blood effectively, therefore must coordinate hemostasis clamp when using, this allows for above-mentioned hemostatic material usually needs surgery to cure when in use
Raw look-ahead amount of bleeding or repeated multiple times use haemostatic clamp.In addition, being loaded with the hemostatic article of thrombin class coagulant makes
During, thrombin class protein molecular is easily outer to be spilt in normal blood vessels, therefore there is induction normal blood flow generation blood coagulation generation
The risk of thrombus, thus independent of body clotting mechanism and can rapid wound closure hemostatic article in surgical operation more by green grass or young crops
Look at.
And the hemostatic article of hemostasis is realized by physical action, such as hemostatic gauze, sponge need to pass through in hemostasis
Physics compressing is carried out to the bleeding surface of a wound, while material by absorbing the water in blood, promotes blood coagulation, anthemorrhagic speed is slow, effect compared with
Difference.Also, this kind of material is only effective to the less bleeding of flow velocity, therefore must coordinate hemostasis clamp when using, and this is allowed for
State hemostatic material usually needs surgeon's look-ahead amount of bleeding or repeated multiple times use haemostatic clamp when in use.Meanwhile,
Traditional hemostatic material (such as cotton yarn, bandage) imitates for the hemostasis of irregular shape, the live common wound of deep, narrow, arteriorrhexis
Fruit is very undesirable.In addition, also there are other shortcomings：1) postoperative suture is influenceed, increases new surface of a wound bleeding；2) it can not drop naturally
Solution, forms residual in vivo；3) hemostasis needs prolonged pressed by external force to realize that physics stops blooding, for fragile nerve or
Brain tissue is not applied to.Another kind of inorganic matter hemostatic material, including zeolite, kaolin etc., this kind of material use natural silicon aluminium acid
The characteristics such as salt material loose structure, high-specific surface area, by absorbing the moisture in blood, concentrate the local hemostatic composition of the surface of a wound,
So as to realize quick-acting haemostatic powder.And the mesoporous silicon material manually prepared equally has the characteristics such as loose structure, high-specific surface area, and
It is same that effectively hemostasis can be achieved compared to the natural aluminosilicate silicate material advantage such as have composition, particle diameter, aperture controllable.However, this kind of
Material is helpless for the big bleeding of blood flow, also limited for large area surface of a wound effect；Although and this kind of material Jie is local
Tissue reaction is benign, but the mesoporous silicon grain for entering vascular and histoorgan can cause serious general toxicity.
And the injectable type self-healing hemostatic material that the present invention is provided is the polymer-network method using belt surface electric charge
Grain, the colloidal gel material obtained by intergranular electrostatic interaction self assembly.Due to the electrostatic interaction between microgel
It is to be physical crosslinking and with invertibity, therefore when this colloidal gel is destroyed by destructive shearing force, micro-gel particles
Between electrostatic interaction destroyed by external force, fluent material of the colloidal gel from rigid solid material to mobility
Transformation, this process is referred to as shear shinning behavior.When external force is cancelled, the fast quick-recovery of interaction between colloid, colloid
Grain is assembled into colloidal gel by re-forming physical crosslinking.Therefore hemostatic material of the present invention has syringeability and oneself
Healing properties, self-healing performance test shows that the selfreparing rate after failure by shear is more than more than 85%.Importantly, this hair
Bright hemostatic material need not introduce chemical crosslink reaction in injection and solidification process, different from chemical macromolecule, also not introduce
Small molecule crosslinking agent, good biocompatibility, degradable absorption has no toxic side effect, and security is good, makes it in biomedical sector
Extensive use is possibly realized.
In addition, the hemostatic material that the present invention is provided has preferable mechanical strength, the modulus of elasticity of gel hemostatic material is big
In 10kPa, quick-acting haemostatic powder can also be realized for the bleeding tissue that amount of bleeding is big, vascular pressure is high.Hemostatic material also of the present invention
The high-specific surface area of the colloidal solid of material, the strength of stability of colloidal gel are conducive to the closing of wound.
2. the advantage in terms of medicament slow release
Hemostatic article is one of essential bio-medical material in surgical operation, to promote hemostasis, control operation wound
Face infection equivalent risk, acceleration catalysis Wound healing, it is to improve clinical hemostasis, promote that hemostatic material and functional drug, which are used in combination,
Enter one of effective means of wound healing.The biology of these blood-clotting agents, antibacterial-anti-inflammatory drug or promotion organization reparative regeneration is living
Property medicine include small-molecule drug and protide macromolecular drug, these medicines during with hemostatic article compound use all
There is insoluble drug release uncontrollable, the problem of molecules like pharmaceuticals easy in inactivation.Therefore, exploitation can load medicine and controllable in drug release
Hemostatic material has important medical application value.Traditional hemostatic material is generally using the good hemostatic article of preprocessing (as only
Blood film, styptic powder, styptic sponge etc.), material is directly mixed in drug solution, adsorbs real in the surface physics of support by medicine
The loading of existing medicine.This mode typically results in hemostatic material when in use, the violent release of contained medicine, for promoting wound repair
Protein medicaments can not realize release lasting for a long time.
The hemostatic material contrast of the present invention has more advantages with conventional carriers material.1) it is micro- solidifying with micro-or nano size
Glue particle be elementary cell, compared to conventional porous timbering material have more high-specific surface area, therefore can adsorption protein content
It is higher；2) loading of growth factor is that the freeze-dried powder of micro-gel particles is directly blended with the growth factor aqueous solution, molten in particle
Protein molecular enters micro-gel particles network internal under osmotic pressure effect during swollen, therefore the release of albumen is mainly by micro-
The degradation rate control of gel；3) the main degradation rate by gelatin microgel of the rate of release of growth factor regulates and controls, therefore
The rate of release of the growth factor of loading is regulated and controled by controlling the degree of cross linking of microgel to realize, further by difference
Growth factor be loaded into the colloidal solids of the different degrees of cross linking, it is possible to achieve the orderly controlled release of a variety of growth factors.
Brief description of the drawings
Fig. 1 is the stereoscan photograph of type A gelatin micro-gel particles prepared by the method for embodiment 1.
Fig. 2 is the compound adhesive being made up of the A types and type B gelatin colloidal solid of oppositely charged prepared by the method for embodiment 1
The rheology test result of the selfreparing behavior of body gel.
Fig. 3 is the calcium alginate and type A gelatin colloidal solid composition of oppositely charged prepared by the method described in embodiment 2
Composite colloid gel selfreparing behavior rheology test result.
Fig. 4 represents that hemostatic material prepared by method described in embodiment 6 realizes the orderly release of biological activity protein medicine.
Fig. 5 represents to confirm the hemostasia effect of hemostatic material of the present invention by zoopery described in embodiment 7.
Embodiment
Following non-limiting examples can make one of ordinary skill in the art be more fully understood the present invention, but not with
Any mode limits the present invention.In following embodiments, unless otherwise specified, used experimental method is conventional method, institute
It can be bought with material, reagent etc. from biological or chemical company.
The freeze-dried powder of each colloidal solid is obtained after colloidal solid is freeze-dried in example below, wherein the freezing is dry
Dry condition is：By colloidal solid -60 DEG C,<It is freeze-dried 2-3 days under 300Pa.
Embodiment 1
Using type A gelatin as raw material, it is set to heat 40 DEG C of dissolvings in deionized water, configuration concentration is 5w/v% gelatin water
Solution, regulation aqueous gelatin solution pH value is 3, then to the ethanol that 3 times of liquor capacities are added in solution, generates gelatin microgel
The dispersion liquid of grain；25wt% glutaraldehyde water solutions are added into dispersion liquid respectively, gelatin micro-gel particles are crosslinked, penta 2 are added
The amount of aldehyde is respectively every gram of μ L of gelatin correspondence 25wt% glutaraldehydes 66, and cross-linking reaction time 12hr, eccentric cleaning obtains surface
The dispersion liquid of the positive type A gelatin micro-gel particles in zeta potentials position.Same method, surface is prepared by raw material of type B gelatin
Zeta potentials are the dispersion liquid for the type B gelatin micro-gel particles born.Measured using laser particle analyzer and prepare micro-gel particles size
It is as shown in table 1 with zeta potential data.The freeze-dried freeze-dried powder (be labeled as GelA) for respectively obtaining type A gelatin particle and
The freeze-dried powder (being labeled as GelB) of type B gelatin particle.The pattern of gelatin particle is observed by SEM,
As a result it is as shown in Figure 1.The particle diameter distribution of the type A gelatin particle prepared is narrow, and size is in the range of 150-300nm.
The performance parameter of the different type gelatin particle of table 1.
Type A gelatin and type B gelatin micro-gel particles are dispersed in 20mM NaOH alkaline aqueous solutions respectively, respectively obtained
It is dispersed with positively charged type A gelatin micro-gel particles and the dispersion liquid of negatively charged type B gelatin micro-gel particles, incites somebody to action both
It is sufficiently mixed, stirs, of the dispersion liquid of two kinds of different micro-gel particles, wherein type A gelatin and type B gelatin mixing must be dispersed with
Grain number amount ratio is 1:1；100mM salt acid for adjusting pH value is added into dispersion liquid to 7.0, stirring mixing, freeze-drying is contained
There is the freeze-dried powder of two kinds of different gelatin gels particles, labeled as GelA+B, hemostasis pulvis can be used as.Said mixture is freezed
Powder is blended with the 1mM of appropriate amount NaCl solution respectively, and is quickly uniformly mixed and obtains injectable type self-healing colloid
Gel, obtains the colloidal gel containing different microgel colloidal solid volume fractions, obtains injectable type hemostasis gel.Prepare gained
The colloidal gel of different component, as rheometer, the viscoelastic property of hemostasis gel material to obtained by is evaluated.As a result such as the institute of table 2
Show, the increase of colloidal volume fraction, the modulus of elasticity increase of gel；When volume fraction is identical, oppositely charged colloidal solid group
Into gel elastomer modulus be significantly stronger than the colloidal gel of one-component.It is 120vol% in microgel colloidal solid volume fraction
When, the colloidal gel modulus of elasticity of GelA+B components is about 38kPa.
The selfreparing behavior of colloidal gel is characterized by rheometer, and specific method of testing is as follows.To colloidal gel
Carry out continuous rheometer test：Carry out concussion time sweep first, sample is applied frequency be 1Hz and strain be 0.5% it is outer
Power, the storage modulus (or modulus of elasticity, G ') and loss modulus (or viscous modulus, G ") of test sample, now gel cut low
The behavioral inelasticity of solid is shown in the case of shear force, therefore storage modulus G ' is more than loss modulus G " and keeps stable.This single order
G ' the values of section are the initial elastic modulus of sample.Then gradually increase the strain of application from 0.1% to 1000%, during this
By applying external force by sample broke, elastic modulus G ' is gradually reduced, terminating below G ", i.e. colloidal dispersion from rigid solid to viscous
Property fluid changes, destructurized.Immediately cancel external force effect, investigate the recovery situation of sample elastic modulus.Will
After external force release, energy storage (elasticity) modulus that sample recovers and percentage (%) quantitative expedition gel of its initial store elastic mould
Self-repair efficiency.The self-repair efficiency of gel is as shown in table 3, the gel elastomer modulus being made up of oppositely charged colloidal solid
It is significantly stronger than the colloidal gel of one-component.The self-repair procedure of GelA+B colloidal gels is as shown in Fig. 2 gel is in failure by shear
Its modulus of elasticity momentary recovery, reviews one's lessons by oneself complex modulus and returns to initial modulus more than 85% afterwards in 5 minutes.And so review one's lessons by oneself
Multiple behavior can repeated：During the failure by shear of multiple circulations is applied to sample, cancel every time after external force, gel
Modulus of elasticity all can quick-recovery, and return to more than the 80% of initial elastic modulus soon.
Gel obtained by the micro-gel particles that the different colloidal gels prepared in the embodiment 1 of table 2. contain different volumes fraction
The elastic modulus G of material '
Gel rubber material obtained by the micro-gel particles of the different colloidal gel different volumes fractions prepared in the embodiment 1 of table 3.
Self-repair efficiency
* note：Self-repair efficiency is to be continued using 1000% strain after your shear gel material 60s, detects stress release
The percentage (%) that the modulus of elasticity in 5min recovers afterwards.
Embodiment 2
Using type A gelatin as raw material, dissolved in the case where heating 40 DEG C, configuration obtains the type A gelatin aqueous solution that concentration is 5w/v%,
It is 11 to adjust pH value, then to the ethanol that 3.5 times of volumes are added in solution, generates type A gelatin micro-gel particles suspension；To outstanding
25wt% glutaraldehyde water solutions are added in supernatant liquid, gelatin micro-gel particles are crosslinked, the amount for adding glutaraldehyde is per g gelatin correspondence
25wt% glutaraldehydes 66 μ L, cross-linking reaction time 12hr, are subsequently added in glycine and unreacted aldehyde radical, and eccentric cleaning is obtained
Type A gelatin micro-gel particles, particle size and surface zeta potential point position parameter are as shown in table 4.
Alginic acid micro-gel particles are prepared by emulsion method, specific preparation method is as follows：By the water of 1wt% sodium alginate
Solution is added dropwise in 50mM calcium chloride waters and continues high-speed stirred (mixing speed>5000rpm), that is, calcium alginate is obtained
Micro-gel particles, particle size and surface zeta potential point position parameter are as shown in table 4.By type A gelatin and calcium alginate micro-gel particles
It is dispersed in respectively in 10mM acetic acid acidic aqueous solution, respectively obtains the dispersion liquid of positively charged type A gelatin micro-gel particles
Ith, the dispersion liquid II of negatively charged alginic acid micro-gel particles, dispersion liquid I and dispersion liquid II are sufficiently mixed, stirred, score
The amounts of particles ratio of dispersion liquid III, wherein type A gelatin and calcium alginate micro-gel particles mixing is 2:1；Add into dispersion liquid III
The sodium hydroxide for entering 100mM adjusts pH value to 7.0, and stirring mixing, freeze-drying obtains being mixed with the jelly of the different colloidal solids of two-phase
Dry powder.Above-mentioned micro-gel particles freeze-dried powder and the 1mM of certain volume NaCl salting liquids are blended, and quickly stirring mixing
Uniformly obtain injectable type self-healing colloidal gel, i.e., hemostatic material of the present invention, wherein microgel colloidal solid volume
Fraction accounts for the 50vol% or 100vol% of colloidal gel volume.The mechanics parameter of colloidal gel is evaluated using rheometer,
As a result as shown in fig. 3 and table 5, the colloidal gel that the calcium alginate of oppositely charged and the blending of type A gelatin colloidal solid are obtained
Modulus of elasticity increases with the increase of colloidal solid volume fraction, during volume fraction 100vol%, storage (elasticity) modulus G '
More than 12kPa.Self-healing efficiency increases, body likewise as the increase of colloidal solid volume fraction in 5 minutes after failure by shear
During fraction 100vol%, elastic modulus G ' self-healing efficiency is more than 80%.
The type A gelatin and the performance parameter of alginic acid microgel prepared in the embodiment 2 of table 4.
Mechanical strength of the self-healing colloidal gel prepared in the embodiment 2 of table 5. in the case of different colloidal volume fractions
(rheometer test elastic modulus G ') and self-healing efficiency.
* note：Self-repair efficiency is to be continued using 1000% strain after your shear gel material 60s, detects stress release
The percentage (%) that the modulus of elasticity in 5min recovers afterwards.
Embodiment 3
Using type B gelatin as raw material, dissolved in the case where heating 40 DEG C, configuration obtains the type B gelatin aqueous solution that concentration is 5w/v%,
It is 3 to adjust pH value, then to the ethanol that 3 times of volumes are added in solution, generates type B gelatin micro-gel particles suspension；To suspension
25wt% glutaraldehyde water solutions are added in liquid, gelatin micro-gel particles are crosslinked, the amount for adding glutaraldehyde is per g gelatin correspondence
25wt% glutaraldehydes 80 μ L, cross-linking reaction time 12hr, are centrifuged repeatedly and are resuspended in deionized water, obtain type B gelatin micro- solidifying
Glue particle, particle size and surface zeta potential point position parameter are as shown in table 6.
Chitosan quaternary ammonium salt is dissolved in deionized water configuration concentration for 2.5w/v% chitosan quaternary ammonium saline solutions.By gelatin
B micro-gel particles are dispersed in 20mM acetic acid acidic aqueous solution, further that chitosan quaternary ammonium saline solution and gelatin B is micro- solidifying
Glue particle dispersion is fully blended, stirring, obtains dispersion liquid III, wherein type B gelatin micro-gel particles and polymer chitosan quaternary ammonium
The mass ratio of salt is 10:1, the sodium hydroxide that 100mM is added into dispersion liquid III adjusts pH value to 7.0, is then freeze-dried,
Obtain the compound freeze-dried powder of type B gelatin colloidal solid and chitosan quaternary ammonium salt.By above-mentioned freeze-dried powder and certain volume
10mM NaCl salting liquids blending, and be quickly uniformly mixed and obtain injectable type self-healing colloidal gel, i.e., institute of the present invention
The hemostatic material stated, wherein gelatin micro-gel particles volume fraction account for self-healing gel volume fraction for 100vol%.Use stream
Become instrument to evaluate the mechanics parameter of colloidal gel, as a result as shown in table 7, the type B gelatin particle and shell of oppositely charged gather
The colloidal gel that sugared quaternary ammonium salt blend is obtained is when colloidal solid volume fraction is 100vol%, and storage (elasticity) modulus G ' is about
48kPa.Self-healing efficiency is when colloidal solid volume fraction is 100vol% in 5 minutes after failure by shear, and elastic modulus G ' is certainly
Healing efficiency is more than 89%.
The performance parameter of the type B gelatin micro-gel particles prepared in the embodiment 3 of table 6.
Self-healing prepared by the type B gelatin micro-gel particles and chitosan quaternary ammonium salt blend prepared in the embodiment 3 of table 7. is coagulated
The mechanical strength (rheometer test elastic modulus G ') and self-healing efficiency of glue.
* note：Self-repair efficiency is to be continued using 1000% strain after your shear gel material 60s, detects stress release
The percentage (%) that the modulus of elasticity in 5min recovers afterwards.
Embodiment 4
Using type A gelatin as raw material, dissolved in the case where heating 40 DEG C, it is water-soluble that configuration obtains the type A gelatin that concentration is 10w/v%
Liquid, regulation pH value is 11, then to the ethanol that 2 times of volumes are respectively added in solution, generates type A gelatin micro-gel particles suspension；
25wt% glutaraldehyde water solutions are added into suspension, gelatin micro-gel particles are crosslinked, the amount for adding glutaraldehyde is per g gelatin
Correspondence 25wt% glutaraldehydes 264 μ L, cross-linking reaction time 12hr, are centrifuged repeatedly and are resuspended in deionized water, obtain type A gelatin
Micro-gel particles, particle size and surface zeta potential point position parameter are as shown in table 8.
Polyethylene glycol (PEG, molecular weight 2kDa) is dissolved in deionized water configuration concentration for the 5w/v%PEG aqueous solution.Will be upper
In the scattered obtained PEG aqueous solution of type A gelatin micro-gel particles for stating method preparation, further by the PEG aqueous solution and type A gelatin
Microgel dispersion is fully blended, stirring, obtains dispersion liquid III, wherein type A gelatin micro-gel particles and PEG mass ratio is 1:
2.Then dispersion liquid III pH value is adjusted to 7.0, is then freeze-dried, the compound of type A gelatin colloidal solid and PEG is obtained
Freeze-dried powder.Above-mentioned freeze-dried powder and the 10mM of certain volume NaCl salting liquids are blended, and are quickly uniformly mixed
To self-healing colloidal gel, i.e., hemostatic material of the present invention, wherein gelatin micro-gel particles volume fraction account for self-healing and coagulated
Colloid fraction is 100vol%.The mechanics parameter of colloidal gel is evaluated using rheometer, as a result as shown in table 9, when
When type A gelatin colloidal solid volume fraction is 100vol% in colloidal gel, (elasticity) modulus G ' about 19kPa are stored.Shearing is broken
Self-healing efficiency is when colloidal solid volume fraction is 100vol% in 5 minutes after bad, and elastic modulus G ' self-healing efficiency is about
83%.
The performance parameter of the type A gelatin micro-gel particles prepared in the embodiment 5 of table 8.
The mechanics of self-healing gel prepared by type A gelatin micro-gel particles and the PEG blending prepared in the embodiment 4 of table 9. is strong
Spend (rheometer test elastic modulus G ') and self-healing efficiency.
* note：Self-repair efficiency is to be continued using 1000% strain after your shear gel material 60s, detects stress release
The percentage (%) that the modulus of elasticity in 5min recovers afterwards.
Embodiment 5
Using type A gelatin as raw material, dissolved in the case where heating 40 DEG C, configuration obtains the type A gelatin aqueous solution that concentration is 5w/v%,
It is 11 to adjust pH value, then to the ethanol that 3.5 times of volumes are respectively added in solution, generates type A gelatin micro-gel particles suspension；With
25wt% glutaraldehyde water solution cross-linked gelatin micro-gel particles are added afterwards, and the amount for adding glutaraldehyde is per g gelatin correspondence 25wt%
The μ L of glutaraldehyde 66, cross-linking reaction time 12hr, eccentric cleaning obtains type A gelatin micro-gel particles, particle size and surface zeta potential
Point position parameter is as shown in table 10.
Alginic acid micro-gel particles are prepared by emulsion method, specific preparation method is as follows：By the water of 1wt% sodium alginate
Solution adds in calcium chloride water and continues high-speed stirred (mixing speed>5000rpm), that is, calcium alginate particle is obtained,
Particle size and surface zeta potential point position parameter are as shown in table 10.
Type A gelatin and calcium alginate micro-gel particles are dispersed in 10mM acetic acid acidic aqueous solution respectively, respectively
The dispersion liquid II of dispersion liquid I to positively charged type A gelatin micro-gel particles, negatively charged alginic acid micro-gel particles,
Dispersion liquid I and dispersion liquid II are sufficiently mixed, stirred, dispersion liquid III is obtained, wherein type A gelatin and alginic acid micro-gel particles is mixed
The amounts of particles ratio of conjunction is 2:1；The sodium hydroxide that 100mM is added into dispersion liquid III adjusts pH value to 7.0, and stirring mixing is cold
It is lyophilized dry, obtain being mixed with the compound freeze-dried powder of the different colloidal solids of two-phase.Polyethylene glycol (PEG, molecular weight 2kDa) is dissolved in
Deionized water configuration concentration is the 5w/v%PEG aqueous solution, is blended with above-mentioned compound freeze-dried powder, and quickly stirring obtains self-healing
Gel, i.e., hemostatic material of the present invention, two of which colloidal solid volume fraction accounts for the 100vol% of colloidal gel volume.
The mechanics parameter of colloidal gel is investigated using rheometer, as a result as shown in table 11, as colloidal solid volume fraction 100vol%,
(elasticity) modulus G ' is stored more than 87kPa.After failure by shear in 5 minutes self-healing efficiency likewise as colloidal solid volume fraction
Increase and increase, during volume fraction 100vol%, elastic modulus G ' self-healing efficiency is more than 91%.
The type A gelatin and the performance parameter of calcium alginate microgel prepared in the embodiment 5 of table 10.
Mechanical strength of the self-healing colloidal gel prepared in the embodiment 5 of table 11. in the case of different colloidal volume fractions
(rheometer test elastic modulus G ') and self-healing efficiency.
* note：Self-repair efficiency is to be continued using 1000% strain after your shear gel material 60s, detects stress release
The percentage (%) that the modulus of elasticity in 5min recovers afterwards.
Embodiment 6
Using type A gelatin as raw material, type A gelatin micro-gel particles are prepared by anti-solvent method described in embodiment 1, glutaraldehyde is handed over
It is that every g gelatin is 66 μ L using the amount of 25wt% glutaraldehydes to join concentration, prepares positively charged type A gelatin microgel
Grain；Using type B gelatin as raw material, type B gelatin micro-gel particles, wherein glutaraldehyde are prepared by the anti-solvent method described in embodiment 1
It is that every g gelatin is 264 μ L using the amount of 25wt% glutaraldehydes to be crosslinked concentration, prepares negatively charged type B gelatin microgel
Particle, preparation parameter and gained micro-gel particles size and zeta points position data such as table 12.
The performance parameter of the different type gelatin micro-gel particles prepared in the embodiment 6 of table 12.
Type A gelatin particle is dispersed in the aqueous solution of the Basic Fibroblast Growth Factor (bFGF) of the concentration containing 100ng/ml
In, type B gelatin particle is dispersed in the aqueous solution of bone morphogenesis protein-2 (BMP-2) of the concentration containing 100ng/ml, respectively
Obtain being loaded with bFGF type A gelatin particle and be loaded with the dispersion liquid of BMP-2 type B gelatin micro-gel particles.Above two is bright
Glue particle dispersion compares 1 according to granule number:1 is sufficiently mixed, and is freeze-dried, and obtains being loaded with two kinds of gelatin of different growth factors
Particle freeze-dried powder.Above-mentioned micro-gel particles freeze-dried powder and the 1mM of certain volume NaCl salting liquids are blended, and quickly stirred
Mix well mixed, obtain being loaded with different growth factors, wherein injectable type self-healing colloidal gel, microgel colloidal solid body
Fraction accounts for the 100vol% of colloidal gel volume.Two kinds of different growth factors release in vitro from colloidal gel carrier material
Dynamic (dynamical) release profiles are as shown in figure 4, be loaded with bFGF type A gelatin because the low degradation rate of the degree of cross linking is very fast, therefore bFGF
Rate of release is very fast, is loaded with BMP-2 type B gelatin because the high degradation rate of the degree of cross linking is slow, therefore BMP-2 rates of release are more delayed
Slowly；As a result show that colloidal gel of the present invention can realize the orderly release of different growth-factor medications.
Embodiment 7
Type A gelatin and type B gelatin micro-gel particles dispersion liquid are obtained by implementing the preparation method in 1, by two kinds of gelatin
Particle is dispersed in 20mM aqueous hydrochloric acid solution respectively, respectively obtain be dispersed with positively charged type A gelatin micro-gel particles and
The dispersion liquid of negatively charged type B gelatin micro-gel particles, both are sufficiently mixed, stirred, and must be dispersed with two kinds of differences micro- solidifying
The amounts of particles ratio of the dispersion liquid of glue particle, wherein type A gelatin and type B gelatin mixing is 1:1；100mM is added into dispersion liquid
Sodium hydroxide adjust pH value to 7.0, stirring mixing, freeze-drying obtains lyophilized containing two kinds of different gelatin gels particles
Powder.Said mixture freeze-dried powder and phosphate buffer solution PBS are blended, and is quickly uniformly mixed and obtains injectable type
Self-healing colloidal gel, i.e., hemostatic material of the present invention, two of which colloidal solid volume fraction accounts for colloidal gel volume
100vol%.Using the new zealand rabbit back backbone Cranial defect bleeding surface of a wound as animal experimental model, manufacture 2.5-4mm bleeding wound
Mouthful, gelatin-based hemostasis gel is gently oppressed into blutpunkte after 1min by common medical syringe direct injection in the bleeding surface of a wound
It is closed, bleeding stopping period is 1min.The process of zoopery is as shown in Figure 5.
Claims (10)
1. a kind of injectable type self-healing hemostatic material, is prepared by the following method：
(1) using gelatin as raw material, it is made to dissolve by heating in deionized water, configuration concentration is water-soluble for 1~10w/v% gelatin
Liquid, tune pH value is 1-6 or 8-14, is added into solution>The polar organic solvent of 2 times of liquor capacities, generates gelatin microgel
Grain dispersion liquid, adds cross-linking agents and reacts 1~12h, centrifugation, cleaning obtain gelatin micro-gel particles；
Wherein, the zeta potentials of the gelatin micro-gel particles be -30~+30mV, the gelatin micro-gel particles it is a diameter of
20nm~2 μm；
(2) the surface zeta potential potential for preparing step (1)>+ 10mV gelatin micro-gel particles, are dispersed in pH<5 acidity
The aqueous solution or pH>In 9 alkaline aqueous solution, the dispersion liquid of positively charged gelatin micro-gel particles must be dispersed with, then it is negative with band
The polymeric particles dispersion liquid of electric charge compares 1 according to granule number:10~10:1 blending, or and isoelectric point<6 negatively charged parent
The aqueous high molecular aqueous solution according to gelatin micro-gel particles and hydrophilic macromolecule mass ratio 1:10~10:1 blending, is adjusted with pH
Save agent and adjust pH to 7.0, freeze-drying obtains gelatin micro-gel particles freeze-dried powder I；
(3) the surface zeta potential potential for preparing step (1)<- 10mV gelatin micro-gel particles, are dispersed in pH<5 acidity
The aqueous solution or pH>In 9 alkaline aqueous solution, obtain the dispersion liquid of gelatin micro-gel particles, then with positively charged polymeric particles
Dispersion liquid compares 1 according to granule number:10~10:1 mixing, or and isoelectric point>The 8 positively charged hydrophilic macromolecule aqueous solution
According to the mass ratio 1 of gelatin micro-gel particles and hydrophilic macromolecule:10~10:1 blending, pH is adjusted to 7.0 with pH adjusting agent, cold
It is lyophilized dry, obtain gelatin micro-gel particles freeze-dried powder II；
(4) the surface zeta potential potential for preparing step (1) is dispersed in neutrality for -10~+10mV gelatin micro-gel particles
In the aqueous solution, then polymeric particles dispersion liquid with another surface zeta potential potential in -10~+10mV according to granule number compares 1:10
~10:1 blending, or with the hydrophilic macromolecule aqueous solution of isoelectric point 6~8 according to gelatin micro-gel particles and hydrophily high score
The mass ratio 1 of son:10~10:1 blending, freeze-drying obtains gelatin micro-gel particles freeze-dried powder III；
(5) gelatin micro-gel particles freeze-dried powder I, gelatin micro-gel particles freeze-dried powder II or gelatin micro-gel particles are freezed
Powder III is blended with aqueous solution respectively, obtains injectable type self-healing hemostatic material；
Wherein, the surface charge of described positively charged polymeric particles is+5~+60mV, negatively charged polymeric particles
Surface charge be -5~-60mV, a diameter of 100nm~20 μm of the polymeric particles, what is obtained in step (5) notes
Colloidal solid percentage of the total volume is 50vol%~150vol% in emitting self-healing hemostatic material；In step (3), (4)
(5) molecular weight of the hydrophilic macromolecule described in is 1k~500kDa.
2. injectable type self-healing hemostatic material according to claim 1, it is characterised in that described positively charged height
Molecule particles dispersion liquid is with chitosan, type A gelatin, polyacrylamide, NIPA, polyethyleneimine
One or more are prepared as raw material, described negatively charged polymeric particles dispersion liquid with hyaluronic acid, alginic acid,
One or more in type A gelatin, type B gelatin or polyacrylic acid are prepared as raw material, described surface zeta potential potential-
10~+10mV polymeric particles dispersion liquid with one kind in collagen, albumin, gelatin, polyvinyl alcohol, polyethylene glycol or
It is several to be prepared as raw material；Described isoelectric point>8 positively charged hydrophilic macromolecule be chitosan, type A gelatin,
One or more in polyacrylamide, NIPA, polyethyleneimine, the isoelectric point<6 it is negatively charged
The hydrophilic macromolecule of lotus is the one or more in hyaluronic acid, alginic acid, type A gelatin, type B gelatin or polyacrylic acid, institute
The macromolecule for the isoelectric point 6~8 stated is the one or more in collagen, albumin, gelatin, polyvinyl alcohol, polyethylene glycol.
3. injectable type self-healing hemostatic material according to claim 1, it is characterised in that described in step (1)
Polar organic solvent is one or more of combinations in methanol, ethanol, isopropanol, butanol, acetone, acetonitrile, tetrahydrofuran；Institute
The crosslinking agent stated is glutaraldehyde, glyceraldehyde, formaldehyde, carbodiimide, saturated dihalide, isocyanates, diisocyanate, glutamine
One or more in transaminase, Geniposide.
4. injectable type self-healing hemostatic material according to claim 1, it is characterised in that described in step (1)
The mol ratio 0.1~10 of crosslinking agent and amino group in gelatin in the reaction system of cross-linking reaction.
5. injectable type self-healing hemostatic material according to claim 1, it is characterised in that in step (2) and step (3)
Described in acidic aqueous solution and alkaline aqueous solution in contained ion concentration be respectively less than 200mM.
6. injectable type self-healing hemostatic material according to claim 1, it is characterised in that in step (2) and step (3)
Described in pH adjusting agent include acidic materials and alkaline matter, the acidic materials be glucolactone, HCl, HNO3、
H2SO4In one or more, the alkaline matter be urea and the combination of urase or sodium hydroxide, calcium hydroxide, hydrogen-oxygen
Change the one or more in potassium, ammoniacal liquor.
7. injectable type self-healing hemostatic material according to claim 1, it is characterised in that described in step (5)
Aqueous solution be 120~200mM of ion concentration, pH value for 6~8 any aqueous solution, aqueous solution of hydrophilic macromolecule, non-aqueous
One or more of combinations in the nanoparticulate dispersion of dissolubility.
8. injectable type self-healing hemostatic material according to claim 1, it is characterised in that contain in the aqueous solution
Bioactive substance.
9. the injectable type self-healing hemostatic material described in any one of claim 1~8 is being prepared for body surface organization, body cavity
Inner tissue's organ have the stopping blooding of the blood surface of a wound, it is anti-be adhered, anti-infection, the hemostatic article of promotion organization healing and/or closing wound
In application.
10. application according to claim 1, it is characterised in that described hemostatic article be styptic powder agent, hemostasis granules,
Hemostatic ball, hemostasis aerosol, styptic sponge, hemostasis gel, haemostatic membrane, tampon or hemostatic adhesive bandage.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710335184.0A CN106983905B (en) | 2017-05-12 | 2017-05-12 | A kind of injectable type self-healing hemostatic material and its preparation method and application |
| PCT/CN2018/086502 WO2018205996A1 (en) | 2017-05-12 | 2018-05-11 | Injectable type self-healing hemostatic material, preparation method therefor and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710335184.0A CN106983905B (en) | 2017-05-12 | 2017-05-12 | A kind of injectable type self-healing hemostatic material and its preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106983905A true CN106983905A (en) | 2017-07-28 |
| CN106983905B CN106983905B (en) | 2019-10-11 |
Family
ID=59419456
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710335184.0A Active CN106983905B (en) | 2017-05-12 | 2017-05-12 | A kind of injectable type self-healing hemostatic material and its preparation method and application |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN106983905B (en) |
| WO (1) | WO2018205996A1 (en) |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108310448A (en) * | 2018-02-07 | 2018-07-24 | 广州迈普再生医学科技有限公司 | A kind of preparation method of fluid gelatin hemostatic material |
| WO2018205996A1 (en) * | 2017-05-12 | 2018-11-15 | 深训华诺生物科技有限公司 | Injectable type self-healing hemostatic material, preparation method therefor and use thereof |
| CN109438727A (en) * | 2018-10-17 | 2019-03-08 | 中国人民解放军总医院 | A kind of self-healing property hydrogel of fluorescence response and preparation method thereof |
| CN110527115A (en) * | 2019-08-01 | 2019-12-03 | 华南理工大学 | Multi-functional selfreparing hydrogel and preparation method thereof and the application in determination of biogenic amines |
| CN110743038A (en) * | 2019-11-06 | 2020-02-04 | 大连理工大学 | Double-network structure composite hydrogel and preparation method and application thereof |
| CN110760076A (en) * | 2019-11-06 | 2020-02-07 | 大连理工大学 | Injectable high-strength composite hydrogel based on colloidal particle-iPRF dual-network structure and preparation method and application thereof |
| CN111068111A (en) * | 2018-10-19 | 2020-04-28 | 胡尚秀 | Injectable self-assembled microsphere gel, application and preparation method thereof |
| CN111386132A (en) * | 2017-10-02 | 2020-07-07 | 阿莱奥生物医学工程有限公司 | Aqueous tissue adhesive |
| CN113134113A (en) * | 2021-04-07 | 2021-07-20 | 赛克赛斯生物科技股份有限公司 | Preparation method of absorbable hemostatic fluid gelatin and absorbable hemostatic fluid gelatin |
| CN114028606A (en) * | 2021-10-26 | 2022-02-11 | 浙江理工大学 | Chitosan and protamine antibacterial hemostatic microsphere and preparation method thereof |
| CN114848668A (en) * | 2021-01-20 | 2022-08-05 | 香港中文大学 | Composition with wound healing promoting and quick hemostasis functions |
| CN115845121A (en) * | 2022-12-07 | 2023-03-28 | 华南理工大学 | Hemostatic, antibacterial and healing promoting microgel assembly powder and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007120818A2 (en) * | 2006-04-12 | 2007-10-25 | Massachusetts Institute Of Technology | Compositions and methods for inhibiting adhesions |
| CN105796478A (en) * | 2016-03-22 | 2016-07-27 | 王华楠 | Nanometer colloid particle-assembled high-strength self-repairing injectable composite colloid gel material and preparation method and application thereof |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102688525B (en) * | 2012-05-07 | 2013-11-27 | 东南大学 | Bio-macromolecular hydrogel and preparation method thereof |
| US8680240B1 (en) * | 2012-11-01 | 2014-03-25 | George David Falus | Tissue sealant for use in non-compressible hemorrhage |
| US20160121017A1 (en) * | 2014-11-04 | 2016-05-05 | Biomedica Management Corp | SINGLE SOLUTION of Gel-LIKE FIBRIN HEMOSTAT |
| CN106983905B (en) * | 2017-05-12 | 2019-10-11 | 深圳华诺生物科技有限公司 | A kind of injectable type self-healing hemostatic material and its preparation method and application |
-
2017
- 2017-05-12 CN CN201710335184.0A patent/CN106983905B/en active Active
-
2018
- 2018-05-11 WO PCT/CN2018/086502 patent/WO2018205996A1/en active Application Filing
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007120818A2 (en) * | 2006-04-12 | 2007-10-25 | Massachusetts Institute Of Technology | Compositions and methods for inhibiting adhesions |
| CN105796478A (en) * | 2016-03-22 | 2016-07-27 | 王华楠 | Nanometer colloid particle-assembled high-strength self-repairing injectable composite colloid gel material and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| HUANA WANG, ET.AL.: "Oppositely Charged Gelatin Nanospheres as Building Blocks for Injectable and Biodegradable Gels", 《ADV. MATER.》 * |
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018205996A1 (en) * | 2017-05-12 | 2018-11-15 | 深训华诺生物科技有限公司 | Injectable type self-healing hemostatic material, preparation method therefor and use thereof |
| CN111386132A (en) * | 2017-10-02 | 2020-07-07 | 阿莱奥生物医学工程有限公司 | Aqueous tissue adhesive |
| CN108310448A (en) * | 2018-02-07 | 2018-07-24 | 广州迈普再生医学科技有限公司 | A kind of preparation method of fluid gelatin hemostatic material |
| CN108310448B (en) * | 2018-02-07 | 2021-03-02 | 广州迈普再生医学科技股份有限公司 | Preparation method of fluid gelatin hemostatic material |
| CN109438727B (en) * | 2018-10-17 | 2020-08-07 | 中国人民解放军总医院 | Fluorescent response self-healing hydrogel and preparation method thereof |
| CN109438727A (en) * | 2018-10-17 | 2019-03-08 | 中国人民解放军总医院 | A kind of self-healing property hydrogel of fluorescence response and preparation method thereof |
| CN111068111A (en) * | 2018-10-19 | 2020-04-28 | 胡尚秀 | Injectable self-assembled microsphere gel, application and preparation method thereof |
| CN110527115A (en) * | 2019-08-01 | 2019-12-03 | 华南理工大学 | Multi-functional selfreparing hydrogel and preparation method thereof and the application in determination of biogenic amines |
| CN110743038A (en) * | 2019-11-06 | 2020-02-04 | 大连理工大学 | Double-network structure composite hydrogel and preparation method and application thereof |
| CN110760076A (en) * | 2019-11-06 | 2020-02-07 | 大连理工大学 | Injectable high-strength composite hydrogel based on colloidal particle-iPRF dual-network structure and preparation method and application thereof |
| CN114848668A (en) * | 2021-01-20 | 2022-08-05 | 香港中文大学 | Composition with wound healing promoting and quick hemostasis functions |
| CN114848668B (en) * | 2021-01-20 | 2024-03-26 | 香港中文大学 | Composition with functions of promoting wound healing and rapidly stopping bleeding |
| CN113134113A (en) * | 2021-04-07 | 2021-07-20 | 赛克赛斯生物科技股份有限公司 | Preparation method of absorbable hemostatic fluid gelatin and absorbable hemostatic fluid gelatin |
| CN113134113B (en) * | 2021-04-07 | 2022-07-01 | 赛克赛斯生物科技股份有限公司 | Preparation method of absorbable hemostatic fluid gelatin and absorbable hemostatic fluid gelatin |
| CN114028606A (en) * | 2021-10-26 | 2022-02-11 | 浙江理工大学 | Chitosan and protamine antibacterial hemostatic microsphere and preparation method thereof |
| CN115845121A (en) * | 2022-12-07 | 2023-03-28 | 华南理工大学 | Hemostatic, antibacterial and healing promoting microgel assembly powder and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106983905B (en) | 2019-10-11 |
| WO2018205996A1 (en) | 2018-11-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106983905B (en) | A kind of injectable type self-healing hemostatic material and its preparation method and application | |
| US20230381288A1 (en) | Hemostatic compositions | |
| US9821025B2 (en) | Hemostatic compositions | |
| US9694101B2 (en) | Flowable collagen-based hemostat and methods of use | |
| CA2851332C (en) | Hemostatic compositions | |
| KR20130083393A (en) | Process for making dry and stable hemostatic compositions | |
| US20130096082A1 (en) | Hemostatic compositions | |
| US20160121017A1 (en) | SINGLE SOLUTION of Gel-LIKE FIBRIN HEMOSTAT | |
| CN112300418A (en) | Adhesive high-efficiency hemostatic microsphere and preparation method thereof | |
| US20230165944A1 (en) | Hemostat reconstitution methods and devices | |
| US11311643B2 (en) | Fibrin and/or dialdehyde starch hydrolysate materials, and preparation and use thereof | |
| US20190388516A1 (en) | Hemostatic products | |
| US20230138347A1 (en) | Synthetic surgical hemostat | |
| CA3033757A1 (en) | Hemostatic products |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20181010 Address after: 518104 Houting Community, Shajing Street, Baoan District, Shenzhen City, Guangdong Province Applicant after: Shenzhen Sino Biological Technology Co., Ltd. Address before: 116000 College of life sciences, Dalian University of Technology, 2 linggong Road, hi tech park, Dalian, Liaoning Applicant before: Wang Huanan |
|
| TA01 | Transfer of patent application right | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |