CN109453427A - A kind of cleaning method before animal tissue's acellular - Google Patents
A kind of cleaning method before animal tissue's acellular Download PDFInfo
- Publication number
- CN109453427A CN109453427A CN201811450410.0A CN201811450410A CN109453427A CN 109453427 A CN109453427 A CN 109453427A CN 201811450410 A CN201811450410 A CN 201811450410A CN 109453427 A CN109453427 A CN 109453427A
- Authority
- CN
- China
- Prior art keywords
- animal tissue
- acellular
- cleaning method
- method before
- tissue
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—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]
-
- 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
-
- 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
-
- 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
- A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Botany (AREA)
- Dermatology (AREA)
- Veterinary Medicine (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- General Chemical & Material Sciences (AREA)
- Biophysics (AREA)
- Urology & Nephrology (AREA)
- Zoology (AREA)
- Chemical Or Physical Treatment Of Fibers (AREA)
Abstract
The present invention provides the cleaning method before a kind of animal tissue's acellular, comprising the following steps: is shredded after removing sundries, striking off fat；Animal tissue is immersed in the soaking solution of sodium carbonate and NaGC ultrasonic then clean with clear water；Ultrasound is impregnated in the PBS buffer solution containing ascorbic acid, then is cleaned with clear water, and this method can not only sterilize effectively, sterilize and inactivation of virus, will not damage the original basic structure of extracellular organization matrix.
Description
Technical field
The present invention relates to technical field of biological materials, more particularly, to the cleaning side before a kind of animal tissue's acellular
Method.
Background technique
Cell free histoorgan matrix has been applied to tissue reparation, the test of various organizational projects and regenerative medicine and has ground
Study carefully.The extracellular matrix of human body and many animal tissues and organ has great similitude and homology.By allogeneic or different
Kind allosome tissue's organ takes off bio-matrix material made of cell, has been successfully used to the repairing of tissue clinical medicine and has repaired
It is multiple.Good histoorgan matrix, after being implanted into host, matrix scaffold material provides initial biomethanics and supports, by with
Host cell interaction adjusts the behavior (as adhered to, migration, proliferation and differentiation) of cell, with growing into for host cell,
Histoorgan matrix itself is gradually converted to new tissue.
After the original cell component for removing animal tissue's organ, human body cell can also be combined in vitro, make existing three-dimensional box
The histoorgan matrix of frame structure is cellularised again and functionalization, production may migrate to the tissue and organ of human body.
Histoorgan matrix is the 3 D stereo frame being made of the structural protein of various complexity and functional protein, and
Containing there are many other active compounds.Main component includes collagenous fibres, glycoprotein, mucin etc., and other compositions have ammonia
Carbohydrates, some lipids and the growth factors such as base glucan (hyaluronic acid, chondroitin sulfate).Prepare the biology of histoorgan matrix
Process flow is sufficiently complex, acquisition, preservation, cleaning, disinfection, de- cell including histoorgan, reduction antigenicity, inactivation of virus
With the processes such as terminal sterilization.Cleaning treatment wherein before the de- cell of animal tissue is very big to the destruction of histoorgan matrix, is going out
While bacterium living and virus, seriously change histoorgan matrix biopolymers chemical composition, breaks ring 3 D stereo frame
Ultra microstructure and reduction biomechanical property.These change the reaction for influencing host to implantation host material, may cause tissue
Substrate products clinical effectiveness is deteriorated, it is difficult to reach the requirement of tissue reparation.
Therefore, the present invention provides the cleaning method before the de- cell of the novel animal tissue of one kind, can either effectively sterilize,
Sterilizing and inactivation of virus, will not damage the original basic structure of extracellular organization matrix, not change major biochemical ingredient
And biomechanical property.
Summary of the invention
The present invention provides the cleaning method before a kind of animal tissue's acellular, comprising the following steps:
S1. it is shredded after removing sundries, striking off fat；
S2. animal tissue is immersed in the soaking solution of sodium carbonate and NaGC ultrasonic then clean with clear water；
S3. ultrasound is impregnated in the PBS buffer solution containing ascorbic acid, then is cleaned with clear water.
Preferably, the mass ratio of animal tissue and soaking solution is 1:1-10 in S2.
Preferably, soaking temperature is 10-40 DEG C in S2, ultrasonic time 1-3h.
Preferably, the concentration of sodium carbonate is 0.1-1.0mol/L in S2, and the concentration of NaGC is 2.5-5.0mol/L.
Preferably, the clear water is the pure water of aseptic filtration.
Preferably, the mass fraction of ascorbic acid is 1-5wt% in S3.
Preferably, soaking temperature is 10-40 DEG C in S3, ultrasonic time 0.5-1h.
Innovative point of the invention is: first shredding animal tissue, is conducive to the infiltration of cleaning solution, uses sodium carbonate and sweet
Ammonia sodium taurocholate, the two, which combines, effectively degreasing to sterilize, and tissue protein will not be caused to be denaturalized, finally using containing anti-bad
The PBS buffer solution of hematic acid can reduce the damage of subsequent de- cell processing, increase the de- biocompatibility knitted of animal.
Obviously, above content according to the present invention is not being departed from according to the ordinary technical knowledge and customary means of this field
Under the premise of the above-mentioned basic fundamental thought of the present invention, the modification, replacement or change of other diversified forms can also be made.
The specific embodiment of form by the following examples remakes further specifically above content of the invention
It is bright.But the range that this should not be interpreted as to the above-mentioned theme of the present invention is only limitted to example below.It is all to be based on above content of the present invention
The technology realized all belongs to the scope of the present invention.
Specific embodiment
Embodiment 1
A kind of cleaning method before animal tissue's acellular, comprising the following steps:
S1. it is shredded after removing sundries, striking off fat；
S2. animal tissue is immersed in the soaking solution containing 0.1mol/L sodium carbonate and 2.5mol/L NaGC
The mass ratio of ultrasound, animal tissue and soaking solution is 1:1, and soaking temperature is 10 DEG C, ultrasonic time 1h, after ultrasound again
It is cleaned with sterile pure water；
S3. ultrasound is impregnated in the PBS buffer solution containing 1wt% ascorbic acid, soaking temperature is 10 DEG C, and ultrasonic time is
0.5h, ultrasound are cleaned with sterile pure water again later.
Pigskin tissue is tested, measurement result shows compared with fresh pig dermis, and cleaning method of the invention does not have
Damage dermal tissue matrix.
Embodiment 2
A kind of cleaning method before animal tissue's acellular, comprising the following steps:
S1. it is shredded after removing sundries, striking off fat；
S2. animal tissue is immersed in the soaking solution of 1.0mol/L sodium carbonate and 5.0mol/L NaGC and is surpassed
The mass ratio of sound, animal tissue and soaking solution is 1:10, and soaking temperature is 40 DEG C, ultrasonic time 3h, after ultrasound again
It is cleaned with sterile pure water；
S3. ultrasound is impregnated in the PBS buffer solution containing 5wt% ascorbic acid, soaking temperature is 40 DEG C, and ultrasonic time is
1h, ultrasound are cleaned with sterile pure water again later.
Pigskin tissue is tested, measurement result shows compared with fresh pig dermis, and cleaning method of the invention does not have
Damage dermal tissue matrix.
Embodiment 3
A kind of cleaning method before animal tissue's acellular, comprising the following steps:
S1. it is shredded after removing sundries, striking off fat；
S2. animal tissue is immersed in the soaking solution of 0.5mol/L sodium carbonate and 3.5mol/L NaGC and is surpassed
The mass ratio of sound, animal tissue and soaking solution is 1:5, and soaking temperature is 25 DEG C, ultrasonic time 2h, and ultrasound is used again later
Sterile pure water is cleaned；
S3. ultrasound is impregnated in the PBS buffer solution containing 3wt% ascorbic acid, soaking temperature is 25 DEG C, and ultrasonic time is
0.8h, ultrasound are cleaned with sterile pure water again later.
Pigskin tissue is tested, measurement result shows compared with fresh pig dermis, and cleaning method of the invention does not have
Damage dermal tissue matrix.
Embodiment 4
A kind of cleaning method before animal tissue's acellular, comprising the following steps:
S1. it is shredded after removing sundries, striking off fat；
S2. animal tissue is immersed in the soaking solution of 0.3mol/L sodium carbonate and 4.0mol/L NaGC and is surpassed
The mass ratio of sound, animal tissue and soaking solution is 1:7, and soaking temperature is 30 DEG C, ultrasonic time 1h, and ultrasound is used again later
Sterile pure water is cleaned；
S3. ultrasound is impregnated in the PBS buffer solution containing 1wt% ascorbic acid, soaking temperature is 30 DEG C, and ultrasonic time is
1h, ultrasound are cleaned with sterile pure water again later.
Pigskin tissue is tested, measurement result shows compared with fresh pig dermis, and cleaning method of the invention does not have
Damage dermal tissue matrix.
Claims (7)
1. the cleaning method before a kind of animal tissue's acellular, which comprises the following steps:
S1. it is shredded after removing sundries, striking off fat；
S2. animal tissue is immersed in the soaking solution of sodium carbonate and NaGC ultrasonic then clean with clear water；
S3. ultrasound is impregnated in the PBS buffer solution containing ascorbic acid, then is cleaned with clear water.
2. the cleaning method before a kind of animal tissue's acellular according to claim 1, which is characterized in that animal in S2
The mass ratio of tissue and soaking solution is 1:1-10.
3. the cleaning method before a kind of animal tissue's acellular according to claim 1, which is characterized in that impregnated in S2
Temperature is 10-40 DEG C, ultrasonic time 1-3h.
4. the cleaning method before a kind of animal tissue's acellular according to claim 1, which is characterized in that carbonic acid in S2
The concentration of sodium is 0.1-1.0mol/L, and the concentration of NaGC is 2.5-5.0mol/L.
5. the cleaning method before a kind of animal tissue's acellular stated according to claim 1, which is characterized in that the clear water is equal
For the pure water of aseptic filtration.
6. the cleaning method before a kind of animal tissue's acellular according to claim 1, which is characterized in that anti-bad in S3
The mass fraction of hematic acid is 1-5wt%.
7. the cleaning method before a kind of animal tissue's acellular according to claim 1, which is characterized in that impregnated in S3
Temperature is 10-40 DEG C, ultrasonic time 0.5-1h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811450410.0A CN109453427A (en) | 2018-11-30 | 2018-11-30 | A kind of cleaning method before animal tissue's acellular |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811450410.0A CN109453427A (en) | 2018-11-30 | 2018-11-30 | A kind of cleaning method before animal tissue's acellular |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109453427A true CN109453427A (en) | 2019-03-12 |
Family
ID=65612075
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811450410.0A Withdrawn CN109453427A (en) | 2018-11-30 | 2018-11-30 | A kind of cleaning method before animal tissue's acellular |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109453427A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080248080A1 (en) * | 2000-08-16 | 2008-10-09 | Duke University | Decelluarlized Tissue Engineered Constructs and Tissues |
| CN102580152A (en) * | 2012-03-09 | 2012-07-18 | 潘银根 | Method for preparing acellular bone |
| CN104771784A (en) * | 2015-05-05 | 2015-07-15 | 北京帝康医药投资管理有限公司 | Tissue accellular solution |
| CN105126169A (en) * | 2015-07-10 | 2015-12-09 | 蒋青 | Allograft bone meal as well as preparation method and application thereof |
-
2018
- 2018-11-30 CN CN201811450410.0A patent/CN109453427A/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080248080A1 (en) * | 2000-08-16 | 2008-10-09 | Duke University | Decelluarlized Tissue Engineered Constructs and Tissues |
| CN102580152A (en) * | 2012-03-09 | 2012-07-18 | 潘银根 | Method for preparing acellular bone |
| CN104771784A (en) * | 2015-05-05 | 2015-07-15 | 北京帝康医药投资管理有限公司 | Tissue accellular solution |
| CN105126169A (en) * | 2015-07-10 | 2015-12-09 | 蒋青 | Allograft bone meal as well as preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 熊前程等: "《有机化学》", 31 August 2018, 西安交通大学出版社 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wilshaw et al. | Production of an acellular amniotic membrane matrix for use in tissue engineering | |
| Hoganson et al. | The retention of extracellular matrix proteins and angiogenic and mitogenic cytokines in a decellularized porcine dermis | |
| US20220160937A1 (en) | Rapid allograft treatment systems and methods | |
| CN106492285A (en) | Injectable Acellular cartilaginous matrix particulate and its application in implant | |
| CN104055795B (en) | A kind of injectable implant and preparation method thereof | |
| JP2021184828A (en) | Compositions comprising mesenchymal stem cells and uses thereof | |
| CN109331228A (en) | A kind of preparation method of anti-infectious submucous layer of small intestine biomaterial | |
| KR101410533B1 (en) | A method for treating material derived from biological tissue | |
| EP3326660B1 (en) | A ready to use biodegradable and biocompatible artificial skin substitute and a method of preparation thereof | |
| CN101773687A (en) | Preparation method of composite soft-tissue patch | |
| CN112870445A (en) | Preparation method and application of soft tissue repair material | |
| CN103861151A (en) | Method for preparing acellular placenta substrate material | |
| CN103418001B (en) | The method for disinfection and sterilization of a kind of animal tissue material and corresponding animal tissue soaking solution | |
| CN109675112B (en) | Preparation method of human-derived acellular dermal matrix | |
| WO2004011631A3 (en) | Methods and compositions for treating tissue defects using pulsed electromagnetic field stimulus | |
| CN108066750A (en) | Stem cell and its secretion are used to treat the new application of skin burn | |
| CN109771694A (en) | The preparation method and application of self assembly polypeptide nano fiber water gel scaffold material | |
| US20120022233A1 (en) | Collagen implant | |
| Scomazzon et al. | An increase in Wharton's jelly membrane osteocompatibility by a genipin-cross-link | |
| CN108066824A (en) | A kind of new method for preparing skin blemish medicine | |
| CN114306755B (en) | Biological patch for posterior scleral reinforcement and preparation method thereof | |
| CN109453427A (en) | A kind of cleaning method before animal tissue's acellular | |
| Kumaresan et al. | Development of Human Umbilical cord based scaffold for tissue engineering application | |
| CN108057131A (en) | A kind of novel agent box containing stem cell | |
| CN105709276B (en) | A kind of chorion de-cell liquid and method for removing cells |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20190312 |