CN106474533A - A kind of it is applied to combine dressing of the surface of a wound and preparation method thereof - Google Patents
A kind of it is applied to combine dressing of the surface of a wound and preparation method thereof Download PDFInfo
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- CN106474533A CN106474533A CN201610580200.8A CN201610580200A CN106474533A CN 106474533 A CN106474533 A CN 106474533A CN 201610580200 A CN201610580200 A CN 201610580200A CN 106474533 A CN106474533 A CN 106474533A
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- A—HUMAN NECESSITIES
- 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
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
- A61L26/0028—Polypeptides; Proteins; Degradation products thereof
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- A—HUMAN NECESSITIES
- 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
- A61L24/0005—Ingredients of undetermined constitution or reaction products thereof
-
- A—HUMAN NECESSITIES
- 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
- A61L24/001—Use of materials characterised by their function or physical properties
- A61L24/0015—Medicaments; Biocides
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- A—HUMAN NECESSITIES
- 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
- A61L24/001—Use of materials characterised by their function or physical properties
- A61L24/0042—Materials resorbable by the body
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- A—HUMAN NECESSITIES
- 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
- 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
- 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
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0057—Ingredients of undetermined constitution or reaction products thereof
-
- A—HUMAN NECESSITIES
- 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
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/0066—Medicaments; Biocides
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- A—HUMAN NECESSITIES
- 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
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/009—Materials resorbable by the body
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4741—Keratin; Cytokeratin
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- A—HUMAN NECESSITIES
- 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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/30—Compounds of undetermined constitution extracted from natural sources, e.g. Aloe Vera
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/418—Agents promoting blood coagulation, blood-clotting agents, embolising agents
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- A—HUMAN NECESSITIES
- 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
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/04—Materials for stopping bleeding
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Abstract
The present invention relates to a kind of be applied to combine dressing of the surface of a wound and preparation method thereof, the composition of the dressing is：Hair-keratin and traditional Chinese medicine ingredients, the hair include any one in human hair and animal hair, and the composition of the traditional Chinese medicine ingredients includes pseudo-ginseng, Paris polyphylla, RADIX PSAMMOSILENE, borneol and Moschus.The preparation method includes the preparation of the preparation of hair-keratin, the preparation of traditional Chinese medicine ingredients and dressing.Chinese tradition traditional Chinese medicine is combined by the present invention with biomaterial, using the carrier that hair-keratin gel etc. is repaired as traditional Chinese medicine ingredients wound, the wound-surface cover with biological dressing property can be formed, which is a kind of wound-surface cover of excellent, simultaneously, grow with autologous skin as wound slowly heals, be that the regeneration for promoting skin provides an environment for being conducive to wound healing.
Description
Technical field
The present invention relates to a kind of dressing, and in particular to a kind of be applied to combine dressing of the surface of a wound and preparation method thereof.
Background technology
The wound of human body and animal body is seen everywhere, various wounds can all cause different degrees of cell degeneration, necrosis and
Tissue defect, it is necessary to which tissue repair is carried out by hyperplasia and the formation of intercellular matrix.Although control at present treatment trauma or
The medicament categories of hemostasis are various, a feast for the eyes, but through internal series of complex after having quite a few haemostatic medicament be administered orally
Physiology course completing, and simply do not directly act on wound blood vessels by its medicinal powder, reach the purpose of hemostasis.Have
Although medicine can directly spread on wound, pharmaceutical drying, movable injury easily involve wound, cause pain or open once again
Split, and as medicine can not be absorbed by the skin, using needing to clear up medicine foam or the chip that wound stays during rear or dressing, cause
The pain of patient, it is also possible to delay the healing of wound.
Traditional Chinese medicine is combined by the dressing of the application with biomaterial, builds the matrix of simulation body tissue and function
And mounting system, rebuild or recover the final goal that skin barrier is injured Wound treating.
Content of the invention
It is an object of the invention to provide a kind of be applied to combine dressing of the surface of a wound and preparation method thereof.The purpose of the present invention is
It is achieved through the following technical solutions：
A kind of combine dressing for being applied to the surface of a wound, the composition of the dressing is：Hair-keratin and traditional Chinese medicine ingredients, the Chinese medicine
The composition of composition includes pseudo-ginseng, Paris polyphylla, RADIX PSAMMOSILENE, borneol and Moschus.
Further, the weight ratio of the hair-keratin and traditional Chinese medicine ingredients is 1～3:3～1；In the traditional Chinese medicine ingredients
The parts by weight of various composition are：20～40 parts of pseudo-ginseng, 10～25 parts of Paris polyphylla, 10～20 parts of RADIX PSAMMOSILENE, 20～40 parts of borneol and
0.001～0.8 part of Moschus.
Further, the hair-keratin is prepared from by hair, and the hair is included in human hair and animal hair
Any one.
Further, the hair-keratin includes that granular hair-keratin or gelation obtain hair-keratin；Institute
Stating animal hair includes wool.
The preparation method of a kind of compound dressing for being applied to the surface of a wound, the method comprising the steps of：
(1) preparation of hair-keratin：The hair-keratin includes that granular hair-keratin or gelation obtain hair
Keratin：Concrete grammar is：
S1：Degreasing and softening：Weighing hair carries out degreasing and softening；
S2：Decolourize：Hair after softening is positioned in de-inking solution and is decolourized, the composition of the de-inking solution is Jiao
3～5 weight portion of sodium phosphate, 1～2 weight portion of potassium peroxydisulfate, 70～90 parts by volume of hydrogen peroxide, 22～33 parts by volume of concentrated ammonia liquor and
350～450 parts by volume of water；
S3：Cleaning, drying：Hair after decolouring is carried out, and is dry；
S4：Prepared by powder：Hair after the drying is shredded, and adds liquid nitrogen frozen to grind, and obtains length for 0.1～2mm
Hair meal；Meal is placed in ball grinder, agate bead is put into, physiological saline is added, rotate, ball milling, until 50 μm of particle diameter
Till～90 μm, then it is homogenized for hair-keratin particle；
S5：Hair-keratin particle：In homogenate, add physiological saline to be centrifuged, abandoning supernatant, and remove precipitation
Top layer and bottom, take the hair-keratin of stage casing even particle size, plus physiological saline mixed and continues centrifugation, repeats above-mentioned behaviour
Make 2～4 times, the hair-keratin of stage casing even particle size is finally still taken, is then obtained as highly purified granular hair
Keratin；
S6：Sterilizing：Packaging, sealing, using cobalt60Irradiation method sterilizes, and preserves, stand-by；
S7：The hair-keratin of gelation：Granular hair-keratin after sterilizing in step S6 is added water, fully
Mix, incubation 2-3 hour in incubator is positioned over, to more homogeneous color and proterties is assumed；Sterilizing, obtains hair angle egg
White gel；
(2) preparation of traditional Chinese medicine ingredients：
Weigh pseudo-ginseng in traditional Chinese medicine ingredients, Paris polyphylla, RADIX PSAMMOSILENE, borneol and Moschus in proportion, pulverized various respectively
End, and mix and obtain traditional Chinese medicine ingredients；
(3) preparation of dressing：
By the hair-keratin of the granular hair-keratin prepared in step (1) or gelation and step (2)
In obtain traditional Chinese medicine ingredients mixing be uniformly dispersed, obtain composite auxiliary material.
Further, in the S1 of step (1), the degreasing and the method for softening are：Soaked 20～60 minutes with ethanol, leaching
During bubble at interval of stirring in 3～10 minutes once, soak and finish the flushing of rear flowing water；After flushing, hair is positioned over hypochlorous acid
Soaking 20～60 minutes in the solution that sodium is prepared with water, stirring in 3～10 minutes being spaced in the immersion process once, immersion is finished
Flowing water is rinsed afterwards；
In the S2 of step (1), the hair after softening is positioned over 2-3 hour in de-inking solution, in the middle of decolorization
Every stirring in 10～30 minutes once, soak and the flushing of rear flowing water is finished, till washing de-inking solution off；The composition of the de-inking solution is
4 weight portion of sodium pyrophosphate, 1.5 weight portion of potassium peroxydisulfate, 80 parts by volume of hydrogen peroxide, 27.5 parts by volume of concentrated ammonia liquor and 400 body of water
Product part；
In the S3 of step (1), the hair after decolouring is soaked in water, soak to finish and rinsed with water, filter, dry in air blast
Dry overnight for 37 DEG C in dry case.
Further, in the S1 of step (1), the sodium hypochlorite is 1～5 with the volume ratio of water:9, the sodium hypochlorite
For 10% sodium hypochlorite；
In the S2 of step (1), crinite de-inking solution will be placed and be placed in 37 DEG C of constant temperature and decolourized；The pyrophosphoric acid
Sodium is 99% sodium pyrophosphate, and potassium peroxydisulfate is 99.5% potassium peroxydisulfate, and hydrogen peroxide is 30% hydrogen peroxide, and concentrated ammonia liquor is 25% dense ammonia
Water.
Further, in the S4 of step (1), ball milling：Meal 2g is placed in ball grinder, is put into agate bead, add physiology
Salt solution 10ml, ball milling 1～3 hour, particle size is examined under a microscope, is added physiological saline 5ml and continues grinding, until shape
Become about 60 μm -80 μm of particle diameter；The quantity of the agate bead：Diameter 1cm agate bead 20, diameter 60mm agate bead 100；
In the S6 of step (1), the highly purified hair-keratin mud drum for obtaining dress is sterilized using 60Coradiation method, dosage
12Kg.
Further, in the S7 of step (1), add water fully to mix in granular hair-keratin, be positioned over 37 DEG C
Incubation 2-3 hour.
Further, the hair-keratin is the hair-keratin of gelation, the hair-keratin of the gelation
Concentration (g/ml) is 0.1～5%.
Further, the concentration of the hair-keratin of the gelation is 0.8～3%.
Further, the preparation method of the hair-keratin gel can also be replaced by the following method：
(1) hair-keratin becomes salt：1. hair is weighed, the mixed solution of second alcohol and water is added in hair, heat, gradually
Rise high-temperature to solution start backflow, while backflow while drip aqueous slkali, to pH value be 6.5～7.5 when, backflow with dropping and meanwhile knot
Bundle, after terminating, continues stirring, makes reaction completely, obtains into salt hair-keratin；2. the obtained salt hair-keratin that becomes is filtered
Go out, washing lotion is rinsed, wash to not in alkalescence, dry, obtain hair-keratin sulfonate；3. by obtained hair-keratin sulphur
Hydrochlorate is crushed to micron order, is lyophilized, obtains hair-keratin sulfonic acid salt powder；(2) preparation of hair-keratin gel：Weigh hair
Keratin sulfonic acid salt powder is sent out, water is added, fully mix, be positioned in incubator, to assuming more homogeneous color and proterties is
Only, sterilized using 60Coradiation method, obtain hair-keratin gel.
The invention provides a kind of be applied to combine dressing of the surface of a wound and preparation method thereof, which mainly has the advantage that
For：
Chinese tradition traditional Chinese medicine is combined by the present invention with biomaterial, hair-keratin biomaterial have ooze beneficial to cell
The three dimensional scaffold structure for entering, adhere to and breeding, with the characteristic for being substantially better than other materials in terms of cell adherence and propagation, with
When there is nerve-inducing, also show that good effect in terms of hemostasis, using hair-keratin gel etc. as traditional Chinese medicine ingredients
The carrier that wound is repaired, can form the wound-surface cover with biological dressing property, and which is that a kind of surface of a wound of excellent covers
Cover material, meanwhile, grow with autologous skin as wound slowly heals, be to promote the regeneration of skin to provide one to be conducive to the surface of a wound to heal
The environment of conjunction.Specifically, (1) its haemostatic effect is very good, and can effectively accelerate wound healing；(2) which effectively prevents bacterium sense
Dye, (3), with good biocompatibility, (4) control and absorb wound exudate, and (5) gas permeability is strong, are suitable for gas and vapor
Through (6) protect cambium, and (7) advantage such as have no toxic side effect, prepare the immunity of dressing nothing using hairs such as human hair or wools
Originality, will not allergy；；(8) only external application is needed, easy to use, effect is significant；Importantly, the engineering dressing voluntarily can dissolve
It is absorbed by organisms, the pain of many and patient that bleeds when eliminating releasing, also will not leave behind chip and delay the healing of wound, it is to avoid
Ordinary adjuvants stick together the damage again of the surface of a wound, are that the regeneration for promoting skin provides an environment for being conducive to wound healing.The work
At the wound face for being applied to human body or animal body that journey dressing can be fabulous.
Description of the drawings
The present invention is described in further detail below according to accompanying drawing.
Fig. 1 is the structural representation in light Microscopic observation hair-keratin described in the embodiment of the present invention；
Fig. 2 is that the structure for observing hair-keratin under ESEM (1.69 × 103) described in the embodiment of the present invention is shown
It is intended to；
Fig. 3 is that the structure for observing hair-keratin under ESEM (1.25 × 103) described in the embodiment of the present invention is shown
It is intended to；
Fig. 4 is the structural representation for observing hair-keratin under transmission electron microscope (1 × 105) described in the embodiment of the present invention
Figure；
Fig. 5 is the structural representation for observing hair-keratin under transmission electron microscope (2 × 105) described in the embodiment of the present invention
Figure；
Fig. 6 is the scanning electron microscope (SEM) photograph of the hair-keratin for being loaded with traditional Chinese medicine ingredients described in the embodiment of the present invention；
Fig. 7 is the animal model schematic diagram that combine dressing is spread on the surface of a wound described in the embodiment of the present invention；
Fig. 8 is the schematic diagram after the surface of a wound recoveries later in 21 days described in the embodiment of the present invention；
Fig. 9 is that the dyeing of the mouse dermal fibroblast described in the embodiment of the present invention shows NTx PE situation；
Figure 10 is the proterties of the hair-keratin of the gelation of 4% described in the embodiment of the present invention.
Specific embodiment
A kind of described in the embodiment of the present invention is applied to combine dressing of the surface of a wound and preparation method thereof.Below with specific experiment
Specific embodiment is described as a example by case, it will be appreciated that specific embodiment described herein only in order to explain the present invention,
It is not intended to limit the present invention.
Embodiment 1
A kind of combine dressing for being applied to the surface of a wound, the composition of the dressing is：Hair-keratin and traditional Chinese medicine ingredients, the Chinese medicine
The composition of composition includes pseudo-ginseng, Paris polyphylla, RADIX PSAMMOSILENE, borneol and Moschus.
The weight ratio of hair-keratin and traditional Chinese medicine ingredients is 1～3:3～1, preferably 1:1～2, such as 1:1.
The parts by weight of the various composition in traditional Chinese medicine ingredients can be：1. 20 parts of pseudo-ginseng, 25 parts of Paris polyphylla, 10 parts of RADIX PSAMMOSILENE,
20 parts of borneol and 0.001 part of Moschus；2. 40 parts of pseudo-ginseng, 10 parts of Paris polyphylla, 20 parts of RADIX PSAMMOSILENE, 40 parts of borneol and 0.8 part of Moschus；3. three
7 30 parts, 18 parts of Paris polyphylla, 15 parts of RADIX PSAMMOSILENE, 30 parts of borneol and 0.1 part of Moschus；4. 25 parts of pseudo-ginseng, 12 parts of Paris polyphylla, RADIX PSAMMOSILENE 18
Part, 35 parts of borneol and 0.03 part of Moschus；5. 33 parts of pseudo-ginseng, 23 parts of Paris polyphylla, 13 parts of RADIX PSAMMOSILENE, 25 parts of borneol and 0.008 part of Moschus；
6. 22 parts of pseudo-ginseng, 15 parts of Paris polyphylla, 11 parts of RADIX PSAMMOSILENE, 38 parts of borneol and 0.08 part of Moschus.
Used as further preferably embodiment, hair-keratin is prepared from by hair, and hair includes human hair and moves
Any one in thing hair；Animal hair can be wool etc..
Used as further preferably embodiment, the hair-keratin includes granular hair-keratin or gelation
Hair-keratin, the preferred hair-keratin of gelation.
Used as further preferably embodiment, the concentration (mass/volume) of the hair-keratin of the gelation is permissible
For：1. 0.1%；2. 5%；3. 0.8%；4. 3%；5. 1.5%；6. 2%.
Embodiment 2
The preparation method of a kind of compound dressing for being applied to the surface of a wound, the method concrete grammar are as follows：
(1) preparation of hair-keratin：
S1：Degreasing：5g hair is weighed, is placed in 1000ml beaker.Add 99.7% soaked in absolute ethyl alcohol 30 minutes, interval
Stirred once with glass bar within 5 minutes, flowing water is rinsed；
S2：Soften：Hair after cleaning is positioned over the >=solution of 10% sodium hypochlorite 10ml and distilled water 90ml configuration
In, soak at room temperature 30 minutes, interval are stirred once for 5 minutes with glass bar, and flowing water is rinsed；
S3：Decolourize：By the weight of 5g hair, 99% sodium pyrophosphate (Na is weighed4P2O7) 4g, 99.5% potassium peroxydisulfate
(K2S2O8) 1.5g, 30% hydrogen peroxide (H2O2) 80ml, 25% concentrated ammonia liquor 27.5ml, distilled water 400ml, de-inking solution is configured to,
The hair of softening is placed in this solution, is placed in 37 DEG C of Water Tanks with Temp.-controlled, was stirred 1 time with glass bar every 20 minutes, viewing head
Color is sent out to light yellow, this process used time general 2-3 hour, shorter than conventional method a lot；Hair after decolouring is placed
On screen cloth, flowing water is rinsed, until washing de-inking solution off.
S4：Cleaning, drying：Remove ionized water and the hair more than 12 hours after above-mentioned decolouring is soaked, remove remaining reagent；Go
Ionized water is rinsed, and is placed in air dry oven 37 DEG C and dries overnight after filtration；
S5：Grinding：It is placed in ceramic mortar after Hair grooming scissors after drying is shredded, liquid nitrogen frozen grinding is added, extremely
Hair becomes 1mm length, obtains hair meal；
S6：Ball milling：Hair meal 2g is weighed, is placed in ball grinder, be put into agate bead, add physiological saline 10ml, 460
Turn/min, ball milling 2 hours, the size of observed under electron microscope particle diameter is inverted, is added physiological saline 5m and continues grinding 1-2
Hour, until the human hair keratin particle homogenate of about 60 μm -80 μm of particle diameter is formed, preferably 70 μm；The quantity of above-mentioned agate bead：
Diameter 1cm agate bead 20, diameter 60mm agate bead 100；
S7：Tubulature：In above-mentioned homogenate add physiological saline 30ml, by the liquid rinse in ball grinder be transferred to 50ml from
In heart pipe；
S8：Centrifugation：Centrifuge tube is placed in the Centrifuge Cup of centrifuge, 3000 turns/min of centrifugal speed, 20min is centrifuged, is abandoned
Remove supernatant, remove the top layer of keratin precipitation and bottom with key, take stage casing even particle size keratin move to new from
In heart pipe, plus physiological saline 40ml is mixed, and continues centrifugation once, abandoning supernatant, then removes the table of keratin precipitation with key
Layer and bottom, the keratin for taking stage casing even particle size moved in new centrifuge tube, then plus physiological saline 40ml mix, again from
The heart, abandons supernatant, then the top layer with key removal keratin precipitation and bottom, and the keratin for taking stage casing even particle size is moved to
In new centrifuge tube, highly purified keratin mud, i.e., granular hair-keratin is obtained；It is temporarily placed in 4 DEG C of preservations；
S9：Sterilizing：The highly purified granular hair-keratin for obtaining is loaded in polybag, good seal, uses carton
Being sent to radiation center after packaging carries out Co60Radiation sterilization, dosage 12Kg, 4 DEG C of refrigerators are placed on, stand-by；
The highly purified keratin mud for preparing (i.e. granular hair-keratin) is seen under om observation (× 20)
Examine, which maintains the original stable fascicular texture of hair, as shown in Figure 1.
Fig. 2～5 are the granular hair-keratin electron microscope, and Fig. 2 behaves and acts in an unreasonable way section SEM (1.69 × 103), Fig. 3
Face SEM (1.25 × 10 is delivered in behaviour3), Fig. 4 is human hair TEM (1 × 105), Fig. 5 is human hair cortex TEM (2 × 105).
From ESEM (SEM), through the Hair-material surface smoother that this patent method is processed, no normal dark hair
Scale structure, the convex patch shape in scrobicula；Cross section hair cortex is still in nothing structure homogeneous shape.By the visible hair of transmission electron microscope (TEM)
Send out melanin in keratin granular materials to be all disintegrated.
(2) preparation of traditional Chinese medicine ingredients：
Weigh pseudo-ginseng in traditional Chinese medicine ingredients, Paris polyphylla, RADIX PSAMMOSILENE, borneol and Moschus in proportion, pulverized various respectively
End, size are 80～200 mesh, preferably 120～150 mesh, and the powder of various composition is mixed obtain traditional Chinese medicine ingredients.
(3) preparation of dressing：
By the traditional Chinese medicine ingredients obtained in the granular hair-keratin prepared in step (1) and step (2) by than
Example mixing is uniformly dispersed, and obtains composite auxiliary material, as shown in fig. 6, Fig. 6 is the ESEM of the hair-keratin for being loaded with traditional Chinese medicine ingredients
Figure SEM (3.88 × 102).
The amount of various composition in the ratio and traditional Chinese medicine ingredients of hair-keratin and traditional Chinese medicine ingredients in step (2) and (3)
As described in Example 1, restriction is not repeated herein.
Embodiment 3
It can be any one of following two methods that gelation in the present invention obtains the preparation method of hair-keratin：
1st, weigh and add in (such as embodiment 2) that the present invention obtains highly purified granular hair-keratin 50mg
In the water of 5ml, fully mix, incubation 2-3 hour in incubator is positioned over, to more homogeneous color and proterties is assumed；Go out
Bacterium, obtains the hair-keratin of gelation, and its concentration is 1%.
2、S1：10g wool or human hair is weighed, the mixed solution being put into equipped with 180ml absolute ethyl alcohol and 20ml distilled water
There-necked flask in, heating 15 minutes after be warming up to 75 DEG C, solution start backflow；Then, the NaOH for dripping 2ml in backflow is molten
Liquid, is 7 to keratic pH value is aoxidized, and flows back and dropping is while terminate, the time continues 90 minutes；After dropping and backflow terminate,
Continue to stir 5 hours at 25 DEG C, make reaction completely, obtain into salt hair-keratin.
S2：Salt hair-keratin will be become to leach, and rinsed with washing lotion, the washing lotion is that absolute ethyl alcohol is mixed with equivalent distilled water
Close, unreacted NaOH is removed, with pH detection paper whether in alkalescence, wash to not in alkalescence, be put into 60 DEG C of air dry ovens
Middle drying 6 hours, obtains dry hair-keratin sulfonate；
S3：It is in fragility through hair-keratin sulfonate obtained from above-mentioned process, crushes in Meteor-style ball grinder
To keratin powder, ball milling condition (ratio of grinding media to material, milling time, rotating speed etc.) is controlled to make the particle of hair-keratin sulfonic acid salt powder
At 50 μm or so, it is lyophilized；
S4：The hair-keratin of gelation：Operate in super-clean bench, take obtain in 100mg step S3 lyophilized after hair
Send out in keratin sulfonic acid salt fines and 5ml sterilized water is added, first material is mixed by hand using agitator, then fully mixed with mediation mixing machine
Even, 37 DEG C of incubations about 2-3 hour is positioned over, then the more homogeneous hair-keratin gel of color and proterties is obtained, then,
Sterilized using 60Coradiation method, the hair-keratin of gelation is obtained, its concentration is 2%.
Figure 10 is the proterties of the hair-keratin of 4% gelation for concentration, the property of the hair-keratin of 2% gelation
Shape is diluter.
Then the thin traditional Chinese medicine ingredients of the hair-keratin of the gelation and the present invention make the combine dressing of the present invention.Hair
The ratio of keratin and traditional Chinese medicine ingredients as described in Example 1, is not repeated herein restriction.
Embodiment 4
The toxotest for being applied to the combine dressing of the surface of a wound is as follows：
(1) prepared by sample leaching liquor：Carry drug application material (i.e. the hair-keratin containing traditional Chinese medicine ingredients) 1g and add 10ml physiology
In salt solution, crush after liquid nitrogen frozen solidification, take 1g and add in 10ml physiological saline, after vibration mixing in cell culture incubator
37 DEG C of placement 7d, suction out supernatant after centrifugation, prepare and carry drug application material leaching liquor stoste (initial 100% concentration Hair-material 100mg
: immersion liquid 1ml=100mg/ml), filtration sterilization, standby.
(2) external acute systemic toxicity：May in drug application material (dressing i.e. of the present invention) preparation process to carrying
The acute toxicity for causing preliminary assessment in addition.
As a result show：According to the standard scheme of acute toxicity test, carry out conventional gavage to experiment mice, and with postoperative not
Observed with time point.As a result show, preferably, in 7 day observation period, nothing is dead, each group in the 72h observation period for mouse ordinary circumstance
Animal has no poisoning symptom or bad reaction, and nothing is dead, and body weight increase is normal, and variance analysis shows two test group and physiological saline
Between group there is no notable difference in Mouse Weight change, and this shows two kinds of the hair-keratin of hair-keratin particle and gelation
Form material leaching liquor is all nothing Acute Toxicity.
(3) cell toxicity test：Mtt assay method determines cell relative number and relative activity：
As a result show：The cell rate of increase relatively that the variable concentrations leaching liquor of medicine dressing materials is measured is carried in three time points
More than 80% is all reached, cytotoxicity is classified as one-level, with good cell compatibility, no cytotoxicity, illustrate processed
In journey and for causing the change of hair self property and compatibility.It is shown in Table 1：
1 powder MTT result statistical analysis of table
PCEMNR (PCE) micronucleus test：Evaluate the possible genetoxic of material.Micronuclear rates refer to containing
The polychromatic erythrocyte number of micronucleus, is represented with permillage (‰).
As a result show：The microkernel incidence of powder homogenate group (hair-keratin i.e. of the present invention) and negative control group
Nothing significant difference (P>0.05), (P notable with positive controls comparing difference<0.01), tested material mouse bone marrow cells are thermophilic incarnadines more
Cell micronucleus experiment is negative, and illustrates that hair-keratin particle group nothing causes chromosome aberration toxic action.It is shown in Table 2
Table 2 Polychromatic erythrocyte (PCE) micronucleus test result
Embodiment 5
Animal experiment study：
1st, the experimental study that different proportion traditional Chinese medicine ingredients human hair keratin material is affected on wound healing
Wound repair is tested
Take healthy SD rat, anaesthetized with pentobarbital (30mg/kg), 1cm is opened by its back backbone side, scraper holostrome is cut
Except skin, diameter 1.8cm, an area 2.54cm is formed2Circular full thickness skin cut off the surface of a wound, offside antimere skin make
For normal own control.The compound dressing of the present invention that the hair-keratin of preparation and fibroblast tissue are composited
The surface of a wound is uniformly spread on, and gauze flap coverage, bolus dressing process wound, as shown in Figure 7.
Observation：After test group of animals wound, the 3d days surface of a wound have reduced, it is seen that new granulation tissue generates.21～28d after wound,
Most of wound healing, as shown in Figure 8.
2nd, the experimental study that different proportion traditional Chinese medicine ingredients human hair keratin pulvis is affected on wound healing
HE staining tissue slides are observed：Survey skin corium to thicken, average thickness about 0.58mm when 4 weeks, it is seen that substantial amounts of angle egg
Assemble under white particle corium, have cell and fibrous tissue growth between keratin, have a small amount of multinucleate giant cell, interior phagocytosis has angle egg
White particle.
Immunohistochemical staining is observed：In section, mouse dermal fibroblast secretes NTx albumen in a large number, and collagen is fine
Closely, brown stain, NTx albumen are interspersed with histocyte for dimension arrangement；Filling position cutaneous immunisation group after 8th week
Weave chemistry dyeing observation figure is as shown in figure 9, Fig. 9 shows NTx PE situation for dyeing.
3. bacterium infection observation：Obvious bacterium infection situation is had no injured to after fully recover.
Conclusion：The combine dressing effect highly significant of the present invention, in the case of without any medication, does not occur substantially sending out
Inflammation, and 21-28d, most wound healings after hindering, hour angle protein gel can be degraded substantially within 12 weeks.
Specifically, hair-keratin biomaterial has the three dimensional scaffold structure for penetrating into beneficial to cell, adhering to and breed,
With the characteristic for being substantially better than other materials in terms of cell adherence and propagation, while with nerve-inducing, in terms of hemostasis
Show good effect；Coordinate the hair-keratin of the present invention again so as to which effect is extremely notable.
Embodiment 6
The clinical practice situation for the present invention being applied to the combine dressing of the surface of a wound is as follows：
(1) experimenter selects：Select the various surface of a wound of adult each 30.
(2) processing method：The conventional treatment surface of a wound, the human hair keratin of the traditional Chinese medicine ingredients being respectively adopted containing the present invention coagulate
Glue/pulvis (pulvis is the particle of the present invention) place's flap coverage, is compared with traditional treatment method.
(3) observation index
The observation cycle：1 month；
Local gross examination of skeletal muscle：The change of depression filling position flatness, injection site skin have or not red and swollen sepage and scleroma
Deng；
The combine dressing effect highly significant containing traditional Chinese medicine ingredients and hair-keratin of the present invention, is not having in volunteer
In the case of any medication, there is not obvious inflammation, and 21-28d after hindering, most surface of a wound recover good.
Note：The studies above is all carried out in the case of Ethics Committee and experimenter's informed consent.
Finally it should be noted that：Above-described each embodiment is merely to illustrate technical scheme, rather than to which
Limit；Although being described in detail to the present invention with reference to the foregoing embodiments, it will be understood by those within the art that：
Which still can be modified to the technical scheme described in previous embodiment, or which part or all technical characteristic are entered
Row equivalent；And these modifications or replacement, do not make the essence of appropriate technical solution depart from various embodiments of the present invention technical side
The scope of case.
Claims (10)
1. a kind of combine dressing for being applied to the surface of a wound, it is characterised in that：The composition of the dressing is：Hair-keratin becomes with Chinese medicine
Point, the hair includes that any one in human hair and animal hair, the composition of the traditional Chinese medicine ingredients and parts by weight include
20～40 parts of pseudo-ginseng, 10～25 parts of Paris polyphylla, 10～20 parts of RADIX PSAMMOSILENE, 20～40 parts of borneol and 0.001～0.8 part of Moschus.
2. the combine dressing for being applied to the surface of a wound according to claim 1, it is characterised in that：The hair-keratin and Chinese medicine
The weight ratio of composition is 1～3:3～1.
3. the combine dressing for being applied to the surface of a wound according to claim 2, it is characterised in that：The hair-keratin includes
Granular hair-keratin or the hair-keratin of gelation；The animal hair includes wool.
4. a kind of preparation method of the compound dressing for being applied to the surface of a wound according to any one of claims 1 to 3, its are special
Levy and be：The method comprising the steps of：
(1) preparation of hair-keratin：The hair-keratin is the hair-keratin of granular hair-keratin or gelation：
Concrete grammar is：
S1：Degreasing and softening：Weighing hair carries out degreasing and softening；
S2：Decolourize：Hair after softening is positioned in de-inking solution and is decolourized, the composition of the de-inking solution is pyrophosphoric acid
3～5 weight portion of sodium, 1～2 weight portion of potassium peroxydisulfate, 70～90 parts by volume of hydrogen peroxide, 22～33 parts by volume of concentrated ammonia liquor and water
350～450 parts by volume；
S3：Cleaning, drying：Hair after decolouring is carried out, and is dry；
S4：Prepared by powder：Hair after the drying is shredded, and adds liquid nitrogen frozen to grind, and obtains the hair that length is 0.1～2mm
Send out meal；Meal is placed in ball grinder, agate bead is put into, physiological saline is added, rotate, ball milling, until 50 μm～90 μ of particle diameter
Till m, then it is homogenized for hair-keratin particle；
S5：Hair-keratin particle：In homogenate, add physiological saline to be centrifuged, abandoning supernatant, and remove the table of precipitation
Layer and bottom, take the hair-keratin of stage casing even particle size, plus physiological saline is mixed and continues centrifugation, repeats aforesaid operations 2
～4 times, the hair-keratin of stage casing even particle size is finally still taken, then obtain as highly purified granular hair angle egg
In vain；
S6：Sterilizing：Packaging, sealing, using cobalt60Irradiation method sterilizes, and preserves, stand-by；
S7：The hair-keratin of gelation：Granular hair-keratin after sterilizing in step S6 is added water, fully mixed
Even, incubation 2-3 hour in incubator is positioned over, to more homogeneous color and proterties is assumed；Sterilizing, obtains hair-keratin
Gel；
(2) preparation of traditional Chinese medicine ingredients：
Pseudo-ginseng in traditional Chinese medicine ingredients, Paris polyphylla, RADIX PSAMMOSILENE, borneol and Moschus are weighed in proportion, by various grind into powders respectively, and
Mix and obtain traditional Chinese medicine ingredients；
(3) preparation of dressing：
To obtain in the hair-keratin of the granular hair-keratin prepared in step (1) or gelation and step (2)
To traditional Chinese medicine ingredients mixing be uniformly dispersed, obtain composite auxiliary material.
5. the preparation method of through engineering approaches dressing according to claim 4, it is characterised in that：The hair-keratin is gel
The hair-keratin of change, the concentration of the hair-keratin of the gelation is 0.1～5%.
6. the preparation method of through engineering approaches dressing according to claim 5, it is characterised in that：The hair angle egg of the gelation
White concentration is 0.8～3%.
7. the preparation method of the compound dressing for being applied to the surface of a wound according to claim 4, it is characterised in that：Step (1)
S1 in, the degreasing and soften method be：Soaked 20～60 minutes with ethanol, at interval of 3～10 minutes in immersion process
Stirring once, is soaked and finishes the flushing of rear flowing water；After flushing, hair is positioned in the solution that sodium hypochlorite is prepared with water and soaks 20
～60 minutes, stirring in 3～10 minutes is spaced in the immersion process once, soak and finish the flushing of rear flowing water；
In the S2 of step (1), the hair after softening is positioned over 2-3 hour in de-inking solution, 10 are spaced in decolorization
Stirring in～30 minutes once, is soaked and finishes the flushing of rear flowing water, till washing de-inking solution off；The composition of the de-inking solution is burnt phosphorus
4 weight portion of sour sodium, 1.5 weight portion of potassium peroxydisulfate, 80 parts by volume of hydrogen peroxide, 27.5 parts by volume of concentrated ammonia liquor and 400 parts by volume of water；
In the S3 of step (1), the hair after decolouring is soaked in water, soak to finish and rinsed with water, filter, in air dry oven
In 37 DEG C dry overnight.
8. the preparation method of the compound dressing for being applied to the surface of a wound according to claim 7, it is characterised in that：Step (1)
S1 in, the volume ratio of the sodium hypochlorite and water is 1～5:9, the sodium hypochlorite is 10% sodium hypochlorite；
In the S2 of step (1), crinite de-inking solution will be placed and be placed in 37 DEG C of constant temperature and decolourized；The sodium pyrophosphate is
99% sodium pyrophosphate, potassium peroxydisulfate are 99.5% potassium peroxydisulfate, and hydrogen peroxide is 30% hydrogen peroxide, and concentrated ammonia liquor is 25% concentrated ammonia liquor.
9. the preparation method of the compound dressing for being applied to the surface of a wound according to claim 4, it is characterised in that：Step (1)
S4 in, ball milling：Meal 2g is placed in ball grinder, is put into agate bead, add physiological saline 10ml, ball milling 1～3 hour,
Basis of microscopic observation particle size, adds physiological saline 5ml and continues grinding, until forming about 60 μm -80 μm of particle diameter；The agate
The quantity of Nao pearl：Diameter 1cm agate bead 20, diameter 60mm agate bead 100；
In the S6 of step (1), the highly purified hair-keratin mud drum for obtaining dress is adopted cobalt60Irradiation method sterilizes, dosage 12Kg；
In the S7 of step (1), add water fully to mix in granular hair-keratin, be positioned over 37 DEG C of incubation 2-3 little
When.
10. the preparation method of through engineering approaches dressing according to claim 5, it is characterised in that：The hair-keratin gel
Preparation method can also be prepared by the following method：
(1) hair-keratin becomes salt：Hair is weighed, the mixed solution of second alcohol and water is added, heat, backflow, drip in backflow
Terminate when aqueous slkali is 6.5～7.5 to pH value, stirring then proceeded to, is made reaction completely, salt hair-keratin is filtrated to get into,
Dry, micron order is broken to, be lyophilized, that is, obtain hair-keratin sulfonic acid salt powder；
(2) preparation of hair-keratin gel：Hair-keratin sulfonic acid salt powder is weighed, water is added, is fully mixed, is positioned over temperature
It is incubated in case, sterilizing, obtains hair-keratin gel.
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| CN108744027A (en) * | 2018-07-04 | 2018-11-06 | 长春工业大学 | A kind of protein-based liquid adhesive bandage and preparation method thereof |
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