CN109364299A - A kind of basin bottom Biological Repair mesh sheet and preparation method thereof - Google Patents
A kind of basin bottom Biological Repair mesh sheet and preparation method thereof Download PDFInfo
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- CN109364299A CN109364299A CN201811436014.2A CN201811436014A CN109364299A CN 109364299 A CN109364299 A CN 109364299A CN 201811436014 A CN201811436014 A CN 201811436014A CN 109364299 A CN109364299 A CN 109364299A
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- basin bottom
- mesh sheet
- bottom biological
- aqueous solution
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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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
- A61L27/3633—Extracellular matrix [ECM]
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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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3691—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
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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
- 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
-
- 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
- A61L27/58—Materials at least partially resorbable by the body
-
- 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
-
- 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
- A61L2300/41—Anti-inflammatory agents, e.g. NSAIDs
-
- 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
- A61L2300/412—Tissue-regenerating or healing or proliferative 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
- A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
- A61L2300/602—Type of release, e.g. controlled, sustained, slow
- A61L2300/604—Biodegradation
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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
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/34—Materials or treatment for tissue regeneration for soft tissue reconstruction
Abstract
The present invention discloses a kind of basin bottom Biological Repair mesh sheet and preparation method thereof, and using animal tissue as raw material, the immunizing compositions such as removal cell, DNA are twisted line after being cut into thin strips shape, are woven into sticking patch the basin bottom biological sticking patch.De- cell material obtains bigger intensity at line by twisting to less be easily broken off, and because there is intertexture between line and line after weaving, the external force for stretching tearing is not acting on a point but on multiple intertwined points, therefore there is good mechanical property, it solves the disadvantage that de- cell material basin bottom biological sticking patch mechanical strength difference and needs introducing crosslinked agent or synthetic material, and the structure of the mesh sheet is made by weaving, the size in mesh sheet aperture can be easy to adjust by braided parameter, meet the needs of practical application to reach.Meanwhile the basin bottom biological sticking patch remains the ingredients such as original three-dimensional structure, non-collagen and growth factor in extracellular matrix, promotes the functional reconstruction and postoperation recovery of tissue.
Description
Technical field
The present invention relates to technical field of biological material, and in particular to a kind of basin bottom Biological Repair mesh sheet and preparation method thereof.
Background technique
Female pelvic dysfunction is mainly shown as pelvic organ prolapse and stress incontinence etc., it has been reported that 50%
Multipara it is different degrees of there are problems that pelvic floor dysfunction.Surgical operation is the more serious pelvic floor dysfunction for the treatment of
Main method, operation method are mainly the neoplasty and suspension of various tissues and physiological structure.It generally requires to be implanted into benefit simultaneously
The materials such as piece are to enhance operative effect.No-station pole canopy basin bottom neoplasty is the chief surgical mode at current basin bottom.With medical life
The fast development of object material, various basin bottoms patching material are widely used in clinic.
Basin bottom sticking patch is broadly divided into synthetic material sticking patch and biomaterial sticking patch two major classes.
1. synthetic material sticking patch
The made basin bottom sticking patch main component of synthetic material is heteropolymer, including two class of degradable and non-degradable.It is non-can
Degradation-type synthetic material sticking patch tensile strength is good, but because that cannot can not be organically combined, be set with body tissue by tissue degradation
Easily cause postoperative local organization inflammation and infection after entering, the structural intergrity after retaining in vivo for a long time is destroyed, transportable
To its hetero-organization, cause chronic inflammation and foreign body reaction, needs to carry out revision procedure；Secondly because in vivo with surrounding tissue power
The difference for learning compliance, can gradually cause the fiberization of surrounding tissue and gradually form fibrous capsule, by prolonged
It uses, erosion increased risk of the product for adjacent tissue organ.
As technology develops, degradable synthetic material sticking patch has also been derived, and degradable synthetic material sticking patch, one
As be that acute inflammatory reaction although mechanical property is fine, is caused using the synthesis such as polylactic acid after implanting, it is followed by chronic
Inflammation ultimately forms granulation tissue, fiber package；And the high concentration cream that the materials such as polylactic acid are partially formed in degradation process
Acid, hydroxyacetic acid easily cause cytotoxicity.
Synthetic material mechanical strength is preferable, but biological property is poor, and is unable to inducing tissue regeneration and healing.Therefore synthesis is mended
Piece developing direction is imitation biochemistry, i.e., using different weaving methods and addition natural biologic material such as collagen, fiber egg
White etc., simulated tissue characteristic to enhance healing potential, and then improves the success rate of basin bottom prosthesis.
2. biomaterial sticking patch
Biomaterial sticking patch can be divided into allogeneic and take off cellular material and the de- cell tissue of xenogenesis mostly from organization material
Material etc..
Allogenic material is mostly derived from the product of human skin dermal tissue, although they, which have, promotes basin bottom neoplasty
The ability of organization healing afterwards, but there are sources to lack, infectivity disease (such as AIDS) risk, so using by certain
Limitation.
Xenogenesis takes off cell material and is mainly derived from the tissue such as corium, small intestine, bladder, pericardium of animal, is by thin
The processing of the immunizing compositions such as born of the same parents, DNA, thus obtain remain in extracellular matrix original three-dimensional structure and collagenous fibres at
The material divided.The ingredients such as three-dimensional structure, collagen, non-collagen and growth factor in extracellular matrix are host cell
Stick, be proliferated, the environment for providing and adapting to is provided, facilitating the functional reconstruction of tissue, and then organize after promoting basin bottom neoplasty
Healing.
Cell material sticking patch is taken off in practical applications because of its own mechanical strength deficiency, needs to introduce ring in its process
The chemical cross-linking agents such as oxide or glutaraldehyde, have had the disadvantage in that potential cytotoxicity, and degradation rate is slower, with tissue
Regeneration mismatches, and can lead to the reaction such as fibrosis, chronic inflammation.Grant number is the Chinese invention patent of CN103800942, pelvis
Bottom patch is that non-degradable and degradable synthetic material are prepared into sticking patch by technologies such as electrostatic spinnings；Application No. is
201610678065.0 Chinese invention patent only keeps multilayer tight although using biomaterial prepares sticking patch with fixture
Closely connected conjunction, or each layer is firmly fit together with adhesive or suture, without inherently increasing biomaterial
Mechanical strength.
For this purpose, on the basis of not introducing synthetic material or crosslinking agent, in order to enhance the mechanics of basin bottom biology patching material
Intensity, the present invention come into being.
Summary of the invention
It is an object of the invention to be directed to shortcoming of the biological sticking patch in the prior art in structural strength, one is provided
Kind basin bottom Biological Repair mesh sheet.
For this purpose, basin bottom Biological Repair mesh sheet provided by the invention, wherein the mesh sheet is compiled by xenogenesis acellular matrix material
It knits.
Further, the xenogenesis acellular matrix material by xenogenesis acellular matrix by cleaning slitting, inactivation of virus,
Degreasing, removes DNA and removes α-Gal antigen, sizing, is lyophilized de- cell.
Further, the acellular matrix includes but is not limited to the submucous layer of small intestine of mammal, mucous membrane of urinary bladder
One or more combinations of lower layer, submucous lamina of stomach, dermal matrix, pericardium, meninx, amnion, internal organs film, peritonaeum.
Further, the acellular matrix is the submucous layer of small intestine of animal.
The present invention also provides a kind of preparation methods of basin bottom Biological Repair mesh sheet, comprising:
(1) it pre-processes,
The animal tissue's cleaning for taking fresh slaughtered animals is placed in acetum immersion, strikes off the mucous membrane for removing animal tissue
Layer, muscle layer, placenta percreta, lymph node isolate submucosa, longitudinal to be uniformly cut into thin strips shape, and are rinsed with purified water, obtain basin
Bottom biological material for repairing；
(2) inactivation of virus,
The mixed aqueous solution that basin bottom biological material for repairing is used to Peracetic acid and ethyl alcohol impregnates under room temperature in ultrasound, into
Row inactivation of virus is simultaneously cleaned by ultrasonic with purified water；
(3) degreasing,
Using ethanol solution, impregnates under ultrasound, normal temperature condition, be cleaned by ultrasonic later using water for injection；
(4) cell is taken off, DNA is removed and removes α-Gal antigen,
Using containing trypsase and containing the mixed aqueous solution of EDTA, impregnates under ultrasound condition, be cleaned by ultrasonic later using PBS；
Using the aqueous solution of the enzyme containing DNA, impregnated under ultrasound condition；Later using PBS drift ultrasonic cleaning；
Using the aqueous solution containing alpha-galactosidase, impregnated under ultrasound condition；It is cleaned by ultrasonic later using PBS；
Using NaOH aqueous solution, under ultrasound condition, PBS ultrasonic cleaning is used after soak at room temperature until neutral；
(5) it is formed, is lyophilized, braiding, sterilizing
By treated, fine strip shape submucosa is twisted line, is fixed on mold, after freeze-drying, is woven into the basin bottom of mesh sheet shape
Biological Repair mesh sheet, PET packaging bag carry out irradiation sterilization after packing.
Further, in pre-treatment step, the concentration of acetic acid is 0.01%-0.5%, soaking time 10-120min, is moved
The ratio of object tissue and acetum is 1:2-1:10.
Further, in viral inaction steps, the concentration of Peracetic acid is 0.5-1.5%, and the concentration of ethyl alcohol is 15-
25%, the ratio of basin bottom biological material for repairing and mixed aqueous solution is 1:2-1:10, soaking time 30-120min.
Further, in defatting step, the concentration of ethyl alcohol is 90-100%, the ratio of basin bottom biological material for repairing and ethyl alcohol
Example is 1:2-1:10, and the soak at room temperature time is 0.5-12h.
Further, in de- cell, remove DNA and go in α-Gal antigen step:
The content of trypsase and EDTA are respectively 0.01-0.10% and 0.01- 0.05% in mixed aqueous solution, and basin bottom biology is repaired
Material and trypsase/EDTA solution ratio are mended as 1:2-1:10, under ultrasound condition, impregnates 15- under the conditions of 36 ± 2 DEG C
40min；
In aqueous solution containing DNA enzymatic the content of DNA enzymatic be 0.05-10U/ml, basin bottom biological material for repairing with containing the water-soluble of DNA enzymatic
The ratio of liquid is 1:2-1:10, and soaking temperature is 36 ± 2 DEG C, soaking time 15-40min；
The content of alpha-galactosidase is 0.05-10U/ml, basin bottom biological material for repairing in aqueous solution containing alpha-galactosidase
Ratio with alpha-galactoside enzyme solutions is 1:2-1:10, and soaking temperature is 20-37 DEG C, soaking time 15-40min；
The concentration of NaOH aqueous solution is 5-40mM, and the ratio of basin bottom biological material for repairing and NaOH solution is 1:5-1:50, room temperature
Soaking time is 20-60min.
Compared with prior art, the present invention have following remarkable advantage and the utility model has the advantages that
The technical problem to be solved by the present invention is to be directed to the shortcoming of basin bottom biological sticking patch, a kind of New type basin bottom biology is provided
Repair mesh.
1. basin bottom Biological Repair mesh sheet provided by the invention is organized with corium, small intestine, pericardium of animal etc. as raw material,
The immunizing compositions such as cell, DNA are removed, line is twisted after being cut into thin strips shape, is woven into mesh sheet.Xenogenesis take off cell material by twisting at
Line obtains bigger mechanical strength to less be easily broken off, and is woven into after the sticking patch of mesh sheet shape because between line and line
There is intertexture, the external force for stretching tearing is not acting on a point but therefore has good mechanical property on multiple intertwined points
Can, solve the disadvantage that xenogenesis takes off cell material basin bottom biological sticking patch mechanical strength difference, and the structure of the mesh sheet is by weaving
Technique is made, and the size in mesh sheet aperture can be easy to adjust by braided parameter, meet the needs of practical application to reach.
2. basin bottom Biological Repair mesh sheet provided by the invention remains original three-dimensional structure, collagen in extracellular matrix
The ingredients such as fiber, non-collagen and growth factor have promoting healing effect, accelerate functional reconstruction and the repairing of basin bottom of tendon
Postoperative healing.
3. basin bottom Biological Repair mesh sheet provided by the invention, ECM three-dimensional structure tool inducing cell and blood vessel grow into function,
It itself can gradually degrade while new tissue is grown into, the polypeptide moiety of catabolite has anti-microbial property, after can reducing implantation
The incidence of inflammation and infection.
4. basin bottom Biological Repair mesh sheet provided by the invention, introducing crosslinked agent and synthetic material, will not have potential
Cytotoxicity less will lead to the reaction such as fibrosis, chronic inflammation.
5. promoting healing can be added according to clinical demand in basin bottom Biological Repair mesh sheet provided by the invention during the preparation process
Substance or antibiotic can also load promoting healing substance or antibiotic by immersion way before implanting, thus into one
Step promotes Wound healing and reduces infection rate.
Detailed description of the invention
Fig. 1 is the pictorial diagram of basin bottom Biological Repair mesh sheet.
Fig. 2 is the structural schematic diagram of basin bottom Biological Repair mesh sheet.
Fig. 3 is that basin bottom Biological Repair mesh sheet is sliced HE colored graph.
Fig. 4 is the microgram of basin bottom Biological Repair mesh sheet.
Fig. 5 is the preparation flow figure of basin bottom Biological Repair mesh sheet.
Specific embodiment
Illustrate embodiments of the present invention below by way of specific specific example, those skilled in the art can be by this specification
Other advantages and efficacy of the present invention can be easily understood for disclosed content.The present invention can also pass through in addition different specific realities
The mode of applying is embodied or practiced, the various details in this specification can also based on different viewpoints and application, without departing from
Various modifications or alterations are carried out under spirit of the invention.
Embodiment 1
The preparation of trees-Osima jacoti, Osima excavata basin bottom Biological Repair mesh sheet, please refers to Fig. 5:
(1) it pre-processes
It takes the chitterlings tissue wash of fresh slaughtered animals clean, is placed in 0.5% acetum and impregnates 30min, chitterlings and acetic acid
The ratio of solution is 1:5, scrapes mucous layer, muscle layer, placenta percreta, the lymph node that division removes chitterlings jejunum, separation using physics
Submucosa out, it is longitudinal to be uniformly cut into thin strips shape, it is rinsed 3 times using purified water, obtains biological material for repairing, i.e. mucous membrane of small intestine
Lower layer, following abbreviation SIS materials.
(2) inactivation of virus
Using the mixed aqueous solution containing 1.0% Peracetic acid and 15% ethyl alcohol, the ratio of SIS material and mixed aqueous solution is 1:
10, under ultrasound condition, soaking at room temperature 100min is virus inactivated.It is cleaned by ultrasonic 3 times using purified water later.
(3) degreasing
The ethyl alcohol for the use of concentration being 95%, the ratio of SIS material and ethyl alcohol are 1:10, under ultrasound condition, soak at room temperature 2h.Later
It is cleaned by ultrasonic 3 times using water for injection.
(4) cell is taken off, DNA is removed and removes α-Gal antigen
Using containing 0.02% trypsase and containing the mixed aqueous solution of 0.02%EDTA, SIS material and trypsase/EDTA solution
Ratio be 1:5, under ultrasound condition, impregnate 30min under the conditions of 37 DEG C.It is cleaned by ultrasonic 3 times using PBS later.
Using the aqueous solution of the enzyme of DNA containing 5U/ml, the ratio of SIS material and DNA enzymatic solution is 1:5, under ultrasound condition,
20min is impregnated under the conditions of 37 DEG C.Later using PBS drift ultrasonic cleaning 3 times.
Using the aqueous solution of the alpha-galactosidase containing 5U/ml, the ratio of SIS material and alpha-galactoside enzyme solutions is 1:
5, under ultrasound condition, 20min is impregnated under the conditions of 30 DEG C.It is cleaned by ultrasonic 3 times using PBS later.
The NaOH aqueous solution for the use of concentration being 25mM, the ratio of SIS material and NaOH solution are 1:20, under ultrasound condition,
Soak at room temperature 50min.Later using PBS ultrasonic cleaning until neutral.
(5) it is formed, is lyophilized, braiding, sterilizing
By treated, band-like fine strip shape submucosa is twisted line, is fixed on mold, after freeze-drying, is woven by braider
At the basin bottom Biological Repair mesh sheet of mesh sheet shape in Fig. 1, irradiation sterilization, basin bottom Biological Repair mesh sheet are carried out after PET packaging bag packaging
Schematic diagram see Fig. 2.
Embodiment 2
The preparation of trees-Osima jacoti, Osima excavata basin bottom Biological Repair mesh sheet
(1) it pre-processes
It takes the chitterlings tissue wash of fresh slaughtered animals clean, is placed in 0.01% acetum and impregnates 120min, chitterlings and vinegar
The ratio of acid solution is 1:10, scrapes mucous layer, muscle layer, placenta percreta, the lymph node that division removes chitterlings jejunum using physics, point
Submucosa is separated out, segment is cut into, is rinsed 3 times using purified water.
(2) inactivation of virus
Using the mixed aqueous solution containing 0.5% Peracetic acid and 25% ethyl alcohol, the ratio of SIS material and mixed aqueous solution is 1:
15, under ultrasound condition, soaking at room temperature 120min is virus inactivated.It is cleaned by ultrasonic 3 times using purified water later.
(3) degreasing
The ethyl alcohol for the use of concentration being 90%, the ratio of SIS material and ethyl alcohol are 1:15, under ultrasound condition, soak at room temperature 4h.Later
It is cleaned by ultrasonic 3 times using water for injection.
(4) cell is taken off, DNA is removed and removes α-Gal antigen
Using containing 0.05% trypsase and containing the mixed aqueous solution of 0.01%EDTA, SIS material and trypsase/EDTA solution
Ratio be 1:10, under ultrasound condition, impregnate 20min under the conditions of 37 DEG C.It is cleaned by ultrasonic 3 times using PBS later.
Using the aqueous solution of the enzyme of DNA containing 1U/ml, the ratio of SIS material and DNA enzymatic solution is 1:10, under ultrasound condition,
30min is impregnated under the conditions of 37 DEG C.Later using PBS drift ultrasonic cleaning 3 times.
Using the aqueous solution of the alpha-galactosidase containing 1U/ml, the ratio of SIS material and alpha-galactoside enzyme solutions is 1:
10, under ultrasound condition, 30min is impregnated under the conditions of 30 DEG C.It is cleaned by ultrasonic 3 times using PBS later.
The NaOH aqueous solution for the use of concentration being 40mM, the ratio of SIS material and NaOH solution are 1:10, under ultrasound condition,
Soak at room temperature 30min.Later using PBS ultrasonic cleaning until neutral.
(5) it is formed, is lyophilized, braiding, sterilizing
By treated, fine strip shape submucous layer of small intestine is twisted line, is fixed on mold, after freeze-drying, is woven by braider
The basin bottom Biological Repair mesh sheet of networking sheet, PET packaging bag carry out irradiation sterilization after packing.
Embodiment 3
For the safety of sample, immunogenic substance detection is carried out to the sample that embodiment 1-2 is prepared.
(1) cell residue quantity measuring method: being fixed, paraffin embedding with 10% neutral formalin, is cut into 0.4 micron thin
Piece, through dimethylbenzene dewaxing, serial dehydration of alcohol, hematoxylin-eosin stains, microscopically observation cell residue situation and matrix
Fibre structure.
(2) DNA content detection method: according to YY/T 0606.25-2014, " animal derived biomaterial DNA residual is measured
Determine method: fluorescence colour " it is detected.
(3) α-Gal antigenic content detection method: after sample is fixed with paraformaldehyde, routine paraffin wax embedded section, piece thickness
3 microns.Immunohistochemical reaction is carried out using the special affinity characteristic of biotin labeling BSI-B4 and α-Gal antigen.Coloration result
Determine: dark brown yellow particle is strong positive (+++), and brown yellow granule is positive (++), and yellow particle is weakly positive (+), not
See and is colored as negative (-).
(4) lipid content detection method: referring to soxhlet extraction methods in " fatty measurement in 5009.6 food of GB/T " into
Row measurement.
As a result it see the table below:
Embodiment 4
Biology performance, histology, bacterial endotoxin and anti-microbial property inspection are carried out to the sample that embodiment 1-2 is prepared
It surveys.
(1) biology performance detects
Method: it is tested referring to GB/T16886 series methods.
As a result: cell-cytotoxic reaction is 1 grade；Without delayed allergy；Intradermal reaction shows test specimen and solvent
The difference of mean score is compareed less than 1.0；Without pyrogenicity；Without hemolytic reaction；Genetic toxicity test is the results show that mouse typhus sramana
Salmonella back mutation (Ames) test is negative, mouse lymphoma assay is negative, dye-free body distortion property；Without urgency
Property systemic toxicity reaction；Without sub- chronic generalized toxicity；Muscular grafting 30 days, 60 days, 90 days tissues with negative controls
Reaction is without significant difference.
(2) histology
1) optical microphotograph sem observation
Method: being fixed, paraffin embedding with 10% neutral formalin, is cut into 0.4 micron of thin slice, through dimethylbenzene dewaxing, series wine
Essence dehydration, hematoxylin-eosin stains, microscopically observation cell residue situation and matrix fiber structure.
As a result: cell-free and cell fragment residual；Collagenous fibres are continuous, no fracture, as shown in Figure 3.
2) Ultrastructural observation
Method: it is scanned using electron scanning mirror.
As a result: material porous structure, collagenous fibres are without fracture, as shown in Figure 4.
(3) detection of bacterial endotoxin
Method: it is detected referring to correlation technique in GB/T 14233.
As a result: being respectively less than 2.15EU/ part.
(4) anti-microbial property detects
Sample obtained by Examples 1 and 2 is ground respectively in the hydrochloric acid of 0.01M with grinding rod, until it is visible by naked eyes particle,
And adjusting its concentration is 100mg/10mL.Pepsin digestion is added, pepsin: sample ratio is 1:10.Continue at 25 DEG C
Stir 48h, after be cooled to 4 DEG C, the 0.1 M sodium hydroxide that 1/10 volume is added adjusts PH to 7.2-7.4.
Hybrid NC machine tool base plate is prepared, picks them separately a little cultured staphylococcus aureus and large intestine with oese
Bacteria suspension is made in sterile saline 5ml in bacillus inclined-plane culture substratess.It takes bacteria suspension 1.0ml and above-mentioned is degraded
Sample 1ml is added in the culture dish of sterilizing-drying, and addition is cooled to 50 DEG C or so of ordinary nutritional broth agar culture medium,
It shakes up, spare after sufficiently condensing, 35~37 DEG C of inversions are cultivated 24 hours, observe bacterial growth situation；Increase simultaneously and does not have to
Anti-biotic material and 5 μ g/mL antibacterial peptides of addition compare, and as a result see the table below:
Embodiment 5
Mechanics properties testing is carried out to the embodiment 1-2 sample being prepared and without the sample 1-2 of cutting and braiding.
(1) suture strength
Method: with the non-absorbing suture of 3-0 in sample both sides center far from being sutured at edge 2mm, by the suture other end and sample
The other end be separately fixed on tensiometer both ends, stretched with the speed of 20mm/min, until stitch points are torn, record
Maximal force.
(2) tensile strength
Method: it is 10mm shape that sample is cut to width respectively in both directions；In relative humidity 40%-60%, temperature after cutting
It is tested after being placed 2 hours in 22 ± 2 DEG C of environment of degree.Fixture spacing is 25mm, and sample both ends are fixed on cupping machine
Collet on, the maximal force with the speed tensile of 100mm/min, when record fracture.
(3) bursting strength
Method: according to the measurement method of " YY 0500-2004 cardiovascular implant artificial blood vessel " 8.3.3.2 probe rupture strength
It chooses 9.5mm diameter probe to be detected, as a result see the table below:
Those skilled in the art can make a variety of variations to the present invention according to the above description.Thus, do not violating this hair
Under the premise of bright claim objective, certain details in embodiment should not constitute limitation of the invention, and the present invention will be with
The range that the appended claims define is as protection scope.
Claims (10)
1. a kind of basin bottom Biological Repair mesh sheet, it is characterised in that: the mesh sheet is woven by xenogenesis acellular matrix material.
2. basin bottom Biological Repair mesh sheet according to claim 1, it is characterised in that: the structure of the mesh sheet is by weaving
It is made, the size in mesh sheet aperture can be adjusted by braided parameter.
3. basin bottom Biological Repair mesh sheet according to claim 1, it is characterised in that: the acellular matrix includes but unlimited
Be formed on the submucous layer of small intestine of mammal, submucous layer of bladder, submucous lamina of stomach, dermal matrix, pericardium, meninx, amnion,
One or more combinations of internal organs film, peritonaeum.
4. basin bottom Biological Repair mesh sheet according to claim 3, it is characterised in that: the acellular matrix is the small of animal
Intestinal submucosa.
5. a kind of basin bottom Biological Repair mesh sheet as described in claim 1 ~ 4 is in the repairing healing purposes of pelvic floor tissue.
6. a kind of preparation method of basin bottom Biological Repair mesh sheet, which comprises the following steps:
(1) it pre-processes,
The animal tissue's cleaning for taking fresh slaughtered animals is placed in acetum immersion, strikes off the mucous membrane for removing animal tissue
Layer, muscle layer, placenta percreta, lymph node isolate submucosa, longitudinal to be uniformly cut into thin strips shape, and are rinsed with purified water, obtain basin
Bottom biological material for repairing；
(2) inactivation of virus,
Basin bottom biological material for repairing is used into the mixed aqueous solution containing Peracetic acid and ethyl alcohol, is soaked under room temperature in ultrasound
Bubble, is virus inactivated and is cleaned by ultrasonic with purified water；
(3) degreasing,
Using ethanol solution, impregnates under ultrasound, normal temperature condition, be cleaned by ultrasonic later using water for injection；
(4) cell is taken off, DNA is removed and removes α-Gal antigen,
Using containing trypsase and containing the mixed aqueous solution of EDTA, impregnates under ultrasound condition, be cleaned by ultrasonic later using PBS；
Using the aqueous solution of the enzyme containing DNA, impregnated under ultrasound condition；Later using PBS drift ultrasonic cleaning；
Using the aqueous solution containing alpha-galactosidase, impregnated under ultrasound condition；It is cleaned by ultrasonic later using PBS；
Using NaOH aqueous solution, under ultrasound condition, PBS ultrasonic cleaning is used after soak at room temperature until neutral；
(5) it is formed, is lyophilized, braiding, sterilizing,
By treated, fine strip shape submucosa is twisted line, is fixed on mold, after freeze-drying, is woven into the basin bottom of mesh sheet shape
Biological Repair mesh sheet, PET packaging bag carry out irradiation sterilization after packing.
7. preparation method according to claim 6, which is characterized in that in pre-treatment step, the concentration of acetic acid is
The ratio of 0.01%-0.5%, soaking time 10-120min, animal tissue and acetum is 1:2-1:10.
8. preparation method according to claim 6, which is characterized in that in viral inaction steps, the concentration of Peracetic acid
For 0.5-1.5%, the concentration of ethyl alcohol is 15-25%, and the ratio of basin bottom biological material for repairing and mixed aqueous solution is 1:2-1:10,
Soaking time is 30-120min.
9. preparation method according to claim 6, which is characterized in that in defatting step, the concentration of ethyl alcohol is 90-
100%, the ratio of basin bottom biological material for repairing and ethyl alcohol is 1:2-1:10, and the soak at room temperature time is 0.5-12h.
10. preparation method according to claim 6, which is characterized in that in de- cell, remove DNA and go α-Gal antigen step
In:
The concentration of trypsase and EDTA are respectively 0.01-0.10% and 0.01- 0.05% in mixed aqueous solution, and basin bottom biology is repaired
Material and trypsase/EDTA solution ratio are mended as 1:2-1:10, under ultrasound condition, impregnates 15- under the conditions of 36 ± 2 DEG C
40min；
In aqueous solution containing DNA enzymatic the content of DNA enzymatic be 0.05-10U/ml, basin bottom biological material for repairing with containing the water-soluble of DNA enzymatic
The ratio of liquid is 1:2-1:10, and soaking temperature is 36 ± 2 DEG C, soaking time 15-40min；
The content of alpha-galactosidase is 0.05-10U/ml, basin bottom biological material for repairing in aqueous solution containing alpha-galactosidase
Ratio with alpha-galactoside enzyme solutions is 1:2-1:10, and soaking temperature is 20-37 DEG C, soaking time 15-40min；
The concentration of NaOH aqueous solution is 5-40mM, and the ratio of basin bottom biological material for repairing and NaOH solution is 1:5-1:50, room temperature
Soaking time is 20-60min.
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