CN106139237A - A kind of new and effective compound hemostatic sponge and preparation method thereof - Google Patents
A kind of new and effective compound hemostatic sponge and preparation method thereof Download PDFInfo
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- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/10—Polypeptides; Proteins
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
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- A61L24/0036—Porous materials, e.g. foams or sponges
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
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- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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Abstract
The present invention relates to a kind of new and effective compound hemostatic sponge and preparation method thereof.The technical scheme is that and Col is dissolved in acetic acid aqueous solution, add appropriate ORC, under room temperature, after magnetic agitation is uniformly dispersed to powder, lyophilization, obtain composite Col ORC.Composite Col ORC is placed in the ethanol/MES buffer of 15 45% immersion, filters, abandon filtrate and obtain filtrate；Press solid-liquid ratio 1:2.5 4.5 again, take filtrate and ethanol/MES buffer, after soaking 3 6h, be slowly added EDC and NHS, after reacting 4 8h, with in 1M disodium-hydrogen and excess EDC and NHS, filter, abandon filtrate, deionized water wash, obtain cross-linking products.Sthptic sponge prepared by the present invention accelerates hemostasis process, and inanimate object toxicity, promotion organization healing.
Description
Technical field
The invention belongs to medical haemostatic sponge field, be specifically related to one and enter with collagen protein and oxidized regenerated cellulose
Compound hemostatic sponge of row crosslinking preparation and preparation method thereof.
Background technology
Stop blooding most important for the success of Oral and Maxillofacial Surgery, particularly with complication and experienced by orthognathous
Surgical site infections patient and there is bleeding tendency such as levels in patients with thrombocytopenia.Along with the development of Hemostasis, much clog
Commonly use in clinic with sewing method.Many hemostasis with packs materials, are used for hands including gelfoam, chitin and chitosan sponge
In art, these materials are respectively arranged with benefit and limitation.Such as, gelfoam spreads on wound, made of soft, insufficient pressure, makes hemostasis
Time is slightly longer, without infecting in the most uncontrollable alveolus of antiinflammatory action.And its biodegradation rate is very fast, it is unsuitable for implanting
Internal.Chitin and chitosan sponge anastalsis research, still in the initial stage, needs more discussion.Therefore continually develop novel
, the hemostatic material that good biocompatibility, haemostatic effect are fast be the problem that this area is constantly researched and developed.
Summary of the invention
It is an object of the invention to provide a kind of new and effective compound hemostatic sponge and preparation method thereof, this compound hemostatic sea
Silk floss, cross-links by a certain percentage by oxidized regenerated cellulose and I-type collagen, and prepared sthptic sponge accelerates hemostasis
Process, and inanimate object toxicity, promotion organization healing.
The technical solution used in the present invention is: a kind of new and effective compound hemostatic sponge: by collagen protein and oxidation regeneration
Cellulose crosslinked prepared；The quality of described oxidized regenerated cellulose is the 0.1-1% of collagen protein quality.
A kind of preparation method of new and effective compound hemostatic sponge, method is as follows:
1) preparation of composite Col-ORC: collagen protein sponge (Col) is dissolved in acetic acid aqueous solution, obtains collagen
Albumen acetic acid aqueous solution, joins in collagen protein acetic acid aqueous solution by appropriate oxidized regenerated cellulose (ORC), under room temperature, and magnetic
After power stirring is uniformly dispersed to powder, lyophilization 24-48h, obtain the composite of collagen protein and oxidized regenerated cellulose
Col-ORC。
Preferably, described collagen protein acetic acid aqueous solution is: every 10ml acetic acid aqueous solution adds 0.03-0.1g collagen egg
Bai Haimian；By volume percentage ratio, the concentration of described acetic acid aqueous solution is 0.5-1.2%.
Preferably, the addition of oxidized regenerated cellulose is the 0.1-1% of collagen protein quality.
2) composite Col-ORC is placed in ethanol/MES buffer that concentration expressed in percentage by volume is 15-45% immersion 30-
45min, filters, abandons filtrate and obtain filtrate；Press solid-liquid ratio 1:2.5-4.5 again, take filtrate and concentration expressed in percentage by volume is 35-45%
Ethanol/MES buffer, soak after 3-6h, be slowly added 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride
(EDC) to concentration is 10-20mg/mL, and adding N-hydroxy-succinamide (NHS) to concentration the most again is 2.25-3.75mg/
ML, after reaction 4-8h, with in 1M disodium-hydrogen and EDC and the NHS 1-3h of excess, filters, abandons filtrate, deionized water wash,
Obtain cross-linking products.
3) by cross-linking products lyophilization, new and effective compound hemostatic sponge is obtained.
The preparation method of above-mentioned new and effective compound hemostatic sponge, the preparation method of described collagen protein sponge is such as
Under:
1) remove fat: fresh cattle heel string is scraped off the fascia on surface, oils and fats, with distilled water immersion, washing, after chopping, put into
In tissue mashing machine, smash homogenate to pieces；It is soaking in sodium carbonate solution 10-18h of 0.05%-0.2% with mass fraction, filters,
Remove the cattle heel string that fat processes.
2) extract: remove the cattle heel string that fat processes, add pepsin and acetic acid aqueous solution, at 0-4 DEG C, stir 3-4
My god, gained crude extract, centrifugal, take supernatant.Preferably, pepsic addition is, every 100 grams of cattle going fat to process with
Tendon, adds pepsin 3-5g.
3) saltouing: in the supernatant after centrifugal, under stirring, the concentration of addition NaCl granule to sodium chloride reaches 2-3M,
Stand 12-18h after precipitation to be precipitated, filter, obtain the precipitation of thick white shape.
4) dialysis: by the precipitation 0.1M swelling 10-18h of Tris-HCl solution of thick white shape, loads afterwards and retains point
In the bag filter that son amount is 7000-14000, use successively the acetic acid solution of 0.5M, 0.3M, 0.1M as extracellular fluid dialysis, every kind
Concentration is dialysed 1.5-3 days respectively, finally with distilled water as extracellular fluid dialysis, dialyses 2-4 days, and the collagen protein after being dialysed is molten
Liquid.
5) it is dried: the collagen solution after dialysis is put in container, after refrigerator pre-freeze, puts in freezer dryer,
Lyophilization 24-48h, obtains collagen protein sponge.
The invention has the beneficial effects as follows: after the present invention extracts collagen solution from cattle heel string, introduce oxidation regeneration fine
Dimension element powder, after nontoxic crosslinking, makes the Col/ORC hemostasis composite of different proportion.Observed by surface texture, physics and chemistry
Performance evaluation, biological safety and efficiency analysis, make New Co l/ORC hemostasis composite reach more effectively to stop blooding, soon
Speed vivo degradation, and promote organization healing.
Accompanying drawing explanation
Fig. 1 a is the SEM figure of the compound hemostatic sponge of collagen protein sponge and difference/oxidized regenerated cellulose content；
Wherein: (A) Control (wound you collagen protein sponge), 500 ×, (a) 200 ×；
(B) Col (self-control collagen protein sponge), 500 ×, (b) 200 ×；
(C) Col-0.25%ORC, 500 ×, (c) 200 ×；
(D) Col-0.5%ORC, 500 ×, (d) 200 ×；
(E) Col-1%ORC, 500 ×, (e) 200 ×.
Fig. 1 b is the pore-size distribution of the compound hemostatic sponge of collagen protein sponge and difference/oxidized regenerated cellulose content
Figure.
Fig. 2 is the infrared spectrum of the compound hemostatic sponge of collagen protein sponge and difference/oxidized regenerated cellulose content.
Fig. 3 is the compound hemostatic sponge cytotoxicity to ACC-M cell for embodiment 1-embodiment 3 preparation.
Fig. 4 a is Col and different content Col/ORC compound hemostatic sponge hot strength comparison diagram.
Fig. 4 b is Col and different content Col/ORC compound hemostatic sponge contact angle determination comparison diagram.
Fig. 4 c is Col and different content Col/ORC compound hemostatic sponge porosity comparison diagram.
Fig. 4 d is Col and different content Col/ORC compound hemostatic sponge sucks in water rate comparison diagram.
Fig. 5 is in rabbit ear tremulous pulse and liver injury model, Col and different content Col/ORC compound hemostatic sponge hemorrhage
Amount.
Fig. 6 is different materials implantation in rabbit leg 1 day respectively, and 7 days, (HE contaminated the local pathological tissue figure after 14 days and 28 days
Color).
Detailed description of the invention
Embodiment 1
(1) preparation of collagen protein sponge
1) remove fat: fresh cattle heel string is scraped off the fascia on surface, oils and fats, with distilled water immersion, washing, after chopping, put into
Tissue mashing machine smashs to pieces homogenate.It is soaking in sodium carbonate solution 12h of 0.1% with mass fraction, the cattle filtered, fat must be gone to process
Heel string.
2) extract: take the cattle heel string 10g going fat to process obtained above and put in there-necked flask, add the stomach egg of 0.45g
White enzyme and 0.3% acetic acid solution of about 1000ml.Stir 3 days at 0-4 DEG C.By gained crude extract in High speed refrigerated centrifuge
Centrifugal, rotating speed 10000rpm is centrifuged 20min, takes supernatant.
3) saltouing: in the supernatant after centrifugal, under stirring, the concentration of addition NaCl granule to sodium chloride reaches 2-3M,
Stand 12-18h after precipitation to be precipitated, filter, obtain the precipitation of thick white shape.
4) dialysis: by the precipitation of thick white shape, with the 0.1M swelling 12h of Tris-HCl solution, load afterwards and retain molecule
Amount is in the bag filter of 10,000, uses the acetic acid solution of 0.5M, 0.3M, 0.1M as extracellular fluid dialysis, every kind of concentration dialysis 2 successively
My god.Finally with distilled water as extracellular fluid dialysis, dialyse 3 days, the collagen solution after being dialysed.
5) it is dried: the collagen solution after dialysis is put in stainless steel disc, refrigerator pre-freeze, is then placed in lyophilization
In machine, lyophilization 24-48h, obtain collagen protein sponge (Col), be stored in-20 DEG C, standby.
(2) preparation of compound hemostatic sponge
1) preparation of composite Col-ORC: add 0.06g collagen protein according to every 10ml acetic acid aqueous solution (1%v/v)
Sponge (Col), prepares collagen protein acetic acid aqueous solution.Take 100ml collagen protein acetic acid aqueous solution, add oxidized regenerated cellulose
(ORC), addition is collagen protein quality the 0.25% of oxidized regenerated cellulose (ORC), under room temperature, magnetic agitation is to powder
After being uniformly dispersed, lyophilization 24-48h, obtain the composite Col-0.25% of collagen protein and oxidized regenerated cellulose
ORC。
2) take 25mg composite Col-0.25%ORC, be placed in 10ml ethanol/MES buffer (40%v/v), soak
30min, filters, abandons filtrate and obtain filtrate；Press solid-liquid ratio 1:3.5 again, take 1mg filtrate and 3.5ml ethanol/MES buffer
(40%v/v), after soaking 4h, being slowly added EDC to concentration is 15mg/mL, and adding NHS the most again to concentration is 3.75mg/mL,
Reaction 4h after, with in 1M disodium-hydrogen and excess EDC and NHS 1h, filter, abandon filtrate, be washed with deionized 30min,
Obtain cross-linking products.
3) cross-linking products is put into and freezer dryer is dried 72h, obtain ORC content be 0.25% new and effective compound
Sthptic sponge Col-0.25%ORC, is stored in-20 DEG C.
Embodiment 2
(1) preparation of collagen protein sponge: with embodiment 1
(2) preparation of compound hemostatic sponge
1) preparation of composite Col-ORC: add 0.06g collagen protein according to every 10ml acetic acid aqueous solution (1%v/v)
Sponge (Col), prepares collagen protein acetic acid aqueous solution.Take 100ml collagen protein acetic acid aqueous solution, add oxidized regenerated cellulose
(ORC), addition is collagen protein quality the 0.5% of oxidized regenerated cellulose (ORC), under room temperature, magnetic agitation is to powder
After being uniformly dispersed, lyophilization 24-48h, obtain the composite Col-0.5%ORC of collagen protein and oxidized regenerated cellulose.
2) take 25mg composite Col-0.5%ORC, be placed in 10ml ethanol/MES buffer (40%v/v), soak
30min, filters, abandons filtrate and obtain filtrate；Press solid-liquid ratio 1:3.5 again, take 1mg filtrate and 3.5ml ethanol/MES buffer
(40%v/v), after soaking 4h, being slowly added EDC to concentration is 15mg/mL, and adding NHS the most again to concentration is 3.75mg/mL,
Reaction 4h after, with in 1M disodium-hydrogen and excess EDC and NHS 1h, filter, abandon filtrate, be washed with deionized 30min,
Obtain cross-linking products.
3) cross-linking products is put into and freezer dryer is dried 72h, obtain ORC content be 0.5% new and effective compound
Sthptic sponge Col-0.5%ORC, is stored in-20 DEG C.
Embodiment 3
(1) preparation of collagen protein sponge: with embodiment 1
(2) preparation of compound hemostatic sponge
1) preparation of composite Col-ORC: add 0.06g collagen protein according to every 10ml acetic acid aqueous solution (1%v/v)
Sponge (Col), prepares collagen protein acetic acid aqueous solution.Take 100ml collagen protein acetic acid aqueous solution, add oxidized regenerated cellulose
(ORC), addition is collagen protein quality the 1.0% of oxidized regenerated cellulose (ORC), under room temperature, magnetic agitation is divided to powder
After dissipating uniformly, lyophilization 24-48h, obtain the composite Col-1%ORC of collagen protein and oxidized regenerated cellulose.
2) take 25mg composite Col-0.25%ORC, be placed in 10ml ethanol/MES buffer (40%v/v), soak
30min, filters, abandons filtrate and obtain product；Press solid-liquid ratio 1:3.5 again, take 1mg filtrate and 3.5ml ethanol/MES buffer (40%
V/v), after soaking 4h, being slowly added EDC to concentration is 15mg/mL, and adding NHS the most again to concentration is 3.75mg/mL, reaction
After 4h, with in 1M disodium-hydrogen and excess EDC and NHS 1h, filter, abandon filtrate, be washed with deionized 30min, obtain friendship
Co-product.
3) cross-linking products is put into and freezer dryer is dried 72h, obtain new and effective being combined that ORC content is 1% and stop
Sea of blood silk floss Col-1%ORC, is stored in-20 DEG C.
Embodiment 4 effect experimental
1, Fig. 1 a and Fig. 1 b is collagen protein sponge and the compound hemostatic sponge of difference/oxidized regenerated cellulose content
SEM schemes.It is network structure from the surface of Fig. 1 a and Fig. 1 b, Control and Col.When adding the short fibre of oxidized regenerated cellulose
After dimension, the oxidized regenerated cellulose fiber in pencil is interspersed in collagen protein sponge, causes the netted knot of a part of collagen protein
Structure ruptures.The average pore size data measuring different sponge show, contrast other seas containing oxidized regenerated cellulose chopped fiber
Continuous material, homemade pure collagen protein sponge Col aperture is maximum, is 209 μm；And commodity collagen protein sponge Control is minimum,
It is 120 μm.The pore size of Col-0.25%ORC, Col-0.5%ORC and Col-1%ORC is respectively 181.5 μm, 168.2 μm
With 145.5 μm.Obviously, along with oxidized regenerated cellulose the increasing of content in Col/ORC complex, the aperture of material gradually subtracts
Little.
2, Fig. 2 is the infrared spectrum of compound hemostatic sponge prepared by embodiment 1-embodiment 3, from Figure 2 it can be seen that 3305cm-1
The peak at place is intermolecular hydrogen bonding O-H stretching vibration.-NH is typically 3540～3180cm-1There are two weak and sharp absworption peaks.
1632cm-1, 1543cm-1And 1235cm-1The peak at place represents contraction vibration respectively 1680～1630cm-1C=O (amide I
Band), shrink vibration 1570～1510cm-1Amide II carry, and shrink vibration 1335～1200cm-1Amide Ⅲ band.
According to chemical molecular formula and FT-IR spectrum, the analysis of ORC functional group is shown: 1. at 3420cm-1The contraction of the characteristic peak-OH at place
Vibration, typically 3400～3450cm-1Between occur；2.1728cm-1The contraction vibration of the peak representation carboxy C=O at place, C=O
Generally at 3000cm-1Left and right has a small peak, at 1700cm-1Have strong peak；3.1016cm-1The peak at place is CH2C-O in OH
Shrink vibration.The FTIR spectrum of contrast Col and Col/ORC, except at 1728cm-1Locate slight carboxyl C=O and absorb outside vibration,
Occur without new crest.Along with the oxidized regenerated cellulose powder increase of content, 1080cm in Col/ORC complex-1Peak, place
Intensity and 1728cm-1The peak shoulder of place carboxyl C=O is gradually increased.This shows to send out between collagen protein and oxidized regenerated cellulose
Give birth to special intermolecular interaction.1080cm-1The peak at place is probably C-O stretching vibration on secondary alcohol, also in composite
It is probably in collagen protein the C-H stretching vibration peak of the carbon being connected with amino.1724cm-1The peak at place is C=O on the ester bond of ORC
Stretching vibration characteristic peak, 1314cm-1The peak at place is probably the bending vibration of methylene, it is also possible to the C-O stretching vibration of ester, and
1080cm-1Peak is probably C-O stretching vibration on secondary alcohol in composite, is then probably and is connected with amino in collagen protein
The C-H stretching vibration peak of carbon.
3, Fig. 3 is the compound hemostatic sponge of enforcement 1, embodiment 2 and the embodiment 3 preparation cytotoxicity to ACC-M cell.
Act on ACC-M cell with each material lixiviating solution and carry out MTT experiment observation in 1,3 and 5 days, be grouped into: Control (negative control,
Commodity collagen protein sponge), Col, the collagen protein composite of the mark ORC containing different quality (0.25%, 0.5% and 1%),
Blank (does not does any process), positive control (64g/L phenol).Result is with time dependent versus cell activity table
Showing, cytoactive when blank group cell is cultivated 1 day is as comparison, n=6.(* P < 0.05 and * * P < 0.01, T-text).
In addition to adding the positive controls of 64g/L phenol, all to cytotoxic when other materials is cultivated 1 day and 3 days with ACC-M cell
Property.Col-1%ORC group cell is when cultivating 5 days, and versus cell activity value has significant difference compared with blank group.Glue
Former protein material avirulence is that the biological safety of the collagen protein owing to extracting from cattle heel string is recognized, the preparation of material
Journey strict sterilization, and it is not added with toxic chemical.And 0.1%ORC group produces mild toxicity to the cell after cultivating 5 days, point
It is more, under long-term sour environment that analysis reason is likely due to its carboxyl-content, it is suppressed that the growth of logarithmic growth after date cell.
4, the compound hemostatic sponge using embodiment 1 preparation carries out acute systemic toxicity, result such as table 1.No matter
Detection group or matched group, all mices all do not find death condition, and do not find any Acute systemic toxicity outbreak pathology disease
Shape, the result of table 1 shows all test mices body weight change situation at 24h and 72h.All Mice Body weight averages have increase, and
Showing no obvious abnormalities reaction, illustrative material is without Acute systemic toxicity.
Table 1 acute toxicity test monitoring result
5, Fig. 4 a-Fig. 4 d is the hot strength contrast of Col and the Col/ORC compound hemostatic sponge of different content, contact angle
Measuring contrast, sponge porosity contrasts, and rate that sponge sucks in water contrasts.From Fig. 4 a-Fig. 4 d, physicochemical property test result shows
The hot strength of Col is maximum, and by contrast, the hot strength of Col-0.25%ORC is less than Col, and along with oxygen in composite
Changing the increase of regenerated fiber cellulose content, the hot strength of material is gradually reduced.This result is also coincide with SEM image data, pencil
Oxidized regenerated cellulose fiber be interspersed between collagen protein, cause part collagen meshwork to rupture.In a word, contrast
Other test material, because Col-0.25%ORC has prominent porosity, surface moist and water absorption, the most resonable
Changing in performance test, the hot strength of Col is optimal, and Col-0.25%ORC takes second place.
6, the anthemorrhagic performance of compound hemostatic sponge is as shown in table 2 and Fig. 5.By to rabbit ear arterial injury models and hepatic injury
Bleeding stopping period and the amount of bleeding of model are evaluated.From table 2 and Fig. 5, in arteria auricularis damage model, four kinds of materials
The bleeding stopping period of (Col, Col-0.25%ORC, Col-0.5%ORC and Col-1%ORC) is significantly lower than Control group 136s
Bleeding stopping period (P < 0.05).The anthemorrhagic speed (22.5 ± 3.99s) of Col-0.25%ORC is the fastest, next to that Col-0.5%ORC,
Bleeding stopping period is 30.5 ± 8.29s.Fig. 5 shows, four kinds of materials (Col, Col-0.25%ORC, Col-0.5%ORC and Col-
Amount of bleeding 1%ORC) is significantly lower than the Control group (P < 0.05) that amount of bleeding is 82.95 ± 2.67g.Col-0.25%ORC
(36.87 ± 9.07g) and the amount of bleeding no significant difference of Col-0.5%ORC (33.28 ± 10.08g).Col-1%ORC and Col
Amount of bleeding be 49.18 ± 8.29g and 63.92 ± 12.52g respectively.
Col and the average bleeding stopping period of different Col/ORC composite sponge in 2 two kinds of trauma models of table
7, Fig. 6 is different materials implantation in rabbit leg 1 day respectively, 7 days, the local pathological tissue figure (HE after 14 days and 28 days
Dyeing).Histology picture when every kind of material that Fig. 6 is representative implants 1,7,14,28 days.Material through HE dyeing
The fragment shape expected in irregular bulk or be dispersed in, color is the darkviolet deeper than normal structure or peony.Some massive material
Do not colour, be owing to the disappearance of material or section and dyeing course existing region of being unstained.After implanting 1 day, some are not
The Col material being colored in muscular tissue in island (Fig. 6 a).Intradermal acute inflammatory reaction seen from Fig. 6 b and being dispersed in
Col-ORC material fragment is swallowed by multinucleated giant cell.Seen from Fig. 6 c, irregular Control material, is likely to be due to part flesh thin
Born of the same parents' caused by necrosis.After implanting 7 days, a large amount of inflammatory cell infiltrations, capillary hemorrhage and fibroblast proliferation see muscle
In tissue (Fig. 6 d).Top at Fig. 6 e, it is seen that inflammatory cell is centered around the Col/ORC surrounding materials not being colored in island.
Shown in Fig. 6 f, the muscle fiber of partial necrosis is replaced by fibrous connective tissue, it is seen that irregular Control material and multinuclear are big and small
Born of the same parents.After implanting 14 days (Fig. 6 g-i), three groups all have inflammatory reaction, Col and Col-ORC group fibres visible hamartoplasia.Finally, residual
Col-ORC and the Control material stayed sees Fig. 5 h and Fig. 6 i respectively.It addition, Control group yet suffers from multinucleated giant cell.
After implanting 28 days, in each group muscular tissue, it is showed no the material fragment (Fig. 6 j-l) of residual.
Shown by result above: FT-IR spectral results shows that oxidized regenerated cellulose contains hydrophilic group, therefore have very
Strong toughness.Oxidized regenerated cellulose powder can increase the surface moist of collagen protein sponge, water absorption, hot strength.Always
It, contrast other test materials, because Col-0.25%ORC has prominent porosity, surface moist and water absorption, with
Time in physics and chemistry performance test, the hot strength of Col is optimal, and Col-0.25%ORC takes second place.
Compared with Control, Col and Col/ORC complex has the anthemorrhagic performance of brilliance.Col-0.25%ORC stops
Blood effect is well owing to superior anthemorrhagic performance includes bleeding stopping period and amount of bleeding than Col.Col-0.25%ORC is when hemostasis
Advantage between is mainly due to two reasons.First, good surface moist can make blood quickly immerse have bigger
The material surface of diffusion area.Secondly, Col-0.25%ORC contains carboxylated hydrophilic group and good physical property, thus
Add Engorged quantity and quick-acting haemostatic powder is provided.
Col-0.25%ORC is possible not only to quickly absorb blood, it is also possible to directly participate in hemostasis by affecting thrombin
Process, the haemostatic effect of this material is substantially improved, it is possible to degradable after implanting 28 days, to surrounding tissue without substantially
Anomalous effects.In sum, this composite can be widely used in Oral and Maxillofacial Surgery as hemostasis obturator, has
Wide application prospect.
Claims (6)
1. a new and effective compound hemostatic sponge, it is characterised in that: cross-linked by collagen protein and oxidized regenerated cellulose and prepare；
The quality of described oxidized regenerated cellulose is the 0.1-1% of collagen protein quality.
2. the preparation method of the new and effective compound hemostatic sponge described in claim 1, it is characterised in that: method is as follows:
1) preparation of composite Col-ORC: be dissolved in acetic acid aqueous solution by collagen protein sponge, obtains collagen protein acetic acid water
Solution, joins appropriate oxidized regenerated cellulose in collagen protein acetic acid aqueous solution, and under room temperature, magnetic agitation to powder is disperseed
After Jun Yun, lyophilization 24-48h, obtain the composite Col-ORC of collagen protein and oxidized regenerated cellulose；
2) composite Col-ORC is placed in ethanol/MES buffer that concentration expressed in percentage by volume is 15-45% immersion 30-
45min, filters, abandons filtrate and obtain filtrate；Press solid-liquid ratio 1:2.5-4.5 again, take filtrate and concentration expressed in percentage by volume is 35-45%
Ethanol/MES buffer, soak after 3-6h ,-3-ethyl-carbodiimide hydrochloride is extremely to be slowly added 1-(3-dimethylamino-propyl)
Concentration is 10-20mg/mL, and adding N-hydroxy-succinamide to concentration the most again is 2.25-3.75mg/mL, after reaction 4-8h,
With in 1M disodium-hydrogen and excess 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride and N-hydroxysuccinimidyl acyl
Imines 1-3h, filters, abandons filtrate, deionized water wash, obtain cross-linking products；
3) by cross-linking products lyophilization, new and effective compound hemostatic sponge is obtained.
3. according to the preparation method of the new and effective compound hemostatic sponge described in claim 1, it is characterised in that: described collagen
The preparation method of protein sponge is as follows:
1) remove fat: fresh cattle heel string is scraped off the fascia on surface, oils and fats, with distilled water immersion, washing, after chopping, put into tissue
In bruisher, smash homogenate to pieces；It is soaking in sodium carbonate solution 10-18h of 0.05%-0.2% with mass fraction, filters, fat must be removed
The cattle heel string processed；
2) extract: remove the cattle heel string that fat processes, add pepsin and acetic acid aqueous solution, stir 3-4 days at 0-4 DEG C, institute
Obtain crude extract, centrifugal, take supernatant；
3) saltouing: in the supernatant after centrifugal, under stirring, the concentration of addition NaCl granule to sodium chloride reaches 2-3M, waits to sink
Precipitation stands 12-18h after going out, and filters, obtains the precipitation of thick white shape；
4) dialysis: by the precipitation 0.1M swelling 10-18h of Tris-HCl solution of thick white shape, load molecular cut off afterwards
For in the bag filter of 7000-14000, use the acetic acid solution of 0.5M, 0.3M, 0.1M as extracellular fluid dialysis, every kind of concentration successively
Dialysis 1.5-3 days respectively, finally with distilled water as extracellular fluid dialysis, dialyse 2-4 days, the collagen solution after being dialysed；
5) it is dried: the collagen solution after dialysis is put in container, after refrigerator pre-freeze, puts in freezer dryer, freezing
It is dried 24-48h, obtains collagen protein sponge.
4. according to the preparation method of the new and effective compound hemostatic sponge described in claim 3, it is characterised in that: step 2) in,
Pepsic addition is, every 100 grams of cattle heel strings going fat to process, and adds pepsin 3-5g.
5. according to the preparation method of the new and effective compound hemostatic sponge described in claim 2, it is characterised in that: described collagen
Albumen acetic acid aqueous solution is: every 10ml acetic acid aqueous solution adds 0.03-0.1g collagen protein sponge；By volume percentage ratio, described
The concentration of acetic acid aqueous solution be 0.5-1.2%.
6. according to the preparation method of the new and effective compound hemostatic sponge described in claim 2, it is characterised in that: oxidation regeneration is fine
The 0.1-1% that addition is collagen protein quality of dimension element.
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