CN102580153B - Method for preparing allograft acellular dermal matrixes - Google Patents
Method for preparing allograft acellular dermal matrixes Download PDFInfo
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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 allograft acellular dermal matrixes.
Background technology
Allograft acellular dermal matrixes is the organizational project project that grows up on the tissue engineering basis, 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 processed through special physics and chemistry, and the composition that may cause the immune rejection after human body is implanted in tissue is removed, and kept simultaneously former organized three-dimensional bracket structure, so it has the effect of induced tissue regeneration.Existing acellular dermal matrix production technology has the following disadvantages: operate more complicatedly, the cycle is longer, and efficient is lower; Easily cause the damage of substrate.Therefore, need to 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 allograft acellular dermal matrixes light to hypodermal cell epimatrix natural structure infringement.
The technical solution used in the present invention is:
A kind of method for preparing allograft acellular dermal matrixes, this tissue patch is prepared from by Corii Sus domestica, Corii Bovis seu Bubali or Corii Caprae seu Ovis, comprises the following steps:
A, preparation split-thickness skin graft technique: with dermatome, animal skin is made the split-thickness skin graft that thickness is 0.1-2.0mm, with card punch, split-thickness skin graft is made difform split-thickness skin graft.
B, virus inactivation technology: split-thickness skin graft was carried out deactivation 30-60 minute with the inactivation of virus solution that the sodium chloride of the peracetic acid of mass concentration 0.1%-2.0% and mass concentration 1.0%-5.0% forms, 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, described surfactant is a kind of in peregal, sodium dodecyl aminopropionitrile, fatty alcohol-polyoxyethylene ether, QULA ketone, and the mass ratio of skin graft and mixed defatting agent is 1:4-7;
D, take off cell technique: 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, described activator is mass concentration 0.1%-0.3% ammonium sulfate, described degreasing agent is mass concentration 0.2%-0.5% peregal, a kind of in 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 through taking off the split-thickness skin graft for preparing after cell technique, after putting into the freezer dryer lyophilizing, needs according to 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 for forming by sheet type through mesh, the mesh aperture of mesh type is 0.1-0.5cm, mesh density 1-5/c ㎡, circular hole is machine-processed for forming through punching by sheet type, the aperture of circular hole is 0.1-0.5cm, circular hole density 1-5/c ㎡, particle type is prepared from through micropulverizer by sheet type, the diameter of microgranule is 0.01-0.3cm,
F, packing and irradiation sterilization technique: formed package, through gamma-ray irradiation sterilization is finished product.
Beneficial effect of the present invention: the inactivation of virus solution that (1) b step adopts peracetic acid and sodium chloride solution to form carries out deactivation, has both reached the effect of inactivation of virus, has avoided again the peracetic acid soln inactivation of viruses and the problem of the split-thickness skin graft swelling that produces; (2) the mixed defatting agent that is comprised of soda, Caustic soda, surfactant in the c step has advantages of that degreasing time is short, efficient is high, 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 technique, 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 illustrate the present invention rather than to the bright further restriction of we.
Embodiment 1
A kind of method for preparing allograft acellular dermal matrixes, this tissue patch is prepared from by Corii Sus domestica, Corii Bovis seu Bubali or Corii Caprae seu Ovis, comprises the following steps:
A, preparation split-thickness skin graft technique: with dermatome, animal skin is made the split-thickness skin graft that thickness is 0.1mm, with card punch, split-thickness skin graft is made difform split-thickness skin graft;
B, virus inactivation technology: split-thickness skin graft was carried out deactivation 30 minutes with the inactivation of virus solution that the sodium chloride of the peracetic acid of mass concentration 0.1% and mass concentration 1.0% forms, 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 technique: 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 through taking off the split-thickness skin graft for preparing after cell technique, after putting into the freezer dryer lyophilizing, needs according to 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 for forming by sheet type through mesh, the mesh aperture of mesh type is 0.1cm, 1/c of mesh density ㎡, circular hole is machine-processed for forming through punching by sheet type, the aperture of circular hole is 0.1cm, 1/c of circular hole density ㎡, particle type is prepared from through micropulverizer by sheet type, the diameter of microgranule is 0.01cm,
F, packing and irradiation sterilization technique: formed package, through gamma-ray irradiation sterilization is finished product.
Embodiment 2
A kind of method for preparing allograft acellular dermal matrixes, this tissue patch is prepared from by Corii Sus domestica, Corii Bovis seu Bubali or Corii Caprae seu Ovis, comprises the following steps:
A, preparation split-thickness skin graft technique: with dermatome, animal skin is made the split-thickness skin graft that thickness is 0.1mm, with card punch, split-thickness skin graft is made difform split-thickness skin graft;
B, virus inactivation technology: split-thickness skin graft was carried out deactivation 30 minutes with the inactivation of virus solution that the sodium chloride of the peracetic acid of mass concentration 0.1% and mass concentration 1.0% forms, 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 technique: 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 through taking off the split-thickness skin graft for preparing after cell technique, after putting into the freezer dryer lyophilizing, needs according to 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 for forming by sheet type through mesh, the mesh aperture of mesh type is 0.1cm, 1/c of mesh density ㎡, circular hole is machine-processed for forming through punching by sheet type, the aperture of circular hole is 0.1cm, 1/c of circular hole density ㎡, particle type is prepared from through micropulverizer by sheet type, the diameter of microgranule is 0.01cm,
F, packing and irradiation sterilization technique: formed package, through gamma-ray irradiation sterilization is finished product.
Embodiment 3
A kind of method for preparing allograft acellular dermal matrixes, this tissue patch is prepared from by Corii Sus domestica, Corii Bovis seu Bubali or Corii Caprae seu Ovis, comprises the following steps:
A, preparation split-thickness skin graft technique: with dermatome, animal skin is made the split-thickness skin graft that thickness is 1.0mm, with card punch, split-thickness skin graft is made difform split-thickness skin graft;
B, virus inactivation technology: split-thickness skin graft was carried out deactivation 45 minutes with the inactivation of virus solution that the sodium chloride of the peracetic acid of mass concentration 1.0% and mass concentration 3.0% forms, 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 technique: 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 through taking off the split-thickness skin graft for preparing after cell technique, after putting into the freezer dryer lyophilizing, needs according to 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 for forming by sheet type through mesh, the mesh aperture of mesh type is 0.3cm, 3/c of mesh density ㎡, circular hole is machine-processed for forming through punching by sheet type, the aperture of circular hole is 0.3cm, 3/c of circular hole density ㎡, particle type is prepared from through micropulverizer by sheet type, the diameter of microgranule is 0.1cm,
F, packing and irradiation sterilization technique: formed package, through gamma-ray irradiation sterilization is finished product.
Embodiment 4
A kind of method for preparing allograft acellular dermal matrixes, this tissue patch is prepared from by animal skin such as pig, cattle, sheep, comprises the following steps:
A, preparation split-thickness skin graft technique: with dermatome, animal skin is made the split-thickness skin graft that thickness is 1.0mm, with card punch, split-thickness skin graft is made difform split-thickness skin graft;
B, virus inactivation technology: split-thickness skin graft was carried out deactivation 45 minutes with the inactivation of virus solution that the sodium chloride of the peracetic acid of mass concentration 1.0% and mass concentration 3.0% forms, 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 technique: 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 through taking off the split-thickness skin graft for preparing after cell technique, after putting into the freezer dryer lyophilizing, needs according to 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 for forming by sheet type through mesh, the mesh aperture of mesh type is 0.3cm, 3/c of mesh density ㎡, circular hole is machine-processed for forming through punching by sheet type, the aperture of circular hole is 0.3cm, 3/c of circular hole density ㎡, particle type is prepared from through micropulverizer by sheet type, the diameter of microgranule is 0.1cm,
F, packing and irradiation sterilization technique: formed package, through gamma-ray irradiation sterilization is finished product.
Embodiment 5
A kind of method for preparing allograft acellular dermal matrixes, this tissue patch is prepared from by Corii Sus domestica, Corii Bovis seu Bubali or Corii Caprae seu Ovis, comprises the following steps:
A, preparation split-thickness skin graft technique: with dermatome, animal skin is made the split-thickness skin graft that thickness is 2.0mm, with card punch, split-thickness skin graft is made difform split-thickness skin graft;
B, virus inactivation technology: split-thickness skin graft was carried out deactivation 60 minutes with the inactivation of virus solution that the sodium chloride of the peracetic acid of mass concentration 2.0% and mass concentration 5.0% forms, 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 technique: 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 through taking off the split-thickness skin graft for preparing after cell technique, after putting into the freezer dryer lyophilizing, needs according to 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 for forming by sheet type through mesh, the mesh aperture of mesh type is 0.5cm, 5/c of mesh density ㎡, circular hole is machine-processed for forming through punching by sheet type, the aperture of circular hole is 0.5cm, 5/c of circular hole density ㎡, particle type is prepared from through micropulverizer by sheet type, the diameter of microgranule is 0.3cm,
F, packing and irradiation sterilization technique: formed package, through gamma-ray irradiation sterilization is finished product.
Embodiment 6
A kind of method for preparing allograft acellular dermal matrixes, this tissue patch is prepared from by animal skin such as pig, cattle, sheep, comprises the following steps:
A, preparation split-thickness skin graft technique: with dermatome, animal skin is made the split-thickness skin graft that thickness is 2.0mm, with card punch, split-thickness skin graft is made difform split-thickness skin graft;
B, virus inactivation technology: split-thickness skin graft was carried out deactivation 60 minutes with the inactivation of virus solution that the sodium chloride of the peracetic acid of mass concentration 2.0% and mass concentration 5.0% forms, 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 technique: 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 through taking off the split-thickness skin graft for preparing after cell technique, after putting into the freezer dryer lyophilizing, needs according to 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 for forming by sheet type through mesh, the mesh aperture of mesh type is 0.5cm, 5/c of mesh density ㎡, circular hole is machine-processed for forming through punching by sheet type, the aperture of circular hole is 0.5cm, 5/c of circular hole density ㎡, particle type is prepared from through micropulverizer by sheet type, the diameter of microgranule is 0.3cm,
F, packing and irradiation sterilization technique: formed package, through gamma-ray irradiation sterilization is finished product.
In c step degreasing process, surfactant can also be QULA ketone.
Beneficial effect of the present invention: the inactivation of virus solution that (1) b step adopts peracetic acid and sodium chloride solution to form carries out deactivation, has both reached the effect of inactivation of virus, has avoided again the peracetic acid soln inactivation of viruses and the problem of the split-thickness skin graft swelling that produces; (2) the mixed defatting agent that is comprised of soda, Caustic soda, surfactant in the c step has advantages of that degreasing time is short, efficient is high, 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 technique, 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 allograft acellular dermal matrixes, this acellular dermal matrix tissue patch is used for for medical institutions soft tissue defects and the 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 Dermal defect and the damaged repairing and treating of sub-dermal soft tissue that cause 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 injury that causes of a variety of causes etc.
Claims (1)
1. method for preparing allograft acellular dermal matrixes, this tissue patch is prepared from by Corii Sus domestica, Corii Bovis seu Bubali or Corii Caprae seu Ovis, it is characterized in that, comprises the following steps:
A, preparation split-thickness skin graft technique: with dermatome, animal skin is made the split-thickness skin graft that thickness is 0.1mm-2.0mm, with card punch, split-thickness skin graft is made difform split-thickness skin graft;
B, virus inactivation technology: split-thickness skin graft was carried out deactivation 30 minutes-60 minutes with the inactivation of virus solution that the sodium chloride of the peracetic acid of mass concentration 0.1%-2.0% and mass concentration 1.0%-5.0% forms, 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 minutes-60 minutes, described surfactant is a kind of in peregal, QULA ketone, and the mass ratio of skin graft and mixed defatting agent is 1:4-7;
D, take off cell technique: 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 minutes-60 minutes, described activator is mass concentration 0.1%-0.3% ammonium sulfate, described degreasing agent is mass concentration 0.2%-0.5% peregal, a kind of in 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 through taking off the split-thickness skin graft for preparing after cell technique, after putting into the freezer dryer lyophilizing, needs according to clinical different indication, with acellular dermal matrix respectively cutting be shaped into: sheet type, mesh type, circular hole, particle type, circular hole is pricked skin mechanism for forming by sheet type through mesh, the mesh aperture of mesh type is 0.1 cm-0.5cm, ㎡-5/c ㎡ of 1/c of mesh density, circular hole is machine-processed for forming through punching by sheet type, the aperture of circular hole is 0.1-0.5cm, ㎡-5/c ㎡ of 1/c of circular hole density, particle type is prepared from through micropulverizer by sheet type, the diameter of microgranule is 0.01cm-0.3cm,
F, packing and irradiation sterilization technique: formed package, through gamma-ray irradiation sterilization is finished product.
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| 复合表面活性剂处理生物源性骨组织作为骨移植材料的生物安全性研究;陈磊等;《中国医药生物技术》;20090810;第4卷(第4期);第270页左栏第2段、右栏第5段1.2方法、第275页左栏第1段 * |
| 陈磊等.复合表面活性剂处理生物源性骨组织作为骨移植材料的生物安全性研究.《中国医药生物技术》.2009,第4卷(第4期),第270页左栏第2段、右栏第5段1.2方法、第275页左栏第1段. |
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