A kind of preparation method of compound hemostatic medical tissue glueTechnical field
The present invention relates to a kind of preparation methods of compound hemostatic medical tissue glue, belong to medical adhesive technical field.
Background technique
All the time, handle that wound is widely used in surgical field is suture.Traditional surgical operation suturingIt is not only operationally time-consuming with the hemostasis on organ-tissue, increase the difficulty of operation and tissue repair, is brought animally to patientSuture pain, while being also easy bacterial infection and causing some inflammatory reactions, infection, hyperplasia, the rupture of tissue, to group occurThe case where knitting the damage, necrosis or even disunion of organ.On the other hand, most of surgery operating wound generally all injures trueSkin and skin corium are hereinafter, the fibre composition of wound skin corium during reparation carries out hyperplasia, in the compact of operation suture threadUnder pulling force, therefore the interstitial space for providing cytothesis and hyperplasia is blocked, fibre composition just by fibrosis and forms granulationTissue, granulation tissue formation while, directly influence the secretion and absorption of collagen, cause collagen, pigment it is heavyProduct, collagen deposition abundant and its it is structural get muddled, be exactly the principal structural component of cicatricial tissue, it finalAs a result scar is shown as.
Scar is for the tissue before damage, an always incomplete replacement.From the point of view of aesthetical point, skin is causedThe destruction of skin is two degree of shades on numerous patients ' psychologicals, eternal mental wound, because having scar confident, entirelyThe world has ten hundreds of people to live in the shade of scar.It flows in nowadays material desire, U.S.A has become essential in lifeNot part, certainly, people to operation also proposed higher more perfect requirement, i.e., to mitigate surgical procedure to the greatest extentIn and postoperative pain, the fastest progresss wound tissue's reparation and while heal, be reduced as far as opening operation knife wound mouth instituteBring scar proliferation problem.
Part of the medical tissue glue in clinic in surgical operation for certain organs is binded and repairing, in bone surgeryThe combination and positioning of bone, joint, gear division operation in be used for tooth repairing etc., in the fields such as caesarean operation and beauty,Medical tissue glue more has the unrivaled superiority of other methods, not only, mitigates the operation stitching pain of patient, reaches promotionWound healing and the effect repaired more reduce the generation of complication, can also effectively reduce the formation of scar, this is for numerousIt is all a perfect for patient especially the gynemetrics's caesarean birth women and the medical professional of surgical operation suturing of liking to be beautifulMessage.
Adhesive effect of the medical tissue glue in human-body biological cell and tissue generally relates to bonding between cell, nothingIt is active with it is active between bonding, three aspects of bonding between inner body and ectosome, be specifically also manifested in soft tissue, ability to speakOn section, bone cement and skin binder, and what tissue glue also exactly put forward on the basis of this, between soft tissueBonding, it is understood that the bonding between cell and cell.As a kind of medical instrument consumptive material, directly with tissue, bloodLiquid, cell and each internal organs are contacted, and everyway should have high security, and ideal medical adhesive should have:
(1) safe and nontoxic, non-stimulated, nothing three causes (carcinogenic, teratogenesis, mutagenesis)；
(2) good biocompatibility, biological absorbable degradation；
(3) rapid curing, bonding action are high；
(4) stability is preferable, easily stored.
Medical adhesive can be generally divided into two major classes: chemical binder and biological adhesive.Wherein chemical binder accounts forMain body comes under chemical binder such as a-cyanoacrylate class, gelatin system class and polyurethanes, and clinically normal at presentSeveral prods；Biological adhesive is then based on fibrinogen (FS).
Summary of the invention
The technical problems to be solved by the invention: it for the problem that existing medical tissue glue bonding action is poor, providesA kind of preparation method of compound hemostatic medical tissue glue.
In order to solve the above technical problems, the technical solution adopted by the present invention is that:
(1) gelatin, fibrin gel are added in deionized water, with 180~200r/min stirring in 37~40 DEG C of water-baths15~20min, heat preservation, obtains colloidal solution；
(2) catechu phenolic group modification of chitosan is slowly added in colloidal solution, with 220~260r/min under 37~40 DEG C of water-bathsRevolving speed stirs 50~60min, obtains mixed solution；
(3) by tannic acid be added mixed solution in, in 37~40 DEG C of water-baths with 100~120r/min revolving speed crosslinking 15~20min stands 1.5~2h, 30~40min of ultraviolet-sterilization, obtains compound hemostatic medical tissue glue.
The gelatin, fibrin gel, catechu phenolic group modification of chitosan, tannic acid, the ratio between deionized waterAre as follows: according to parts by weight, 30~40 parts of gelatin, 10~20 parts of fibrin gels, 5~10 share tea phenolic group are weighed respectively and is changedProperty chitosan, 1~3 part of tannic acid, 80~100 parts of deionized waters.
The specific preparation step of fibrin gel described in step (1) are as follows:
(1) fibrinogen is added in physiological saline, 10~15min is stirred with 150~160r/min revolving speed under room temperature, is obtained fineFibrillarin original solution；
(2) by fibrin ferment, anhydrous calcium chloride be added deionized water in, under room temperature with 120~140r/min revolving speed stirring 20~30min obtains thrombin solution；
(3) fibrinogen solution is slowly dropped on six orifice plates with disposable needle tubing, keeps solution evenly laid out in bottom, then willThrombin solution is added dropwise in fibrinogen solution, is come into full contact with, uniformly mix, be placed on 36~38 DEG C of shaking tables stand 20~30min obtains fibrin gel.
The fibrinogen, physiological saline, fibrin ferment, anhydrous calcium chloride, the ratio between deionized water are as follows: by weightNumber meter is measured, weighs 1~3 part of fibrinogen, 10~20 parts of physiological saline, 3~5 parts of fibrin ferments, 2~4 parts of anhydrous chlorine respectivelyChange calcium, 20~30 parts of deionized waters.
The content of fibrinogen described in step (1) is 2~4g/L.
The specification of fibrin ferment described in step (2) is 500~2000IU.
The specific preparation step of catechu phenolic group modification of chitosan described in step (2) are as follows:
(1) 1- ethyl-(3- dimethylaminopropyl) carbodiimide hydrochloride, n-hydroxysuccinimide, dehydrated alcohol are addedEnter in deionized water, with 100~120r/min revolving speed 20~30min of stir-activating under room temperature, obtains mixed solution；
(2) chitosan solution is added in mixed solution, 15~20min is stirred with 160~180r/min revolving speed under room temperature, is obtained mixedClose liquid；
(3) 3,4-Dihydroxyphenylacetic acid solution is slowly added dropwise into mixed liquor, dropwise addition 5~10min of the world, with 200 under room temperature~220r/min revolving speed 1~2h of stirring, standing reaction 22~for 24 hours, it is freeze-dried 5~7 days, obtains catechu phenolic group modification of chitosan.
The chitosan solution, 3,4-Dihydroxyphenylacetic acid solution, 1- ethyl-(3- dimethylaminopropyl) carbon two are sub-Amine hydrochlorate, n-hydroxysuccinimide, dehydrated alcohol, the ratio between deionized water are as follows: according to parts by weight, weigh respectively20~30 parts of chitosan solutions, 20~30 parts of 3,4-Dihydroxyphenylacetic acid solution, 5~10 parts of 1- ethyls-(3- dimethylamino thirdBase) carbodiimide hydrochloride, 5~10 parts of n-hydroxysuccinimides, 20~30 parts of dehydrated alcohols, 40~50 parts of deionized waters.
The specific preparation step of chitosan solution described in step (2) are as follows: deionized water is added in chitosan by mass ratio 1: 5In, 30~40min is stirred with 200~220r/min revolving speed under room temperature, obtains chitosan solution.
The specific preparation step of 3,4-Dihydroxyphenylacetic acid solution described in step (3) are as follows: in mass ratio 1: 5 by 3,4- dihydroxyBase phenylacetic acid is placed in deionized water, is stirred 15~20min under room temperature with 120~140r/min revolving speed, is obtained 3,4- dihydroxy benzenesAcetic acid solution.
The present invention is compared with other methods, and advantageous effects are:
(1) present invention prepares compound hemostatic medical tissue glue, gelatin is a kind of collagen from Animal Skin using gelatin as raw materialThe fat-free component protein matter obtained after partially hydrolysed, gelfoam has loose porous structure, absorbable to overweight itself10~12 times of blood, and attachment and the coagulation factor of blood platelet can be activated, release and aggreation are generated, sludged blood is promotedIt is formed, forms nerve and blood vessel tight structure, realize quick-acting haemostatic powder, gelfoam is neutral in pH, can be 4~6 in human bodyWeek can degrade；
(2) present invention prepares compound hemostatic medical tissue glue using tannic acid as crosslinking agent, and tannic acid is a kind of common chemical combinationObject can degrade in vivo, in generally existing bark and fruit, due to containing unique polyphenol hydroxyl structure, Ke Yiyu in tannic acidThe large biological molecules such as protein-polysaccharide, which combine, complexing occurs with metal ion, there is good reproducibility, brightThe association reaction of glue and tannic acid is that multiple spot hydrophobic bond and hydrogen bond are coefficient as a result, the tannic acid containing hydrophobic group point between the twoIn the form of hydrophobic reactant in conjunction with gelatin, two o'clock Hydrogenbond, phenol occur son for the phenolic hydroxyl group of tannic acid and the polar group of gelatinHydroxyl is as hydrogen bond donor, and the ketonic oxygen on peptidyl is as receptor.Multiple spot hydrophobic bond and hydrogen bond is common between gelatin and tannic acidEffect forms gelatin-tannin acid complex, to improve the gumminess of compound hemostatic medical tissue glue；
(3) present invention prepares compound hemostatic medical tissue glue, chitosan is a kind of band by addition catechu phenolic group modification of chitosanPositive electricity alkaline polysaccharide has good biocidal property, has hemostatic function, can degrade in vivo, since chitosan surface is positively charged,The rapid aggregation of blood platelet and erythrocyte can be effectively facilitated into blood clot or certain polymerization effect itself can also occurIt answers, generates network structure to assemble free red blood cell, chitosan has certain viscosity after meeting blood, is woven with certain glue to groupAttached property, wound closure promote hemostasis to complete；
(4) present invention is grafted catechol group in chitosan molecule structure by coupling reaction, prepares the modified shell of catechu phenolic groupGlycan, catechu phenolic group are a kind of very strong polar groups, can be with the protein and polysaccharide polymer formation hydrogen bond in organismInteraction, catechu phenolic group are easily oxidized, and can form ortho position quinoid structure, quinoid structure in alkalinity or under conditions of containing oxidantVery not quietly, it can continue to react between any two with active hydrogen reaction, the quinoid structure after oxidation, be formed very strongThe chemical bond of active force, ultimately forms cross-linked structure, further enhances compound hemostatic medical tissue gluing knotting strength；
(5) present invention prepares compound hemostatic medical tissue glue, fibrin gel is fiber egg by addition fibrin gelBai Danti polymerize the biological macromolecule material of formation, the fiber egg in fibrin gel under fibrin ferment and catalytic factor effectWhite original can quickly form insoluble fibrin polymer, and the networking that further interweaves under the action of calcium ion and fibrin fermentBlood platelet and hemadsorption are formed blood clot by shape, complete hemostasis seals effect.Its anastalsis is small independent of blood simultaneouslyPlate or coagulation factor are applicable in the capillary hemorrhage patient of coagulation disorders or substantial viscera, fibrin gel very muchAs a kind of Biodegradable material, the treatment of wound healing can be widely used in.Since fibrin gel can promote carefullyThe adherency and proliferation of born of the same parents, and be natural biologic material, good biocompatibility can be used as drug, cell or cell factorSlow-released carrier.
Specific embodiment
In mass ratio 1: 5 by chitosan be added deionized water in, under room temperature with 200~220r/min revolving speed stirring 30~40min obtains chitosan solution, and in mass ratio 1: 5 is placed in 3,4-Dihydroxyphenylacetic acid in deionized water, under room temperature with 120~140r/min revolving speed stirs 15~20min, obtains 3,4-Dihydroxyphenylacetic acid solution, according to parts by weight, weighs 20~30 respectivelyPart chitosan solution, 20~30 parts of 3,4-Dihydroxyphenylacetic acid solution, 5~10 parts of 1- ethyls-(3- dimethylaminopropyl) carbonDiimmonium salt hydrochlorate, 5~10 parts of n-hydroxysuccinimides, 20~30 parts of dehydrated alcohols, 40~50 parts of deionized waters, by 1- secondBase-(3- dimethylaminopropyl) carbodiimide hydrochloride, n-hydroxysuccinimide, dehydrated alcohol are added in deionized water,With 100~120r/min revolving speed 20~30min of stir-activating under room temperature, mixed solution is obtained, it is molten that mixing is added in chitosan solutionIn liquid, 15~20min is stirred with 160~180r/min revolving speed under room temperature, mixed liquor is obtained, 3,4-Dihydroxyphenylacetic acid solution is delayedSlowly it is added dropwise in mixed liquor, dropwise addition 5~10min of the world, 1~2h is stirred with 200~220r/min revolving speed under room temperature, stands reaction22~for 24 hours, it is freeze-dried 5~7 days, obtains catechu phenolic group modification of chitosan, then according to parts by weight, weigh 1~3 part of fiber respectivelyProteinogen, 10~20 parts of physiological saline, 3~5 parts of fibrin ferments, 2~4 parts of anhydrous calcium chlorides, 20~30 parts of deionized waters, by fiberProteinogen is added in physiological saline, stirs 10~15min under room temperature with 150~160r/min revolving speed, obtains fibrinogen solution,Fibrin ferment, anhydrous calcium chloride are added in deionized water, 20~30min is stirred with 120~140r/min revolving speed under room temperature, is obtained solidifyingFibrinogen solution is slowly dropped on six orifice plates with disposable needle tubing, keeps solution evenly laid out in bottom by hemase solution, thenThrombin solution is added dropwise in fibrinogen solution, is come into full contact with, is uniformly mixed, is placed on 36~38 DEG C of shaking tables and stands 20~30min obtains fibrin gel, then according to parts by weight, respectively the gelatin, 10~20 parts of fibrins of 30~40 parts of weighingGel, 5~10 share tea phenolic group modification of chitosan, 1~3 difference tannic acid, 80~100 parts of deionized waters, by gelatin, fiber eggWhite gel is added in deionized water, stirs 15~20min in 37-40 DEG C of water-bath with 180~200r/min, and heat preservation obtains colloidCatechu phenolic group modification of chitosan is slowly added in colloidal solution by solution, is turned under 37~40 DEG C of water-baths with 220-260r/minSpeed 50~60min of stirring, obtains mixed solution, tannic acid is added in mixed solution, in 37~40 DEG C of water-baths with 100~120r/min revolving speed is crosslinked 15~20min, stands 1.5~2h, obtains compound hemostatic medical tissue glue.
Gelatin, fibrin gel are added in deionized water, 15min is stirred with 180r/min in 37 DEG C of water-baths, is protectedTemperature obtains colloidal solution；Catechu phenolic group modification of chitosan is slowly added in colloidal solution, is turned under 37 DEG C of water-baths with 220r/minSpeed stirring 50min, obtains mixed solution；Tannic acid is added in mixed solution, with the crosslinking of 100r/min revolving speed in 37 DEG C of water-baths15min stands 1.5h, ultraviolet-sterilization 30min, obtains compound hemostatic medical tissue glue.Gelatin, fibrin gel, catechu phenolic groupModification of chitosan, tannic acid, the ratio between deionized water are as follows: according to parts by weight, weigh respectively 30 parts gelatin, 10 parts of fibresFibrillarin gel, 5 share tea phenolic group modification of chitosan, 1 part of tannic acid, 80 parts of deionized waters.Fibrin gel is specifically preparedStep are as follows: fibrinogen is added in physiological saline, 10min is stirred with 150r/min revolving speed under room temperature, obtains fibrinogenSolution；Fibrin ferment, anhydrous calcium chloride are added in deionized water, 20min is stirred with 120r/min revolving speed under room temperature, obtains fibrin fermentSolution；Fibrinogen solution is slowly dropped on six orifice plates with disposable needle tubing, keeps solution evenly laid out in bottom, then will coagulateHemase solution is added dropwise in fibrinogen solution, is come into full contact with, and is uniformly mixed, is placed on 36 DEG C of shaking tables and stands 20min, obtain fineFibrillarin gel.Fibrinogen, physiological saline, fibrin ferment, anhydrous calcium chloride, the ratio between deionized water are as follows: by weightNumber meter weighs 1 part of fibrinogen, 10 parts of physiological saline, 3 parts of fibrin ferments, 2 parts of anhydrous calcium chlorides, 20 parts of deionizations respectivelyWater.The content of fibrinogen is 2g/L.The specification of fibrin ferment is 500IU.The specific preparation step of catechu phenolic group modification of chitosanAre as follows: 1- ethyl-(3- dimethylaminopropyl) carbodiimide hydrochloride, n-hydroxysuccinimide, dehydrated alcohol addition are goneIn ionized water, with 100r/min revolving speed stir-activating 20min under room temperature, mixed solution is obtained；It is molten that mixing is added in chitosan solutionIn liquid, 15min is stirred with 160r/min revolving speed under room temperature, obtains mixed liquor；By 3,4-Dihydroxyphenylacetic acid solution be slowly added dropwise toIn mixed liquor, world 5min is added dropwise, is freeze-dried 5 days, obtains with 200r/min revolving speed stirring 1h, standing reaction 22h under room temperatureTea phenolic group modification of chitosan.Chitosan solution, 3,4-Dihydroxyphenylacetic acid solution, 1- ethyl-(3- dimethylaminopropyl) carbonDiimmonium salt hydrochlorate, n-hydroxysuccinimide, dehydrated alcohol, the ratio between deionized water are as follows: according to parts by weight, respectivelyWeigh 20 parts of chitosan solutions, 20 parts of 3,4-Dihydroxyphenylacetic acid solution, 5 parts of 1- ethyls-(3- dimethylaminopropyl) carbon twoInferior amine salt hydrochlorate, 5 parts of n-hydroxysuccinimides, 20 parts of dehydrated alcohols, 40 parts of deionized waters.Chitosan solution specifically prepares stepSuddenly are as follows: in mass ratio 1: 5 chitosan is added in deionized water, stirs 30min under room temperature with 200r/min revolving speed, obtains chitosanSolution.The specific preparation step of 3,4-Dihydroxyphenylacetic acid solution are as follows: in mass ratio 1: 5 by 3,4-Dihydroxyphenylacetic acid be placed in fromIn sub- water, 15min is stirred with 120r/min revolving speed under room temperature, obtains 3,4-Dihydroxyphenylacetic acid solution.
Gelatin, fibrin gel are added in deionized water, 18min is stirred with 190r/min in 39 DEG C of water-baths, is protectedTemperature obtains colloidal solution；Catechu phenolic group modification of chitosan is slowly added in colloidal solution, is turned under 39 DEG C of water-baths with 240r/minSpeed stirring 55min, obtains mixed solution；Tannic acid is added in mixed solution, with the crosslinking of 110r/min revolving speed in 39 DEG C of water-baths18min stands 1.8h, ultraviolet-sterilization 35min, obtains compound hemostatic medical tissue glue.Gelatin, fibrin gel, catechu phenolic groupModification of chitosan, tannic acid, the ratio between deionized water are as follows: according to parts by weight, weigh respectively 35 parts gelatin, 15 parts of fibresFibrillarin gel, 8 share tea phenolic group modification of chitosan, 2 parts of tannic acid, 90 parts of deionized waters.Fibrin gel is specifically preparedStep are as follows: fibrinogen is added in physiological saline, 12min is stirred with 155r/min revolving speed under room temperature, obtains fibrinogenSolution；Fibrin ferment, anhydrous calcium chloride are added in deionized water, 25min is stirred with 130r/min revolving speed under room temperature, obtains fibrin fermentSolution；Fibrinogen solution is slowly dropped on six orifice plates with disposable needle tubing, keeps solution evenly laid out in bottom, then will coagulateHemase solution is added dropwise in fibrinogen solution, is come into full contact with, and is uniformly mixed, is placed on 37 DEG C of shaking tables and stands 25min, obtain fineFibrillarin gel.Fibrinogen, physiological saline, fibrin ferment, anhydrous calcium chloride, the ratio between deionized water are as follows: by weightNumber meter weighs 2 parts of fibrinogens, 15 parts of physiological saline, 4 parts of fibrin ferments, 3 parts of anhydrous calcium chlorides, 25 parts of deionizations respectivelyWater.The content of fibrinogen is 3g/L.The specification of fibrin ferment is 1250IU.The specific preparation step of catechu phenolic group modification of chitosanAre as follows: 1- ethyl-(3- dimethylaminopropyl) carbodiimide hydrochloride, n-hydroxysuccinimide, dehydrated alcohol addition are goneIn ionized water, with 110r/min revolving speed stir-activating 25min under room temperature, mixed solution is obtained；It is molten that mixing is added in chitosan solutionIn liquid, 18min is stirred with 170r/min revolving speed under room temperature, obtains mixed liquor；By 3,4-Dihydroxyphenylacetic acid solution be slowly added dropwise toIn mixed liquor, world 8min is added dropwise, is freeze-dried 6 days, obtains with 210r/min revolving speed stirring 1h, standing reaction 23h under room temperatureTea phenolic group modification of chitosan.Chitosan solution, 3,4-Dihydroxyphenylacetic acid solution, 1- ethyl-(3- dimethylaminopropyl) carbonDiimmonium salt hydrochlorate, n-hydroxysuccinimide, dehydrated alcohol, the ratio between deionized water are as follows: according to parts by weight, respectivelyWeigh 25 parts of chitosan solutions, 25 parts of 3,4-Dihydroxyphenylacetic acid solution, 8 parts of 1- ethyls-(3- dimethylaminopropyl) carbon twoInferior amine salt hydrochlorate, 8 parts of n-hydroxysuccinimides, 25 parts of dehydrated alcohols, 45 parts of deionized waters.Chitosan solution specifically prepares stepSuddenly are as follows: in mass ratio 1: 5 chitosan is added in deionized water, stirs 35min under room temperature with 210r/min revolving speed, obtains chitosanSolution.The specific preparation step of 3,4-Dihydroxyphenylacetic acid solution are as follows: in mass ratio 1: 5 by 3,4-Dihydroxyphenylacetic acid be placed in fromIn sub- water, 18min is stirred with 130r/min revolving speed under room temperature, obtains 3,4-Dihydroxyphenylacetic acid solution.
Gelatin, fibrin gel are added in deionized water, 20min is stirred with 200r/min in 40 DEG C of water-baths, is protectedTemperature obtains colloidal solution；Catechu phenolic group modification of chitosan is slowly added in colloidal solution, is turned under 40 DEG C of water-baths with 260r/minSpeed stirring 60min, obtains mixed solution；Tannic acid is added in mixed solution, with the crosslinking of 120r/min revolving speed in 40 DEG C of water-baths20min stands 2h, ultraviolet-sterilization 40min, obtains compound hemostatic medical tissue glue.Gelatin, fibrin gel, catechu phenolic group changeProperty chitosan, tannic acid, the ratio between deionized water are as follows: according to parts by weight, weigh respectively 40 parts gelatin, 20 parts of fibersProtein gel, 10 share tea phenolic group modification of chitosan, 3 parts of tannic acid, 100 parts of deionized waters.Fibrin gel is specifically preparedStep are as follows: fibrinogen is added in physiological saline, 15min is stirred with 160r/min revolving speed under room temperature, obtains fibrinogenSolution；Fibrin ferment, anhydrous calcium chloride are added in deionized water, 30min is stirred with 140r/min revolving speed under room temperature, obtains fibrin fermentSolution；Fibrinogen solution is slowly dropped on six orifice plates with disposable needle tubing, keeps solution evenly laid out in bottom, then will coagulateHemase solution is added dropwise in fibrinogen solution, is come into full contact with, and is uniformly mixed, is placed on 38 DEG C of shaking tables and stands 30min, obtain fineFibrillarin gel.Fibrinogen, physiological saline, fibrin ferment, anhydrous calcium chloride, the ratio between deionized water are as follows: by weightNumber meter weighs 3 parts of fibrinogens, 20 parts of physiological saline, 5 parts of fibrin ferments, 4 parts of anhydrous calcium chlorides, 30 parts of deionizations respectivelyWater.The content of fibrinogen is 4g/L.The specification of fibrin ferment is 2000IU.The specific preparation step of catechu phenolic group modification of chitosanAre as follows: 1- ethyl-(3- dimethylaminopropyl) carbodiimide hydrochloride, n-hydroxysuccinimide, dehydrated alcohol addition are goneIn ionized water, with 120r/min revolving speed stir-activating 30min under room temperature, mixed solution is obtained；It is molten that mixing is added in chitosan solutionIn liquid, 20min is stirred with 180r/min revolving speed under room temperature, obtains mixed liquor；By 3,4-Dihydroxyphenylacetic acid solution be slowly added dropwise toIn mixed liquor, world 10min is added dropwise, 2h is stirred with 220r/min revolving speed under room temperature, stands reaction for 24 hours, is freeze-dried 7 days, obtainsCatechu phenolic group modification of chitosan.Chitosan solution, 3,4-Dihydroxyphenylacetic acid solution, 1- ethyl-(3- dimethylaminopropyl)Carbodiimide hydrochloride, n-hydroxysuccinimide, dehydrated alcohol, the ratio between deionized water are as follows: according to parts by weight, pointAlso known as amount 30 parts of chitosan solutions, 30 parts of 3,4-Dihydroxyphenylacetic acid solution, 10 parts of 1- ethyls-(3- dimethylaminopropyl) carbonDiimmonium salt hydrochlorate, 10 parts of n-hydroxysuccinimides, 30 parts of dehydrated alcohols, 50 parts of deionized waters.Chitosan solution is specifically madeStandby step are as follows: in mass ratio 1: 5 chitosan is added in deionized water, stirs 40min under room temperature with 220r/min revolving speed, obtains shellGlycan solution.The specific preparation step of 3,4-Dihydroxyphenylacetic acid solution are as follows: in mass ratio 1: 5 is placed in 3,4-Dihydroxyphenylacetic acidIn deionized water, 20min is stirred with 140r/min revolving speed under room temperature, obtains 3,4-Dihydroxyphenylacetic acid solution.
Reference examples: the compound hemostatic medical tissue glue of Dongguan company production.
The compound hemostatic medical tissue glue that example and reference examples are prepared is detected, specific detection is as follows:
Bonding action: it is one layer thin to be coated on sheet glass formation by structure and ingredient in order to simulate bonding wound for gelatin solutionThin collagen film, with the Optical instrument of test organization glue.The specific method is as follows: the filtered hot gelatin of a certain concentration is moltenDrop is added on glass sheet surface, and glass bar is uniformly smeared and opened, the specification of sheet glass 6cm × 2cm, surpasses under natural conditions in cleanNet workbench is dry for 24 hours, and it is several must to be covered with a thin layer of gelatin film sheet glass.The area band of test be center S=2cm ×2cm, the tissue sol solution of 50uL is added dropwise to one piece of sheet glass middle ground band, and 20uL is added dropwise in another piece of sheet glass center 2cm × 2cmCrosslinking agent carbodiimides solution, central 2cm × 2cm contact surface bonding of two blocks of sheet glass avoids intermediate residue gas as far as possibleBubble, presses 10min or 5min(a certain regular time in 37 DEG C of baking ovens of 100g counterweight), weight beam tests adhesiveOptical instrument, the maximum, force F, F/S that writing down weight beam two blocks of sheet glass of stretching can bear then are to organize on unit areaThe bonding action of glue.
Blood compatibility: subject material is one group of experiment of four ratios: taking 6 Boiling tubes, it is numbered 1-4, aAnd b, wherein 1-4 is respectively 9/1,8/1,7/1,6/1 experimental group of gelatin/carboxymethyl chitosan mass ratio, and a is negative control, and b isPositive control, three parallel groups of every group of setting.Freeze-dried material sample 0.05g, the physiological saline of each ratio is added in 1-4 test tube10ml；A test tube adds physiological saline 10ml；B adds deionized water 10ml；37 DEG C of water bath with thermostatic control 0.5h；It is again 10-200uL with specificationLiquid-transfering gun be added to every test tube and dilute fresh anticoagulation 200uL, mix well, be transferred to 37 DEG C of warmed-up constant temperature at this timeWater-bath vibrator, with speed water bath with thermostatic control 1h appropriate；Each test tube solution is successively transferred to centrifuge tube, in centrifuge 1000r/Min is centrifuged 5min, and the supernatant 200uL after taking centrifugation is in 96 orifice plates, wherein 96 orifice plate peripheries make a circle and remove ionized water200uL, absorbance value of the measurement at 540nm in microplate reader.Take the mean value of three parallel groups as experimental result, according to public affairsFormula calculates hemolysis rate.
Specific test result such as table 1.
1 performance characterization contrast table of table
| Detection project | Example 1 | Example 2 | Example 3 | Reference examples |
| Bonding action/N | 2.50 | 2.45 | 2.55 | 0.70 |
| Hemolysis rate/% | 0.33 | 0.43 | 0.35 | 1.55 |
As shown in Table 1, compound hemostatic medical tissue glue prepared by the present invention has good bonding action and blood compatibility, isThe medical tissue glue of function admirable.