CN104254347A - Method of forming dextran and thrombin sheets - Google Patents

Abstract

A method of forming hemostatic sheets. Dextran and water are mixed to form a dextran-water mixture. The dextran-water mixture is formed into a first support layer. Fibrinogen and thrombin are mixed to form a fibrinogen and thrombin mixture. The fibrinogen and thrombin mixture is dispersed on the first support layer to form a hemostatic sheet.

Description

Form the method for dextran and thrombin sheet material

Technical field

Relate generally to of the present invention has the product of haemostatic properties.More particularly, the present invention relates to the method being formed and be suitable for dextran (dextran) and thrombin (thrombin) sheet material used in hemostat application.

Background technology

For stoping from wound bleeding, the natural reaction of health is the complex process startup blood coagulation by being called as coagulation cascade (coagulation cascade).Described cascade comprises two approach, finally causes producing enzyme thrombin, and its catalysis fibre proteinogen (fibrinogen) is converted into fibrin (fibrin).

Then, fibrin is cross-linked to form sludged blood (clot), causes hemostasis.For not having wound not serious in the individuality of antagonism situation (countervening condition), health can carry out this process to stop from the mode of wound excessive blood loss usually effectively.But, when serious wounds, or clotting mechanism be destroyed individuality when, this may be really not so.

But, individual for this type of, can by using coagulation cascade composition directly to wound, especially thrombin and Fibrinogen, and realize hemostasis.Bleeding wounds wrapping is also common way, and isolate to a certain extent and protect wound area, and also providing and execute stressed means to wound, this also can help Bleeding control.

Although at mild trauma or when injuring (as operation) in " controlled ", can implement these methods satisfactorily, many situations needing most this kind of process are also those the most difficult situations providing this kind of process.The example of this kind of wound comprises, and such as, those are during the struggle or as the unexpected wound of thunder bolt result.In this case, the existence of injured individual may depend on that prevention is by the hemostasis reached in the hemorrhage and injured rear first few minutes of wound.Unfortunately, due to this kind of injured situation, possibly suitable medical intervention cannot be used immediately.

Particularly, it is problematic for processing perforating wound (penetrating wound) such as bullet wounds or wound that some are caused by shell fragment.This is owing to being difficult to place binder and/or therapeutic agent at accurate injury site, described injury site comprise below just body surface, be difficult to the region that routine techniques maybe can not be used to arrive.

The people such as Jiang teach Electrospun dextran fiber (electrospun dextran fibers) at Biomacromolecules, v. 5, p. in 326-333 (2004).The direct Electrospun of medicament be combined with fiber (as BSA, lysozyme) enters in fiber.Described fiber also may comprise other polymer with the spinning of dextran Static Spinning.

The people such as Jiang are at Journal of Biomedical Materials Research Part B:Applied Biomaterials, p. disclose electricity spinning fibre in 50-57 (2006), it is as shell with using dextran as the compositions of core using polycaprolactone (poly (c-caprolactone)).These fibers provide the slow releasing of medicament (bovine serum albumin, BSA), and described medicament is also entered in fiber by Electrospun.

The United States Patent (USP) the 6th, 753 of the people such as Smith, No. 454 disclose electricity spinning fibre, and it comprises the substantially uniform mixture of hydrophilic polymer and at least weak hydrophobic polymer, and it can be used for forming binder.Described binder can comprise activating agent (as dextran).But disclosed fiber is not easy to be dissolved in liquid.

The United States Patent (USP) the 6th, 762 of the people such as MacPhee, No. 336 teach hemostasis multilamellar binder, and it comprises the thrombin layer between two fibrinogen layer.Described binder can also contain other absorbable material, as glycolic or based on the polymer of lactic acid or copolymer.No matter be electricity spinning fibre or dextran fiber are not all used as binder constituent by instruction.

The United States Patent (USP) the 6th, 821 of the people such as Smith, No. 479 teach a kind of method of preserving biomaterial in the protectiveness substrate of drying, and described substrate comprises fiber as electricity spinning fibre.A kind of composition of described fiber can be dextran, but does not describe homogeneous phase dextran fiber.

The United States Patent (USP) the 7th, 101 of the people such as Cochrum, No. 862 teach hemostatic composition for Bleeding control and method.Described compositions comprises cellulose goods (as gauze), covalency or ionomer polysaccharide on it.Described cross-linked polysaccharides can be dextran.But described compositions is not Electrospun, and does not comprise extrinsic coagulation agent in described compositions.

No. 2004/0018226th, the U.S. Patent Publication of the people such as Wnek, discloses the fiber produced as Electrospun by EDM Technology.Described fibrous inside comprises inner cover, for comprising not miscible with fiber material.Dextran is not instructed as fibre composition.

No. 2007/0160653rd, the U.S. Patent Publication of the people such as Fisher, teach a kind of hemostatic textile, it comprises hemostatic factor (as thrombin, Fibrinogen), but described fiber adds the second fiber (as silk, pottery, bamboo, fiber crops, artificial silk etc.) by Electrospun glass is formed.

No. 2008/0020015th, the U.S. Patent Publication of the people such as Carpenter, teaches wound dressing, and it comprises the hydrogel that various biodegradable polymers and having is scattered in allos in described polymer or autologous precursor (such as stem cell).Described polymer is prepared by Electrospun, and a kind of polymers compositions can be dextran.But described polymer can not contact with liquid one and namely dissolve, and provides scaffold because they are necessary for along with passage of time delivery of cells, although the final in situ biodegradation of described polymer.

No. 2008/0265469th, the U.S. Patent Publication of the people such as Li, describes Electrospun nano-fibers, and it can comprise dextran.But, do not describe described nanofiber and be soluble in liquid.

No. 2009/0053288th, the U.S. Patent Publication of the people such as Eskridge, teach a kind of braiding hemostatic textile, it comprises the glass wire of about 65% and the bamboo fibre line of about 35%.Described fiberglass component can Electrospun, and hemostatic factor such as thrombin can be combined with fabric, such as, by being immersed in thrombin solution by material.The document shows that dextran can be used as hygroscopic agent and adds.

Need to continue to provide the ways and means of improvement to start the blood coagulation in wound, lose blood to stop or at least to slow down.Particularly, need the Continual Improvement mode of being easy to be easy to impel the ability of stopping blooding in serious wounds, particularly limited when processing immediately of being implemented by medical worker or can not.

No. 2011/0150973rd, the U.S. Patent Publication of the people such as Bowlin, discloses a kind of method of sending one or more target agent to target location.Described method comprises target location application or the hemostasia products sending hemostasis.The hemostasia products of described hemostasis comprises the Electrospun dextran fiber dissolved with liquid comes into contact.The hemostasia products of described hemostasis also comprises one or more target agent be combined with described Electrospun dextran fiber.Application or send causes at Electrospun dextran fibrolysis described in target location in liquid, thus discharges one or more target agent in liquid.

Summary of the invention

One embodiment of the invention relate to the method forming hemostasis sheet material.Dextran and water are mixed to form dextran-aqueous mixtures.Described dextran-aqueous mixtures is formed the first supporting layer (support layer).Fibrinogen and thrombin are mixed to form Fibrinogen and Thrombin mixture.Described Fibrinogen and Thrombin mixture are dispersed on described first supporting layer and form hemostasis sheet material.

Accompanying drawing explanation

Accompanying drawing is included to provide understands embodiment further, and it is merged in and forms the part of this description.Accompanying drawing illustrates embodiment, and together with the description for explaining the principle of embodiment.Owing to can understand them better by reference to detailed description below, many expection advantages of other embodiment and embodiment will be readily appreciated that.Element in accompanying drawing is not necessarily by scale relative to each other.Similar label represents corresponding similar portion.

Fig. 1. the schematic diagram of electric spinning device.The key element of electrospinning system comprises high voltage power supply, for the raw material storage device of polymer and ground connection axle (grounded mandrel).This system adopts cylindrical target axle; But electrospinning process can be suitable for producing more complicated shape.Single and/or multiple polymer can be transported to electric field from one or more raw material storage device independently or side by side.Can use and produce laminated construction from the different and polymer of uniqueness of independently source Electrospun in chronological order.

Fig. 2 A and B. A, air brush base dextran process schematic representation; B, by the dextran fiber of electric mist explained hereafter.The quantity of material described is possible the amount of enough about 2 hemostasia products.Note the setting-out of material.Use electric field by dextran targeting to axle.

Fig. 3. the scanning electron micrograph of Electrospun dextran fiber.The nominal average cross sectional diameter of single fiber is 1 micron, provides a large amount of surface areas.

Fig. 4 A-E. is by the schematic diagram of the exemplary hemostasia products of Electrospun dextran processbearing astrocyte.A, with the hemostasia products of non-permeability support material as backing (backing); B, with the hemostasia products of netted support material; C, with non-permeability backing and hemostasia products electricity spinning fibre being maintained the netted support material on backing; D, for by therapeutic agent delivery to the hemostasia products of deep layer wound; E, for by therapeutic agent delivery to the alternative of the hemostasia products of deep layer wound.

Fig. 5 A and B. is exposed to the change of animal body based intracellular cvtokine (cytokine) level of salmon Fibrinogen/thrombin hemostasia products.(A) log ratio of the Blood Cytokines level determined when the level of IL-1 β, IL-6, TNF-α, IFN-γ, IL-4 and IL-10 is illustrated as initial surgery implantable intravascular mouth and the peak level after exposing.All change is seen in proinflammatory reaction (IL-1 β, IL-6, TNF-α, IFN-γ) and humoral response (IL-4 and IL-10).(B) the change display of individual animals based intracellular cvtokine is first exposes (the first arrow) and venoclysis albumen subsequently (the second arrow) induces reaction, and this reaction can detect upper once getting in the sample of blood.

The immunoglobulin that Fig. 6 A-F. is produced by western blotting qualitative evaluation pig response salmon protein.(A) salmon (Sal), people (Hu) and pig (Sw) fibrinogen preparation and with the PAGE of respective egg white matter trace coming from two kinds of animal (B and C) serum.These groups show the serum profiles from the blood exposed after front and final euthanasia.By specificity HRP anti-pig IgG second antibody and according to detecting the visual IgG isotype appeared in serum of protein be attached in gel sample.The position of IgG heavy chain and light chain composition in arrow instruction pig albumen swimming lane, also can by second antibody identification.(kDal × 10, left side ^-3) show molecular weight.(D) salmon (Sal), people (Hu) and pig (Sw) thrombin preparation and with the PAGE of respective egg white matter trace coming from the same animals serum that (C and D) illustrates.In these animals, in E, recognize thrombin, but at F(arrow) salmon protein swimming lane in have faint reaction.Camera calibration in detection system is the white ribbon in F to the heavy chain pig thrombiase albumen on film.

The time course that the animal internal antibody that Fig. 7 A-D. is exposed to salmon thrombin/Fibrinogen hemostasia products via transdermal patches program occurs.Anti-igg reagent is used to carry out ELISA.Following further antigens is used as the target in ELISA: (A) salmon Fibrinogen, (B) salmon thrombin, (C) human fibrinogen and (D) human thrombin.After the absorbance increase observed at sample sets A subsequently, B, C betides venoclysis salmon protein.Each curve representative is from the data of different animals.

The time course that the animal internal antibody that Fig. 8 A-D. is exposed to salmon thrombin/Fibrinogen hemostasia products via abdominal part paster program occurs.Anti-igg reagent is used to carry out ELISA.Following further antigens is used as the target in ELISA: (A) salmon Fibrinogen, (B) salmon thrombin, (C) human fibrinogen and (D) human thrombin.

Skin healing process after Fig. 9 A-D. through thickness wound.Hinder the image of sampling in the 7th day from contrast (A) and (B) of salmon hemostasia products process, show necrotic fibres concretion and be filled with wound defect (*) and project along with the epithelial cell at all oriented wound center in two cases of the wound healing process after initial blood coagulation.(H & E dyes, scale=100 um).The sampling display of the 28th day is hindered by Hypertrophic and hyperkeratotic competing property of epidermis re-epithelialization from (D) of contrast (C) and the process of salmon hemostasia products.(H & E dyes, scale=100 um).

Figure 10. the schematic diagram of coagulation cascade.

Detailed description of the invention

One embodiment of the invention relate to the method being formed hemostasia products by multiple sheet material hemostatic material.As the initial step forming hemostasia products, prepare the sheet material of hemostatic material.

Sheet material has relatively uniform thickness and outward appearance.Form the sheet material with relatively uniform thickness and outward appearance and strengthen the accurate hemostasia products that uses to treat the ability of particular injury.

When hemostasia products is applied to injury site, for the manufacture of hemostasia products material dissolves thus discharge described material to injury site, and provide haemostatic effect.

In one embodiment, action site is wound bed (wound bed), and the activating agent sent by this hemostasia products is the factor or the medicament that participate in coagulation cascade, as thrombin and Fibrinogen.Use this hemostasia products to wound to cause dissolving in the blood of dextran fiber in wound bed, this causes again activating agent discharge or be delivered to or enter this site.

The thrombin be combined with hemostasia products and fibrinogenic form have biological activity when they touch blood.Therefore, when dissolving, thrombin action, in Fibrinogen, is translated into fibrin, then in wound, forms sludged blood, stops blood to flow out.

The present invention uses layer, and it makes the easy commercialization of product, and the composition in hemostasia products is more uniformly disperseed.

The invention provides dextran fiber, particularly Electrospun dextran fiber.Described Electrospun dextran fiber can make the various hemostasia products being applicable to various uses.Usually, the Electrospun dextran fiber in one or more targets (interest) material and described hemostasia products is combined, such as, for sending the object of one or more target substances to target liq.When with liquid comes into contact, Electrospun dextran fiber dissolves within the time of one relatively short, and the material be combined is released to liquid environment.

In one embodiment of the invention, be hemostasia products by Electrospun dextran processbearing astrocyte.Hemostasia products generally includes the activating agent be combined with described Electrospun dextran fiber, by using hemostasia products to action site, described bioactive agent delivery is delivered to action site (such as wound).

Action site contains maybe will containing liquid, and when hemostasia products is applied to action site, Electrospun dextran fibrolysis in hemostasia products is in liquid, and the activating agent be combined with dextran fiber or be trapped in dextran fiber mat or activating agent is around discharged in liquid.

In one embodiment, action site is wound bed, and the activating agent sent by this hemostasia products is the factor or the medicament that participate in coagulation cascade, as thrombin and Fibrinogen.Use this hemostasia products to wound to cause dissolving in the blood of dextran fiber in wound bed, this causes again activating agent discharge or be delivered to or enter this site.

The thrombin be combined with hemostasia products and fibrinogenic form have biological activity when they touch blood.Therefore, when dissolving, thrombin action, in Fibrinogen, is translated into fibrin, then in wound, forms sludged blood, therefore stops blood to flow out.

In some embodiments of the present invention, only use and spin dextran fiber, after therefore sludged blood is formed, just there is no need interference sludged blood to remove hemostasia products composition, because what does not leave in this site.In other embodiments, as described below, hemostasia products can comprise other material, as supported or back lining materials, initial use hemostasia products fast after, may remove with further by conventional method process wound subsequently.

Electrospun is a kind of on-mechanical processing policy, and can expand to adapt to Large Copacity demand, meets the needs of commercialization process.Fig. 1 provide for Electrospun a type schematic diagram is set.In the process, with pulse current injectingt polymer solution or melt to set up charge unbalance.Then, charged solution is placed near ground connection target (in FIG, ground connection axle).

At critical voltage place, charge unbalance starts the surface tension overcoming source of polymer, forms electrified jet.In the electric field, jet is directed to ground connection target and carrier solvent evaporation.As shown in Figure 1, depend on reaction condition and polymer used in this process, Electrospun can be utilized to produce the meticulous aerosol of material or the continuous print non-woven mat of fibrous material.

For many polymer, the character of electro-spinning process essentially provides the Altitude control to gained fibre diameter.By regulating the initial concentration being present in polymer in described Electrospun solution, micron can be realized to nano level diameter by selectivity simply.By controlling the motion relative to the ground connection target of described source solution, fibril can be deposited as random matrix or be deposited as the array limiting axis orientation arrangement along.

Fig. 2 A shows the second schematic diagram of electric spinning device.The key element of electrospinning system comprises high voltage power supply, for the raw material storage device of polymer and ground connection target center axle.The system described adopts cylindrical target axle; But electrospinning process can be suitable for producing more complicated shape.

Single and/or multiple polymer can be transported to electric field from one or more raw material storage device independently or side by side.In addition, can use and produce laminated construction from the different and polymer of uniqueness of independently source Electrospun in chronological order.Fig. 2 B shows Electrospun about 10 grams and is dissolved in the result of the dextran in deionized water to circular spindle target, as what describe in embodiment 1 below.Fig. 3 shows the scanning electron micrograph of the Electrospun dextran fiber of the average cross-sectional diameter about 1 micron of single fiber.

Person of skill in the art will appreciate that, Electrospun is not the unique channel of producing dextran fiber.Such fiber can be produced by other method being atomized (aerosolization).But, electric field contribute to effective collection of fiber and Electrospun may produce evenly fiber.Also can be used for other technology spinning dextran fiber, comprise those United States Patent (USP)s people such as Luo the 7th, 067, No. 444, the United States Patent (USP) the 6th, 116 of the people such as Bogue, the United States Patent (USP) the 5th of the people such as No. 880 and Fuisz, those technology described in 447, No. 423, its respective full content is incorporated to herein all by reference.

Especially, so-called " cotton candy machine (cotton-candy machines) " (electrostatic force with or without applying) can be suitable for manufacturing dextran fiber of the present invention use.In embodiment 2 below, provide the more detailed description of the method preparing dextran fiber of the present invention.

Other method is included between two plates or other flat surfaces compresses dextran solution, and by two plates or surperficial drawn apart from one another, usually repeats.Dextran fiber is formed between two surfaces.

In some embodiments, other material instead of dextran is used to form fiber in hemostasia products of the present invention, especially when (but being not limited to) use cotton candy machine.The example of this kind of material includes but not limited to saccharide, as glucose, sucrose etc.

For the production of the commercially available dextran of electricity spinning fibre of the present invention, by the enzyme catalysis on some lactobacillus cell surface, from Sucrose synthesis, the most famous lactobacillus is Leuconostoc mesenteroides (Leuconostoc mesenteroides) and Streptococcus mutans (Streptococcus mutans).Dextran is a kind of complexity, branched chain glucans (glucan) (by the molecular polysaccharide of many D-Glucoses), is made up of the chain of various length (such as from 10 to 200 kilodaltons).Straight chain is made up of α-1,6 glycosidic bond between glucose molecule, and side chain is started by α-Isosorbide-5-Nitrae glycosidic bond (and also having α-1,2 and α-1,3 glycosidic bond in some cases).

The dextran be available commercially within the scope of wide molecular weight, such as, from about 10 kilodaltons (kDa) to about 200kDa.Commercial formulation is the mixture of the dextran of different molecular weight, usually in narrower weight range, and can according to such as, the dextran of " low " or " height " molecular weight provides.Such as, " Dextran 40 " has the mean molecule quantity of 40 kDa, and " Dextran 75 " has the mean molecule quantity of 75 kDa, etc.

In enforcement of the present invention, the typical molecular weight ranges of the dextran for Electrospun from about 10 to about 200kDa, or from about 25 to about 200kDa, or from about 50 to about 200kDa, or from about 75 to 200 kDa, and be generally from about 60 to 90 kDa, or from about 100 to about 200kDa.

In addition, those skilled in the art will appreciate that, in dextran preparation, the median magnitude (median size) of dextran molecule also has such effect: if intermediate value weight is height by specified quantitative, can use the fiber of less dextran Formation period desired amount.

Generally speaking, the condition for Electrospun dextran is as follows: ambient temperature is from about 60 to about 75 °F, and relative humidity is from about 30% to about 40%, and typically at least about 20%.The fiber obtained typically in the scope that cross-sectional diameter is nanometer or millimeter, usually from about 0.75 micron to about 1.25 microns.

Described electricity spinning fibre is " drying ", should avoid being exposed to humidity, to prevent Premature disintegration.But some water and fiber are combined, and fiber composition containing the water from about 7 to about 8%, but when by x-ray irradiation sterilization fibre, can must be less than about 5%.

Hemostasia products of the present invention is formed by uniform Electrospun dextran substantially usually.The dextran amount of each hemostasia products can change widely, and this depends on the size of manufactured hemostasia products, and typical hemostasia products formula is that each hemostasia products uses from about 5-10 gram dextran (usual 100,000-200,000 Mr).

But this scope can be expanded widely, such as, from being low to moderate about 0.5 gram or less (for little hemostasia products) to as high as each hemostasia products 100 or more gram (for large hemostasia products).In some embodiments of the present invention, it is useful for using the dextran (such as, each hemostasia products about 0.1 is to about 0.5 gram of dextran) of small amount that the activating agent sent by hemostasia products is concentrated into smaller size smaller.

The more important thing is the concentration of the dextran being spun into fiber in solution by it.Usually, the concentration for the solution dextran of Electrospun is in the scope of every milliliter of (ml) solvent from about 0.1 to about 10 gram, or from about 0.5 to about 5 gram every milliliter, and usually such solution concentration be about 1 gram every milliliter ± about 0.15 milligram (mg).Preferred scope be every milliliter will by the solution of spinning from about 0.9 to about 1.1 gram of dextran.

Person of skill in the art will appreciate that, due to the change of dextran preparation middle-molecular-weihydroxyethyl scope, and due to claim the commercial preparations of specific molecular weight range criticize between intrinsic change, typically need to test and often criticize the performance of dextran about Electrospun.This test is within the general knowledge of those skilled in the art, and be usually directed to the Electrospun test of the dextran of a series of concentration being dissolved in suitable solvent, to determine which concentration results has the fiber properties expected most, such as stability (such as to heat, wet etc.), uniformity, cross-sectional diameter etc.

Person of skill in the art will appreciate that, when attempting the dextran of new lot, successful important indicator is obviously.In spinning solution, dextran causes very little from " splash (spitting) " pin, but dextran causes the generation of dry drop too much, or is difficult to spinning at all.

Equally, when humidity is too low, similar result can be there is, namely can not form fiber and in some cases can not effectively targeting in ground connection mandrel.These characteristics can be assessed according to method known in those skilled in the art, but be not limited to Visual Observations Observations, fibre strength and flexible test, by electron microscope observation, dissolubility test, to resistance that is hot and/or that irradiate, color and tendency of fading, etc.One skilled in the art will understand that all these tests can be carried out in the situations such as the heat of various condition, humidity.Also zoopery can be used to assess formula.

The area (length and width) of hemostasia products of the present invention can change widely, and can adjust by regulating spinning parameter.In addition, after spinning, dextran fiber mat can cut into desired size.Usually, the length of a hemostasia products and/or width from about 0.5 centimetre or less to about 30 centimetres or more, but also can consider greater or lesser size.

Depend on the desired use of hemostasia products, the height of hemostasia products or thickness can be very different equally.In certain embodiments, hemostasia products has the thickness of about 1 millimeter to about 5 centimetres.

The thickness (relevant to volume) of hemostasia products may affect the rate of dissolution with dextran during liquid comes into contact.Such as, under the comparable condition of fibrous face layer, thin hemostasia products (such as about 2 millimeters) will dissolve faster by thicker hemostasia products.

In most of embodiment, the dissolving of dextran fiber is extremely rapidly, such as, after being exposed to liquid, about 5 minutes or less, or about 4 minutes or less, or about 3 minutes or less, or about 2 minutes or less, or about 1 minute or less, such as, hemostasia products typically only needs dissolve a little second (such as from about 1 to about 20 second).

Dissolve so rapidly and can be called " instantaneous " or " immediately " dissolving in this article.Regulate rate of dissolution by compression Electrospun dextran pad, speed described in the compression negatively influencing of higher level, namely compression degree is larger, and rate of dissolution is slower.Rate of dissolution has superiority rapidly, particularly in time sending bioactivator (as hemorrhage) to action site (as wound).When disposing hemostasia products, the rapid dissolving of carrier dextran fiber provide to wound extremely rapidly hemorrhage send.

Person of skill in the art will appreciate that the existence of too much liquid flux may dissolve dextran.But, for the better result of Electrospun dextran realization normally solvent be water, particularly deionized water or distilled water or deionization, distilled water (ddH ₂or the relatively pure water of other forms O).In addition, there is few environmental effect relevant to the use of water.

Have been found that in general, the salt of high concentration in solvent, should be avoided.But often use salt to promote the spinning of some electrospun polymer, this is not suitable for dextran.The concentration of salt in spinning solution should be remained on bottom line, successfully to form dextran fiber.

One or more activating agents be combined with the dextran fiber of described hemostasia products can be any activating agents, and described activating agent is desirable for the site be delivered to use or application Electrospun dextran fiber plant or has superiority.In one embodiment of the invention, described Electrospun dextran fiber plant is hemostasia products and is used to send beneficial agent, such as, to wound.

This wound comprises the wound of health or tissue integrity or breaks, its result as wound (such as accident trauma, the wound caused as gunshot wound, knife injury etc. by conflict) and occurring, and also comprises the wound of deliberately generation, as operative incision, health perforating wound, etc.

Usually, described medicament has bioactivator that is useful or therapeutic effect at wound site.In one embodiment, described site is bleeding wounds, needs to form blood clot to stop or to slow down hemorrhage.In the present embodiment, target therapeutic material can comprise, and such as, thrombin and Fibrinogen, although promoting other medicament in hemostasis with active function, including but not limited to capscian, also can be included.

In addition, Electrospun or non-Electrospun collagen protein, absorb the medicament of moisture, when the various dry salt be placed in tend to absorbing fluid during such as contacting blood, engineering thrombin or thrombin Counterfeit Item, technical fiber proteinogen, cause vasospasm to help to shrink and seal medicament (such as ADP, the 5-hydroxy tryptamine (5-hydroxytryptamine of bleeding vessel, 5-HT) with thromboxane (thromboxane, TXA-2)) etc., also can be included.

In addition, other composition of coagulation cascade can be added to hemostasia products, such as: those tissue factors being usually only expressed in the tissue factor on damaged cell surface and starting normal coagulation cascade; Strengthen platelet aggregation and promote vasoconstrictive serotonin; With other medicament being used for replacing the coagulation cascade composition lacked in hemophilia, such as, factor 7(activates so-called outside extrinsic clotting cascade) and platelet crude extract.

These medicines are cascade by initiating reaction network downstream far away and " jump open " blood coagulation in essence, as shown in Figure 10.In Fig. 10, the various factor (and their alternative terms and/or characteristic and/or activity) is as follows:

Factor Ⅴ II(Proconvertin, proconvertin): serine protease, vitamin K dependent synthesis in liver;

Factor IX: the vWF of Glycoprotein binding, is produced by endotheliocyte and liver;

Factors IX (Christmas-Eve Factor, Christmas factor): serine protease;

The factor X(Stuart-Prowler factor, Stuart factor): prothombin is thrombin by serine endopeptidase; With

Factor XI, plasma thromboplastin antecedent (plasma throml oplastin antecedant): serine protease, plasma protein;

Factor XI, plasma thromboplastin antecedent I(Hageman factor, Hageman factor (HF)): serine protease, plasma protein incorporating collagen albumen;

FXIII (fibrin stabilized enzyme): stable fibers protein polymers, plasma protein, is also present in platelet and monocytic series.

In Fig. 10, italic path represents suppression, and shows the central role of thrombin in activation blood coagulation and inactivation blood coagulation, wherein:

The cofactor of the Xa factor that VI=makes thrombinogen change to thrombin;

APC=activated protein C, extracellular signaling molecule, suppresses FVI(to be equivalent to FVa, the cofactor of the XA factor that thrombinogen is changed to thrombin by proteolytic event) and FVIIIa; With

The fibrinolysis inhibitor of TAFI=thrombin activation, the inhibitor that sludged blood dissipates.

In addition, the activating agent that its effect is the final stage promoting wound healing can also be comprised, such as, to promote cell migration and to reinvent.The introducing of collagen protein (collagen) is the example of such activating agent.

In the embodiment of this invention, one or more these activating agents arbitrary can be used.Therapeutic agent must be suitable for drying, and is combined with other composition of hemostasia products in the dry state, because at least one composition used in liquid possibility negative effect hemostasia products.Such as, activating agent can be dehydration or lyophilizing, or anhydrated by some other means.

Generally, when them and Electrospun dextran fiber is combined time, be present in the water yield in described material lower than about 5%, and preferably lower than about 2%.These materials keep all or part of activity as during rehydration in blood.The therapeutic substance be combined with device of the present invention is when with liquid comes into contact, before their dry or dehydrations of general maintenance at least about 25% or about 50% or even about 75 to 100% activity, compared with the standard preparation of described material, code test well known by persons skilled in the art is used to detect.

In some embodiments, by thrombin or Fibrinogen or the two, be combined with hemostasia products.In some embodiments, thrombin and Fibrinogen are salmon thrombin and Fibrinogen.Use salmon includes but not limited to as the advantage of these material sources the propagation need not considering pathogen (such as virus), and when using thrombin or Fibrinogen (the such as cattle) in the mankind or other mammals source, this propagation may occur.

When in pig model (pig model), salmon thrombin and Fibrinogen are highly effective and with no harmful side-effects, and pig model is generally acknowledged animal model, and being considered to has predictive result to the mankind.

Add the fibrinogenic amount of hemostasia products to normally in the scope of about 10 milligrams to about 3 grams.In certain embodiments, in each hemostasia products fibrinogenic amount between about 20 milligrams to about 1 gram.

Add the amount of the thrombin of each hemostasia products to normally between about 10 to 10000 NIH units.In certain embodiments, in each hemostasia products the amount of thrombin between about 20 to 6000 NIH units.

In some embodiments, can Electrospun therapeutic agent itself separately.Such as, therapeutic agent is dissolved in solution and from solvent spinning.In some embodiments, therapeutic agent can be electrospun to fiber.In other embodiments, activating agent can be electrospun to other forms, as microdroplet (droplet), pearl (bead) etc.

In some implementations, can by activating agent as thrombin electron spray together with sucrose form sucrose microdroplet, this thrombin that tends towards stability, and other target substance also " can be caught " be delivered to hemostasia products.

For thrombin and Fibrinogen; in most of embodiment; these (or other) activating agents are dry particles or the particle form of fine dispersion, as fine powder (fine powder) or dust (dust), because Electrospun may tend to the activity reducing them.In other words, activating agent itself can not Electrospun.

Donor material provides the large contact surface area being beneficial to dissolving when these materials and fluid contact in fine powder form.Generally, such granule will have about 1 to 10, the average diameter of 000 micron, and, in certain embodiments, between about 10 to 1000 microns.

Form such dried solid particle by any one in several means, include but not limited to grinding, pulverizing, crushing etc.But, person of skill in the art will appreciate that, in hemostasia products, also can comprise these activating agents of other form, such as: thin slice, thin film, sheet material, line etc.In addition, in some embodiments, when with excipient (excipient) or carrier is combined time, thrombin and Fibrinogen are the forms of Electrospun microdroplet.

By in many suitable technology well known by persons skilled in the art, any one carrys out combining of realize target material and excipient or carrier, and will partly depend on exact form and the means on hand of described material.Such as, for the microgranular thrombin pulverized and Fibrinogen, by spilling, jolting, air blast etc. by described drug combination on excipient or carrier layer.

Depend on the density of fiber mat, described target substance may be scattered in the wrought mat of whole fiber relatively uniformly or may be mainly limited to the top part of fiber mat.If do not have backing, a rear embodiment is preferred, drop to prevent target particles material permeance mat or drop out mat.

The density of (such as increasing) fiber mat can be adjusted, such as, by adjusting its thickness and/or by compressing this pad under stress, making fiber be close together.Also there is other technology for combining, such as by dry but dissolve in the sheet material of the material of liquid or material strip is placed in carrier layer or between carrier layer, as discontinuity layer or the material that adds with the form Electrospun of discontinuity layer, etc., and any such technology can be used.

Can be that craft or mechanization are implemented for assembling the technology of hemostasia products of the present invention, or the combination of available manual operations and mechanization.Special in thrombin, the thrombin of 5000 NIH units is powder of relatively small amount.Therefore, inert carrier (inert carrier) or filler (bulking agent) can be added as glucose, to ensure that activating agent disperses more completely in hemostasia products.

Combining of target substance and excipient can be carried out according to many different arrangements.Such as, the ground floor of excipient can be formed, and one or more described material can be combined with ground floor.Then the second layer of another excipient can be formed on target substance.And identical or other target substance can be combined with the second layer, the rest may be inferred.

The last of excipient or outermost layer can be added, evicted from by from layer below to prevent target substance.The variable amountsization of the figuration oxidant layer that hemostasia products of the present invention uses is very large, and from few to 1-2 to nearly several, or even hundreds of is individual, depends on the desirable characteristics of hemostasia products.

Typically, hemostasia products will comprise 1-2 layer.In other embodiments, hemostasia products can comprise between 2-20 layer.The humidity of very slight amount is present in ready hemostasia products, and thrombin and Fibrinogen can be contributed to catch and remain on hemostasia products surface.

In some embodiments of the present invention, hemostasia products also comprises one or more supporting structures or support material of being combined in wherein.Such as, backing combination can be entered hemostasia products.

Can from various Electrospun material, as polyglycolic acid (polyglycolic acid, PGA), polylactic acid (polylactic acid, PLA) and their copolymer (PLGAs) form support material; Also has charged nylon etc.In one embodiment, described support material is the Electrospun dextran fiber of compression.With " the Electrospun dextran fiber of compression ", we refer to that Electrospun dextran fiber is pressed together under stress.

Such as, under pressure between two plates (such as is secondary), carry out the compression of Electrospun dextran fiber, and the fiber mat of compressible about 3 inch height (thickness) is to the sheet material of about 0.5 inch height or even less (such as about 0.1 to about 0.4 inch).In some embodiments, by the direct Electrospun of Electrospun dextran fiber in previous Electrospun support material, but in other embodiments, respectively after Electrospun, support material and Electrospun dextran fiber are such as joined together by respective one or more layers being connected.

In other embodiments, support material is not Electrospun material, but the material of some other (usually lightweight).The example of such material includes but not limited to gauze, various plastics, and therefore hydrogel and other penetration enhancement blood also form the absorbent material of blood clot, etc.

Support material can be or can not dissolve in liquid, can be maybe slowly be dissolved in liquid, and can be or can not be to liquid permeability.The material of slow dissolving comprises those are formed Absorbable rod or dissolving (biodegradable) suture (stitche) or suture (suture) material by it, comprises the polymer of PGA, polylactic acid and caprolactone.

In certain embodiments, support material can rapid solution relatively, as being less than dissolving in about 1 hour.In other embodiments, support material can be dissolved within 8 weeks from about 10 days.In either case, support material provides does not need to remove hemostasia products and there is not the advantage destroying sludged blood risk.

But under any circumstance, support material should not disturbed dissolving immediately of excipient and send the activating agent that is combined with it in the liquid of excipient dissolving.Therefore, support material may only in the side of Electrospun dextran fiber plant, when being (such as) hemostasia products with this device of box lunch and putting on wound, the orientation of hemostasia products is convenient to the contacting blood in excipient instead of support material and wound bed, namely, when being placed on wound, support material is hemostasia products " top " or outmost surface.

Such as, describe this embodiment in Figure 4 A, wherein Electrospun dextran fiber 10 is illustrated as being deposited in the support material 20 of non-porous, liquid impenetrability.When being applied to wound, excipient 10 will down towards wound, and the support material 20 of non-porous will towards away from wound.

This arrangement has an advantage, can apply pressure by support material to wound, is beneficial to interruption hemorrhage.As selection, support material can comprise liquid can be allowed to reach the hole of excipient in hemostasia products, opening or space, even if when support material exists.Such as, support material can be net (net) or netted (web) material, and it can not dissolve (or slowly dissolving) but the target substance that permission liquid freely reaches excipient and is combined.

Schematically illustrate this embodiment in figure 4b, wherein show Electrospun dextran fiber 10 to be deposited on net 40 (or may below, or above and below, or run through fabric), wherein net 40 is partly with the display of diplopia (phantom) form, and diplopia part is covered by Electrospun dextran fiber 10.Again in other embodiments, " backing " or " top " support material and the second netted support material can be there is in said device.

Schematically illustrate this embodiment in figure 4 c, wherein show Electrospun dextran fiber 10 to be deposited in non-porous support material 50 and to cover with Web materials 60, namely Electrospun dextran fiber 10 " is sandwiched " between non-porous support material 50 and Web materials 60.

Those skilled in the art can be contemplated to other combination and shapes of many figuration oxidant layer and support material, and it will provide advantage according to special setting.Such as, the holder that excipient winding or winding can be extended is as filament or cord, and maybe excipient can be had the given configuration winding of empty shape around one, may place hemostasia products in described cavity, described reciprocal of duty cycle is as shell hole etc.

In such embodiment, provide Fibrinogen in the form of a powder and by powder dispense on the surface of one deck of hemostasia products.In other embodiments, Fibrinogen is mixed with thrombin, then fibrinogen-thrombin mixture is scattered on one deck of hemostasia products.According to the concentration of each hemostasia products up to about 10 grams, Fibrinogen can be provided on the surface of each hemostasia products.

Directly manufacture in another embodiment of hemostasia products in the present invention, dextran is mixed with water until prepare full and uniform mixture.Can provide dextran in powder form, it has the microgranule of opposite fine.Relatively highly purified dextran and water can be selected, as the purity typically for medical usage.The example of a this applicable water is distilled water.

In a configuration, to have an appointment the dextran of 3 grams to about 9 grams and about 6 ml waters.In other embodiments, have an appointment 6 grams of dextrans and about 6 ml waters mixing.It will be apparent to one skilled in the art that and multiple technologies can be used dextran and water to be mixed, with from dextran and the full and uniform mixture of aquatic product.A nonrestrictive example that can be used for technology dextran and water mixed is Electrospun.

For preparing the full and uniform mixture of dextran and water and the incorporation time needed may depend on various factors, as carrying out mixing and the device type used.In some configuration, this mixing is performed for more than about 30 minutes.Described mixing can be carried out to the temperature about between 70oF at about 40oF.

Secondly, thrombin is mixed with dextran-aqueous mixtures.Thrombin is added dextran-aqueous mixtures, and to prepare hemostasia products from it, wherein the concentration of thrombin is about 20 to 6, between 000 NIH unit.

Can provide thrombin in the form of a powder, it has the microgranule of opposite fine.Relatively highly purified thrombin can be selected, as the purity typically for medical usage.

Be similar to the method prepared dextran-aqueous mixtures and use, can Electrospun be used in time mixing thrombin and dextran-aqueous mixtures.The mixing of thrombin and dextran-aqueous mixtures can be carried out to the temperature about between 70oF at about 40oF.

For preparing the full and uniform mixture of thrombin, dextran and water and the incorporation time needed may depend on various factors, as carrying out mixing and the device type used.In some configuration, this mixing carries out about 10 to 30 minutes.In other configuration, this mixing carries out about 1 hour.

Once complete mixing, mixture is introduced in Electrospun machine.Electrospun machine is configured to produce the sheet material had up to about 1 meter of width.But depending on needs size for hemostasia products, also can use shorter width.

Be placed in below Electrospun machine by a releasing piece, fiber is placed on releasing piece.During processing and shearing, releasing piece is that fiber provides support.Should select not with the releasing piece manufacturing in hemostasia products the interaction between component used.Releasing piece can be configured to after shearing and/or other technique complete and product separation.

Due to the factor as challenged, described challenge merges be combined with by Fibrinogen with forming other composition utilized in hemostasia products, Fibrinogen can be provided on the surface of one deck in hemostasia products, and can not be merged in several layers in one or more layers.

It is close that the problem crux that thorn hinders site is thoroughly that wounded tissue is difficult to relatively.Such as, for bullet wound (such as in lower limb or stock), can not occur as many hemorrhage surface, but relatively deeply in tissue, cavity is formed by bullet in it, cavity is difficult to the hemostasia products process by being simply covered in exterior trauma place (as the inlet point of bullet, cutter, shrapnel, sword, bayonet etc., or other wound caused).

This respect of the present invention solves hinders relevant problem to stinging, and described thorn is thoroughly hindered and caused bleeding profusely in deep tissues, and the present invention utilizes the characteristic of dextran hemostasia products highly dissoluble.Complicated factor in this wound types relates to ability highly dissoluble hemostatic material being delivered to this site.

Hemorrhage and other mobility event may be had at the inlet point of wound, and hemostasia products be used to this surface site hemostasia products may be caused to dissolve completely on surface, there is no hemorrhage source basic in delivery of active substances to wound cavity.The present invention solves this event by providing active agent delivery to go deep into wound.Existing hemostasia products can not fully address this problem.

The present invention can be suitable for use in the hemostasia products of this wound by providing its shape and application and solve this problem.Such as, provide cylindric " cigar shape " hemostasia products of extension, it contains thrombin and Fibrinogen, and can contain support material.

Hemostasia products can be stored in protectiveness overcover, packaging or pipe.This pipe protection hemostasia products departs from surrounding.The all preferred sterilizing of hemostasia products and pipe.These parts can be invested further in the outer package of (such as) paper, polymer, blister bag, be similar to for disposable syringe such, to prevent from losing sterilizing state.

During use, tear the hemostasia products that outer package acquisition sterile tube comprises.In some embodiments, remove one end of pipe and be placed in the accessible part of outermost of wound.Pipe also can comprise " piston " or similar device, and it makes user hemostasia products be discharged pipe and enter wound, and result is that " injection " hemostasia products enters wound.

Can use such as that those send the device of (vaginal delivery) such as cotton balls (i.e. " cylinder in cylinder ") for sheath, or syringe-like device can be used.Therefore hemostasia products is in depth introduced in tissue along wound track, and the therapeutic agent in hemostasia products is delivered to the place needing most them, and namely wound is inner.

In other embodiments, do not comprise piston in essence, but the style of pipe is two ends all energy openings, and by applying pressure at the Rather, openings end of pipe, hemostasia products is pushed in wound from one end, any object being applicable to entering at least in part pipe can be used to apply pressure, fully hemostasia products is released pipe and enter in wound.The example of this type objects comprises finger and club.

This object can be included in hemostasia products of the present invention.Person of skill in the art will appreciate that, due to the pliability that some configurations are relatively high, the present embodiment of described hemostasia products can comprise support material (such as biocompatibility net, bar etc. in hemostasia products surrounding or inside, it will be decomposed by biodegradation), when making hemostasia products go deep into wound, firmer and less pliability.

In addition, the outermost end of hemostasia products, namely (such as using piston) to it applies pressure hemostasia products is discharged pipe and enters that end of wound, available support material is strengthened, enough power can be transmitted, to be shifted out from pipe by hemostasia products for use in the piston or other objects that promote hemostasia products.

In fig. 4d, provide an illustrative diagram of this embodiment of the present invention, wherein hemostasia products 100 is illustrated as and is encapsulated in pipe 200, and hemostasia products 100 comprises spinning dextran fiber 110 and (optionally) support material 120, and has first end 130 and the second end 140.

Hemostasia products 100 is encapsulated in pipe 200, but for the sake of clarity, does not show with the form of diplopia.Pipe 200 has opening 210 and 220, uses the first two opening all can cover (not shown lid) or all can open, if especially whole instrument bag is wrapped in sterilization packaging 400.Before using, remove or break sterilization packaging 400, to take out described apparatus.In order to use described apparatus, opening 210 and 220 must be opened.

In order to hemostasia products 100 being delivered to thorn thoroughly wound, by object as piston 300 inserts opening 210 end of pipe.When piston 300 touches device end 130, apply pressure to hemostasia products 100, hemostasia products 100 finally via opening 220(with by arrow indicated direction) be pushed out pipe 200, and enter thorn and thoroughly hinder (not shown).Second schematic diagram of this hemostasia products has been shown in Fig. 4 E.

In this schematic diagram, do not comprise support material, and the hemostasia products material of the drying be combined with therapeutic agent, sterilizing (such as dextran fiber) is located in or is placed in cylinder that is little, that encapsulate, and there is lid one end of described cylinder, and the other end has piston.During use, abandon lid, the breach opening of cylinder being placed in wound can insert wound, and forces piston, transfer or injection hemostasia products material go deep into wound.

Similar designs can be used to send hemostasia products to hole or passage, as nasal passage, syrinx, vagina, anus, intravasation, etc.The parts used in this applications will be formed about 1 to about 6 inch long and from the shape of about inch to 1 inch diameter, namely the size of hemostasia products will be suitable for inserted through outside opening and stretch in hole or wound cavity in order.

The invention is intended to comprise all this kind of arrangement of carrier layer described here and support material, shape and embodiment.

Can hemostasia products described in sterilizing before using, usually by using electromagnetic radiation, such as, X-ray, gamma ray, ultraviolet etc.If described hemostasia products comprises thrombin, the moisture reducing hemostasia products (such as binder or gauze) may be needed to being less than about 5%, to protect thrombin activity during sterilizing.

By the hemostasia products that drying manufactures, such as under vacuo, or by using the manufacture method from starting just reduction moisture, realize the reduction of moisture.Typically, hemostasia products described in X-ray sterilizing is used with the dosage between about 5 to 25 kilograys (kGray).Anyly do not destroy carrier or all can be used for sterilizing hemostasia products of the present invention with the be combined method of activity of material of fiber.

When hemostasia products is binder, thrombin and Fibrinogen can be comprised with the target substance of the joint fiber of described hemostasia products, and described hemostasia products can be used for stop hemorrhage.But the scope of active component can change along with the embody rule of hemostasia products.

Such as, the hemostasia products only comprising thrombin can be used for little wound or intervenes with other combinationally using.In addition, other therapeutic benefit material also can be combined with described hemostasia products, includes but not limited to: antibiotic, the medicine eased the pain, somatomedin, bone morphogenetic protein, vaso-active substance (as causing angiospastic material), reduces the steroid of inflammation, and their combination.

In other embodiments, device of the present invention does not comprise the medicament promoting blood coagulation.Person of skill in the art will appreciate that, device height of the present invention is suitable for sending many target substances to the liquid environment of needs or position.Such as, this device can be designed to delivery treatments or the benefit materials any wet environment to health, as long as have enough liquid to dissolve Electrospun dextran fiber and release of active agent, and wherein to dissolve dextran be not in-problem.

Oral administration, per nasal, through trachea, per anum, to include but not limited to, as antimicrobial, analgesics, nutrient etc. through the example of the material of lung and Transvaginal delivery.Oral application comprises sends the material useful to tooth treatment, such as, and antibiotic, analgesic, whitening agent etc.Can comprise with the described treatment of device conbined usage or the example of benefit materials, antibiotic, the medicine eased the pain, somatomedin, bone morphogenetic protein, vaso-active substance (as causing angiospastic medicine), inflammation reduce sex steroid and their combination.

But, in some embodiments, there is no body fluid (if or leptochymia existed), the hemostasia products used can by moistening " activation ", such as by spraying, or apply moisture source (such as, by hemostasia products is exposed to moist material as sponge) by alternate manner, or hemostasia products is thrown in liquid (such as water body), to cause the release of the target agent be combined with dextran fiber.

Due to little, the lightweight characteristic of the area of hemostasia products, they are desirable for the space of equipment and the very valuable situation of weight.The example of this situation includes but not limited to: military operation, and wherein the weight of soldier's equipment parts and size are major issues; In first-aid kit; Critical care (such as space flight, camping etc.) during travel; Deng.

Described hemostasia products can be used for not needing the various situation of consideration space and weight and various object.Such as, in traditional operating room, hemostasia products of the present invention provides to be used thrombin to surgical incision and fibrinogenicly facilitates means.

When needs packaging and final one or more dry matters of release, but wherein said dry matter is difficult to or does not need straightforward manipulation, and such as wherein measuring minimum or material is poisonous situation, also can advantageously utilize hemostasia products of the present invention.

In this case, Electrospun dextran fiber of the present invention can serve as " scaffold (scaffolding) " or carrier for holding, storing and/or transport described material, until use time, namely until with dissolve described Electrospun dextran fiber liquid comes into contact and with discharge in described material to liquid time.This kind of material can comprise, such as, enzyme or their precursor (such as proenzyme (pro-enzymes) or proenzyme (zymogens)) and their substrate, i.e. the material of activator protein or enzyme (such as protease, cofactor etc.), etc.

The invention still further relates to the use of the stabilizing agent of the composition premature breakdown that the described hemostasia products of opposing utilizes.Described stabilizing agent can also improve the available shelf-life of described hemostasia products.In certain embodiments, described stabilizing agent provides hemostasia products to have shelf-life at least about 2 years.In other embodiments, described hemostasia products shows the shelf-life of at least 3 years.

As used herein, term can be by the implication of shelf-life, and when amplifying observation or amplifying observation be not as magnifier or microscopic examination, described hemostasia products does not show obvious degraded.

A kind of this stabilizing agent is suitable for being combined with thrombin.It is believed that, thrombin stabilizing agent enters in the structure of thrombin, thus reduces the zymolytic speed of blood coagulation.Before thrombin is mixed with other composition for the manufacture of described hemostasia products, at least one thrombin stabilizing agent can be mixed with thrombin.

In one embodiment, described thrombin stabilizing agent contains sugar, as sucrose.In certain embodiments, sucrose by thrombin weight to be used in thrombin stabilizing agent up to the concentration of about 5%.In other embodiments, sucrose concentration counts about 1% by thrombin weight.

Before thrombin stabilizing agent is mixed with thrombin, thrombin stabilizing agent can be mixed with dextran.It is believed that, dextran strengthens the ability that sucrose enters thrombin structure.

In certain embodiments, dextran by thrombin weight with the concentration up to about 5% in thrombin stabilizing agent.In other embodiments, dextran concentration counts about 1% by thrombin weight.

Similarly, stabilizing agent is combined with Fibrinogen.Before Fibrinogen being applied in other composition of tourniquet bandage, Fibrinogen stabilizing agent is mixed with Fibrinogen.It is believed that, Fibrinogen stabilizing agent enters in fibrinogenic structure, thus reduces the speed of fibrinogenolysis.

In one embodiment, described Fibrinogen stabilizing agent contains sugar, as sucrose.In certain embodiments, sucrose by Fibrinogen weighing scale to be used in Fibrinogen stabilizing agent up to the concentration of about 5%.In other embodiments, the concentration of sucrose presses Fibrinogen weighing scale between about 2% to 3%.Again in other embodiments, sucrose concentration counts about 1% by Fibrinogen weight.

Before composite fibre proteinogen stabilizing agent and Fibrinogen, Fibrinogen stabilizing agent can be mixed with solubilizing agent.It is believed that, solubilizing agent strengthens the ability that sucrose enters fibrin original structure.In certain embodiments, solubilizing agent is detergent.In other embodiments, solubilizing agent is general stream Buddhist nun gram (Pluronic).

In certain embodiments, solubilizing agent by Fibrinogen weight to be used in Fibrinogen stabilizing agent up to the concentration of about 1%.In other embodiments, the concentration of solubilizing agent counts about 0.002% by Fibrinogen weight.

In another embodiment of the present invention, Fibrinogen and thrombin are placed in dissolving film matrix surface and/or are integrated in dissolving film matrix.Use Fibrinogen and thrombin to make described hemostasia products can be positioned in the hemorrhage of health with this structure and therefore need on the position of hemostasis.

Dissolving film can be configured to dissolve relatively rapidly as during blood when being exposed to liquid.In certain embodiments, described film be less than in about 30 seconds dissolve.In other embodiments, described film be less than in about 5 seconds dissolve.A kind of example of suitable dissolving film is sold by Hughes Medical Corp.

The example of another kind of dissolving film is dissolving paper, and it will cause the material manufacture of health hazard to form to patient after dissolving.In certain embodiments, the material manufacture that described dissolving paper can realize hemostatic capability by fortifying fibre proteinogen and thrombin at least one forms.A kind of example of this dissolving paper is sold by Daymark Technologies.

In another embodiment, between two-layer soluble material, Fibrinogen and thrombin is provided.A kind of example of this type of suitable dissolving film is sold by Hughes Medical Corp, and it is stated as discussed above.

Use spirit of the present invention, Fibrinogen and thrombin can be provided with various configurations.In a this configuration, provide at least one of Fibrinogen and thrombin in powder form, hold it in the interlayer of soluble material.

Described soluble material should have enough structural intergrities to remain on wherein by Fibrinogen and thrombin, can not interact with Fibrinogen and thrombin simultaneously.When being exposed to liquid as blood, described soluble material also should be dissolved relatively rapidly, to make Fibrinogen and thrombin discharge from it.

As used herein, " rapid solution " means described soluble material and decomposes sufficient degree, makes the pith of Fibrinogen and thrombin in less than about 30 seconds and contacting blood.In other embodiments, described soluble material is decomposed in less than about 10 seconds.

Described soluble material also should be convenient to easily engage, so that two-layer soluble material can around its edge conjunction together, to form the enclosure being suitable for Fibrinogen and thrombin being remained on wherein.

A kind of example of the applicable technology for the soluble material that is connected to each other is, uses a small amount of liquid at least one of the material pieces being intended to be bonded together.Water causes the slight degradation of soluble material, and make when being placed in adjacent one another are by two-layer soluble material, the layer of soluble material combines.

Described soluble material can be manufactured by multiple material.Described soluble material should not cause negative effect to the stability of Fibrinogen and thrombin.Material for the manufacture of dissolvable layer also should be selected not on the material being intended to use the human or animal of described product to cause any adverse health and affecting.

In certain embodiments, the material manufacturing hemostasia products use can be combined and improve speed of stopping blooding separately or with Fibrinogen or thrombin.The example that can be used for the composition of soluble material comprises cellulose derivative materials.

In one configuration, the desired locations using independently device described hemostasia products to be maintained patient to be correlated with.The example of this kind of device is a gauze, and it is wrapped in the bleeding part of health.

In another arrangement, described hemostasia products be coated with binding agent at least partially.Described binding agent can around the arrangement at least partially at hemostasia products edge.In an alternative configuration, the substantial portion of described adhesive coverage hemostasia products inner surface.

In the configuration that this binding agent may contact with the bleeding sites of patient body, should biocompatible adhesive be selected, contact by described binding agent the potential patient experience complication caused to reduce with bleeding tissue.

In another embodiment, Fibrinogen and thrombin can be placed in the inner and/or outer shell surface of shell, not dissolve as blood when described shell is exposed to liquid.This structure can be convenient to form blood clot thereon, makes to remove described blood clot from patient.

This structure can be similar to traditional tea-bag, its housing can be connected on cord, described cord and housing combination use with realize described product to be arranged in closest to the region needing to stop blooding or from patient removes described product and the blood clotting be combined at least one.

Can be manufactured described cord by enough strong material, no matter make is when the described product of layout or in time removing described product from patient, cord all can not rupture.Also described cord should be manufactured by unlikely producing bad biology interactional material.

Shell can be configured within seclected time, discharge Fibrinogen or thrombin.The speed that Fibrinogen or thrombin are discharged can adjust based on the speed of the blood of discharging from patient and/or volume.

In the selectable configuration of one, shell can be configured to degrade in an elongated segment time range.When described shell is degraded, Fibrinogen and thrombin can be discharged from described product.

By controlling the speed of speed that Fibrinogen and thrombin discharge from product and/or shell degraded, the formation with relative large scale sludged blood drops to minimum by described product, and may be beneficial to the more sludged blood with reduced size of formation.If compare the sludged blood causing formation relatively large, this less sludged blood is more easily degraded in vivo.

This structure of hemostasia products may be particularly suitable for being combined with hemorrhage in patient, and wherein hemorrhage in the hole of health, the hole of this health can arrive from health outside.The example of the surgery operating technology that can be combined with described hemostasia products comprises hole and operation on tonsils.

In another embodiment, Fibrinogen and thrombin are pressed into tablet.Except Fibrinogen and thrombin, described tablet also can comprise at least one excipient.Described excipient not only should be beneficial to Fibrinogen and thrombin combines, and also should promote that tablet dissolves relatively rapidly.

As used herein, term " relatively rapidly " mean when be placed in tablet described in liquid be less than in about 30 seconds dissolve.Other configuration in, tablet be less than in about 10 seconds dissolve.Solution tablet makes Fibrinogen and thrombin discharge rapidly from tablet rapidly, makes these materials to provide quick-acting haemostatic powder.

In addition, in certain embodiments, the excipient for preparing described tablet should not reduce stability and/or the dissolubility of Fibrinogen and thrombin.In certain embodiments, the excipient for preparing described tablet should increase the stability of Fibrinogen and thrombin.

The example of a this excipient is Sorbitol (sorbitol), and it forms granule, such as, by using spraying dry.In a this configuration, described granule has common spherical and have usually uniform size.

Spray-dired Sorbitol granule not only provides the flow behavior of advantage, and when using direct compress technique to form tablet, shows desirable compactingproperties.

In addition, spray-dired Sorbitol granule for discharging Fibrinogen and thrombin provides good dissolubility from tablet.The example of a this spray-dired Sorbitol granule specified by SORBITAB SD 250 under SPI Pharmaceutical sell.

Another kind of can be mannitol (mannitol) manufacturing the excipient used in tablet, and it forms granule, such as, can use spraying dry.Can form the granule with narrow particle size distribution, it reduces the Potential feasibility of the composition isolation when forming tablet.

An advantage of mannitol is this material is nonhygroscopic, and described mannitol is not moisturized to other composition used in tablets, and can not contribute to formed tablet technique during or tablet formed after absorb moisture.Thus the Fibrinogen of described mannitol protection water sensitivity and thrombin.

Spray-dired mannitol particles not only provides the flow behavior of advantage, and when using direct compress technique to form tablet, shows desirable compactingproperties.

Spray-dired mannitol particles also impels described tablet fater disintegration or dissolving, and Fibrinogen and thrombin can be discharged from tablet rapidly.The example of a this spray-dired mannitol particles is that the SPI Pharmaceutical of being specified by MANNOGEM EZ is sold.

Other material that can be used as excipient when preparing tablet comprises fructose and maltose.Be similar to other excipient discussed above, aforesaid excipient can be formed as granule before using, and mixes with other composition used in tablets.

The another kind of excipient that can be combined with Fibrinogen and thrombin is instant (platform) being sold by the SPI Pharmaceutical under PHARMABURST 500 specifies.The ability that this material provides tablet to dissolve rapidly, also provides compacting and the fragility characteristic of expectation simultaneously.

Depending on the excipient used in tablets, also may need lubricant when preparing described tablet.Described lubricant can improve the physical property of tablet.The example of this kind of physical property comprises brittleness, frangible degree and hardness.The example of a this lubricant is sodium stearyl fumarate (sodium stearyl fumarate), can buy from the SPI Pharmaceutical LUBRIPHARM appointment.

May many factors be depended in the concentration manufacturing in tablet the lubricant used, as the type of excipient that uses.In certain embodiments, the concentration by weight of described lubricant is up to about 5%.In other embodiments, the concentration by weight of described lubricant is about 2% to 3%.Again in other embodiments, the concentration by weight of described lubricant is about 2.5%.

Once composition mixes, mixture is compressed, thus cause composition to form tablet.In certain embodiments, compression stress is at least 5000 pounds/square inch (psi).In other embodiments, compression stress is about 10,000 psi to about 12, between 000 psi.

When using previous process to prepare tablet, unnecessaryly dextran may be comprised.Even if dextran may be unnecessary, but dextran may be used together with other composition for preparing tablet.

In another embodiment of the invention, Fibrinogen and thrombin can be merged into rapid solution tablet, as the technology by using the Catalent sold under being specified by Zydis.

Described rapid solution tablet be less than in 30 seconds dissolve, and some configuration in, be less than in about 5 seconds dissolve.Tablet described in rapid solution is important, because when tablet dissolved, wherein contained Fibrinogen and thrombin are released, thus can produce anastalsis.

The amount of Fibrinogen and the thrombin used in tablets can be selected based on amount of bleeding.In certain embodiments, have in each tablet up to about 1 gram of Fibrinogen and thrombin.In other embodiments, to have an appointment in each tablet 500 microgram Fibrinogen and thrombins.

In another embodiment of the invention, Fibrinogen and thrombin be applied to the surface of applicator or mix in applicator (applicator).This applicator makes Fibrinogen and thrombin be sent the region needing hemostasis exactly.

In a this configuration, described applicator has can by the extension just using the people of hemostasia products to grasp.Described applicator can have the structure being similar to swab (swab).This structure of hemostasia products is particularly suitable for the position being difficult to directly arrive.An example that can use the situation of this hemostasia products process is epistaxis.

At least Fibrinogen and in thrombin can separately Electrospun or with another composition as Electrospun together with dextran.Use the fiber of this explained hereafter can reel around the far-end of applicator.

Described applicator arrangement can be become once hemostasia products runs into blood namely discharge Fibrinogen and thrombin.Make in this way, Fibrinogen and thrombin will cause sludged blood to be formed.Described sludged blood can remove with it from patient.If sludged blood is enough little, can be allowed to be retained in patient body described sludged blood can be finally degraded.

In the another kind configuration of this hemostasia products, can at least one be configured to maintenance and connects and be bordering on applicator or be limited to applicator by Fibrinogen and thrombin, make when Fibrinogen and thrombin cause at least one sludged blood to be formed, this sludged blood remains adhered on applicator.This structure is beneficial to and removes sludged blood from patient, and may be enough large so that do not wish to need the place that keeps in vivo to remove from patient at sludged blood.

Do not discharged by from applicator for ease of Fibrinogen and thrombin, Fibrinogen and thrombin can be incorporated in the material being attached to described applicator one end.The example of a this material is foam.Described foam can be open celled foam or closed-cell foam.Described foam should have enough large hole to receive Fibrinogen and thrombin.Described foam should not have strong affinity to Fibrinogen or thrombin, is exposed to water with convenient Fibrinogen and thrombin, and these compositions are discharged by from foam.

In another embodiment of the invention, Fibrinogen and thrombin are mixed foam.The example of a this suitable foam is absorbable gelatin sponge, as what buy from the Novartis under VETSPON appointment.

Depend on the application needing to use sthptic sponge, may need before the region described sthptic sponge being applied to needs hemostasis, described sthptic sponge of prewetting.

Use another advantage of foam to be can be configured to flexible by described foam, described hemostatic foam can be bent to suit the requirements the shape in the region of hemostasis.Once described hemostatic foam is bent to required structure, it just can maintain that structure, even if do not use fixture to maintain described hemostatic foam in required shape and/or position.

Similar in appearance to above-described foam, this foam can be open celled foam or closed-cell foam.Described foam should not have strong affinity to Fibrinogen or thrombin, is exposed to water with convenient Fibrinogen and thrombin, and these compositions are discharged by from foam.

Fibrinogen and thrombin can be mixed in the composition being used to manufacture foam, make not to be surface Fibrinogen and thrombin being applied to described foam, but Fibrinogen and thrombin are scattered in whole foam matrix.

This structure is beneficial to Fibrinogen and thrombin discharges from foam duration, and when needing when experiencing hemorrhage body region again and form sludged blood, may be useful especially.

Rapid solution tablet is suitable for use in various application.The example of a this application is oral hemorrhage.If intend described product to be combined in oral hemorrhage, described tablet may be the tablet of seasoning.

Be combined in various surgery operating technology, be necessary to form otch at patient body.Once complete operation, be necessary to use stitching thread (suture) or nail (staple) to carry out closed described otch.Although stitching thread or nail are effective for being kept together by tissue, these closed mechanism can not always stop blood to be flowed out by described otch effectively.

Described hemostasia products can be placed in the sutural of blood process to go up at least partially.Described Fibrinogen contacts this blood with thrombin, and therefore provides hemostasis.

With multiple medical treatment, as given patient delivery's chemotherapeutics, be combined vascular access device.In one configuration, vascular access device is that surgical operation is implanted in the large vein of patient's heart.Described vascular access device may stay the appropriate location a very long time, as the time more than a year.

A challenge of surgical operation implantable intravascular access to plant is that it is hemorrhage to stop around vascular access device.Such as, blood may ooze out stitching thread or surrounding catheter.Seepage can not stoped in some cases, be necessary patient to be brought into operating room to make great efforts to stop seepage.

Hemostasia products can be placed around vascular access device, thus cause hemostasis.Depend on the shape of vascular access device, hemostasia products may have multiple structure.

In other cases, the conduit be combined with vascular access device may be porous.Hemostasia products may be used to provide hemostasis, thus anti-Hemostatic Oral Liquid is through void channels.

One or more previous constructions of described hemostasia products may be applicable to stop from patient's bleeding, time as removed conduit from the thigh tremulous pulse in patient.In such configuration, described hemostasia products may comprise binding agent, and when occurring when stopping blooding, it maintains hemostasia products in position.

Aortic root operation is used to treat aortal expansion or expansion.Due to aortal character, stop blooding plays an important role in the success of this program.Hemostasia products discussed above structure can be used to provide hemostasis, and aorta can not be caused to shrink, described aorta shrinks and is caused by the medicament of prior art continually, and described medicament is used for being combined to provide hemostasis with such operation.

For the wound more goed deep in patient, may need to use described hemostasia products with the form of oakum, it be inserted in wound at least in part, thus cause hemostasis in wound.

Again in another arrangement, hemostasia products can be provided with the structure of elongating, as the shape of pad-type of stopping blooding.Also hemostasia products can be provided with the shape of anthelmintic or rope.Such structure can be used for the hemostasis after vagina operation.Such structure also can be combined the extension hole in patient body, as being the hole of being caused by gunshot wound.

The cylindrical structural of described hemostasia products is also suitable for use in tissue to be had in the situation of big uncork.In this case, multiple cylindrical hemostasia products can be inserted big uncork, thus fill a part for this opening.

For the benefit of use the hemostasia products of elongation, described hemostasia products may be needed to be stored in applicator, as in piston.Described piston can be used to the hemostasia products elongated to insert in the wound in patient body.

In another structure of hemostasia products, hemostasia products is often planted composition and is packed separately, such as, in syringe.This structure makes with each composition of different rate-allocation, and making can according to specific patient or the wound customization hemostasis for particular type.Described applicator can allow optionally or side by side to send often kind of composition.

Described hemostasia products can substantially by Fibrinogen, thrombin be suitable for being formed by the carrier of bio-absorbable.As selection, described Fibrinogen and thrombin can be placed in the outer surface of hemostasia products, after having made hemostasis, hemostasia products will be taken out by patient body.

Again in another configuration, send described Fibrinogen and thrombin by spray.Can provide described Fibrinogen and thrombin with the form of microsphere (microsphere), it can use spray container to distribute.

Also described hemostasia products may be combined with robotic surgical.Although robot is provided the ability of carrying out surgical procedure by the surgeon remote with patient location, there is certain limitation in robot.Hemostasia products can be combined with described robotic surgery program, to provide hemostasis, thus overcome this limitation.

Except being used for producing hemostasis in the mankind, spirit of the present invention is applicable to and is combined with other animals.The example of this kind of animal that the present invention can be used for comprises Canis familiaris L. and cat.

In another embodiment of the invention, the water of effective dose is mixed with dextran form moisture dextran solution.After this, moisture dextran solution Electrospun is formed dextran sheet material.

Described dextran sheet material can be stored, until it needs for the manufacture of hemostasia products.In a configuration, dextran sheet material is rolled.Roll dextran sheet material and not only reduce the region that dextran sheet material takies when storing described dextran sheet material, and the Potential feasibility that before reducing the described hemostasia products of manufacture, described dextran sheet material is destroyed.

When rolling dextran sheet material, should be careful, not by dextran sheet material overwind, because this process will increase the density of dextran sheet material.As selection, may need before manufacturing described hemostasia products to roll up dextran sheet material tightly to increase the density of dextran sheet material.

Only before needs manufacture described hemostasia products, described thrombin and Fibrinogen are mixed in the ratio discussed in other parts of present patent application.Described mixing should make thrombin in the mixture and the relatively uniform dispersion of Fibrinogen.

In certain embodiments, thrombin is dispersed on dextran sheet material, with the concentration of thrombin providing square centimeter dextran sheet material to have 2 to 200 NIH units.

In certain embodiments, Fibrinogen is dispersed on dextran sheet material, with the fibrinogen concentration providing square centimeter dextran sheet material to have 20 to 60 grams.

Dextran sheet material is untied, and thrombin and fibrinogen mixture is dispersed on the surface of dextran sheet material.In certain embodiments, thrombin and fibrinogen mixture are dispersed on the surface of dextran sheet material in a substantially even way.Needing this uniform dispersion, is because it makes every part of hemostasia products have basic styptic activity.

Repeat this process, described sheet material is stacked, until the aequum of hemostasia products performance styptic activity.In certain embodiments, hemostasia products comprises about 2 to 20 layers of dextran layer.

Thrombin and fibrinogen mixture are not placed in the superiors of described dextran sheet material.Use this configuration, thrombin and Fibrinogen are positioned in the interior location of hemostasia products.Manufacture hemostasia products by this way and enhance ability thrombin and Fibrinogen being remained on hemostasia products inside, even if thrombin and Fibrinogen are sprinkled upon on the surface of dextran sheet material.

Although thrombin and Fibrinogen may be placed on the outside of described hemostasia products, before the use, thrombin and a fibrinogenic part also may dissociate out from described hemostasia products.In view of thrombin and fibrinogenic cost, it is desirable to all thrombins and Fibrinogen substantially and all keep being combined with described hemostasia products until need to use described hemostasia products, to maximize effect of described hemostasia products.

Although thrombin and Fibrinogen are mixed before thrombin and Fibrinogen are placed in dextran sheet material, described thrombin and Fibrinogen are dispersed on dextran sheet material fully, to make before hemostasia products being placed in the position needing hemostasis, thrombin and Fibrinogen do not react.

Then hemostasia products is cut into slices.In certain embodiments, described can be formed with usual foursquare shape.Can based on the size of the expection application choice sheet of hemostasia products.Such as, when hemostasia products expection is used in surgical operation application, if than hemostasia products, described can be expected that the application being used for wound has less size.

Can use cutter that hemostasia products is cut into required size.Except being cut into slices by described hemostasia products, described cutter may cause the layer of the contiguous cutter of described dextran sheet material to be compressed in together.This compression causes dextran sheet material to be stayed together.

In certain embodiments, hemostasia products sheet is vacuum-packed.Except keeping hemostasia products aseptic, each layer of vacuum packaging also in compressed tampon product, the ability of anti-interlaminar separation after taking out from packaging before this enhancement layer hemostasia products uses.

The method is because this enhancing the use ability of hemostasia products, because each layer resistant to separation in hemostasia products.Use an advantage of the method to be, do not need extra step to be kept together by each layer.In addition, it does not need to use added ingredient and/or the additional process steps that may affect hemostasia products effect.

Embodiment

The water of dextran and effective dose is mixed to form dextran aqueous solution.Dextran aqueous solution is electrospun to Electrospun dextran sheet material.

Thrombin and Fibrinogen are mixed, is then dispersed on Electrospun dextran sheet material.According to the ratios diverge thrombin of every square centimeter of Electrospun dextran sheet material about 1.3 to 2.7 NIH unit.According to the ratios diverge Fibrinogen of every square centimeter of Electrospun dextran sheet material about 3.6 to 7.4 milligrams.

Repeat this process, until there is the Electrospun dextran sheet material of 8 layers of stacking construction.Thrombin and fibrinogen mixture are not dispersed on the surface of the superiors.The thickness of Electrospun dextran sheet material is about 1 to 3 millimeter.

Then with cutter, hemostasia products is cut into the sheet of width about 4.8 centimetres, length about 4.8 centimetres.Except forming the sheet of required size, cutting makes Electrospun dextran layer be pressed together.This process makes Electrospun dextran layer resistant to separation.By the vacuum packaging of hemostasia products sheet to store until use.Except preventing the pollution of hemostasia products, vacuum packaging makes Electrospun dextran be urged together.

TACHOSIL is used to specify the performance being purchased the hemostasia products that absorbable fibre protein blocking agent product and present patent application describe of lower Nycomed sale to compare.

TACHOSIL product comprises and to be coated in its side with thrombin to start collagen sponge together with Fibrinogen.Human thrombin containing the human fibrinogen of 3.6 to 7.4 milligrams and containing 1.3 to 2.7 NIH units on every square centimeter of product on every square centimeter of product of TACHOSIL product.

Utilize the aforementioned information about thrombin and fibrinogen concentration on TACHOSIL product, TACHOSIL product is cut into square sheets, thrombin and fibrinogenic amount on hemostasia products thrombin and fibrinogenic amount on every sheet TACHOSIL product being approximated prepare above.

Use the pig liver damage model assessment hemostasia products of present patent application and the hemostasia effect of TACHOSIL product, this model had previously been used to evaluate hemostasia effect.Pig liver damage model makes it possible to carry out multiple test on single animal.

Adult, domestic, non-breeding, female pigs that operating weight is about 50-70 kilogram are tested.Preliminary inquiry is carried out to every animal, to confirm that animal health condition is good.Before program, animal is isolated three days, period is optionally for animal provides food and water.

Before start program, implement anesthesia to animal.During program, implement anesthesia and fluid maintenance to animal.During program, below animal, place heating cushion to help the body temperature keeping animal during program.

Carry out midline laparotomy to reach abdomen organ, carry out liver living tissue's cut sections for microscopic examination (hepatic biopsie) herein.The biopsy punch of about 8 millimeters of diameter is used to create the section of each liver living tissue.Each liver living tissue slice width about 8 millimeters, the degree of depth about 4 millimeters.The section of cylindric liver living tissue surgically is removed with shears.

The existence that the continuous freedom of about 20-25 second is hemorrhage is observed in biopsy site.After this, hemostasia products is administered to biopsy site.The hemostasia products of hemostasis was maintained appropriate location in about 20 seconds with hand.

Hemostasia products is stayed on biopsy site by the pressure then removing hands simultaneously.For the obvious sign of active hemorrhage, observe biopsy site about 2 minutes.

If hemorrhage sustainable existence, an extra hemostasia products is placed on biopsy site.Hemostasia products was maintained appropriate location in about 20 seconds with hand.Hemostasia products is stayed on biopsy site by the pressure then removing hands simultaneously.For the obvious sign of active hemorrhage, observe biopsy site about 2 minutes.Previously do not stop if hemorrhage, and repeated this process and reach 4 times.

In contrast, biopsy punch is used to create liver living tissue's section of width about 8 millimeters, the degree of depth about 4 millimeters.Observe biopsy about 2 minutes.During this period of time, biopsy does not stop hemorrhage.

Carry out using the detection of the hemostasia products of present patent application, TACHOSIL product and contrast with random order, dropping to minimum by the potential impact using the order of hemostasia products to cause during assessment.

Carry out the order of hemorrhage assessment and the results are shown in table 1 below of being obtained by this assessment.

Table 1

5 extra animals are used to repeat described assessment.5 extra animals use the order of hemostasia products be also random.

By list in table 1 first time assessment result and the result of 5 extra animals combine.18 samples are had for from each of the hemostasia products of present patent application and TACHOSIL product.

For these 18 samples, the hemostasia products from present patent application causes be 2.9 minutes the average time of hemostasis, has the standard deviation of 1.7 minutes.For these 18 samples, TACHOSIL product causes be 4.7 minutes the average time of hemostasis, has the standard deviation of 1.4 minutes.Therefore for realizing hemostasis, the hemostasia products from present patent application provides in time close to the reduction of 2 minutes.

Before biopsy and after realizing hemostasis, have evaluated the change that animal blood is potential on hematology.For from the hemostasia products of present patent application and TACHOSIL product, Hematology results there is no obvious difference.

The sludged blood that the hemostasia products using pathology to compare present patent application provides and the sludged blood that TACHOSIL product provides.In the biopsy of the hemostasia products process of all use present patent application, equal identifiable design is to fibrin.On the other hand, 5 in 18 biopsies using TACHOSIL product treatment unidentified to fibrin.

For the hemostasia products of present patent application, the bioptic sum needing extra pressing is 5.For TACHOSIL product, the bioptic sum needing extra pressing is 17.Therefore, compared with TACHOSIL product, the hemostasia products of present patent application shows the bioptic reduction more than 70% needing extra pressing.

For the hemostasia products of present patent application, the sum of the extra pressing needed for providing hemostasis is 8.For TACHOSIL product, the sum of the extra pressing needed for providing hemostasis is 24.Therefore, compared with TACHOSIL product, the hemostasia products of present patent application shows the reduction additionally pressed more than 60%.

" basket (basketweave) " structure of the hemostasia products of present patent application allows the erythrocyte trapping being scattered in whole biopsy site.On the other hand, TACHOSIL product causes region between biopsy site base portion or amorphous embedded material that the trapping of erythrocyte sheet occurs.

Because the thrombin that uses in TACHOSIL product and Fibrinogen obtain from human plasma, therefore exist may risk containing infectious pathogen for TACHOSIL product.The advantage that hemostasia products of the present invention is better than TACHOSIL is, hemostasia products of the present invention shows and keeps stable at relatively high temperatures than TACHOSIL product.

Although directly do not compare, when with fluid contact, the Electrospun dextran base used in hemostasia products produced according to the invention dissolves rapidly.Described dextran is being absorbed in health in the relative short time section of a couple of days from several minutes.The rapid loss of dextran carrier provides the beneficial effect relevant with cicatrix risk to reducing inflammation.

In contrast, the collagen sponge used in TACHOSIL product is degraded within the time period of one period longer, such as, up to about 4 months.This manufactured business of one side of TACHOSIL product is indicated as being a characteristic, because maintain fibrin sludged blood in wound surface to realize hemostasis according to indicating collagen sponge.The slow absorption of collagen can increase relevant to inflammation and cicatrix risk.

The Electrospun dextran used in hemostasia products, compared to the collagen sponge used in TACHOSIL product, for hemostasia products provides higher levels of pliability.The pliability of this enhancing makes the hemostasia products surface that adapt to wound easier than TACHOSIL product described in the present patent application.

In detailed description above, with reference to accompanying drawing, these accompanying drawings form its part, and by illustrating that the mode that specific embodiments illustrates also can implement the present invention with this wherein.In this, direction term, such as " top ", " bottom ", "front", "rear", " head ", " tail " etc. relative to the figure be described orientation and use.Owing to can locate the assembly of embodiment with multiple different orientation, direction term is never restrictive for illustration of property object.Should be understood that, under not departing from the scope of the present invention, other embodiment can be adopted and structure or logical changes can be made.Therefore, detailed description above should be considered as there is limited significance, and scope of the present invention is defined by the appended claims.

It is contemplated that, can mix and mate feature disclosed in the application and those are incorporated to feature in above-described application by reference, to adapt to concrete condition.Various other is modified and change will be apparent for those of ordinary skill.

Claims (23)

1. form a method for hemostasis sheet material, it is characterized in that, comprising:

Mixing dextran and water form dextran-aqueous mixtures;

Described dextran-aqueous mixtures is formed the first supporting layer;

Composite fibre proteinogen and thrombin form the mixture of Fibrinogen and thrombin; With

Disperse the mixture of described Fibrinogen and thrombin on described first supporting layer to form hemostasis sheet material.

2. method according to claim 1, is characterized in that, with the concentration of about 3 to 9 grams of dextrans to about 6 ml waters, is mixed by dextran with water.

3. method according to claim 1, is characterized in that, before dextran-aqueous mixtures is formed the first supporting layer, also comprises being mixed into thrombin at least partially in dextran-aqueous mixtures.

4. method according to claim 1, is characterized in that, described first supporting layer has relatively uniform thickness.

5. method according to claim 1, is characterized in that, thrombin is salmon thrombin and wherein Fibrinogen is salmon Fibrinogen.

6. method according to claim 1, is characterized in that, forms the first supporting layer and comprises dextran described in Electrospun-aqueous mixtures formation fiber and deposit described fiber at least in part on top of each other.

7. method according to claim 1, it is characterized in that, also comprise described first supporting layer of compression to the height being less than described first supporting layer elemental height, wherein said compression makes the reduction at least 75% compared with described first supporting layer elemental height of the height of described first supporting layer.

8. method according to claim 1, it is characterized in that, also comprise maintaining body is placed on be dispersed with Fibrinogen and Thrombin mixture hemostasis sheet material on the surface, described Fibrinogen and Thrombin mixture remain on the surface of described hemostasis sheet material by wherein said maintaining body.

9. method according to claim 8, is characterized in that, described maintaining body has the second supporting layer being similar to described first supporting layer structure.

10. method according to claim 1, is characterized in that, also comprises and distributes at least one bioactivator on described hemostasis sheet material.

11. methods according to claim 1, it is characterized in that, described first supporting layer is formed in support material, and described support material comprises the material being selected from the group be made up of the Electrospun dextran of gauze, compression, polyglycolic acid polymer, polylactic acid polymer, caprolactone polymers and charged nylon.

12. 1 kinds formed hemostasis sheet material method, it is characterized in that, comprising:

Mixing dextran and water form dextran-aqueous mixtures;

Thrombin is mixed into described dextran-aqueous mixtures and forms dextran-thrombin-aqueous mixtures;

Described dextran-thrombin-aqueous mixtures is formed the first supporting layer; With

Loosen collagen fibre proteinogen on described first supporting layer with formed hemostasis sheet material.

13. methods according to claim 12, is characterized in that, with the concentration of about 3 to 9 grams of dextrans to about 6 ml waters, are mixed by dextran with water.

14. methods according to claim 12, is characterized in that, before dextran-aqueous mixtures is formed the first supporting layer, also comprise being mixed into thrombin at least partially in dextran-aqueous mixtures.

15. methods according to claim 12, is characterized in that, described first supporting layer has relatively uniform thickness.

16. methods according to claim 12, is characterized in that, thrombin is salmon thrombin and wherein Fibrinogen is salmon Fibrinogen.

17. methods according to claim 12, is characterized in that, form the first supporting layer and comprise dextran-thrombin described in Electrospun-aqueous mixtures formation fiber and deposit described fiber at least in part on top of each other.

18. methods according to claim 12, it is characterized in that, also comprise described first supporting layer of compression to the height being less than described first supporting layer elemental height, wherein said compression makes the reduction at least 75% compared with described first supporting layer elemental height of the height of described first supporting layer.

19. methods according to claim 12, is characterized in that, also comprise maintaining body to be placed on to be dispersed with fibrinogenic hemostasis sheet material on the surface, and described Fibrinogen remains on the surface of described hemostasis sheet material by wherein said maintaining body.

20. methods according to claim 19, is characterized in that, described maintaining body has the second supporting layer being similar to described first supporting layer structure.

21. methods according to claim 12, is characterized in that, also comprise and distribute at least one bioactivator on described hemostasis sheet material.

22. methods according to claim 12, it is characterized in that, described first supporting layer is formed in support material, and described support material comprises the material being selected from the group be made up of the Electrospun dextran of gauze, compression, polyglycolic acid polymer, polylactic acid polymer, caprolactone polymers and charged nylon.

23. 1 kinds formed hemostasis sheet material method, it is characterized in that, comprising:

Mixing dextran and water form dextran-aqueous mixtures;

Described in Electrospun, dextran-aqueous mixtures forms the first supporting layer; With

The steroid of antibiotic, the medicine eased the pain, somatomedin, bone morphogenetic protein, vaso-active substance, minimizing inflammation and their combination are dispersed on described first supporting layer to form hemostasis sheet material.