CN111450320A - Biological tissue abdominal wall defect repairing material - Google Patents
Biological tissue abdominal wall defect repairing material Download PDFInfo
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- CN111450320A CN111450320A CN202010210435.4A CN202010210435A CN111450320A CN 111450320 A CN111450320 A CN 111450320A CN 202010210435 A CN202010210435 A CN 202010210435A CN 111450320 A CN111450320 A CN 111450320A
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- abdominal wall
- dermis
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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
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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
- A61L27/362—Skin, e.g. dermal papillae
-
- 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
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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
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/04—Materials for stopping bleeding
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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
- A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
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- Biomedical Technology (AREA)
- Engineering & Computer Science (AREA)
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- Medicinal Chemistry (AREA)
- Transplantation (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
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- Materials For Medical Uses (AREA)
Abstract
The invention discloses a biological tissue abdominal wall defect repairing material. The material includes a placental tissue implant and decellularized xenogenic dermis. The invention can repair abdominal wall defect peritoneally by placenta tissue implant, and can avoid skin-intestinal loop adhesion, intestinal obstruction, intestinal fistula and long-term pain complications existing in non-peritonealization repair; the combination of acellular xenogenic dermis overcomes the defect of low tension of the repair of the musculus muscularis with muscle.
Description
Technical Field
The invention belongs to the technical field of medical material preparation, and particularly relates to a biological tissue abdominal wall defect repairing material.
Background
Abdominal cavity infection and abdominal trauma are accompanied by blood coagulation and dysfunction of multiple organs (liver, central nervous system) of the whole body, and therefore, abdominal cavity open therapy is essential. The abdominal cavity open therapy can reduce the intra-abdominal pressure, prevent the abdominal cavity clearance syndrome, prevent and treat multiple organ dysfunction, and is convenient for surgeons to timely clear infected necrotic tissues, clear drainage and find complications such as hemorrhage, intestinal fistula and the like at any time. Complications of abdominal cavity patency therapy follow and are difficult to avoid. The best result of the abdominal cavity opening is that the abdominal cavity is closed by full-layer fascia suture in the early stage, namely, after the treatment purpose is realized in the short period of the abdominal cavity opening, all layers of the incision are successfully sutured in the full layer, and the abdominal cavity is closed. However, patients of this type often have abdominal infection, malnutrition, air fistulas, abdominal wall defects, and the like, and early full-thickness abdominal closing is difficult to achieve. The repair of abdominal wall defect needs to be performed after temporary abdominal closing. For the treatment of abdominal wall defects after abdominal cavity opening, peritoneal abdominal wall defect repair is still recommended due to complications of skin-intestinal loop adhesion, intestinal obstruction, intestinal fistula and long-term pain in the absence of peritoneal repair (such as flap transplantation).
The applicant has used the previously self-developed multi-layer placental tissue implant (201710412764.5 a method for manufacturing a medical multi-layer placental tissue implant) and the decellularized xenodermal (201510373277.3 a decellularized xenodermal and a method for manufacturing the same) to repair the abdominal wall defect, and found that the abdominal wall defect is not sufficiently tensioned, the healing days are long, and the repair effect is not ideal.
Disclosure of Invention
The invention aims to provide a biological tissue abdominal wall defect repairing material which can promote the proliferation of wound cells, has self-repairing and self-degrading capabilities, has hemostatic, antibacterial and anti-inflammatory activities, and is used for repairing abdominal wall defects in the later stage of abdominal cavity patency.
A material for repairing the abdominal wall defect of biological tissue is composed of the placenta tissue implant and the acellular heterogeneous corium.
The repair material is prepared by laminating and drying a placenta tissue implant and acellular xenogenic dermis.
The preparation method of the placenta tissue implant is prepared according to the method of '201710412764.5 a preparation method of medical multilayer placenta tissue implant'.
The method for producing the acellular heterogeneous dermis is prepared according to the method described in "201510373277.3 a acellular heterogeneous dermis and its preparation method".
The invention has the beneficial effects that: the invention can repair abdominal wall defect peritoneally by placenta tissue implant, and can avoid skin-intestinal loop adhesion, intestinal obstruction, intestinal fistula and long-term pain complications existing in non-peritonealization repair (such as flap transplantation); the combination of acellular xenogenic dermis overcomes the defect of low tension of the repair of the musculus muscularis with muscle. The product of the invention is used as a repairing material for huge abdominal wall defects, reduces immunogenicity, simultaneously reserves other composition structures of placenta tissues, is used as a material for finally deterministically repairing abdominal defects, greatly reduces the incidence rate of rejection reaction, has the characteristics of better adhesion effect on deep wound surfaces, strong toughness, difficult tearing and the like, and effectively prevents abdominal wall hernia after repair; meanwhile, acellular xenogenic dermis is used, a foundation is laid for final skin grafting of the abdominal wall, and for large skin defects, skin flap grafting is not needed, and only medium-thick or even thin skin is needed to be grafted, so that surgical wounds can be reduced, and postoperative recovery of a patient is promoted; in addition, compared with the traditional single acellular xenogeneic dermis, the material only needs one layer of acellular xenogeneic dermis, so that the repair strength and thickness of the abdominal wall can be ensured, the treatment cost can be reduced, and the burden of a patient can be reduced; finally, the material is a composite material, and the localization of the multi-layer placenta tissue implant and the acellular heterogeneous dermis can be realized through the preparation of the material, so that the material has higher social and economic benefits.
Detailed Description
In order that the invention may be more fully understood, reference will now be made to the following description. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Example 1 preparation of placental tissue implants
Separating the amniotic membrane and chorion tissue from the placental tissue material: the obtained placental tissue is sent to a controlled processing environment under aseptic conditions and soaked in an aseptic basin containing 2.0 wt% aqueous sodium chloride solution for rinsing, and the amniotic membrane and chorion tissue are separated from other placental tissue.
Treating the amniotic membrane and chorionic tissue to remove blood stains, cells and DNA, and sterilizing the decellularized tissue: firstly, repeatedly washing the separated amnion and chorion tissues by using 2.0 wt% sodium chloride aqueous solution to remove blood stains; then, the blood-stained tissue was soaked in a 3.0 wt% aqueous sodium chloride solution and mixed with shaking at room temperature for 120 minutes to remove cells; thereafter, the cell-removed tissue was placed in a DNA-removing solution (consisting of HEPES solution, MgCl)2Solution, CaCl2Solution, purified water, DNase mixed) at room temperature for 5 hours until the residual DNA content in the tissue is not largeAt 250 ng/mg tissue; finally, the DNA-removed tissue was sterilized by soaking in 70% isopropyl alcohol for 50 minutes.
Sequentially stacking multiple layers of sterilized amnion and/or chorion tissues on a mold, and then putting the mold into an oven to dry for 60 minutes at the temperature of 40 ℃ to obtain a dried tissue;
the sequence of the multiple layers is as follows:
1) in the case of a multi-layer amniotic membrane,
a first layer of amnion: basal layer facing down, fibroblast layer facing up;
the middle layer of amnion: the same layer of amnion;
and (3) a last layer of amnion: the fibroblast layer faces down and the basal layer faces up;
2) in the case of a multi-layer chorion,
first layer of chorion: basal layer facing down, fibroblast layer facing up;
intermediate layer chorion: the same layer of chorion;
and (3) a last layer of chorion: the fibroblast layer faces down and the basal layer faces up;
3) when a mixed structure of multiple layers of amniotic and chorionic membranes,
a first layer of amnion: basal layer facing down, fibroblast layer facing up;
intermediate layer chorion: the fibroblast layer faces down and the basal layer faces up;
the middle layer of amnion: the same layer of amnion;
and (3) a last layer of amnion or chorion: the fibroblast layer faces down and the basal layer faces up;
in the case of such a mixed structure, the orientation of the dry tissue must be noted.
Example 2 decellularized heterogeneous dermis
Preparing a pig dermis: cleaning the back skin of a mature white pig (Yorkshire pig breed), removing hairs by a hair removing knife, cutting a skin piece with the thickness of 0.25 mm by using a skin taking drum, discarding the skin piece (for removing epidermis, hair and hair follicles), and taking a skin piece with the thickness of 0.65 mm by using the skin taking drum for later use.
And (3) disinfection: putting the pork skin sheet into a sterile bottle, and adding 0.5 mass percent of peroxyacetic acid disinfectant to soak for 40 minutes.
Cleaning and disinfecting: the cells were washed 3 times with phosphate buffered saline and shaken on a shaker for 30 minutes each time.
And (3) cell removal: adding a cell removal solution, fully mixing and reacting the pig dermis slice with the cell removal solution, mixing the cell removal solution on a shaking table for 10 hours, wherein the cell removal solution is a 4-hydroxyethylpiperazine ethanesulfonic acid buffer solution, the pH value of the cell removal solution is 8, each liter of the 4-hydroxyethylpiperazine ethanesulfonic acid buffer solution contains 20 millimoles of 4-hydroxyethylpiperazine ethanesulfonic acid, the cell removal solution contains sodium deoxycholate, the mass volume percentage solubility of the sodium deoxycholate is 2%, and the ratio of the volume of the cell removal solution to the volume of the pig dermis slice is 30: 1.
the cell-free solution was aspirated, and then phosphate buffered saline was added to wash the pig dermis pieces thoroughly, 3 times with phosphate buffered saline, each time shaking on a shaker for 30 minutes.
Example 3 use of placental tissue implants and acellular xenodermal composites
The method is divided into two using methods:
(1) when the repair material is used, the placenta tissue implant is applied to the position close to the innermost layer of the abdominal wall defect, and the acellular xenogenic dermis is covered on the placenta tissue implant and is sutured.
(2) When the repair material is used, the placenta tissue implant is applied to the position close to the innermost layer of the abdominal wall defect, the acellular xenogenic dermis is covered on the placenta tissue implant, and a layer of placenta tissue implant is arranged on the acellular xenogenic dermis and is sewn.
Example 4 Material tensile testing experiment
According to the method specified in GB/T3923.1-1997, the experimental materials were divided into 3 groups: group 1: placental tissue implants (bilayers); group 2: acellular xenogenic dermis (bilayer); group 3: the composite of placental tissue implant and decellularized xenodermal was subjected to a tensile test, the results of which are shown in table 1:
TABLE 1
Note: represents P <0.05 compared to group 1.
As can be seen from table 1, the tensile strength of the double-layered acellular heterogeneous dermis is the greatest, but the compatibility and the implantability of the acellular heterogeneous dermis are poor, and the tensile strength of the placenta tissue implant alone is too low, so that the scheme of compounding the placenta tissue implant and the acellular heterogeneous dermis is the best, and the composite placenta tissue implant has both high tensile strength and compatibility and implantability.
Example 5 cell proliferation assay experiment
Rat fibroblasts were planted in the experimental materials of groups 1 to 3 prepared in example 4, CCK-8 cell proliferation experiments were performed for 5 days and 7 days, respectively, and the CCK-8 cell proliferation experiments were performed using the CCK-8 detection cell proliferation kit produced by shenzhen, tutopu biotechnology limited, according to the instruction manual, with the test results shown in table 2:
TABLE 2
Note: represents P <0.05 compared to group 1.
As can be seen from table 2, the proliferation amount of cells using a single acellular xenogenic dermis and a single placental tissue implant is significantly less than that of a scheme of combining two materials, and after the materials are combined, the proliferation speed and quantity of cells are increased.
Example 6 wound healing test experiment
SD female rats were subjected to 10% chloral hydrate intraperitoneal injection anesthesia (0.4ml/100g), conventional abdominal skin preparation, sterile drape, median longitudinal incision of the abdomen was made to a length of about 5cm, subcutaneous tissue of the skin was incised, subcutaneous tissue was bluntly separated bilaterally along the incision, rectus abdominis was excised together with the lower transverse abdominal muscle, transverse abdominal fascia and peritoneum, and a preparation size of about 2 × 2cm2The full-thickness abdominal wall defect model of rats was repaired with the materials prepared in groups 1-3 of example 4 (5 rats per group) using 4-0 silk thread interruptedSuturing, and intermittently suturing skin and subcutaneous tissue with 3-0 silk thread. After suturing, rats were observed for wound healing: the wounds were not inflamed, new capillaries formed, collagen formed, healing of the wounds was marked when no pain was felt, the healing time was calculated for each group of rats and the average healing time was measured and is shown in table 3:
TABLE 3
Note: represents P <0.05 compared to group 1.
As can be seen from table 3, the healing time of the wound using the single placental tissue implant and the single acellular heterogeneous dermis is significantly longer than that of the composite material of placental tissue implant and acellular heterogeneous dermis, and the healing time of the wound is shortened after the composite material is used.
The multilayer biological tissue material comprises a multilayer placenta tissue implant and acellular heterogeneous dermis. The multi-layer placenta tissue implant is composed of the amnion and the chorion of a human placenta, and the cell structure is destroyed and DNA components are removed after the process treatment, so that the tissue antigenicity is completely removed, and the immunological rejection reaction after the transplantation is avoided. The reticular fiber membranous structure and extracellular matrix are highly superposed with other membrane tissues in a human body, and the reticular fiber membranous structure and extracellular matrix are rich in various substances such as growth factors, functional proteins and the like, so that the reticular fiber membranous structure and extracellular matrix have excellent effects of inducing cell differentiation and proliferation, resisting inflammation and preventing adhesion. In addition, the multi-layer mechanism consisting of the amnion and the chorion has great enhancement on the mechanical property, and is more suitable for the conditions with certain requirements on the tensile strength, including abdominal wall defect repair.
The acellular xenodermal is prepared by treating pig dermis with a process to destroy intradermal cells, thoroughly remove antigenic DNA and α -galactose, and retain the fibrous framework structure of the dermis layer.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (3)
1. A biological tissue abdominal wall defect repair material is characterized by comprising a placenta tissue implant and acellular xenogenic dermis.
2. The material for repairing a defect in the abdominal wall of a biological tissue as claimed in claim 1, wherein said material for repairing is used by applying a placental tissue implant to a site close to the innermost layer of the defect in the abdominal wall, covering it with acellular xenogeneic dermis and suturing.
3. The material for repairing a defect in the abdominal wall of a biological tissue as claimed in claim 1, wherein said material for repairing a defect in the abdominal wall is used by applying a placental tissue implant to a position close to the innermost layer of the defect in the abdominal wall, covering a layer of acellular xenogeneic dermis, and placing a layer of placental tissue implant on the acellular xenogeneic dermis and suturing.
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| CN202010210435.4A CN111450320A (en) | 2020-03-24 | 2020-03-24 | Biological tissue abdominal wall defect repairing material |
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| CN202010210435.4A CN111450320A (en) | 2020-03-24 | 2020-03-24 | Biological tissue abdominal wall defect repairing material |
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Citations (5)
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| US20160000968A1 (en) * | 2014-07-07 | 2016-01-07 | Mimedx Group, Inc. | Placental tissue grafts and improved methods of preparing and using the same |
| CN106310384A (en) * | 2015-07-01 | 2017-01-11 | 江西瑞诺健医学科技有限公司 | Xenogeneic acellular dermis and preparation method thereof |
| CN107335095A (en) * | 2017-06-05 | 2017-11-10 | 江西瑞诺健医学科技有限公司 | A kind of preparation method of medical multilayer placenta tissue implant |
| CN108066821A (en) * | 2016-11-10 | 2018-05-25 | 上海中医药大学附属曙光医院 | A kind of compound bio sticking patch for abdominal-wall defect reparation |
| CN109224132A (en) * | 2018-09-17 | 2019-01-18 | 上海大学 | MULTILAYER COMPOSITE stomach wall repairs sticking patch bracket and preparation method thereof |
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2020
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| US20160000968A1 (en) * | 2014-07-07 | 2016-01-07 | Mimedx Group, Inc. | Placental tissue grafts and improved methods of preparing and using the same |
| CN106310384A (en) * | 2015-07-01 | 2017-01-11 | 江西瑞诺健医学科技有限公司 | Xenogeneic acellular dermis and preparation method thereof |
| CN108066821A (en) * | 2016-11-10 | 2018-05-25 | 上海中医药大学附属曙光医院 | A kind of compound bio sticking patch for abdominal-wall defect reparation |
| CN107335095A (en) * | 2017-06-05 | 2017-11-10 | 江西瑞诺健医学科技有限公司 | A kind of preparation method of medical multilayer placenta tissue implant |
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Application publication date: 20200728 |