CN106310352A - Preparation method of antibacterial acellular dermal matrix dressing material - Google Patents
Preparation method of antibacterial acellular dermal matrix dressing material Download PDFInfo
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- CN106310352A CN106310352A CN201610934055.9A CN201610934055A CN106310352A CN 106310352 A CN106310352 A CN 106310352A CN 201610934055 A CN201610934055 A CN 201610934055A CN 106310352 A CN106310352 A CN 106310352A
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- 230000002500 effect on skin Effects 0.000 title claims abstract description 94
- 239000011159 matrix material Substances 0.000 title claims abstract description 87
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 230000000844 anti-bacterial effect Effects 0.000 title abstract description 7
- 239000000463 material Substances 0.000 title abstract description 5
- 230000001954 sterilising effect Effects 0.000 claims abstract description 30
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 30
- 229920001661 Chitosan Polymers 0.000 claims abstract description 19
- 238000010257 thawing Methods 0.000 claims abstract description 19
- 238000005553 drilling Methods 0.000 claims abstract description 17
- 238000004108 freeze drying Methods 0.000 claims abstract description 10
- 238000004806 packaging method and process Methods 0.000 claims abstract description 10
- 238000002791 soaking Methods 0.000 claims abstract description 8
- 241000124008 Mammalia Species 0.000 claims abstract description 5
- 230000003115 biocidal effect Effects 0.000 claims description 31
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 24
- 238000000034 method Methods 0.000 claims description 22
- 230000008569 process Effects 0.000 claims description 19
- 230000009514 concussion Effects 0.000 claims description 11
- 238000003860 storage Methods 0.000 claims description 11
- 239000000872 buffer Substances 0.000 claims description 8
- 238000004140 cleaning Methods 0.000 claims description 8
- 238000000465 moulding Methods 0.000 claims description 8
- 238000002604 ultrasonography Methods 0.000 claims description 8
- 239000003945 anionic surfactant Substances 0.000 claims description 7
- BTBJBAZGXNKLQC-UHFFFAOYSA-N ammonium lauryl sulfate Chemical compound [NH4+].CCCCCCCCCCCCOS([O-])(=O)=O BTBJBAZGXNKLQC-UHFFFAOYSA-N 0.000 claims description 4
- 229940063953 ammonium lauryl sulfate Drugs 0.000 claims description 4
- 238000005516 engineering process Methods 0.000 claims description 4
- 239000004094 surface-active agent Substances 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 3
- QZRMKGRPCTVXBS-UHFFFAOYSA-N 2,6,8-trimethylnonan-4-yl dihydrogen phosphate Chemical compound CC(C)CC(C)CC(CC(C)C)OP(O)(O)=O QZRMKGRPCTVXBS-UHFFFAOYSA-N 0.000 claims description 2
- 241001465754 Metazoa Species 0.000 claims description 2
- -1 fatty alcohol sodium isethionate Chemical class 0.000 claims description 2
- 229940045998 sodium isethionate Drugs 0.000 claims description 2
- 239000003513 alkali Substances 0.000 claims 2
- GNFTZDOKVXKIBK-UHFFFAOYSA-N 3-(2-methoxyethoxy)benzohydrazide Chemical compound COCCOC1=CC=CC(C(=O)NN)=C1 GNFTZDOKVXKIBK-UHFFFAOYSA-N 0.000 claims 1
- FGUUSXIOTUKUDN-IBGZPJMESA-N C1(=CC=CC=C1)N1C2=C(NC([C@H](C1)NC=1OC(=NN=1)C1=CC=CC=C1)=O)C=CC=C2 Chemical compound C1(=CC=CC=C1)N1C2=C(NC([C@H](C1)NC=1OC(=NN=1)C1=CC=CC=C1)=O)C=CC=C2 FGUUSXIOTUKUDN-IBGZPJMESA-N 0.000 claims 1
- 230000003213 activating effect Effects 0.000 claims 1
- 238000010309 melting process Methods 0.000 claims 1
- 102000008186 Collagen Human genes 0.000 abstract description 2
- 108010035532 Collagen Proteins 0.000 abstract description 2
- 239000012620 biological material Substances 0.000 abstract description 2
- 229920001436 collagen Polymers 0.000 abstract description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 230000010355 oscillation Effects 0.000 abstract description 2
- 238000005238 degreasing Methods 0.000 abstract 1
- 210000003491 skin Anatomy 0.000 description 39
- 238000012360 testing method Methods 0.000 description 8
- 230000005855 radiation Effects 0.000 description 6
- 238000003325 tomography Methods 0.000 description 6
- 230000035939 shock Effects 0.000 description 5
- 239000000758 substrate Substances 0.000 description 5
- 206010052428 Wound Diseases 0.000 description 4
- 208000027418 Wounds and injury Diseases 0.000 description 4
- 230000029663 wound healing Effects 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 2
- 210000003850 cellular structure Anatomy 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 210000002615 epidermis Anatomy 0.000 description 2
- 238000004080 punching Methods 0.000 description 2
- 238000009864 tensile test Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241000222122 Candida albicans Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 208000028990 Skin injury Diseases 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 229940095731 candida albicans Drugs 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000003431 cross linking reagent Substances 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 210000003725 endotheliocyte Anatomy 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 239000000834 fixative Substances 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 208000026278 immune system disease Diseases 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 239000010985 leather Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 235000013622 meat product Nutrition 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 206010033675 panniculitis Diseases 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000008591 skin barrier function Effects 0.000 description 1
- 231100000075 skin burn Toxicity 0.000 description 1
- 230000036560 skin regeneration Effects 0.000 description 1
- 230000036548 skin texture Effects 0.000 description 1
- 210000004304 subcutaneous tissue Anatomy 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
Classifications
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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
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/40—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing ingredients of undetermined constitution or reaction products thereof, e.g. plant or animal extracts
-
- 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
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
- A61L15/28—Polysaccharides or their derivatives
-
- 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
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
-
- 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
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
- A61L15/425—Porous materials, e.g. foams or sponges
-
- 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
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
- A61L15/46—Deodorants or malodour counteractants, e.g. to inhibit the formation of ammonia or bacteria
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Animal Behavior & Ethology (AREA)
- Hematology (AREA)
- Public Health (AREA)
- Materials Engineering (AREA)
- Dispersion Chemistry (AREA)
- Botany (AREA)
- Chemical Kinetics & Catalysis (AREA)
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- Materials For Medical Uses (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention belongs to the technical field of biological materials of tissue engineering, and particularly relates to a preparation method of an antibacterial acellular dermal matrix dressing material. The preparation method comprises the following steps: selecting a healthy mammal, stripping the skin of the mammal, removing hair, preparing split-thickness skin, performing laser drilling, performing low temperature freeze thawing, performing ultrasonic oscillation, performing degreasing treatment, performing acellular treatment, soaking water-soluble chitosan, performing freeze drying, packaging and performing irradiation sterilization, thus obtaining the antibacterial acellular dermal matrix dressing material. The antibacterial acellular dermal matrix dressing material disclosed by the invention has a collagen three-dimensional entire structure, is thorough in decellularization and short in manufacture cycle, and has the characteristic of antibacterial function.
Description
Technical field
The present invention relates to technical field of biological materials, be specifically related to the preparation of a kind of antibiotic property acellular dermal matrix dressing
Method.
Background technology
Epidermis, corium, subcutaneous tissue together constitute the skin texture of human body, and skin is as organ most important in human body
One of, cover at human body surface, there is defense function, perceptive function, immunologic function, absorption function, regulation body temp. function, excretion
The function such as function and secretion.When human body skin sustains damage, particularly skin burn, burnt degree easily causes the various damage of body
Evil, such as immune system disorder, moisture and protein and other cell component excessively lost, metabolism aggravation etc., burn is tight
Heavy goes back entail dangers to life.Therefore, with wound dressing, wound surface must be covered after wound, play skin barrier protective effect temporarily,
And energy wound healing, large-area burns are even more important by this.
Acellular dermal matrix dressing is exactly a kind of dermal substitute that may be used for full thickness dermal and for burning
The medical dressing of the skin injury wound healings such as wound, it is that natural skin is in a series of process eliminate epidermis and corium
Cell component, but remain dermal extracellular matrix composition and its three-D space structure, such structure can be skin regeneration
There is provided " dermal template ", after inducing transplantation host cell contents such as fibroblast, endotheliocyte etc. break up in supporting structure,
Growth.But acellular dermal matrix (ADM) dressing that traditional method prepares exists, and poor permeability, anti-microbial property be low, bio-compatible
Property the shortcoming such as difference, this severely limits acellular dermal matrix dressing extensive application clinically.
Summary of the invention
The present invention is directed to disadvantages mentioned above present in prior art, propose a kind of acellular dermal prepared and there is antibiotic property
The preparation method of substrate dressing, its feature is that using healthy mammal skin is raw material, sequentially passes through rejecting hair, preparation
Split-thickness skin graft, laser punching, sterilization, low temperature freeze thawing, supersonic oscillations, ungrease treatment, de-cell process, soak water solublity
Antibiotic property acellular dermal matrix dressing is obtained after chitosan, lyophilization, packaging and irradiation sterilization.There is collagen three dimensional structure
Completely, de-cell is thorough, and fabrication cycle is short and has the feature of antibacterial functions.
The preparation method of above-mentioned a kind of antibiotic property acellular dermal matrix dressing provided by the present invention, including following step
Rapid:
The mammal skin peeled off carries out rejecting the process of hair；
Cutisector is utilized the animal skin handled well to be made thickness for 0.5mm ~ 1.5mm split-thickness skin graft；
Using laser drilling that split-thickness skin graft carries out drilling, gained hole is uniformly distributed, and its mesoporous 0.1mm ~
0.5mm, pitch-row 1.5mm ~ 2.5mm；
Use irradiation sterilization technology the dermal matrix of apertures to be carried out disinfection sterilizing, wherein 60Coradiation dosage be 25kGy ~
50kGy, exposure time is 10h ~ 15h；
Dermal matrix after sterilization is placed in precooling 2 ~ 3h in the ultra cold storage freezer of-30 DEG C ~-50 DEG C, then-70
DEG C ~ ultra cold storage freezer of-80 DEG C in take out after precooling 2 ~ 3h, be placed in defrosting 1h in 37 DEG C of water-baths, 2 ~ 4 times repeatedly；
The dermal matrix of freeze thawing treatment is added in buffer and carry out concussion with ultrasound wave and clean, wherein ultrasonic wave concussion
The frequency processed is 3 ~ 7 times, each 30 minutes；
It is 0.8% ~ 2.0% NaOH solution by mass concentration and mass concentration is 0.1% ~ 0.6% surfactant mixed liquor pair
Dermal matrix after cleaning carries out ungrease treatment；
It is that 0.2% ~ 3.0% TritonX-100 carries out de-cell process to the dermal matrix after defat by concentration；
Acellular dermal matrix is soaked in the water-soluble chitosan solution that concentration is 0.5mg/ml ~ 2.5mg/ml；
The above-mentioned acellular dermal matrix soaking chitosan solution is put into freezer dryer and carries out lyophilization, molding
Packaging, the most irradiated sterilization, antibiotic property acellular dermal matrix dressing finished product.
It is an advantage of the current invention that:
It is thorough that obtained antibiotic property acellular dermal matrix dressing takes off cell, and antigenicity is low；
The micropore dermal matrix that is uniformly distributed utilizing laser punching technology to be obtained is beneficial to de-cell, defat and soaks water
Soluble chitosan subsequent treatment；
Preparation process does not uses fixative, cross-linking agent etc. so that the acellular dermal matrix prepared retains complete net
Network space structure；
The gradient precooling program taked in freezing dry process ensures that the acellular dermal matrix mechanical property obtained is good
And cyberspace structure can well be maintained；
Ungrease treatment uses anion surfactant, can preferably reach to slough the purpose of fat；
The compound chitosan with antibacterial functions in acellular dermal matrix, not only increases acellular dermal matrix and applies
Material antibiotic property and also can strengthen acellular dermal matrix anthemorrhagic performance and promote wound healing performance.
Detailed description of the invention
Embodiment one
Peel off healthy neonatal pig skin, reject hair；With cutisector, the fresh porcine skin handled well being made thickness is 0.8mm tomography
Skin graft；With laser drilling, split-thickness skin graft carried out drilling, aperture 0.3mm, pitch-row 2mm；With Co 60, split-thickness skin graft is entered again
Row radiation sterilization, dosage is 30kGy, and exposure time is 12h；Dermal matrix after sterilization is placed in-40 DEG C super
Precooling 3h in cryogenic refrigerator, then take out after precooling 3h in the ultra cold storage freezer of-75 DEG C, it is placed in 37 DEG C of water-baths defrosting
1h, multigelation 3 times；The dermal matrix of freeze thawing treatment is added in buffer and carry out concussion with ultrasound wave and clean, Qi Zhongchao
The frequency that sound wave shock processes is 5 times, each 30 minutes；It is 1.0% NaOH solution by mass concentration again and mass concentration is
0.4% ammonium lauryl sulfate anion surfactant mixed liquor carries out ungrease treatment to the dermal matrix after cleaning；Again with dense
Degree is that 1.0% TritonX-100 carries out de-cell process to the dermal matrix after defat；Again acellular dermal matrix is soaked in
Concentration is in the water-soluble chitosan solution of 1.5mg/ml；Finally by the above-mentioned acellular dermal matrix soaking chitosan solution
Put into freezer dryer and carry out lyophilization, molding packaging, the most irradiated sterilization, antibiotic property acellular dermal matrix
Dressing finished product.
Embodiment two
Peel off healthy neonatal pig skin, reject hair；With cutisector, the fresh porcine skin handled well being made thickness is 0.5mm tomography
Skin graft；With laser drilling, split-thickness skin graft carried out drilling, aperture 0.1mm, pitch-row 1.5mm；Again with Co 60 to split-thickness skin graft
Carrying out radiation sterilization, dosage is 25kGy, and exposure time is 10h；Dermal matrix after sterilization is placed in-30 DEG C
Precooling 2h in ultra cold storage freezer, then take out after precooling 2h in the ultra cold storage freezer of-70 DEG C, it is placed in 37 DEG C of water-baths defrosting
1h, multigelation 2 times；The dermal matrix of freeze thawing treatment is added in buffer and carry out concussion with ultrasound wave and clean, Qi Zhongchao
The frequency that sound wave shock processes is 3 times, each 30 minutes；It is 1.0% NaOH solution by mass concentration again and mass concentration is
0.1% fatty alcohol sodium isethionate anion surfactant mixed liquor carries out ungrease treatment to the dermal matrix after cleaning；Again
It is that 0.2% TritonX-100 carries out de-cell process to the dermal matrix after defat by concentration；Again acellular dermal matrix is soaked
Steep in the water-soluble chitosan solution that concentration is 0.5mg/ml；Finally by the above-mentioned acellular dermal soaking chitosan solution
Substrate is put into freezer dryer and is carried out lyophilization, molding packaging, the most irradiated sterilization, antibiotic property acellular dermal
Substrate dressing finished product.
Embodiment three
Peel off healthy neonatal pig skin, reject hair；With cutisector, the fresh porcine skin handled well being made thickness is 1.5mm tomography
Skin graft；With laser drilling, split-thickness skin graft carried out drilling, aperture 0.5mm, pitch-row 2.5mm；Again with Co 60 to split-thickness skin graft
Carrying out radiation sterilization, dosage is 50kGy, and exposure time is 15h；Dermal matrix after sterilization is placed in-50 DEG C
Precooling 3h in ultra cold storage freezer, then take out after precooling 3h in the ultra cold storage freezer of-80 DEG C, it is placed in 37 DEG C of water-baths defrosting
1h, multigelation 4 times；The dermal matrix of freeze thawing treatment is added in buffer and carry out concussion with ultrasound wave and clean, Qi Zhongchao
The frequency that sound wave shock processes is 7 times, each 30 minutes；It is 2.0% NaOH solution by mass concentration again and mass concentration is
0.6% 1-isobutyl-3,5-dimethylhexylphosphoric acid anion surfactant mixed liquor carries out ungrease treatment to the dermal matrix after cleaning；Again with dense
Degree is that 3.0% TritonX-100 carries out de-cell process to the dermal matrix after defat；Again acellular dermal matrix is soaked in
Concentration is in the water-soluble chitosan solution of 2.5mg/ml；Finally by the above-mentioned acellular dermal matrix soaking chitosan solution
Put into freezer dryer and carry out lyophilization, molding packaging, the most irradiated sterilization, antibiotic property acellular dermal matrix
Dressing finished product.
Reference examples one
Peel off healthy neonatal pig skin, reject hair；With cutisector, the fresh porcine skin handled well being made thickness is 0.8mm tomography
Skin graft；With laser drilling, split-thickness skin graft carried out drilling, aperture 0.3mm, pitch-row 2mm；With Co 60, split-thickness skin graft is entered again
Row radiation sterilization, dosage is 30kGy, and exposure time is 12h；Dermal matrix after sterilization is placed in-75 DEG C super
Cryogenic refrigerator takes out after precooling 3h, is placed in defrosting 1h in 37 DEG C of water-baths, multigelation 3 times；By the corium base of freeze thawing treatment
Matter adds in buffer and carries out concussion with ultrasound wave cleans, and the frequency that wherein ultrasonic wave concussion processes is 5 times, each 30 points
Clock；It is 1.0% NaOH solution by mass concentration again and mass concentration is 0.4% ammonium lauryl sulfate anion surfactant
Mixed liquor carries out ungrease treatment to the dermal matrix after cleaning；It is that 1.0% TritonX-100 is to the corium after defat by concentration again
Substrate carries out de-cell and processes；Acellular dermal matrix is soaked in the water-soluble chitosan solution that concentration is 1.5mg/ml again
In；Finally the above-mentioned acellular dermal matrix soaking chitosan solution is put into freezer dryer and carry out lyophilization, molding
Packaging, the most irradiated sterilization, antibiotic property acellular dermal matrix dressing finished product.
Reference examples two
Peel off healthy neonatal pig skin, reject hair；With cutisector, the fresh porcine skin handled well being made thickness is 0.8mm tomography
Skin graft；With laser drilling, split-thickness skin graft carried out drilling, aperture 0.3mm, pitch-row 2mm；With Co 60, split-thickness skin graft is entered again
Row radiation sterilization, dosage is 30kGy, and exposure time is 12h；Dermal matrix after sterilization is placed in-40 DEG C super
Precooling 3h in cryogenic refrigerator, then take out after precooling 3h in the ultra cold storage freezer of-75 DEG C, it is placed in 37 DEG C of water-baths defrosting
1h, multigelation 3 times；The dermal matrix of freeze thawing treatment is added in buffer and carry out concussion with ultrasound wave and clean, Qi Zhongchao
The frequency that sound wave shock processes is 5 times, each 30 minutes；It is 1.0% NaOH solution by mass concentration again and mass concentration is
0.4% quaternary cationics mixed liquor carries out ungrease treatment to the dermal matrix after cleaning；It is 1.0% by concentration again
TritonX-100 carries out de-cell and processes the dermal matrix after defat；Acellular dermal matrix is soaked in concentration again is
In the water-soluble chitosan solution of 1.5mg/ml；Finally the above-mentioned acellular dermal matrix soaking chitosan solution is put into cold
Lyophilizer carries out lyophilization, molding packaging, the most irradiated sterilization, antibiotic property acellular dermal matrix dressing become
Product.
Reference examples three
Peel off healthy neonatal pig skin, reject hair；With cutisector, the fresh porcine skin handled well being made thickness is 0.8mm tomography
Skin graft；With laser drilling, split-thickness skin graft carried out drilling, aperture 0.3mm, pitch-row 2mm；With Co 60, split-thickness skin graft is entered again
Row radiation sterilization, dosage is 30kGy, and exposure time is 12h；Dermal matrix after sterilization is placed in-40 DEG C super
Precooling 3h in cryogenic refrigerator, then take out after precooling 3h in the ultra cold storage freezer of-75 DEG C, it is placed in 37 DEG C of water-baths defrosting
1h, multigelation 3 times；The dermal matrix of freeze thawing treatment is added in buffer and carry out concussion with ultrasound wave and clean, Qi Zhongchao
The frequency that sound wave shock processes is 5 times, each 30 minutes；It is 1.0% NaOH solution by mass concentration again and mass concentration is
0.4% ammonium lauryl sulfate anion surfactant mixed liquor carries out ungrease treatment to the dermal matrix after cleaning；Again with dense
Degree is that 1.0% TritonX-100 carries out de-cell process to the dermal matrix after defat；Again acellular dermal matrix is put into cold
Lyophilizer carries out lyophilization, molding packaging, the most irradiated sterilization, acellular dermal matrix dressing finished product.
Embodiment four: Mechanics Performance Testing
The antibiotic property acellular dermal matrix dressing that Example one and reference examples one obtain carries out Mechanics Performance Testing, by QB/T
2710-2005 " leather physics and mechanical test tensile strength and the mensuration of percentage elongation " detects, by tensile testing machine
Distance between lower clamp is adjusted to 50mm(standard specimen) or 100mm(large size sample), clamp sample with fixture, it is ensured that sample
Grain at grade, and measures distance L between upper lower clamp0(accurately to 0.5mm), start tensile testing machine, until
Sample fracture, and maximal force when recording fracture is as the distance between fixture when disruptive force F and sample fracture, as sample
Length L during fracture1.The hot strength of sample, is wherein the width of sample, for the thickness of sample.The elongation at break of sample.
Result such as following table:
Embodiment five: fat content is tested
The antibiotic property acellular dermal matrix dressing that Example two and reference examples two obtain carries out fat content test, by GB/T
The method that 9695.7-2008 " meat measures with meat products total lipid content " specifies detects, result such as following table:
Embodiment six: antibiotic property is tested
The acellular dermal matrix dressing that Example three and reference examples three obtain carries out antibiotic property test, and concrete grammar is as follows:
Antibiotic property acellular dermal matrix dressing embodiment three obtained is cut into 2.0cm × 2.0cm size, is placed in aseptic taper
As print group in Ping, and substrate dressing reference examples three obtained as stated above is as comparison print group.2 groups are added respectively
Enter 70mL phosphate buffer (0.03mol L-1) and 5mL staphylococcus aureus (or Candida albicans or escherichia coli)
Bacterium solution, after liquid penetrates in culture medium, cultivates 12-24 hour, carries out colony counting for 37 DEG C.
Bacteriostasis rate computing formula:, wherein X is bacteriostasis rate, and A is average colony number before test agent vibration, and B is test agent vibration
Rear average colony number.Result such as following table:
Knowable to the test result analysis of embodiment 4 ~ 6, antibiotic property acellular dermal matrix dressing of the present invention compared with matched group,
Its mechanical property antibiotic property sloughed thoroughly and there is excellence good, fatty.
It should be pointed out that, the detailed description of the invention more representational example that is the present invention, it is clear that the skill of the present invention
Art scheme is not limited to above-described embodiment, it is also possible to have many variations.Those of ordinary skill in the art, the most public with present invention institute
What what that open or according to file written description was undoubted obtained, all it is considered as this patent scope of the claimed.
Claims (10)
1. the preparation method of an antibiotic property acellular dermal matrix dressing, it is characterised in that comprise the following steps:
A., the mammal skin peeled off carries out rejecting the process of hair；
B. utilize cutisector that the animal skin handled well is made certain thickness split-thickness skin graft；
C. using laser drilling that split-thickness skin graft carries out drilling, gained hole is uniformly distributed；
D. irradiation sterilization technology is used the dermal matrix of apertures to be carried out disinfection sterilizing；
E. the dermal matrix after sterilization is placed under low temperature and carries out cold melting process；
F. the dermal matrix of freeze thawing treatment is added in buffer and carry out concussion with ultrasound wave and clean；
G. with alkali liquor and surfactant mixed liquor, the dermal matrix after cleaning is carried out ungrease treatment；
H. with TritonX-100, the dermal matrix after defat is carried out de-cell to process；
I. acellular dermal matrix is soaked in water-soluble chitosan solution；
J. the above-mentioned acellular dermal matrix soaking chitosan solution is put into freezer dryer and carry out lyophilization, molding
Packaging, the most irradiated sterilization, antibiotic property acellular dermal matrix dressing finished product.
The preparation method of antibiotic property acellular dermal matrix dressing the most according to claim 1, it is characterised in that step b
Described split-thickness skin graft includes epidermal area and skin corium, and the thickness of split-thickness skin graft is 0.5mm ~ 1.5mm.
The preparation method of antibiotic property acellular dermal matrix dressing the most according to claim 1, it is characterised in that step c
The aperture in the hole that described laser prepares is 0.1mm ~ 0.5mm, and pitch-row is 1.5mm ~ 2.5mm.
The preparation method of antibiotic property acellular dermal matrix dressing the most according to claim 1, it is characterised in that step d
Described irradiation sterilization be 60Coradiation dosage be 25kGy ~ 50kGy, exposure time is 10h ~ 15h.
The preparation method of antibiotic property acellular dermal matrix dressing the most according to claim 1, it is characterised in that step e
Described freeze thawing treatment includes dermal matrix is placed in precooling 2 ~ 3h in the ultra cold storage freezer of-30 DEG C ~-50 DEG C, then at-70 DEG C
The ultra cold storage freezer of ~-80 DEG C takes out after precooling 2 ~ 3h, is placed in defrosting 1h in 37 DEG C of water-baths, 2 ~ 4 times repeatedly.
The preparation method of antibiotic property acellular dermal matrix dressing the most according to claim 1, it is characterised in that step f
The frequency that described ultrasonic wave concussion processes is 3 ~ 7 times, each 30 minutes.
The preparation method of antibiotic property acellular dermal matrix dressing the most according to claim 1, it is characterised in that step g
Described alkali liquor and surfactant mixed liquor be 0.8% ~ 2.0% NaOH solution by mass concentration and mass concentration be 0.1% ~
0.6% surfactant composition.
The preparation method of antibiotic property acellular dermal matrix dressing the most according to claim 7, it is characterised in that described table
Face activating agent is in ammonium lauryl sulfate, fatty alcohol sodium isethionate and 1-isobutyl-3,5-dimethylhexylphosphoric acid anion surfactant
One.
The preparation method of antibiotic property acellular dermal matrix dressing the most according to claim 1, it is characterised in that step h
The mass concentration of described TritonX-100 is 0.2% ~ 3.0%.
The preparation method of antibiotic property acellular dermal matrix dressing the most according to claim 1, it is characterised in that step i
The concentration of described water-soluble chitosan is 0.5mg/ml ~ 2.5mg/ml.
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| CN111569151B (en) * | 2020-05-12 | 2021-12-17 | 上海亚朋生物技术有限公司 | Acellular dermal matrix tissue engineering scaffold and preparation method thereof |
| CN114272429A (en) * | 2021-12-16 | 2022-04-05 | 成都汉丁新材料科技有限公司 | Fetal bovine dermal matrix dressing and preparation method thereof |
| CN115068661A (en) * | 2022-06-21 | 2022-09-20 | 杭州倍荣生物科技有限公司 | A kind of calcium alginate composite porous biological matrix dressing, its preparation method and application |
| CN114949359A (en) * | 2022-06-27 | 2022-08-30 | 西安德诺海思医疗科技有限公司 | Acellular matrix particle filler and preparation method thereof |
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