CN105617449B - A kind of multi-functional micropore styptic powder and preparation method thereof - Google Patents
A kind of multi-functional micropore styptic powder and preparation method thereof Download PDFInfo
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- CN105617449B CN105617449B CN201610158215.5A CN201610158215A CN105617449B CN 105617449 B CN105617449 B CN 105617449B CN 201610158215 A CN201610158215 A CN 201610158215A CN 105617449 B CN105617449 B CN 105617449B
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- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
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Abstract
The invention discloses a kind of multi-functional micropore styptic powders and the preparation method and application thereof.This method comprises the following steps：1) starch is mixed with water, obtains starch solution or gelatinized corn starch liquid；2) carboxymethyl chitosan is mixed with water, obtains carboxymethyl chitosan solution；3) starch solution or gelatinized corn starch liquid are mixed with carboxymethyl chitosan solution, obtains blend solution；4) pore-foaming agent is dispersed in blend solution, obtains blending dispersion liquid；5) under stirring condition, ion crosslinking agent is added dropwise and is blended in dispersion liquid, obtained mixed system is mixed with dispersant and emulsifier, is filtered after crosslinked reaction；6) removal step 5) pore-foaming agent in product and drying；7) microwave treatment is carried out to get the styptic powder to step 6) product.The present invention uses the dual cross-linking method of ionomer and microwave treatment；Agents useful for same is environmentally protective, noresidue, and is conducive to the bioactivity for retaining amino in carboxymethyl chitosan.
Description
Technical field
The present invention relates to a kind of multi-functional micropore styptic powders and preparation method thereof, belong to natural biological field of medical materials.
Background technology
With the fast development of basic medical theory and surgical technic and equipment, range of operation and type is made constantly to expand
Greatly.Bleeding is problem common in surgical operation, and excessive bleeding even can be with threat to life, and transfuses blood that not only there is costs
The problems such as height, inadequate blood source, also has the risk of the infection hematologic diseases such as hepatitis C or HIV.Therefore, how quickly and effectively
Stop blooding for the success rate for improving operation, reduces patient's medical treatment cost and alleviation blood source insufficient supply etc. and be of great significance.
However traditional Hemostasis is such as oppressed, is ligatured, suturing, electricity irons, far can not meet become increasingly complex, fine hand
Art and doctors and patients are to safety and validity increasingly higher demands.
With bio-medical material study and apply gradually deeply, various novel biological hemostatic materials also meet the tendency of and
It is raw.If the hemostatic materials such as gelatin class, oxycellulose, polysaccharide and collagen class are in different surgical fields by doctor institute
Using.Oxycellulose class material degradation product is acidity, can cause the inflammatory reaction of the surface of a wound；A kind of foreign matter of gelatin class may increase
Add the infection rate of wound, especially contaminated wound；Collagen class material is animal derived hemostatic material, has and propagates zoonosis
Risk, European and American areas has carried out stringent limitation to the medical instrument of ox source property.
Its anthemorrhagic speed of hemostasis material requirements is fast in ideal art, absorbable or metabolizable, non-human and it is animal derived,
No immune response has no toxic side effect, without postoperative complications in art etc..And polysaccharide hemostatic material, such as starch and chitosan and its spread out
Biological species hemostatic material, good biocompatibility is nontoxic, non-stimulated, no allergic reaction.Therefore recent years, polysaccharide material are fast
Speed development, it is also increasingly wider in the research and application of hemostasis field and other biological medical field.
Starch hemostatic material, especially starch hemostatic powder, because its it is raw materials used for pure plant source, it is renewable, can
Degradation, good biocompatibility, therefore have a wide range of applications in hemostasis field.Starch hemostatic powder currently on the market, such as
AristaTM AH、PerClot、HaemoCerTM, glad speed listen, hemostatic mechanism is single, only relies on spherex and absorbs moisture, dense
Contracting blood forms sludged blood and blocks capillary, to play the effect of hemostasis, does not have biological hemostatic activity, therefore to material
Adhesiveness, absorbent and material water suction after formed gel stereochemical structure have higher requirement.For oozing of blood or exudation
The more surface of a wound of liquid, above-mentioned styptic powder cannot be used directly, and haemostatic effect is undesirable, before Clinical practice medical worker need by
Other materials fully removes the extra blood or moisture of the surface of a wound；And often so that styptic powder is failed because diffusate is more when in use,
Styptic powder need to be added, this undoubtedly brings inconvenience to the operation of medical worker, also increases the medical treatment cost of patient.
A kind of medical hemostatic polysaccharide spherex and preparation method thereof, this method are disclosed in CN201410621755.3
Spherex is prepared using the method that epoxychloropropane is chemically crosslinked.In existing patent and document, the preparation side of spherex
Fado is chemical crosslinking.Spherex structural stability prepared by chemical crosslink technique is preferable, but the crosslinking agent used such as table chlorine
Alcohol, epoxychloropropane, glutaraldehyde, formaldehyde etc. can cause security risk to producer, environment and patient.
A kind of converted starch Absorbable hemostatic material and preparation method thereof, CN200810032631.6 are disclosed in CN200710141944.0
A kind of public biocompatible hemostatic, it is anti-be adhered, promoting healing, the closed modified starch material of surgery, described in the two patents
Content it is roughly the same, be same applicant or inventor.Patent is using various converted starches as raw material, using water as binder, leads to
It crosses spray granulation and prepares spherex.This method is environmentally protective, safe, but spherex structural stability is poor, with
The volatilization of moisture, microballoon can disintegration at powder, easily there is glue packet powder phenomenon in hemostasis, absorbent drastically reduces, very
It unites to moisture absorption is easy, haemostatic effect weakens significantly.
Carboxymethyl chitosan because of it with good biocompatibility, adhesiveness is good, antibacterial anti-inflammatory, to promote wound healing etc. excellent
Point, therefore had a wide range of applications in clinic includes chitosan-based absorbable hemostatic nonwoven fabric, surgical operation at present with preventing adhesion
Nearly 10 section of 3 class medical device product such as liquid obtains CFDA approval listings.Amino in carboxymethyl chitosan is positively charged, can be with table
The electronegative haemocyte in face combines, and promotes the rapid adherency and aggregation of haemocyte, in addition being capable of shape after carboxymethyl chitosan imbibition
At layer of gel film uniform adhesion on wound tissue surface, to play comprehensive hemostatic function.
Converted starch and carboxymethyl chitosan are compounded, not only increase the haemostatic effect of starch hemostatic powder, simultaneously
Also there are many bioactive functions such as good, the promotion wound healing of antibacterial anti-inflammatory, adhesiveness.It is disclosed in CN200910016401.5
A kind of medical compound micropore polysaccharide and application thereof.This method prepares starch and carboxymethyl chitosan in the method being chemically crosslinked
Complex microsphere, equally exist crosslinking agent lower tape come risk.Its crosslinking agent aldehydes and epoxychloropropane used can be with carboxylic first
Side reaction can occur for the amino in base enclosure glycan, and it is living in hemostasis and the biology of antibacterial anti-inflammatory etc. to reduce carboxymethyl chitosan
Property.Low using compound micropore polysaccharide particle porosity made of cross-linking method in patent, imbibition effect is limited, is only applicable to oozing of blood
Property or the surface of a wound of diffusivity bleeding, are not suitable for the larger surface of a wound of amount of bleeding.
Invention content
The object of the present invention is to provide a kind of multi-functional micropore styptic powder and preparation method thereof, this method uses ionomer
With the dual cross-linking method of microwave treatment, the bioactivity of amino in carboxymethyl chitosan is remained, microwave treatment can be further
The stability of polyoses grain structure is improved, the rate of liquid aspiration to improve styptic powder significantly optimizes haemostatic effect, while also advantageous
In reduction initial contaminating bacteria level to reduce sterilizing dose；In addition the method for carrying out pore by adding pore-foaming agent, is prepared into
To styptic powder surface and inside all have a large amount of microcellular structure, so that it is had a specific surface area of opposite bigger, enhancing three
The molecular sieve function of dimension space promotes imbibition ability, greatly improves anthemorrhagic speed.
A kind of preparation method of styptic powder provided by the invention, it includes the following steps：
(1) starch is mixed with water, starch solution is prepared after dissolving or gelatinized corn starch liquid is prepared after swelling；
(2) carboxymethyl chitosan is mixed with water, carboxymethyl chitosan solution is prepared after insoluble matter is removed by filtration；
(3) by starch solution described in step (1) or the gelatinized corn starch liquid and step (2) described carboxymethyl chitosan solution
Mixing, obtains blend solution；
(4) pore-foaming agent is dispersed in the blend solution described in step (3), obtains that dispersion liquid is blended；
(5) under agitation, ion crosslinking agent is added dropwise in the blending dispersion liquid described in step (4), will
The mixed system arrived is mixed with dispersant and emulsifier, is filtered after crosslinked reaction；
(6) pore-foaming agent in the product obtained in removal step (5) and drying；
(7) microwave treatment is carried out to the product obtained in step (6), you can obtain the styptic powder.
Above-mentioned preparation method, in step (1), the mass ratio of the starch and water can be 1：(10~50), concretely
1：(25~40), 1：25 or 1：40；The starch meets following conditions：At 25 DEG C, mass percentage be 2% starch solution
Or the viscosity of gelatinized corn starch liquid is 200~1000mpas, such as 200~500mpas, 200mpas or 500mpas；People in the art
Member can select the type of starch, the present invention select converted starch according to common sense in the field, the converted starch refer to by
Processing, the property of starch meet the present invention claims starch, concretely carboxymethyl starch, crosslinked carboxymethyl fecula, pregel
At least one of starch, pregelatinized starch, cationic starch and pre-gelatinized hydroxypropyl PASELLI EASYGEL；If selecting carboxymethyl
Starch and/or crosslinked carboxymethyl fecula, degree of substitution require to be 0.5~1.2, and such as 0.5.The dissolving or swelling are at 20 DEG C~90 DEG C
Water bath condition under carry out.
Above-mentioned preparation method, in step (2), the mass ratio of the carboxymethyl chitosan and water can be 1：(10~100),
Concretely 1：(30~60), 1：30 or 1：60；The carboxymethyl chitosan meets following conditions：At 25 DEG C, quality percentage contains
Amount for 1% carboxymethyl chitosan aqueous solution viscosity not less than 100mpas (such as 100~200mpas, 100mpas or
200mpas), other physical and chemical indexes should meet the standard of YY0953-2015 medical carboxymethyl chitosans.
Above-mentioned preparation method, in step (3), the starch solution or the gelatinized corn starch liquid and the carboxymethyl chitosan
The mass ratio of solution can be 1：(0.1~10), 1：0.1、1：1 or 1：10.
In above-mentioned preparation method, in step (4), it is a large amount of that the addition of pore-foaming agent may make that the styptic powder being prepared has
Micropore, impart styptic powder very strong capillary effect in hemostasis so that styptic powder of the present invention have it is very strong
Imbibition ability and drain ability, reduce the dosage of styptic powder；The pore-foaming agent quality and the starch and the carboxymethyl shell
The ratio of the quality sum (quality sum of the starch and the carboxymethyl chitosan, referred to as " polysaccharide solid content ") of glycan can
It is 1：(5~500), concretely 1：(100~200), 1：100 or 1：200；The pore-foaming agent includes but are not limited to corn
Protein, CaCO3And MgCO3Grain size Deng, the pore-foaming agent can be 1~10 μm, concretely 2~5 μm, 2 μm or 5 μm；
In above-mentioned preparation method, in step (5), be conducive to retain amino in carboxymethyl chitosan using ion crosslinking agent
Bioactivity, the ion crosslinking agent is added in form of an aqueous solutions, the aqueous solution and the dispersion liquid that is blended
Mass ratio can be 1：(5~100), concretely 1：(30~50), 1：30 or 1：50；The matter of the aqueous solution of the ion crosslinking agent
It can be 0.5~40%, concretely 2~10%, 2% or 10% to measure percentage composition；Those skilled in the art can be according to ability
The type of domain common sense selection ion crosslinking agent, the present invention select the ion crosslinking agent of biological nontoxic, including but not limited to calcium salt, zinc
At least one of salt, aluminium salt, molysite and magnesium salts；And/or
The dispersion liquid and the mass ratio of the dispersant of being blended can be 1：(1~30), concretely 1：(15~20), 1：
15 or 1：20；The mass ratio of the emulsifier and the dispersant can be 1：(10~100), concretely 1：(20~50)；Institute
It can be at least one of petroleum ether, atoleine, dichloromethane, hexamethylene and aviation kerosine to state dispersant；The emulsifier
Can be tween (such as polysorbate60) and/or Span (such as Span 80 or Span 60)；And/or
The temperature of the cross-linking reaction can be 0~30 DEG C, concretely 15~30 DEG C, 15 DEG C or 30 DEG C, the time can be 2~
48h, concretely 30~36h, 30h or 36h.
In above-mentioned preparation method, in step (6), the minimizing technology of the pore-foaming agent can be it is following 1) or 2)：
1) pore-foaming agent is zein；The method is following a1) or a2)：
A1) product of step (5) is scattered in the ethanol water that mass fraction is 80%~92%, in 40~75 DEG C
Lower ultrasonic 5~20min is simultaneously filtered or is centrifuged, and repeat the above steps washing 3~5 times, you can removes the pore-foaming agent；
A2 the product of step (5)) is scattered in the aqueous acetone solution that mass fraction is 70%~80%, at 40~50 DEG C
5~20min of ultrasound is simultaneously filtered or is centrifuged, and repeat the above steps washing 3~5 times, you can removes the pore-foaming agent；
2) pore-foaming agent is CaCO3And/or MgCO3；The method is as follows：The product of step (5) is scattered in containing matter
It is water-soluble to measure the ethyl alcohol that score is the acetic acid of 0.5%~10% (such as 1%) or hydrochloric acid, mass fraction are 65%~95% (such as 70%)
In liquid, 5~20min of ultrasound and suction filtration or centrifugation at 40~75 DEG C, repeat the above steps washing 3~5 times, you can described in removing
Pore-foaming agent.
In above-mentioned preparation method, in step (7), the condition of the microwave treatment is as follows：Microwave power is 500~900W
(such as 500~800W, 500W or 800W), time per treatment are 10s~3min (such as 20s~1min, 20s or 1min), place
It is 3~10 times (such as 6~10 times, 6 times or 10 times) to manage number；Microwave treatment can further improve the stability of styptic powder structure,
Enhance haemostatic effect；The step has certain sterilizing function simultaneously, improves the stability of post sterilization processing quality.
In above-mentioned preparation method, the grain size of the styptic powder is preferably 100~500 μm.
The product being prepared by preparation method described in any one of the above embodiments, is within the scope of the invention.
The said goods are following 1) -5) at least one of in application or have following 1) -5 preparing) at least one
Application in the product of kind function, also within the scope of the present invention：
1) stop blooding；
2) surface of a wound anti-inflammation and sterilization；
3) promote cell Proliferation；
4) promote fibroblastic growth；
5) surface of a wound prevents adhesion.
Compared to existing technology and product, advantage of the invention is that：
(1) a kind of dual cross-linking method of polyoses grain is provided.This method is simple, and controllability is strong；Agents useful for same green
Environmental protection, removal is convenient, the risk of noresidue, and is conducive to the bioactivity for retaining amino in carboxymethyl chitosan.Microwave treatment
The stability that polyoses grain structure can be further increased, to enhance haemostatic effect；It is also beneficial to improve sterilization effect simultaneously.
(2) anthemorrhagic speed is fast, this has benefited from the design of the multiple hemostatic function of this product, is mainly reflected in the following：
There are a large amount of microcellular structure in grain surface and inside, and impart styptic powder has very strong capillary effect, knot in hemostasis
Close polysaccharide molecule itself have good hydrophily, therefore the hemostasis granules rate of liquid aspiration in the present invention faster and water absorbent rate more
Greatly；Amino in carboxymethyl chitosan can also promote the adhesion and aggregations such as haemocyte；Ca2+It is important coagulation factor, if with Ca2+
For ion crosslinking agent, polyoses grain can constantly discharge Ca in hemostasis2+Promote blood coagulation.
(3) styptic powder prepared by the present invention is applicable not only to capillary hemorrhage, veinlet bleeding and diffusivity bleeding,
The surface of a wound larger suitable for amount of bleeding, the clinical scope of application are wider.Styptic powder usage amount is small in hemostasis, reduces patient's
Medical treatment cost.
(4) the composite microporous polyoses grain has antibacterial anti-inflammatory, promotes that wound healing, adhesiveness be good, good biocompatibility,
The advantages that being absorbed by the body or being metabolized, have no toxic side effect.
Description of the drawings
Fig. 1 is the photo of the scanning electron microscope for the multi-functional micropore styptic powder being prepared in embodiment 3.
Fig. 2 is the photo of the zoopery in stopping blooding time test, and wherein Fig. 2-1 is the Hemorrhage Model figure on liver；Figure
2-2 is that styptic powder is applied to the bleeding surface of a wound；Fig. 2-3 is styptic powder wet processes；Fig. 2-4 is styptic powder gelation picture.
Specific implementation mode
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Embodiment 1 prepares multi-functional micropore styptic powder
(1) preparation of carboxymethyl starch paste liquid：By carboxymethyl starch (at 25 DEG C 2% solution viscosity be 500mpas,
Degree of substitution is 0.5) with water according to 1：40 mass ratioes mix, and are placed in 30 DEG C of stirred in water bath and it is made fully to dissolve obtained carboxylic first
Base gelatinized corn starch liquid.
(2) preparation of carboxymethyl chitosan solution：By carboxymethyl chitosan, (at 25 DEG C, viscosity is when 1% aqueous solution
100mpas, degree of substitution 80%) with water according to 1：60 mass ratioes are mixed and are fully dissolved, and carboxymethyl shell is obtained after filtering insoluble matter
Glycan solution for standby.
(3) carboxymethyl starch is pasted into liquid and carboxymethyl chitosan solution according to 1：1 is blended, and obtains blend solution.
(4) disperse pore-foaming agent：The zein that grain size is 5 μm is dispersed in the blend solution of step (3), pore-foaming agent：It is more
Sugared solid content g/g=1：100, it obtains that dispersion liquid is blended.
(5) ionomer：By CaCl2It is configured to the aqueous solution that mass fraction is 2%, under quick stirring, is added dropwise
Into the blending dispersion liquid (referred to as " polysaccharide blend liquid ") of step (4)；By obtained mixed system and the liquid containing Span 80
Paraffin is mixed, and is dispersed with stirring strongly, and after reacting 36h at 15 DEG C, is filtered and is fully washed.Wherein CaCl2It is molten
Liquid：Polysaccharide blend liquid g/g=1：50；Span 80：Atoleine g/g=1：20；Polysaccharide blend liquid：Atoleine g/g=1：15.
(6) pore-foaming agent is removed：The ethanol water that mass fraction is 80%, ultrasound 20min at 40 DEG C, repeats the above steps
Wash 5 times, drying.
(7) microwave treatment：The product of step (6) is placed in microwave and is handled：Microwave power is 500W, time 1min；
After being cooled to room temperature, same operation is handled 10 times repeatedly.
(8) it sieves, preferable particle size ranging from 100-500 μm of powder.
Embodiment 2 prepares multi-functional micropore styptic powder
(1) preparation of pregelatinized starch paste liquid：By pregelatinized starch (2% solution viscosity is 200mpas at 25 DEG C)
With water according to 1：25 mass ratioes mix, and are placed in 50 DEG C of stirred in water bath and are fully swollen obtained pregelatinized starch paste liquid.
(2) preparation of carboxymethyl chitosan solution：By carboxymethyl chitosan, (at 25 DEG C, viscosity is when 1% aqueous solution
200mpas, degree of substitution 80%) with water according to 1：30 mass ratioes are mixed and are fully dissolved, and carboxymethyl is made after filtering insoluble matter
Chitosan solution is spare.
(3) by gelatinized corn starch liquid and carboxymethyl chitosan solution according to 1：1 ratio is blended, and obtains blend solution.
(4) disperse pore-foaming agent：The CaCO that grain size is 2 μm3It is dispersed in the blend solution of step (3), pore-foaming agent：Polysaccharide is solid
Content g/g=1：200, it obtains that dispersion liquid is blended.
(5) ionomer：By FeCl3It is configured to the aqueous solution that mass fraction is 10%, under quick stirring, is added dropwise
Into the blending dispersion liquid (referred to as " polysaccharide blend liquid ") of step (4)；By obtained mixed system and the oil containing Span 80
Ether is mixed, and is dispersed with stirring strongly, and after reacting 30h at 30 DEG C, is filtered and is fully washed.Wherein FeCl3Solution：
Polysaccharide blend liquid g/g=1：30；Span 80：Atoleine g/g=1：50；Polysaccharide blend liquid：Atoleine g/g=1：20.
(6) pore-foaming agent is removed：The ethanol solution that mass fraction is 70%, wherein the acetic acid for being 1% containing mass fraction, in
Ultrasound 10min and suction filtration or centrifugation at 55 DEG C, the washing 4 times that repeats the above steps, drying.
(7) microwave treatment：The product of step (6) is placed in microwave and is handled：Microwave power is 800W, time 20s；It is cold
But to after room temperature, same operation is handled 6 times repeatedly.
(8) it sieves, preferable particle size ranging from 100-500 μm of powder.
The performance test of 2 multi-functional micropore styptic powder of embodiment 3, embodiment 1 and embodiment
(1) water absorbent rate is tested
Water absorbent rate is tested：Weigh 0.1g samples (W0) be added in the purified water of 10.0g or so, wait for that styptic powder is swollen 5min
After left and right causes water suction saturation, 10min is centrifuged under 500rpm rotating speeds, weigh to obtain W1.Water absorbent rate=(W1-W0)/W0× 100%.
Sample is respectively the styptic powder obtained of embodiment 1, styptic powder made from embodiment 2 and certain compound hemostatic powder (Shandong on the market
Sai Kesaisi medicine companies Science and Technology Ltd. " instantaneous " compound micropore polysaccharide styptic powder) and certain starch hemostatic powder (Hangzhou on the market
Synergism medical supplies Co., Ltd " glad speed is listened " MPH absorbable hemostatics microballoon).
Table 1, water absorbent rate result
| Sample | 5min water absorbent rates |
| " instantaneous " styptic powder | 1.5 |
| " glad speed is listened " styptic powder | 2 |
| Embodiment 1 | 3.5 |
| Embodiment 2 | 3 |
Remarks：Table 1 is liquid absorption amount (g) of the 0.1g styptic powders in each period
Compared with styptic powder on the market, styptic powder prepared by the present invention has higher water absorbent rate and in the unit interval
Absorption speed faster.This not only makes because of polysaccharide molecule good hydrophilic property, and there are a large amount of micropore in styptic powder surface and inside, such as
Shown in Fig. 1, this give styptic powders to have very strong capillary effect in hemostasis, to have very strong imbibition ability
With drain ability.Good drain ability can adherency excessive to avoid wound bed fluid amount and interfering material and wound tissue, from
And enhance its haemostatic effect, reduce the usage amount of styptic powder.
(2) bacteriostatic experiment
Bacteriostatic activity samples purifying water dissolution using growth rate method determination sample to pathogen, original concentration are
10mg/ml.Determination sample concentration is in 500 μ g/ml to the inhibition of staphylococcus aureus.
2ml sample solutions are added in the bacteria culture media of 60 DEG C of 38ml, it is 500 μ g/ to be made into styptic powder granule density
The culture medium of ml, using purified water as negative control.Culture medium is uniformly added into the culture medium of 2 each a diameter of 9cm, is waited for completely
After solidification, in the bacteria cake of 2 pieces of a diameter of 5mm of each inoculation of medium.After cultivating 48h or 72h at 25 DEG C, it is straight to measure bacterium colony
Diameter calculates the bacteriostasis rate of sample.
The biocidal property experimental result of table 2, multi-functional micropore styptic powder
As shown in Table 2, the styptic powder bacteriostasis rate that prepared by the present invention can reach 32%-42%, this anti-inflammatory for the surface of a wound
Sterilization is of great significance.
(3) bleeding stopping period is tested
Liver is most easy wound and causes the abdomen organ of lethal hemorrhage, therefore liver trauma Hemorrhage Model is in hemostatic material
It is generally used in material functional study.The present invention, which stops blooding to test, to be as follows：
According to the dosage of 1ml/kg after new zealand rabbit ear edge injects the anesthesia of 3% Nembutal sodium solution, rabbit backstroke is consolidated
Due on operating table, abdomen depilation, 75% alcohol wipe sterilizes.Abdomen is splitted, it is about 2cm that area is cut on liver2, thickness
The bleeding surface of a wound of about 2mm.Rapid paving spills about upper styptic powder, and presses 1min.The surface of a wound is observed after pressing in 5min to go out
Blood situation records bleeding stopping period, if recording and stopping blooding successfully without bleeding is repeated in 5min.
15 rabbits are randomly divided into 3 groups, respectively multi-functional micropore styptic powder group, certain commercially available starch hemostatic powder, it is commercially available certain
Complex polysaccharide styptic powder, every group 5.
Table 3, multi-functional micropore styptic powder are averaged bleeding stopping period
As shown in Table 3, the multi-functional micropore styptic powder that prepared by the present invention is compared with styptic powder in the market, the anthemorrhagic speed time
50% or more is shortened, therefore there is better haemostatic effect.
Fig. 2 is zoopery figure, from Fig. 2-3 as can be seen that when soaking styptic powder surface with water when surface of a wound amount of bleeding is few,
Water is imported into inside styptic powder soon, thus it is also seen that the styptic powder of the present invention has good drain and imbibition energy
Power.Swelling forms gel piece after styptic powder imbibition, and the gel piece itself has certain cohesive force it can be seen from Fig. 2-4；And
When gel piece is stripped, still there are many styptic powders to be adhered on the surface of a wound, illustrate that the styptic powder of the present invention has fine adhesiveness,
These are conducive to the effect that styptic powder plays closing hemostasis.
(4) haemolysis is tested
New blood is taken through New Zealand White Rabbit auricular vein, with 3.8% liquor sodii citratis with 9:1 mixing, is made anti-freezing
Blood.The fresh anticoagulated bloods of 8mL are taken, 10mL normal saline dilutions are added.Take 10mL styptic powders (be respectively 1 styptic powder of embodiment,
2 styptic powder of embodiment, " glad speed is listened " styptic powder as a contrast 1 and the physiological saline leaching liquor of " instantaneous " styptic powder as a contrast 2)
In in test tube, is stood in 37 DEG C and 0.2mL dilute bloods, 37 DEG C of water bath with thermostatic control 1h are added afterwards for 24 hours.Positive controls and negative control
0.2mL dilute bloods are respectively added with 10mL deionized waters and 10mL physiological saline respectively in group, with sample operation repetitive.All groups
Supernatant is taken after 800r/min centrifugations 5min, absorbance is measured in 540nm wavelength.Every group do 5 times it is parallel, take mean value.Hemolysis rate
It calculates according to the following formula：
Hemolysis rate %=(experimental group A540nmNegative control group A540nm)/(positive controls A540nmNegative control group
A540nm) × 100%；
Table 4, multi-functional micropore styptic powder hemolysis rate
Multi-functional micropore styptic powder hemolysis rate prepared by the present invention is 2.3%, is less than 5%, in national Biomaterial assay
Within defined safe range, illustrate that the styptic powder, without destruction, meets the requirement of bio-medical material to red blood cell.
(5) cytotoxicity experiment
The l cell L929 of logarithmic growth phase, after trypsin digestion adjust cell concentration be 4.0 ×
104200 μ l cells are accurately added with liquid-transfering gun per hole by the cell suspension inoculation after adjusting in 96 porocyte culture plates by a/ml
Suspension.In CO2Incubator (37 DEG C, 5% concentration C O2) in culture for 24 hours after, discard original fluid, be changed to the cell culture of styptic powder
Liquid leaching liquor (herein be the styptic powder being prepared in embodiment 2), control group are added normal incubation medium, set up 6
Tissue culture plate is placed in incubator and continues to cultivate by Duplicate Samples.Tissue culture plate is taken out respectively at 48h, 96h, 168h, is placed in
After microscopically observation cellular morphology, the 20 μ l of MTT of 5mg/ml are added per hole, react 4h.Culture solution is abandoned in suction, and DMSO is added
150 μ l, 37 DEG C of constant temperature 10min shake mixing 40s, OD value of each hole at 490nm are surveyed with microplate reader.It calculates according to the following formula thin
The opposite proliferation rate of born of the same parents：
Opposite proliferation rate=(experimental group mean light absorbency/control group mean light absorbency) × 100%
Table 5, mouse fibroblast cell L929 cultivate different in the degradable multi-functional micropore styptic powder leaching liquor of the present invention
Cell proliferation rate after time
Influence of the multi-functional micropore styptic powder leaching liquor to L929 mouse fibroblast cells is as shown in table 5.When cultivating 48h
When, it is 91.9% that styptic powder has slight adverse effect, cell opposite proliferation rate to the increment of cell, and cytotoxicity is 1 grade.When
When culture is to 96h and 168h, it is respectively 112.4% He that styptic powder cell proliferation, which has certain facilitation, opposite proliferation rate,
130%, cytotoxicity is 0 grade.Illustrate that multi-functional micropore hemostasis provided by the invention can promote fibroblastic growth.
Claims (10)
1. a kind of preparation method of styptic powder, it includes the following steps：
(1) starch is mixed with water, starch solution is prepared after dissolving or gelatinized corn starch liquid is prepared after swelling；
The starch is carboxymethyl starch, crosslinked carboxymethyl fecula, pregelatinized starch, pregelatinized starch, cationic starch and pre-
It is gelatinized at least one of hydroxypropyl PASELLI EASYGEL；
(2) carboxymethyl chitosan is mixed with water, carboxymethyl chitosan solution is prepared after insoluble matter is removed by filtration；
(3) starch solution described in step (1) or the gelatinized corn starch liquid and step (2) described carboxymethyl chitosan solution is mixed
It closes, obtains blend solution；
(4) pore-foaming agent is dispersed in the blend solution described in step (3), obtains that dispersion liquid is blended；
(5) under agitation, ion crosslinking agent is added dropwise in the blending dispersion liquid described in step (4), by what is obtained
Mixed system is mixed with dispersant and emulsifier, is filtered after crosslinked reaction；
The ion crosslinking agent is at least one of calcium salt, zinc salt, aluminium salt, molysite and magnesium salts；
(6) pore-foaming agent in the product obtained in removal step (5) and drying；
(7) microwave treatment is carried out to the product obtained in step (6), you can obtain the styptic powder.
2. preparation method according to claim 1, it is characterised in that：In step (1), the mass ratio of the starch and water is
1：(10~50)；The starch meets following conditions：At 25 DEG C, starch solution or gelatinized corn starch liquid that mass percentage is 2%
Viscosity be 200~1000mPas.
3. preparation method according to claim 1 or 2, it is characterised in that：In step (2), the carboxymethyl chitosan with
The mass ratio of water is 1：(10~100)；The carboxymethyl chitosan meets following conditions：At 25 DEG C, mass percentage 1%
Carboxymethyl chitosan aqueous solution viscosity be not less than 100mPas.
4. preparation method according to claim 1 or 2, it is characterised in that：In step (3), the starch solution or described
The mass ratio of gelatinized corn starch liquid and the carboxymethyl chitosan solution is 1：(0.1~10).
5. preparation method according to claim 1 or 2, it is characterised in that：In step (4), the pore-foaming agent quality and institute
The ratio for stating the quality sum of starch and the carboxymethyl chitosan is 1：(5~500)；The pore-foaming agent is zein, CaCO3
Or MgCO3；The grain size of the pore-foaming agent is 1~10 μm.
6. preparation method according to claim 1 or 2, it is characterised in that：In step (5), the ion crosslinking agent is with water
The form of solution is added, and the aqueous solution is 1 with the mass ratio that dispersion liquid is blended：(5~100)；The ion is handed over
The mass percentage for joining agent aqueous solution is 0.5%~40%；And/or
The dispersion liquid and the mass ratio of the dispersant of being blended is 1：(1~30)；The matter of the emulsifier and the dispersant
Amount is than being 1：(10~100)；The dispersant be petroleum ether, atoleine, dichloromethane, hexamethylene and aviation kerosine in extremely
Few one kind；The emulsifier is tween and/or Span；And/or
The temperature of the cross-linking reaction is 0~30 DEG C, and the time is 2~48h.
7. preparation method according to claim 1 or 2, it is characterised in that：In step (6), the removal side of the pore-foaming agent
Method be it is following 1) or 2)：
1) pore-foaming agent is zein；The method is following a1) or a2)：
A1) product of step (5) is scattered in the ethanol water that mass fraction is 80%~92%, is surpassed at 40~75 DEG C
It filters or centrifuges after 5~20min of sound, the washing that repeats the above steps can remove the pore-foaming agent 3~5 times；
A2 the product of step (5)) is scattered in the aqueous acetone solution that mass fraction is 70%~80%, it is ultrasonic at 40~50 DEG C
It filters and centrifuges after 5~20min, the washing that repeats the above steps can remove the pore-foaming agent 3~5 times；
2) pore-foaming agent is CaCO3And/or MgCO3；The method is as follows：The product of step (5) is scattered in containing quality point
In the ethanol water that the acetic acid or hydrochloric acid, mass fraction that number is 0.5%~10% are 65%~95%, surpass at 40~75 DEG C
5~20min of sound is filtered and is centrifuged, and the washing that repeats the above steps can remove the pore-foaming agent 3~5 times.
8. preparation method according to claim 1 or 2, it is characterised in that：The condition of the microwave treatment is as follows：Microwave work(
Rate is 500~900w, and the time per treatment is 10s~3min, and number of processes is 3~10 times；And/or the styptic powder
Grain size is 100~500 μm.
9. the product that the preparation method described in any one of claim 1-8 is prepared.
10. product described in claim 9 has following 1) -5 preparing) at least one of application in the product of function：
1) stop blooding；
2) surface of a wound anti-inflammation and sterilization；
3) promote cell Proliferation；
4) promote fibroblastic growth；
5) surface of a wound prevents adhesion.
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| CN106421881B (en) * | 2016-11-17 | 2019-08-02 | 杭州浙海企业管理有限公司 | A kind of chitosan anti hemorrhagic healing traumatic material and preparation method thereof |
| CN106729950B (en) * | 2016-12-07 | 2019-08-16 | 成都迪康中科生物医学材料有限公司 | A kind of hemostatic material and preparation method thereof |
| CN106730047B (en) * | 2016-12-07 | 2018-05-11 | 成都迪康中科生物医学材料有限公司 | A kind of bleeding stopping and adherence preventing film and preparation method thereof |
| CN106955370A (en) * | 2017-02-10 | 2017-07-18 | 江苏华能药业有限公司 | Composite starch styptic powder |
| CN107261193B (en) * | 2017-04-20 | 2021-02-09 | 首都医科大学附属北京同仁医院 | Hemostatic material and preparation method thereof |
| CN107501597B (en) * | 2017-08-07 | 2020-06-05 | 广州大学 | Preparation method of microporous starch |
| CN109010905A (en) * | 2018-10-10 | 2018-12-18 | 史跃 | A kind of preparation method of multi-functional emergency care of trauma styptic powder |
| CN109529095B (en) * | 2018-11-20 | 2021-07-27 | 福建农林大学 | Lotus seed starch-chitosan composite hemostatic powder and preparation method thereof |
| CN109498833B (en) * | 2018-12-12 | 2021-05-28 | 济南格莱威医疗科技有限公司 | Medical absorbable polysaccharide composite material and application thereof |
| CN109908396B (en) * | 2019-01-08 | 2021-11-02 | 中国人民解放军军事科学院军事医学研究院 | Calcium ion exchange porous starch hemostatic material and preparation method and application thereof |
| CN110538344B (en) * | 2019-10-09 | 2020-08-04 | 北京诺康达医药科技股份有限公司 | Medical degradable hemostatic material and preparation method thereof |
| CN112773928B (en) * | 2021-01-05 | 2021-09-21 | 珠海原妙医学科技股份有限公司 | Starch-based fluffy particles and preparation method and application thereof |
| 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 |
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| CN105056283A (en) * | 2015-07-29 | 2015-11-18 | 陕西博与再生医学有限公司 | Preparation method of composite styptic powder |
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| CN105056283A (en) * | 2015-07-29 | 2015-11-18 | 陕西博与再生医学有限公司 | Preparation method of composite styptic powder |
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