CN102580153A - Method for preparing allograft acellular dermal matrixes - Google Patents
Method for preparing allograft acellular dermal matrixes Download PDFInfo
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- CN102580153A CN102580153A CN2012100606718A CN201210060671A CN102580153A CN 102580153 A CN102580153 A CN 102580153A CN 2012100606718 A CN2012100606718 A CN 2012100606718A CN 201210060671 A CN201210060671 A CN 201210060671A CN 102580153 A CN102580153 A CN 102580153A
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Abstract
The invention disclose a method for preparing allograft acellular dermal matrixes, comprising the following step of processing animal skins into the allograft acellular dermal matrixes by using processes of preparation of segmented skin, viral inactivation, degreasing, decellularization, freeze-drying, cutting, molding, packaging and irradiation disinfection. The method for the preparing allograft acellular dermal matrixes has the advantages of short degreasing time, high efficiency, complete decellularization and small damage to natural structures of dermal extracellular matrixes.
Description
Technical field
The present invention relates to a kind of method for preparing acellular dermal matrix medical tissue sticking patch.
Background technology
Acellular dermal matrix medical tissue sticking patch is the organizational project project that on the tissue engineering basis, grows up, and purpose is exactly to utilize the extracellular matrix of corium to carry out tissue repair.The natural skin histology that has drawn from is handled through special physics and chemistry, and the composition that possibly cause the immune rejection after human body is implanted in the tissue is removed, and kept former three-dimensional bracket structure in a organized way simultaneously, so it has the regenerated effect of induced tissue.Not enough below existing acellular dermal matrix production technology exists: operation is complicated, and the cycle is longer, and efficient is lower; Cause the damage of substrate easily.Therefore, need provide a kind of new technical scheme to solve the problems referred to above.
Summary of the invention
The purpose of this invention is to provide a kind of with short production cycle, efficient is high, the method that prepare acellular dermal matrix medical tissue sticking patch light to hypodermal cell epimatrix natural structure infringement.
The technical scheme that the present invention adopts is:
A kind of method for preparing acellular dermal matrix medical tissue sticking patch, this tissue patch is prepared from Corii Sus domestica, Corii Bovis seu Bubali or Corii Caprae seu Ovis, may further comprise the steps:
A, preparation split-thickness skin graft technology: use dermatome that animal skin is processed the split-thickness skin graft of thickness as 0.1-2.0mm, split-thickness skin graft is processed difform split-thickness skin graft with card punch.
B, virus inactivation technology: the inactivation of virus solution that split-thickness skin graft is formed with the sodium chloride of the peracetic acid of mass concentration 0.1%-2.0% and mass concentration 1.0%-5.0% carried out deactivation 30-60 minute, and the mass ratio of skin graft and inactivation of virus solution is 1:4-7;
C, degreasing process: the mixed defatting agent of adopting mass concentration 1.0%-4.0% soda, mass concentration 1.0%-4.0% Caustic soda and mass concentration 0.1%-1.0% surfactant to form was carried out defat 30-60 minute; Said surfactant is a kind of in peregal, sodium dodecyl aminopropionitrile, fatty alcohol-polyoxyethylene ether, the QULA ketone, and the mass ratio of skin graft and mixed defatting agent is 1:4-7;
D, take off cell technology: the Cell sap that takes off that adopts mass concentration 0.1%-0.5% pancreatin, mass concentration 0.1%-0.3% activator, mass concentration 0.2%-0.5% degreasing agent to form took off cell 30-60 minute; Said activator is a mass concentration 0.1%-0.3% ammonium sulfate; Said degreasing agent is a mass concentration 0.2%-0.5% peregal; A kind of in the mass concentration 0.2%-0.5% sodium lauryl sulphate, skin graft is 1:4-7 with the mass ratio that takes off Cell sap;
E, lyophilizing, cutting, setting process: will be after take off the split-thickness skin graft for preparing after the cell technology, putting into the freezer dryer lyophilizing, according to the needs of clinical different indication; With acellular dermal matrix respectively cutting be shaped into: sheet type, mesh type, circular hole, particle type, mesh type is pricked skin mechanism by sheet type through mesh and is formed fully, the mesh aperture of mesh type is 0.1-0.5cm; Mesh density 1-5/c ㎡; Circular hole is formed through the mechanism of punching by sheet type fully, and the aperture of circular hole is 0.1-0.5cm, circular hole density 1-5/c ㎡; Particle type is prepared from through micropulverizer sheet type, and the diameter of microgranule is 0.01-0.3cm;
F, packing and irradiation sterilization technology: formed package, through gamma-ray irradiation sterilization is finished product.
Beneficial effect of the present invention: (1) b step adopts the inactivation of virus solution of peracetic acid and sodium chloride solution composition to carry out deactivation, has both reached the effect of inactivation of virus, has avoided the peracetic acid soln inactivation of viruses again and the problem of the split-thickness skin graft swelling that produces; (2) the mixed defatting agent of being made up of soda, Caustic soda, surfactant in the c step has that degreasing time is short, efficient is high, the advantage strong to the protective effect of skin graft organizational structure; (3) Cell sap that takes off that adopts enzyme, activator and degreasing agent to form in the d step takes off cell technology, has that the cell of taking off is thorough, the time is short, efficient is high, to the light advantage of hypodermal cell epimatrix natural structure infringement.
The specific embodiment
Following examples are intended to explain the present invention rather than to the bright further qualification of we.
Embodiment 1
A kind of method for preparing acellular dermal matrix medical tissue sticking patch, this tissue patch is prepared from Corii Sus domestica, Corii Bovis seu Bubali or Corii Caprae seu Ovis, may further comprise the steps:
A, preparation split-thickness skin graft technology: use dermatome that animal skin is processed the split-thickness skin graft of thickness as 0.1mm, split-thickness skin graft is processed difform split-thickness skin graft with card punch;
B, virus inactivation technology: the inactivation of virus solution that split-thickness skin graft is formed with the sodium chloride of the peracetic acid of mass concentration 0.1% and mass concentration 1.0% carried out deactivation 30 minutes, and the mass ratio of skin graft and inactivation of virus solution is 1:4;
C, degreasing process: the mixed defatting agent of adopting mass concentration 1.0% soda, mass concentration 1.0% Caustic soda and mass concentration 0.1% peregal to form was carried out defat 30 minutes, and the mass ratio of skin graft and mixed defatting agent is 1:4;
D, take off cell technology: the Cell sap that takes off that adopts mass concentration 0.1% pancreatin, mass concentration 0.1% activator ammonium sulfate, mass concentration 0.2% degreasing agent peregal to form took off cell 30 minutes, and skin graft is 1:4 with the mass ratio that takes off Cell sap;
E, lyophilizing, cutting, setting process: will be after take off the split-thickness skin graft for preparing after the cell technology, putting into the freezer dryer lyophilizing, according to the needs of clinical different indication; With acellular dermal matrix respectively cutting be shaped into: sheet type, mesh type, circular hole, particle type, mesh type is pricked skin mechanism by sheet type through mesh and is formed fully, the mesh aperture of mesh type is 0.1cm; 1/c of mesh density ㎡; Circular hole is formed through the mechanism of punching by sheet type fully, and the aperture of circular hole is 0.1cm, 1/c of circular hole density ㎡; Particle type is prepared from through micropulverizer sheet type, and the diameter of microgranule is 0.01cm;
F, packing and irradiation sterilization technology: formed package, through gamma-ray irradiation sterilization is finished product.
Embodiment 2
A kind of method for preparing acellular dermal matrix medical tissue sticking patch, this tissue patch is prepared from Corii Sus domestica, Corii Bovis seu Bubali or Corii Caprae seu Ovis, may further comprise the steps:
A, preparation split-thickness skin graft technology: use dermatome that animal skin is processed the split-thickness skin graft of thickness as 0.1mm, split-thickness skin graft is processed difform split-thickness skin graft with card punch;
B, virus inactivation technology: the inactivation of virus solution that split-thickness skin graft is formed with the sodium chloride of the peracetic acid of mass concentration 0.1% and mass concentration 1.0% carried out deactivation 30 minutes, and the mass ratio of skin graft and inactivation of virus solution is 1:4;
C, degreasing process: the mixed defatting agent of adopting mass concentration 1.0% soda, mass concentration 1.0% Caustic soda and mass concentration 0.1% peregal to form was carried out defat 30 minutes, and the mass ratio of skin graft and mixed defatting agent is 1:4;
D, take off cell technology: the Cell sap that takes off that adopts mass concentration 0.1% pancreatin, mass concentration 0.1% activator ammonium sulfate, mass concentration 0.2% degreasing agent sodium dodecyl aminopropionitrile to form took off cell 30 minutes, and skin graft is 1:4 with the mass ratio that takes off Cell sap;
E, lyophilizing, cutting, setting process: will be after take off the split-thickness skin graft for preparing after the cell technology, putting into the freezer dryer lyophilizing, according to the needs of clinical different indication; With acellular dermal matrix respectively cutting be shaped into: sheet type, mesh type, circular hole, particle type, mesh type is pricked skin mechanism by sheet type through mesh and is formed fully, the mesh aperture of mesh type is 0.1cm; 1/c of mesh density ㎡; Circular hole is formed through the mechanism of punching by sheet type fully, and the aperture of circular hole is 0.1cm, 1/c of circular hole density ㎡; Particle type is prepared from through micropulverizer sheet type, and the diameter of microgranule is 0.01cm;
F, packing and irradiation sterilization technology: formed package, through gamma-ray irradiation sterilization is finished product.
Embodiment 3
A kind of method for preparing acellular dermal matrix medical tissue sticking patch, this tissue patch is prepared from Corii Sus domestica, Corii Bovis seu Bubali or Corii Caprae seu Ovis, may further comprise the steps:
A, preparation split-thickness skin graft technology: use dermatome that animal skin is processed the split-thickness skin graft of thickness as 1.0mm, split-thickness skin graft is processed difform split-thickness skin graft with card punch;
B, virus inactivation technology: the inactivation of virus solution that split-thickness skin graft is formed with the sodium chloride of the peracetic acid of mass concentration 1.0% and mass concentration 3.0% carried out deactivation 45 minutes, and the mass ratio of skin graft and inactivation of virus solution is 1:5.5;
C, degreasing process: the mixed defatting agent of adopting mass concentration 2.5% soda, mass concentration 2.5% Caustic soda and mass concentration 0.5% sodium dodecyl aminopropionitrile to form was carried out defat 45 minutes, and the mass ratio of skin graft and mixed defatting agent is 1:5.5;
D, take off cell technology: the Cell sap that takes off that adopts mass concentration 0.3% pancreatin, mass concentration 0.2% activator ammonium sulfate, mass concentration 0.3% degreasing agent peregal to form took off cell 45 minutes, and skin graft is 1:5.5 with the mass ratio that takes off Cell sap;
E, lyophilizing, cutting, setting process: will be after take off the split-thickness skin graft for preparing after the cell technology, putting into the freezer dryer lyophilizing, according to the needs of clinical different indication; With acellular dermal matrix respectively cutting be shaped into: sheet type, mesh type, circular hole, particle type, mesh type is pricked skin mechanism by sheet type through mesh and is formed fully, the mesh aperture of mesh type is 0.3cm; 3/c of mesh density ㎡; Circular hole is formed through the mechanism of punching by sheet type fully, and the aperture of circular hole is 0.3cm, 3/c of circular hole density ㎡; Particle type is prepared from through micropulverizer sheet type, and the diameter of microgranule is 0.1cm;
F, packing and irradiation sterilization technology: formed package, through gamma-ray irradiation sterilization is finished product.
Embodiment 4
A kind of method for preparing acellular dermal matrix medical tissue sticking patch, this tissue patch is prepared from animal skin such as pig, cattle, sheep, may further comprise the steps:
A, preparation split-thickness skin graft technology: use dermatome that animal skin is processed the split-thickness skin graft of thickness as 1.0mm, split-thickness skin graft is processed difform split-thickness skin graft with card punch;
B, virus inactivation technology: the inactivation of virus solution that split-thickness skin graft is formed with the sodium chloride of the peracetic acid of mass concentration 1.0% and mass concentration 3.0% carried out deactivation 45 minutes, and the mass ratio of skin graft and inactivation of virus solution is 1:5.5;
C, degreasing process: the mixed defatting agent of adopting mass concentration 2.5% soda, mass concentration 2.5% Caustic soda and mass concentration 0.5% sodium dodecyl aminopropionitrile to form was carried out defat 45 minutes, and the mass ratio of skin graft and mixed defatting agent is 1:5.5;
D, take off cell technology: the Cell sap that takes off that adopts mass concentration 0.3% pancreatin, mass concentration 0.2% activator ammonium sulfate, mass concentration 0.3% degreasing agent sodium lauryl sulphate to form took off cell 45 minutes, and skin graft is 1:5.5 with the mass ratio that takes off Cell sap;
E, lyophilizing, cutting, setting process: will be after take off the split-thickness skin graft for preparing after the cell technology, putting into the freezer dryer lyophilizing, according to the needs of clinical different indication; With acellular dermal matrix respectively cutting be shaped into: sheet type, mesh type, circular hole, particle type, mesh type is pricked skin mechanism by sheet type through mesh and is formed fully, the mesh aperture of mesh type is 0.3cm; 3/c of mesh density ㎡; Circular hole is formed through the mechanism of punching by sheet type fully, and the aperture of circular hole is 0.3cm, 3/c of circular hole density ㎡; Particle type is prepared from through micropulverizer sheet type, and the diameter of microgranule is 0.1cm;
F, packing and irradiation sterilization technology: formed package, through gamma-ray irradiation sterilization is finished product.
Embodiment 5
A kind of method for preparing acellular dermal matrix medical tissue sticking patch, this tissue patch is prepared from Corii Sus domestica, Corii Bovis seu Bubali or Corii Caprae seu Ovis, may further comprise the steps:
A, preparation split-thickness skin graft technology: use dermatome that animal skin is processed the split-thickness skin graft of thickness as 2.0mm, split-thickness skin graft is processed difform split-thickness skin graft with card punch;
B, virus inactivation technology: the inactivation of virus solution that split-thickness skin graft is formed with the sodium chloride of the peracetic acid of mass concentration 2.0% and mass concentration 5.0% carried out deactivation 60 minutes, and the mass ratio of skin graft and inactivation of virus solution is 1:7;
C, degreasing process: the mixed defatting agent of adopting mass concentration 4.0% soda, mass concentration 4.0% Caustic soda and mass concentration 1.0% fatty alcohol-polyoxyethylene ether to form was carried out defat 60 minutes, and the mass ratio of skin graft and mixed defatting agent is 1:7;
D, take off cell technology: the Cell sap that takes off that adopts mass concentration 0.5% pancreatin, mass concentration 0.3% activator ammonium sulfate, mass concentration 0.5% degreasing agent peregal to form took off cell 60 minutes, and skin graft is 1:7 with the mass ratio that takes off Cell sap;
E, lyophilizing, cutting, setting process: will be after take off the split-thickness skin graft for preparing after the cell technology, putting into the freezer dryer lyophilizing, according to the needs of clinical different indication; With acellular dermal matrix respectively cutting be shaped into: sheet type, mesh type, circular hole, particle type, mesh type is pricked skin mechanism by sheet type through mesh and is formed fully, the mesh aperture of mesh type is 0.5cm; 5/c of mesh density ㎡; Circular hole is formed through the mechanism of punching by sheet type fully, and the aperture of circular hole is 0.5cm, 5/c of circular hole density ㎡; Particle type is prepared from through micropulverizer sheet type, and the diameter of microgranule is 0.3cm;
F, packing and irradiation sterilization technology: formed package, through gamma-ray irradiation sterilization is finished product.
Embodiment 6
A kind of method for preparing acellular dermal matrix medical tissue sticking patch, this tissue patch is prepared from animal skin such as pig, cattle, sheep, may further comprise the steps:
A, preparation split-thickness skin graft technology: use dermatome that animal skin is processed the split-thickness skin graft of thickness as 2.0mm, split-thickness skin graft is processed difform split-thickness skin graft with card punch;
B, virus inactivation technology: the inactivation of virus solution that split-thickness skin graft is formed with the sodium chloride of the peracetic acid of mass concentration 2.0% and mass concentration 5.0% carried out deactivation 60 minutes, and the mass ratio of skin graft and inactivation of virus solution is 1:7;
C, degreasing process: the mixed defatting agent of adopting mass concentration 4.0% soda, mass concentration 4.0% Caustic soda and mass concentration 1.0% fatty alcohol-polyoxyethylene ether to form was carried out defat 60 minutes, and the mass ratio of skin graft and mixed defatting agent is 1:7;
D, take off cell technology: the Cell sap that takes off that adopts mass concentration 0.5% pancreatin, mass concentration 0.3% activator ammonium sulfate, mass concentration 0.5% degreasing agent sodium lauryl sulphate to form took off cell 60 minutes, and skin graft is 1:7 with the mass ratio that takes off Cell sap;
E, lyophilizing, cutting, setting process: will be after take off the split-thickness skin graft for preparing after the cell technology, putting into the freezer dryer lyophilizing, according to the needs of clinical different indication; With acellular dermal matrix respectively cutting be shaped into: sheet type, mesh type, circular hole, particle type, mesh type is pricked skin mechanism by sheet type through mesh and is formed fully, the mesh aperture of mesh type is 0.5cm; 5/c of mesh density ㎡; Circular hole is formed through the mechanism of punching by sheet type fully, and the aperture of circular hole is 0.5cm, 5/c of circular hole density ㎡; Particle type is prepared from through micropulverizer sheet type, and the diameter of microgranule is 0.3cm;
F, packing and irradiation sterilization technology: formed package, through gamma-ray irradiation sterilization is finished product.
In the c step degreasing process, surfactant can also be QULA ketone.
Beneficial effect of the present invention: (1) b step adopts the inactivation of virus solution of peracetic acid and sodium chloride solution composition to carry out deactivation, has both reached the effect of inactivation of virus, has avoided the peracetic acid soln inactivation of viruses again and the problem of the split-thickness skin graft swelling that produces; (2) the mixed defatting agent of being made up of soda, Caustic soda, surfactant in the c step has that degreasing time is short, efficient is high, the advantage strong to the protective effect of skin graft organizational structure; (3) Cell sap that takes off that adopts enzyme, activator and degreasing agent to form in the d step takes off cell technology, has that the cell of taking off is thorough, the time is short, efficient is high, to the light advantage of hypodermal cell epimatrix natural structure infringement.
The application of this acellular dermal matrix medical tissue sticking patch, this acellular dermal matrix tissue patch supply medical institutions to be used for soft tissue defects and weak repairing and treating that a variety of causes causes, and for example: 1, the wound surface that causes of a variety of causes covers treatment; 2, the repairing and treating that corium is damaged and sub-dermal soft tissue is damaged that causes of a variety of causes; 3, the repairing and treating of the hernia that causes of a variety of causes; 4, the repairing and treating of the dura defect that causes of a variety of causes; 5, the mucosa and the damaged repairing and treating of submucous tissue that cause of a variety of causes; 6, the soft tissue defects that causes of a variety of causes and weak shaping and beauty repairing and treating; 7, the repairing and treating of the nerve that causes of a variety of causes, tendon, blood vessel, adhesion; 8, repairing and treating of the urinary tract damage that causes of a variety of causes or the like.
Claims (1)
1. method for preparing acellular dermal matrix medical tissue sticking patch, this tissue patch is prepared from Corii Sus domestica, Corii Bovis seu Bubali or Corii Caprae seu Ovis, it is characterized in that, may further comprise the steps:
A, preparation split-thickness skin graft technology: use dermatome that animal skin is processed the split-thickness skin graft of thickness as 0.1-2.0mm, split-thickness skin graft is processed difform split-thickness skin graft with card punch;
B, virus inactivation technology: the inactivation of virus solution that split-thickness skin graft is formed with the sodium chloride of the peracetic acid of mass concentration 0.1%-2.0% and mass concentration 1.0%-5.0% carried out deactivation 30-60 minute, and the mass ratio of skin graft and inactivation of virus solution is 1:4-7;
C, degreasing process: the mixed defatting agent of adopting mass concentration 1.0%-4.0% soda, mass concentration 1.0%-4.0% Caustic soda and mass concentration 0.1%-1.0% surfactant to form was carried out defat 30-60 minute; Said surfactant is a kind of in peregal, sodium dodecyl aminopropionitrile, fatty alcohol-polyoxyethylene ether, the QULA ketone, and the mass ratio of skin graft and mixed defatting agent is 1:4-7;
D, take off cell technology: the Cell sap that takes off that adopts mass concentration 0.1%-0.5% pancreatin, mass concentration 0.1%-0.3% activator, mass concentration 0.2%-0.5% degreasing agent to form took off cell 30-60 minute; Said activator is a mass concentration 0.1%-0.3% ammonium sulfate; Said degreasing agent is a mass concentration 0.2%-0.5% peregal; A kind of in the mass concentration 0.2%-0.5% sodium lauryl sulphate, skin graft is 1:4-7 with the mass ratio that takes off Cell sap;
E, lyophilizing, cutting, setting process: will be after take off the split-thickness skin graft for preparing after the cell technology, putting into the freezer dryer lyophilizing, according to the needs of clinical different indication; With acellular dermal matrix respectively cutting be shaped into: sheet type, mesh type, circular hole, particle type, mesh type is pricked skin mechanism by sheet type through mesh and is formed fully, the mesh aperture of mesh type is 0.1-0.5cm; Mesh density 1-5/c ㎡; Circular hole is formed through the mechanism of punching by sheet type fully, and the aperture of circular hole is 0.1-0.5cm, circular hole density 1-5/c ㎡; Particle type is prepared from through micropulverizer sheet type, and the diameter of microgranule is 0.01-0.3cm;
F, packing and irradiation sterilization technology: formed package, through gamma-ray irradiation sterilization is finished product.
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