CN105381503A - Artificial skin with high activity - Google Patents
Artificial skin with high activity Download PDFInfo
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- CN105381503A CN105381503A CN201510915412.2A CN201510915412A CN105381503A CN 105381503 A CN105381503 A CN 105381503A CN 201510915412 A CN201510915412 A CN 201510915412A CN 105381503 A CN105381503 A CN 105381503A
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- China
- Prior art keywords
- nano chitosan
- lactic acid
- artificial skin
- temperature
- aminoacid
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- 230000000694 effects Effects 0.000 title description 3
- 229920001661 Chitosan Polymers 0.000 claims abstract description 42
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 22
- 229920001184 polypeptide Polymers 0.000 claims abstract description 20
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 20
- 238000000034 method Methods 0.000 claims abstract description 18
- 229920000642 polymer Polymers 0.000 claims abstract description 17
- 238000004519 manufacturing process Methods 0.000 claims abstract description 12
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 48
- 239000000463 material Substances 0.000 claims description 29
- 239000004310 lactic acid Substances 0.000 claims description 24
- 235000014655 lactic acid Nutrition 0.000 claims description 24
- 235000001014 amino acid Nutrition 0.000 claims description 16
- 150000001413 amino acids Chemical class 0.000 claims description 16
- 239000002775 capsule Substances 0.000 claims description 16
- 238000003756 stirring Methods 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 238000002360 preparation method Methods 0.000 claims description 13
- 238000002604 ultrasonography Methods 0.000 claims description 10
- 239000004094 surface-active agent Substances 0.000 claims description 9
- 229920002125 Sokalan® Polymers 0.000 claims description 8
- 230000002301 combined effect Effects 0.000 claims description 8
- 239000008367 deionised water Substances 0.000 claims description 8
- 229910021641 deionized water Inorganic materials 0.000 claims description 8
- 150000007524 organic acids Chemical class 0.000 claims description 8
- 239000004584 polyacrylic acid Substances 0.000 claims description 8
- 239000002994 raw material Substances 0.000 claims description 6
- 241000972773 Aulopiformes Species 0.000 claims description 4
- 230000009471 action Effects 0.000 claims description 4
- 235000019515 salmon Nutrition 0.000 claims description 4
- 241000894007 species Species 0.000 claims description 2
- 241000894006 Bacteria Species 0.000 abstract description 5
- 230000002401 inhibitory effect Effects 0.000 abstract description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 10
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 8
- 206010052428 Wound Diseases 0.000 description 7
- 208000027418 Wounds and injury Diseases 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- 239000010408 film Substances 0.000 description 5
- 230000002378 acidificating effect Effects 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 235000011187 glycerol Nutrition 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 208000028990 Skin injury Diseases 0.000 description 2
- 206010053615 Thermal burn Diseases 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 239000011230 binding agent Substances 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000035699 permeability Effects 0.000 description 2
- 229920000191 poly(N-vinyl pyrrolidone) Polymers 0.000 description 2
- 230000036558 skin tension Effects 0.000 description 2
- 230000029663 wound healing Effects 0.000 description 2
- 108090000526 Papain Proteins 0.000 description 1
- 241000233805 Phoenix Species 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 241000194017 Streptococcus Species 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 229910052785 arsenic Inorganic materials 0.000 description 1
- RQNWIZPPADIBDY-UHFFFAOYSA-N arsenic atom Chemical compound [As] RQNWIZPPADIBDY-UHFFFAOYSA-N 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000000589 cicatrix Anatomy 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000002500 effect on skin Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000003925 fat Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000002949 hemolytic effect Effects 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000005065 mining Methods 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000005789 organism growth Effects 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 229920002635 polyurethane Polymers 0.000 description 1
- 239000004814 polyurethane Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- QQONPFPTGQHPMA-UHFFFAOYSA-N propylene Natural products CC=C QQONPFPTGQHPMA-UHFFFAOYSA-N 0.000 description 1
- 125000004805 propylene group Chemical group [H]C([H])([H])C([H])([*:1])C([H])([H])[*:2] 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 238000001542 size-exclusion chromatography Methods 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000002352 surface water Substances 0.000 description 1
- 239000010409 thin film Substances 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
Classifications
-
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/227—Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
-
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/16—Macromolecular materials obtained by reactions only involving carbon-to-carbon unsaturated bonds
-
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/20—Polysaccharides
-
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
-
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/60—Materials for use in artificial skin
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- 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
- A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
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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
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/404—Biocides, antimicrobial agents, antiseptic 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
- 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
- A61L2300/416—Anti-neoplastic or anti-proliferative or anti-restenosis or anti-angiogenic agents, e.g. paclitaxel, sirolimus
-
- 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
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/34—Materials or treatment for tissue regeneration for soft tissue reconstruction
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Transplantation (AREA)
- Epidemiology (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Dermatology (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Materials For Medical Uses (AREA)
- Cosmetics (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses an artificial skin and a manufacturing method thereof. The artificial skin has the characteristics of good surface tension, plump and elastic texture, and bacterium inhibiting performance, and the tissue of the artificial skin is a polymer of nano chitosan and polypeptide. The artificial skin has the advantages of good biomedical characteristics, high medical value, short technical procedure, simple operation, less equipment, and reduced cost. The artificial skin has a good application prospect and application value.
Description
Technical field
The present invention relates to medical ancillary technique field, especially a kind of skin-grafting artificial skin.
Background technology
Artificial skin has medical application prospect widely in the treatment of burn, scald etc., therefore receives the extensive concern of Chinese scholars and deep research.The skin of burn patient faces tissue fluid and oozes out, infects series of problems such as (particularly charrin's diseases) in recovery process.At present, the high water vapor permeability thin film of many uses is as dressing clinically, comprises the Tegaderm of Minnesota Mining and Manufacturing Company, Britain OpSite etc., comprise polyurethane and add propylene type binding agent, the firm adhesion chaeta of these binding agents in these dressing, easily cause pain when taking off, discomfort is even damaged; And these dressing can not effectively solve problems such as tissue fluid is oozed out, infection.
In clinical treatment, be accelerating wound, promote body cell growth, skin injury need keep certain wetting state, namely requires that wound can not overdrying, again can not excessive moisture.And the artificial skin that several process programs of prior art are made over the course for the treatment of, due to the restriction of itself structure, build the environment of an applicable skin, Organism Growth often can not to skin injury, thus affect the treatment.Therefore, present clinical unit, in the practice for the treatment of burn, still has larger demand to artificial skin.
Summary of the invention
Object of the present invention is exactly the deficiency that will overcome the existence of above-mentioned prior art, one is provided to have some strength and elasticity, permeability is good, closely can attach with wound surface, can wound surface be observed again, aseptic, nontoxic, no antigen or antigenicity little, non-stimulated or zest is less to wound surface, and have stronger bacteria resistance function; Safe and convenient to use, wound surface can be protected not infected, do not hinder autologous skin and skin-grafting plate growth, wound healing; cicatrix is less, and raw material sources are extensive, can manufacture; cost is lower, preserves easily for a long time, again just with the manufacture method of the artificial skin of transport and this artificial skin.
The object of the invention is to: for treatment skin trauma in the urgent need to, for patient provides a kind of bacteria growing inhibiting, and there is better active artificial skin simultaneously.
Another object of the present invention is to provide the process of producing this artificial skin.
A manufacture method for artificial skin, measures by weight, adopts following proportioning raw material: Nano chitosan 120, polypeptide 5, lactic acid 5, organic acid 75, surfactant 30, and absolute viscosity is the polyacrylic acid 60 of 4764mpa.s, deionized water 950, And manufactures in the steps below: the first step, the preparation of A material: remove ionized water 950 weight portion, polyacrylic acid 60 weight portion that organic acid 75 weight portion, absolute viscosity are 4764mpa.s, be placed in reactor, 50 DEG C, dissolve 1.5 hours under 3000 turns/min stirring action, become the A material that pH value is 3.5-4.0, weight portion is 1085, second step, the preparation of B material: after A material is made, keep the temperature of A material in reactor at 50 DEG C and under 3000 turns/min, it stirred, gradually the Nano chitosan of 120 weight portions is added in A material with the time of 30min, until Nano chitosan is dissolved almost completely, become Nano chitosan solution, be called that B expects, 3rd step, after the preparation of the polymer capsules of Nano chitosan and aminoacid or lactic acid: B expects to make, by the B material in reactor under 3000 turns/min stirring and the combined effect of 40-100W ultrasound wave, in 1.5 hours, its temperature is heated to 65 DEG C gradually from 45 DEG C, then, add the surfactant of 30 weight portions again, continue under aforementioned stirring and ultrasound wave combined effect, add the lactic acid of 5 weight portions and the polypeptide (shown in SEQIDNO:1-15) of 5 weight portions gradually, do not stopping stirring and carrying out polyreaction 2 hours under ul-trasonic irradiation, then 1.5 hours are used by the temperature in reactor, 90 DEG C are risen to gradually from 65 DEG C, and keep polyreaction 1 hour at this temperature, namely polyreaction is accused and is finished, filtered while hot, namely the polymer capsules of light salmon Nano chitosan and aminoacid or lactic acid is obtained, 4th step, the polymer capsules of Nano chitosan and aminoacid or lactic acid is processed into artificial skin prepard from Nano chitosan: by the polymer capsules of Nano chitosan and aminoacid or lactic acid, neutrality is washed till with the deionized water of 40 DEG C, and shear film forming, then by this film at 50 DEG C of temperature dry 1.5 hours, namely artificial skin is obtained.
This nanometer artificial skin, has translucent film body, soaks have soft and full elastic form, biomedicine has the characteristic of good anti-bacteria and antitumor cell growth, have good skin smoothness and moist degree simultaneously in normal saline.
By dry nanometer artificial skin, strictly carry out disinfection on request, then seal, pack, put preservation in storage.
Surfactant used can be tween 20, peregal 0S-10, the efficient Uniform stain of peregal 0S-15, JFZ-1, JFZ penetrating agent or other fats polyvingl ether;
Nano chitosan used, for having the Nano chitosan product of following physical and chemical index: powder granule >80 order, powder body volume density is 0.56-0.62g/ml, powder body moisture San 5.6%, the granularity of Nano chitosan particle is 20-60nm, beavy metal impurity content: plumbous <0.lmg/kg, hydrargyrum <0.lmg/kg, arsenic <0.2mg/kg, dissolve in weak acid, diluted acid or six acetone, aseptic;
Ultrasound wave used in technique is the ultrasound wave of the 40W-100W that supersonic generator produces.
The present invention's Nano chitosan used can be 02133792.6 by application number, and the applying date is JIUYUE in 2002 23 days, and the method provided in the Chinese invention patent application file that name is called " a kind of manufacture method of nano-scale chitosan " creates.The present invention's other raw material used is commercially available prod.
The invention has the advantages that: due to this product artificial skin, there is the good biological medical characteristics of bacteria growing inhibiting and antitumor cell growth, maintain the moist degree of good skin and elasticity simultaneously, therefore the skin-grafting of burn, scald, ulcerous skin can be widely used in, wound hemostasis, antibacterial, antiinflammatory also promotes cell regeneration, accelerates wound healing, its rete is thick, good springiness, practical; And technological process is short, processing ease, saving equipment.
Detailed description of the invention
The acquisition of embodiment 1 polypeptide
The tissue extraction preparation method of polypeptide, comprises the following steps:
1) get sea dog cadaver skin, mince, PBS dissolves, and add papain and trypsin treatment 4h, the acidic ethanol being then placed in 4 DEG C digests 16h, and rear centrifugalize is got supernatant spin concentration, lyophilization, obtained lyophilized products;
2) size exclusion chromatography be separated: by step 1) obtained freeze-drying thing be placed in concentration be 2mol/L acetic acid process after, load in sephadex G 50 chromatographic column, take concentration as the acetic acid balance of 2mol/L, pillar flow velocity 10mL/h, collect chromatographic peak 60 component, lyophilizing, obtains described polypeptide;
Described acidic ethanol is the hydrochloric acid being 0.7mol/L by ethanol and concentration is the proportions of 4:1 by volume; The consumption of described acidic ethanol is: every g skin histology adds 8mL acidic ethanol;
Step 1) described in the temperature of centrifugalize be 4 DEG C, centrifugal force is 3000g, and the time is 30min.
Through detection, described polypeptide has found that 15 its sequences of polypeptide are as shown in SEQIDNO:1-15, have bacteria growing inhibiting and keep skin gloss and elastic effect.Therefore, may be used for the application of artificial skin.
SEQIDNO：1：KDQQSKCQITFMDLSHNHE；SEQIDNO：2：WRVMPMFWGLNWWSQKWQ；
SEQIDNO：3：TCPTHCEDNEWGKYESW；SEQIDNO：4：RKPHHPWTGSGGSYPE；
SEQIDNO：5：QWHWNHSFWSDTRH；SEQIDNO：6：KPKNDDSITETLH；
SEQIDNO：7：HMKDVDHHPGPPLNPAPMH；SEQIDNO：8：PMHKSHKIQMIVRWSCRC；
SEQIDNO：9：FNHFCTTEVFLQQFSRF；SEQIDNO：10：QDSWLRIQHAEETRHT；
SEQIDNO：11：VDQYWFPSLGLQRR；SEQIDNO：12：RVKWSVPHSISRQ；
SEQIDNO：13：PVEFKHGMSRFFWSIECIM；
SEQIDNO：14：LWPQVSHPSDPWCRRLCW；SEQIDNO：15：TMQIHSGTWCRMTIPTV。
The manufacture method of the artificial skin of embodiment 2 containing polypeptide
A manufacture method for artificial skin, measures by weight, adopts following proportioning raw material: Nano chitosan 120, polypeptide 5, lactic acid 5, organic acid 75, surfactant 30, and absolute viscosity is the polyacrylic acid 60 of 4764mpa.s, deionized water 950, And manufactures in the steps below: the first step, the preparation of A material: remove ionized water 950 weight portion, polyacrylic acid 60 weight portion that organic acid 75 weight portion, absolute viscosity are 4764mpa.s, be placed in reactor, 50 DEG C, dissolve 1.5 hours under 3000 turns/min stirring action, become the A material that pH value is 3.5-4.0, weight portion is 1085, second step, the preparation of B material: after A material is made, keep the temperature of A material in reactor at 50 DEG C and under 3000 turns/min, it stirred, gradually the Nano chitosan of 120 weight portions is added in A material with the time of 30min, until Nano chitosan is dissolved almost completely, become Nano chitosan solution, be called that B expects, 3rd step, after the preparation of the polymer capsules of Nano chitosan and aminoacid or lactic acid: B expects to make, by the B material in reactor under 3000 turns/min stirring and the combined effect of 40-100W ultrasound wave, in 1.5 hours, its temperature is heated to 65 DEG C gradually from 45 DEG C, then, add the surfactant of 30 weight portions again, continue under aforementioned stirring and ultrasound wave combined effect, add the lactic acid of 5 weight portions and a peptide species (one of SEQIDNO:1-15) of 5 weight portions gradually, do not stopping stirring and carrying out polyreaction 2 hours under ul-trasonic irradiation, then 1.5 hours are used by the temperature in reactor, 90 DEG C are risen to gradually from 65 DEG C, and keep polyreaction 1 hour at this temperature, namely polyreaction is accused and is finished, filtered while hot, namely the polymer capsules of light salmon Nano chitosan and aminoacid or lactic acid is obtained, 4th step, the polymer capsules of Nano chitosan and aminoacid or lactic acid is processed into artificial skin prepard from Nano chitosan: by the polymer capsules of Nano chitosan and aminoacid or lactic acid, neutrality is washed till with the deionized water of 40 DEG C, and shear film forming, then by this film at 50 DEG C of temperature dry 1.5 hours, namely artificial skin is obtained, totally 15 kinds.Wherein with do not add polypeptide for contrast.
The activity of embodiment 3 skin and Function detection
(1) skin surface moisture content scatters and disappears and tests
The artificial skin that Example 2 prepares, is respectively cut into 5cm diameter circular, is immersed in (glycerol is conventional wetting agent) in 10% glycerine water solution.Then remove, apply human body skin 30 minutes.Take off artificial skin, then after 15 minutes, with CKElectronic Multi-functional skin detector, MPA580 type (Cutometer) detection of skin surface water loses.Above-mentioned steps is carried out equally, as a control group (A) with the skin that exists together not being covered with any material.Result shows, and after spreading on skin with the artificial skin not adding polypeptide of embodiment 2, the moisture content of this place's skin speed of scattering and disappearing reduces 18.7%, and prepares with embodiment 2 its moisture loss speed of artificial skin that with the addition of polypeptide and reduce about about 80%.In other words, glycerol can be transferred to skin with the artificial skin that the embodiment 2 of the present invention with the addition of described in polypeptide, the moisture loss of skin be reduced, there is good skin moisture-keeping characteristic, skin can be prevented to greatest extent too dry.
(2) antibacterial skin detects
Artificial skin prepared by embodiment 2 is chosen 10cm respectively
2cultivate in human serum culture medium, do not have artificial skin as well with culture medium, simulation human body environment.Staphylococcus aureus, Hemolytic streptococcus and bacillus pyocyaneus are cultured to 5*10 according to the cultural method that this area is conventional
8cFU/mL, then adds respectively and gets 1 and drip to skin surface, cultivates 5 days for 37 degrees Celsius.Observed result is as follows:
Embodiment 4 skin tension is tested
Using male adult skin as positive control, with the addition of 15 kinds of skins of polypeptide as experimental subject in embodiment 2, there is no the skin adding many embodiments 2 as instruction, by the skin tension measuring method of this area routine, measure the surface tension of this skin in 17.Find, less than normal adult skin surface tension by about 10% with the skin not adding polypeptide in embodiment 2, and 15 kinds of its surface tension of skin that with the addition of polypeptide increase about about 20% than the surface tension of adult skin, show good skin effects.
Based on above artificial skin of the present invention, there is good application prospect.
Sequence table
< 110 > turns round and look at silver-colored phoenix
< 120 > highly active artificial skin
〈210〉1
〈211〉19
〈212〉PRT
< 213 > artificial sequence
KDQQSKCQITFMDLSHNHE；
〈210〉2
〈211〉18
〈212〉PRT
< 213 > artificial sequence
WRVMPMFWGLNWWSQKWQ；
〈210〉3
〈211〉17
〈212〉PRT
< 213 > artificial sequence
TCPTHCEDNEWGKYESW；
〈210〉4
〈211〉16
〈212〉PRT
< 213 > artificial sequence
RKPHHPWTGSGGSYPE；
〈210〉5
〈211〉14
〈212〉PRT
< 213 > artificial sequence
QWHWNHSFWSDTRH；
〈210〉6
〈211〉13
〈212〉PRT
< 213 > artificial sequence
KPKNDDSITETLH；
〈210〉7
〈211〉19
〈212〉PRT
< 213 > artificial sequence
HMKDVDHHPGPPLNPAPMH；
〈210〉8
〈211〉18
〈212〉PRT
< 213 > artificial sequence
PMHKSHKIQMIVRWSCRC；
〈210〉9
〈211〉17
〈212〉PRT
< 213 > artificial sequence
FNHFCTTEVFLQQFSRF；
〈210〉10
〈211〉16
〈212〉PRT
< 213 > artificial sequence
QDSWLRIQHAEETRHT；
〈210〉11
〈211〉14
〈212〉PRT
< 213 > artificial sequence
VDQYWFPSLGLQRR；
〈210〉12
〈211〉13
〈212〉PRT
< 213 > artificial sequence
RVKWSVPHSISRQ；
〈210〉13
〈211〉19
〈212〉PRT
< 213 > artificial sequence
PVEFKHGMSRFFWSIECIM；
〈210〉14
〈211〉18
〈212〉PRT
< 213 > artificial sequence
LWPQVSHPSDPWCRRLCW；
〈210〉15
〈211〉17
〈212〉PRT
< 213 > artificial sequence
TMQIHSGTWCRMTIPTV；
Claims (4)
1. a manufacture method for artificial skin, is characterized in that, measures by weight, adopts following proportioning raw material: Nano chitosan 120, polypeptide 5, lactic acid 5, organic acid 75, surfactant 30, absolute viscosity is the polyacrylic acid 60 of 4764mpa.s, deionized water 950, and manufacture in the steps below: the first step, the preparation of A material: remove ionized water 950 weight portion, polyacrylic acid 60 weight portion that organic acid 75 weight portion, absolute viscosity are 4764mpa.s, be placed in reactor, 50 DEG C, dissolve 1.5 hours under 3000 turns/min stirring action, become the A material that pH value is 3.5-4.0, weight portion is 1085, second step, the preparation of B material: after A material is made, keep the temperature of A material in reactor at 50 DEG C and under 3000 turns/min, it stirred, gradually the Nano chitosan of 120 weight portions is added in A material with the time of 30min, until Nano chitosan is dissolved almost completely, become Nano chitosan solution, be called that B expects, 3rd step, after the preparation of the polymer capsules of Nano chitosan and aminoacid or lactic acid: B expects to make, by the B material in reactor under 3000 turns/min stirring and the combined effect of 40-100W ultrasound wave, in 1.5 hours, its temperature is heated to 65 DEG C gradually from 45 DEG C, then, add the surfactant of 30 weight portions again, continue under aforementioned stirring and ultrasound wave combined effect, add the lactic acid of 5 weight portions gradually, do not stopping stirring and carrying out polyreaction 2 hours under ul-trasonic irradiation, then 1.5 hours are used by the temperature in reactor, 90 DEG C are risen to gradually from 65 DEG C, and keep polyreaction 1 hour at this temperature, namely polyreaction is accused and is finished, filtered while hot, namely the polymer capsules of light salmon Nano chitosan and aminoacid or lactic acid is obtained, 4th step, the polymer capsules of Nano chitosan and aminoacid or lactic acid is processed into artificial skin prepard from Nano chitosan: by the polymer capsules of Nano chitosan and aminoacid or lactic acid, neutrality is washed till with the deionized water of 40 DEG C, and shear film forming, then by this film at 50 DEG C of temperature dry 1.5 hours, namely artificial skin is obtained.
2. a manufacture method for artificial skin, is characterized in that, measures by weight, adopts following proportioning raw material: Nano chitosan 120, polypeptide 5, lactic acid 5, organic acid 75, surfactant 30, absolute viscosity is the polyacrylic acid 60 of 4764mpa.s, deionized water 950, and manufacture in the steps below: the first step, the preparation of A material: remove ionized water 950 weight portion, polyacrylic acid 60 weight portion that organic acid 75 weight portion, absolute viscosity are 4764mpa.s, be placed in reactor, 50 DEG C, dissolve 1.5 hours under 3000 turns/min stirring action, become the A material that pH value is 3.5-4.0, weight portion is 1085, second step, the preparation of B material: after A material is made, keep the temperature of A material in reactor at 50 DEG C and under 3000 turns/min, it stirred, gradually the Nano chitosan of 120 weight portions is added in A material with the time of 30min, until Nano chitosan is dissolved almost completely, become Nano chitosan solution, be called that B expects, 3rd step, after the preparation of the polymer capsules of Nano chitosan and aminoacid or lactic acid: B expects to make, by the B material in reactor under 3000 turns/min stirring and the combined effect of 40-100W ultrasound wave, in 1.5 hours, its temperature is heated to 65 DEG C gradually from 45 DEG C, then, add the surfactant of 30 weight portions again, continue under aforementioned stirring and ultrasound wave combined effect, one peptide species of the lactic acid and 5 weight portions that add 5 weight portions is gradually any one in SEQIDNO:1-15, do not stopping stirring and carrying out polyreaction 2 hours under ul-trasonic irradiation, then 1.5 hours are used by the temperature in reactor, 90 DEG C are risen to gradually from 65 DEG C, and keep polyreaction 1 hour at this temperature, namely polyreaction is accused and is finished, filtered while hot, namely the polymer capsules of light salmon Nano chitosan and aminoacid or lactic acid is obtained, 4th step, the polymer capsules of Nano chitosan and aminoacid or lactic acid is processed into artificial skin prepard from Nano chitosan: by the polymer capsules of Nano chitosan and aminoacid or lactic acid, neutrality is washed till with the deionized water of 40 DEG C, and shear film forming, then by this film at 50 DEG C of temperature dry 1.5 hours, namely artificial skin is obtained.
3. the artificial skin that method prepares as described in any one of claim 1-2.
4. a polypeptide for artificial skin, is characterized in that, sequence is for SEQIDNO:1-15 is shown in any one.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106421930A (en) * | 2016-11-03 | 2017-02-22 | 常州亚环环保科技有限公司 | Preparation method of nonirritating artificial skin |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1795935A (en) * | 2004-12-28 | 2006-07-05 | 广汉恒宇新材料有限公司 | Artificial skin prepard from Nano chitosan and preparing method |
| CN104971385A (en) * | 2015-08-04 | 2015-10-14 | 杨国林 | Artificial skin |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1795935A (en) * | 2004-12-28 | 2006-07-05 | 广汉恒宇新材料有限公司 | Artificial skin prepard from Nano chitosan and preparing method |
| CN104971385A (en) * | 2015-08-04 | 2015-10-14 | 杨国林 | Artificial skin |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106421930A (en) * | 2016-11-03 | 2017-02-22 | 常州亚环环保科技有限公司 | Preparation method of nonirritating artificial skin |
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