CN102406956A - Starch hemostatic microsphere and preparation method thereof - Google Patents
Starch hemostatic microsphere and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a starch hemostatic microsphere and a preparation method thereof, wherein bean starch, potato starch and cereal starch are used as raw materials, the starch hemostatic microsphere is prepared by an inverse suspension polymerization method, the molecular weight is 50, 000-5000, 000, and the particle size is 0.1-1000 μm, and the preparation method comprises the following steps: mixing three kinds of native starch, alkalifying, dispersing with surfactant and vegetable oil under the condition of pH8.5-10.5, cross-linking with cross-linking agent, washing, spray drying, packaging, and sterilizing with cobalt 60. The starch hemostatic microsphere prepared by the invention has good biocompatibility, is used for the hemostatic process of surgical operation, has the water absorption rate which is several times that of the existing hemostatic material, and has the water absorption speed which is obviously improved, thereby enhancing the hemostatic effect. The invention has the advantages of easily obtained raw materials, simple and convenient process, low production cost and wide application prospect.
Description
Technical field
The invention belongs to the hemorrhage field, relate to the starch hemorrhage, especially a kind of starch hemostasis microsphere and preparation method thereof.
Background technology
Surgical hemostasis is one of core of surgical technic, and good hemostatic technique is the key that guarantees that operation is successful.Along with of the raising of various countries' medical circle to hemostatic material effectiveness and security requirement, seek the good biomaterial of occurring in nature, develop haemostatic effect better, have no side effect, nonirritant, be easy to the hemostatic material of machine-shaping, imperative.Being used for surgery and wound at present has the absorbable hemostasia material of blood wound surface mainly to comprise:
1. Fibrin Glue
Fibrin Glue is claimed adhesive fibrin again, is a kind of biological product that from human plasma, extract, and mainly is made up of preparations such as thrombin and Fibrinogens, as the extensive use clinically of wound surface hemostasis adhesive material for many years.
Fibrin Glue is made up of Fibrinogen (containing the XVIII factor), thrombin, aprotinin and calcium chloride.Fibrinogen is under the effect of thrombin and calcium ion; Form stable insoluble fibrin polymer, after polymer formed, bring into play hemostasis, sealing process through following mechanism: 1. polymer was woven into netted; Can adsorb erythrocyte and platelet, form blood clot.2. after fibrin polymer formed, mechanical strength significantly strengthened, but adhesion organization and seal damagedly simultaneously can stimulate capillary endothelial cell and fibroblast growth, and was that support forms granulation tissue with the fibrin net, thereby promoted wound healing.
Fibrin Glue has good hemostasis, bond properties and histocompatibility, can reduce intraoperative blood loss and transfusion volume; Be applicable to that the organa parenchymatosum who is associated with the coagulation function disorder is hemorrhage, be particularly suitable for rough deep wound hemostasis.
Must be noted that Fibrin Glue belongs to the bioprotein preparation when using Fibrin Glue, when depositing and use, all should avoid high temperature, to avoid degeneration, influence result of use; Wound surface for big is hemorrhage in addition, arterial hemorrhage especially, because can not hemostasis by compression, must be aided with other materials.Must be noted that when using the absorbable fibre albumin glue can not intravasation in, in case thrombosis.The art district keeps dry as far as possible, and local temperature remains on about 37 ℃, use this agent after the part should oppress 3～5min, in order to avoid Fibrin Glue is washed away by blood flow before polymerization and influences haemostatic effect.Fibrin Glue is not suitable for tremulous pulse and the hemostasis hemorrhage than large vein.
The said material weak point is that cost is high, possibly bring human body or animal blood borne disease to infect, use inconvenience, anthemorrhagic speed slow, and poor to the trunk haemostatic effect.
2. oxidized cellulose and oxidized regenerated cellulose
Oxidized cellulose is a kind of absorbability local hemostasis material, and a kind of as cellulose derivative has excellent biological compatibility, biodegradability and characteristics such as nontoxic.Oxidized regenerated cellulose is a cellulose becomes the tulle shape or the cotton shape absorbable hemostatic material of cellulosic acid through oxidation processes, has the outward appearance and the quality of cotton yarn, is easy to bag, applies, operation such as filling.Its hemostatic mechanism is that concentrate blood starts clotting mechanism through the characteristic of material suction; Simultaneously, acid carboxyl combines with Hemoglobin F e, makes blood produce acid hematin, forms brown blob of viscose, and the sealing blood capillary is terminal and stop blooding.That present use clinically is the widest is the soluble stanching gauze Surgicel that Johnson Co. produces, and claims again that at home speed is yarn, is a kind of oxidized regenerated cellulose knitting yarn piece that contains thrombinogen, belongs to carboxymethyl cellulose class hemostatic material.
Surgicel can reduce damaged tissue pH value on every side, causes erythrocyte splitting, generates acid hematin, therefore with after blood contacts can form brown stickiness blob of viscose.This peracidity environment also has certain antibacterial ability, and is obvious to G+ and G-bacterium effect.The peracidity environment that Surgicel produces can cause nerve injury through a kind of diffusivity chemical mechanism, should avoid externally perineural directly a large amount of the use.When two or more biological hemostatic material was united use, the sour environment of Surgicel can reduce the haemostatic effect of other materials.More have report to show that Surgicel can strengthen damaged tissue inflammatory reaction on every side, the delay that cicatrizes a wound produces the pressuring nerve symptom at bone cavity and intracranial owing to absorbing blood makes volumetric expansion.
3. natural biological polysaccharide hemostatic material
3.1 chitosan class hemostatic material
Chitosan; Have different physiological roles such as antibacterial, anticancer, blood fat reducing, enhance immunity, the favorable tissue compatibility and higher biological activity, and nontoxic, non-stimulated, can natural degradation; Can be made into powder, liquid, fiber, gel and porous material and be used for various types of wound hemostasis; Also can be compound with one or more other macromolecular materials, or preparing derivant through chemical modification strengthens its anthemorrhagic performance, promotes wound healing.
The anastalsis of chitosan is because of it can stick the erythrocyte generation with erythrocyte surface is crosslinked, and certain repolymerization reaction takes place in blood, causes erythrocyte aggregation, thereby reaches the hemostatic purpose.In addition, chitosan and derivant thereof all have the good restraining effect to microorganisms such as antibacterial, yeast, funguses.
But the hemostatic material that uses chitosan to process merely, its haemostatic effect still is limited, and for bigger hemorrhage, its haemostatic effect is not very desirable.Therefore both at home and abroad generally all adopts and the method for the compound use of other hemorrhages solves.The hemorrhage majority of compound use is a thrombin, and collagen and calcium chloride etc. are also arranged.
3.2 starch type polysaccharide hemostatic material
Derive from the absorbability polysaccharide hemostatic material of modified starch, have many-sided functions such as hemostasis, anti, promotion organization healing.Its representative products PerClot
TM(Starch Medical Inc company) but degradation in vivo; The good compatibility is arranged; Can form simultaneously sludged blood in the part rapidly and performance mechanical barrier effect, in animal abdominal operation experiment, observe its hemostatic effective prevention of postoperative intestinal adhesion again simultaneously in art in the part.
Arista by the production of U.S. Medafor company
TMBe a kind of be the hemostatic material of substrate with the potato starch, its effective ingredient is many micropore polysaccharides.This many micropore polysaccharides extract from natural plant starch, nonhazardous, have the microsphere of biocompatibility, and the micropore of microsphere surface plays the effect of molecular sieve; Have strong absorptive, can the visible component in the blood (platelet, thrombin, fibrin etc.) be accumulated in particle surface at the moisture in the transient absorption blood; Produce " instant gel ", play the effect of mechanicalness shutoff blood vessel cut, simultaneously because the increase of the concentration of local thrombin; Accelerated intrinsic coagulation; Impel Fibrinogen to be converted into fibrin, solidify blood clot, reach solid and reliable hemostasis.But also there is deficiency in the haemostatic effect aspect of this effective hemostatic material, and its water absorption is strong inadequately, and the suction multiple is low, thereby influences the hemostatic effect.
Summary of the invention
The present invention has overcome the weak point of prior art, provides that a kind of good biocompatibility, water absorbent rate are higher, absorption speed starch hemostasis faster microsphere and preparation method thereof.
For realizing the foregoing invention purpose, the present invention adopts following technical scheme:
A kind of starch hemostasis microsphere is characterized in that: be to be raw material with the native starch, starch material is configured to the starch mixed solution; Alkali liquor activation then, the composite starch microsphere of processing through the inverse suspension polymerization method again, its molecular weight are 50; 000～5000,000, grain diameter is 0.1～1000 μ m; Said native starch comprises bean starch, potato starch and cereal kind of starch, and said bean starch, potato starch, the amyloid parts by weight ratio of cereal are 1: 2～5: 2～5.
And said bean starch, potato starch, the amyloid part by weight of cereal are 1: 2: 2.
And said bean starch comprises pea starch, Semen Viciae fabae starch, green starch.
And said potato starch comprises potato starch, sweet potato starch, tapioca.
And said cereal kind of starch comprises corn starch, rice starch, wheaten starch.
A kind of method for preparing of starch hemostasis microsphere, it is characterized in that: step is following:
(1) preparation starch mixed solution: bean starch, potato starch and the cereal kind of starch ratio according to ratio of weight and number 1: 2～5: 2～5 is dissolved in the distilled water of 5～20 times of weight; Alkali liquor is regulated pH 8.5～10.5; Stir, obtain starch alkaline solution after the activation;
(2) preparation vegetable oil-starch mixed solution: get surfactant: vegetable oil=1: 50～500g/ml; Mix; At 60～80 ℃ of stirred in water bath 0.5～2h, join then in the starch alkaline solution of step (1), mix 0.5～4h; The volume ratio of vegetable oil and starch alkali liquor is 5～20: 1, makes vegetable oil-starch mixed solution;
(3) crosslinking Treatment: in step (2), drip cross-linking agent, the amount of cross-linking agent is 1: 5～10g/ml with the ratio of starch, and the dropping time is 1～4h, forms reaction system, reaction 2～10h, and 40～60 ℃ of temperature, mixing speed are 300～1000r/min;
(4) microsphere is refining: the product sucking filtration with step (3), obtain solid, and after solids wash 2-5 time, dry, screening, sterilization make the starch microsphere that stops blooding.
And said vegetable oil comprises soybean oil, olive oil, Oleum Arachidis hypogaeae semen.
And said surfactant comprises span60, span80, tween80, and said cross-linking agent comprises epoxychloropropane, POCl3 ester, sodium trimetaphosphate.
And the washing step of said step (4) is: at first in solid, add 8～10 times of volumes of acetic acid ethyl esters and stir 4～8h, mixing speed is 200～500r/min, sucking filtration; In solid, add 5～10 times of volume dehydrated alcohol then and clean sucking filtration; Use 5～10 times of volume distilled water washs at last.
And the sterilization of said step (4) is to sterilize with cobalt 60.
The present invention has following advantage and good effect is:
1, the starch hemostasis microsphere that makes of the present invention at first mixes composite through the ative starch of multiple separate sources; And then crosslinked with cross-linking agent, the hemostasis microsphere of this method for preparing preparation organically combines the starch molecule of different sizes, forms the complex microsphere of variety classes starch; Effectively improved the water absorbent rate of spherex; Be the several times of existing hemostatic material, its absorption speed also obviously increases, and has improved haemostatic effect.
2, the starch hemostasis microsphere that makes of the present invention; Can be absorbed by human body, good biocompatibility can be used for operating hemostasis; The blood wound surface that has that can directly be sprayed on people, mammal etc. stops blooding, and also can be used as the surgery anti, promotes the organization healing material.
3, to prepare the used oil phase of starch hemostasis microsphere be crude vegetal in the present invention, wide material sources, to human non-toxic side effect, environmentally safe, environmental protection.
4, the starch hemostasis microsphere of the present invention's preparation has the advantage that raw material sources are extensive, cheap, production cost is low, technology is easy, have a extensive future.
Description of drawings:
Fig. 1 is 1# and AristaTM water absorbent rate comparison diagram;
Fig. 2 is 1# and AristaTM absorption speed comparison diagram;
Fig. 3, Fig. 4 are respectively the electromicroscopic photograph of the spherex of the present invention's preparation.
The specific embodiment
Below in conjunction with embodiment, the present invention is further specified, following embodiment is illustrative, is not determinate, can not limit protection scope of the present invention with following embodiment.
A kind of starch hemostasis microsphere is to be raw material with the native starch, and starch material is configured to the starch mixed solution; Alkali liquor activation then, the composite starch microsphere of processing through the inverse suspension polymerization method again, its molecular weight are 50; 000～5000,000, grain diameter is 0.1～1000 μ m; Said native starch comprises bean starch, potato starch and cereal kind of starch, and said bean starch, potato starch, the amyloid part by weight of cereal are 1: 2～5: 2～5, and wherein starch is the commercially available prod.
Inverse suspension polymerization is to be disperse medium with the oil phase; Monomer or macromolecule are as water; Rely on the effect of suspension stabilizer to be dispersed in the oil phase; Form water in oil suspension, the synthetic method that adopts water soluble starter or catalyst to react at aqueous phase, this method is a kind of common method in the spherex preparation.
Embodiment 1
A kind of method for preparing of starch hemostasis microsphere, step is following:
(1) preparation starch alkaline solution: claim pea starch 1g, potato starch 2g, corn starch 2g; Distilled water 30～100ml; Starch dissolution in distilled water, is regulated pH 8.5～10.5 with 1mol/L NaOH, stir 2h; Mixing speed is 400-600r/min, obtains starch alkaline solution after the activation;
(2) preparation vegetable oil-starch mixed solution: after getting surfactant span601-3g, vegetable oil 200-1000ml mixing; At 60～80 ℃ of stirred in water bath 0.5～2h; Join then in the starch alkali liquor of step (1), mix 2～4h, obtain vegetable oil-starch mixed solution;
(3) crosslinking Treatment: to vegetable oil-starch mixed solution and dripping cross-linking agent epoxychloropropane 1～5ml that step (2) obtains, the dropping time is 1～2h, forms reaction system, reaction 4～6h, and 50 ℃ of temperature, mixing speed are 400-600/min;
(4) microsphere is refining: the product sucking filtration with step (3), obtain solid, and add 8～10 times of volumes of acetic acid ethyl esters and stir 4～8h, mixing speed is 200～500r/min, sucking filtration; In solid, add 5～10 times of volume dehydrated alcohol then and clean sucking filtration; Use 5～10 times of volume distilled water washs at last, spray drying obtains pressed powder, obtains product after screening, the co-60 radiation sterilization.
Embodiment 2
A kind of method for preparing of starch hemostasis microsphere, step is following:
(1) preparation starch alkaline solution: claim green starch 1g, tapioca 2.5g, corn starch 3g; Distilled water 30～100ml; Starch dissolution in distilled water, is regulated pH 8.5～10.5 with 1mol/L NaOH, stir 2h; Mixing speed is 400-600r/min, obtains starch alkaline solution after the activation;
(2) preparation vegetable oil-starch mixed solution: after getting surfactant span601-3g, vegetable oil 200-1000ml mixing; At 60～80 ℃ of stirred in water bath 0.5～2h; Join then in the starch alkali liquor of step (1), mix 2～4h, obtain vegetable oil-starch mixed solution;
(3) crosslinking Treatment: to vegetable oil-starch mixed solution and dripping cross-linking agent epoxychloropropane 1～5ml that step (2) obtains, the dropping time is 1～2h, forms reaction system, reaction 4～6h, and 50 ℃ of temperature, mixing speed are 400-600/min;
(4) microsphere is refining: the product sucking filtration with step (3), obtain solid, and add 8～10 times of volumes of acetic acid ethyl esters and stir 4～8h, mixing speed is 200～500r/min, sucking filtration; In solid, add 5～10 times of volume dehydrated alcohol then and clean sucking filtration; Use 5～10 times of volume distilled water washs at last, spray drying obtains pressed powder, obtains product after screening, the co-60 radiation sterilization.
It below is the performance test of the starch hemostasis microsphere of the foregoing description 1 preparation
The water absorbing properties contrast
Water absorbing properties of the present invention adopts the capillary tube method determinator to record, and water filling in acid buret makes acid buret zero graduation liquid level concordant with sand core funnel filter plate lower end.Filter paper is put into sand core funnel, contact fully with filter plate.Open piston, absorb water fully to filter paper, read the burette calibration data, as initial zero graduation.Closure piston takes by weighing the 0.1g powder, evenly is layered on the filter paper, opens piston simultaneously, begin to descend from liquid level, and every 20s, 40s, the liquid level dropping distance is observed in the 60s timing, calculation sample absorption speed and the suction saturated conditions in the unit interval.
Absorbable hemostasia microsphere 1# and Arista in the embodiment of the invention
TM(medafor company, the U.S.) water absorbing properties compares, and it is as shown in table 1 to measure the result
Table 1 water absorbing properties synopsis
| Arista TM | 1# | |
| Water absorbent rate (ml/g) | 6 | 16.5 |
| Absorption speed (ml/g) (first 20s) | 0.1917 | 0.3167 |
| Absorption speed (ml/g) (second 20s) | 0.0333 | 0.1333 |
| Absorption speed (ml/g) (the 3rd 20s) | 0.025 | 0.0833 |
Water absorbent rate refers to the maximum that the 1g sample can absorb water.
Water absorbent rate (ml/g)=water absorption (ml)/sample size (g)
Consulting Fig. 1 is 1# and Arista
TMShown in the water absorbent rate comparison diagram, and visible by table 1, and starch hemostasis microsphere 1# of the present invention is with respect to Arista
TM, its water absorbent rate obviously improves, and is about Arista
TM3 times.
Absorption speed is respectively in the first, the second and the 3rd 20s, the water absorption (ml)/20 (s) in the average speed of suction, V20s=20s
Consulting Fig. 2 is 1# and Arista
TMShown in the absorption speed comparison diagram, and visible by table 1, and the absorption speed of 1# of the present invention in three 20s of difference is all obviously greater than Arista
TM, explain that 1# compares Arista
TMIt is faster, more effective to absorb water.
Control experiment
Research to the dirty hemorrhage model haemostatic effect experiment of Hepar Mus
Experiment purpose:
Observe the haemostatic effect of 1# product to the dirty hemorrhage model of Hepar Mus
Receive the reagent thing:
Title: 1# product
Animal: white mice, Medical University Of Tianjin zoopery center provides
5 of every treated animals, totally 10, body weight 250-300g, male and female are not limit
Experimental technique
Get 10 of white mouse, random packet, is divided into 1# (Tianjin Aileyi Medicine Materials Co., Ltd. provides), matched group (AristaTM) (medafor company, the U.S.) by 5 every group.White mouse is anaesthetized with 3% barbital injection (30-40mg/kg) intravenous injection, on testing stand, fixes white mouse, and the sterile working successively opens abdomen; Expose the white mouse liver, do cutting at liver at random with knife blade, when treating that blood flows out in a large number; Spray hemostatic material immediately; With gauze push 10,20,30s, observe the hemostasis situation of respectively organizing, the record successfully routine number that stops blooding is seen table 2
The table 2 successfully routine number statistical table of stopping blooding
| 1# | Arista TM | |
| Push the 10s routine number that successfully stops blooding | 2/5 | 1/5 |
| Push the 20s routine number that successfully stops blooding | 5/5 | 3/5 |
| Push the 30s routine number that successfully stops blooding | 5/5 | 4/5 |
The electromicroscopic photograph of the starch hemostasis microsphere of the present invention's preparation such as Fig. 3, Fig. 4; From figure, find out that this spherex is the comparatively slick sphere structure in surface, the form rule, particle size distribution is comparatively concentrated; Mostly at 50-300 μ m; Wherein to react overall structure all be regular spheroid to Fig. 3, and particle diameter is comparatively even, and Fig. 4 is the smooth surface of spheroid.
Claims (10)
1. a starch hemostasis microsphere is characterized in that: be to be raw material with the native starch, starch material is configured to the starch mixed solution; Alkali liquor activation then, the composite starch microsphere of processing through the inverse suspension polymerization method again, its molecular weight are 50; 000～5000,000, grain diameter is 0.1～1000 μ m; Said native starch comprises bean starch, potato starch and cereal kind of starch, and said bean starch, potato starch, the amyloid parts by weight ratio of cereal are 1: 2～5: 2～5.
2. starch hemostasis microsphere according to claim 1, it is characterized in that: said bean starch, potato starch, the amyloid parts by weight ratio of cereal are 1: 2: 2.
3. starch hemostasis microsphere according to claim 1, it is characterized in that: said bean starch comprises pea starch, Semen Viciae fabae starch, green starch.
4. starch hemostasis microsphere according to claim 1, it is characterized in that: said potato starch comprises potato starch, sweet potato starch, tapioca.
5. starch hemostasis microsphere according to claim 1, it is characterized in that: said cereal kind of starch comprises corn starch, rice starch, wheaten starch.
6. the method for preparing of a starch as claimed in claim 1 hemostasis microsphere, it is characterized in that: step is following:
(1) preparation starch mixed solution: bean starch, potato starch and the cereal kind of starch ratio according to ratio of weight and number 1: 2～5: 2～5 is dissolved in the distilled water of 5～20 times of weight; Alkali liquor is regulated pH 8.5～10.5; Stir, obtain starch alkaline solution after the activation;
(2) preparation vegetable oil-starch mixed solution: get surfactant: vegetable oil=1: 50～500g/ml; Mix; At 60～80 ℃ of stirred in water bath 0.5～2h, join then in the starch alkaline solution of step (1), mix 0.5～4h; The volume ratio of vegetable oil and starch alkali liquor is 5～20: 1, makes vegetable oil-starch mixed solution;
(3) crosslinking Treatment: in step (2), drip cross-linking agent, the amount of cross-linking agent is 1: 5～10g/ml with the ratio of starch, and the dropping time is 1～4h, forms reaction system, reaction 2～10h, and 40～60 ℃ of temperature, mixing speed are 300～1000r/min;
(4) microsphere is refining: the product sucking filtration with step (3), obtain solid, and after solids wash 2-6 time, dry, screening, sterilization make the starch microsphere that stops blooding.
7. the method for preparing of starch hemostasis microsphere according to claim 6, it is characterized in that: said vegetable oil comprises soybean oil, olive oil, Oleum Arachidis hypogaeae semen.
8. the method for preparing of starch hemostasis microsphere according to claim 6, it is characterized in that: said surfactant comprises span60, span80, tween80, said cross-linking agent comprises epoxychloropropane, POCl3 ester, sodium trimetaphosphate.
9. the method for preparing of starch hemostasis microsphere according to claim 6, it is characterized in that: the washing step of said step (4) is: at first in solid, add 8～10 times of volumes of acetic acid ethyl esters and stir 4～8h, mixing speed is 200～500r/min, sucking filtration; In solid, add 5～10 times of volume dehydrated alcohol then and clean sucking filtration; Use 5～10 times of volume distilled water washs at last.
10. the method for preparing of starch hemostasis microsphere according to claim 6, it is characterized in that: the sterilization of said step (4) is to sterilize with cobalt 60.
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