CN114177358A - Biological scaffold, and forming method and application thereof - Google Patents
Biological scaffold, and forming method and application thereof Download PDFInfo
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- CN114177358A CN114177358A CN202111519206.1A CN202111519206A CN114177358A CN 114177358 A CN114177358 A CN 114177358A CN 202111519206 A CN202111519206 A CN 202111519206A CN 114177358 A CN114177358 A CN 114177358A
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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
- A61L27/60—Materials for use in artificial skin
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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
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/24—Collagen
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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
- A61L27/14—Macromolecular materials
- A61L27/26—Mixtures of macromolecular compounds
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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
- A61L27/56—Porous materials, e.g. foams or sponges
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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
- A61L2430/10—Materials or treatment for tissue regeneration for reconstruction of tendons or ligaments
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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
- A61L2430/32—Materials or treatment for tissue regeneration for nerve reconstruction
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Abstract
The invention relates to a biological scaffold, a forming method and application thereof. The forming method of the biological scaffold comprises the following steps: mixing the degradable biological material with glycosaminoglycan, and swelling in acetic acid aqueous solution to obtain raw material slurry; pre-freezing the raw material slurry by using a refrigerant, controlling the refrigeration effect according to the distance between the raw material slurry and the refrigerant, and obtaining a pre-frozen biological scaffold after completion; and drying the pre-frozen biological scaffold to obtain the biological scaffold. The preparation method of the biological scaffold can achieve the effect of controlling the pre-freezing rate only by adjusting the distance, thereby obtaining the biological scaffold with different pore shapes, meeting different application scenes, being extremely simple and convenient to operate, and reducing the production cost and the production difficulty; and all performance indexes of the obtained product meet the requirements.
Description
Technical Field
The invention belongs to the field of biological materials, and particularly relates to a biological scaffold, a forming method and application thereof.
Background
With the development of science and technology, biological materials are changing day by day. Chinese patent CN101264350B discloses a method for preparing a spinal canal membrane, which specifically comprises the following steps: preparing collagen into a collagen suspension with a certain concentration, injecting the collagen suspension into an open die, vertically freezing the suspension within minus 80 to minus 100 ℃ at a low speed of 1 to 2mm/min, and performing secondary freezing to form a product. In addition, chinese patent document CN101332134B discloses a collagen scaffold product with single longitudinally arranged pores by using a freeze-drying process, which comprises preparing a suspension of type I collagen at a certain concentration, loading into a specific mold, vertically freezing at a speed of 1-2 mm/min in a freezing container at-60 to-110 ℃, and performing secondary conventional freeze-drying to obtain the collagen scaffold with single longitudinally arranged pores.
In the methods, the product moving speed and the container temperature are required to be accurately controlled, and the methods have the defects of high energy consumption, complex device, complex flow and the like and are poor in popularization.
Therefore, the technical scheme of the invention is provided.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a biological scaffold, a forming method and application thereof. The preparation method of the biological scaffold can achieve the effect of controlling the pre-freezing rate only by adjusting the distance, thereby obtaining the biological scaffold with different pore shapes, meeting different application scenes, being extremely simple and convenient to operate, and reducing the production cost and the production difficulty; and all performance indexes of the obtained product meet the requirements.
The scheme of the invention is to provide a method for molding a biological scaffold, which comprises the following steps:
(1) mixing the degradable biological material with glycosaminoglycan, and swelling in acetic acid aqueous solution to obtain raw material slurry;
(2) pre-freezing the raw material slurry by using a refrigerant, controlling the refrigeration effect according to the distance between the raw material slurry and the refrigerant, and obtaining a pre-frozen biological scaffold after completion;
(3) and drying the pre-frozen biological scaffold to obtain the biological scaffold.
Preferably, the degradable biomaterial is collagen or chitosan; the collagen is acid soluble collagen or insoluble collagen fiber. As is generally understood, collagen is soluble in acidic or alkaline solutions when present in a single molecule, oligomeric form; whereas when collagen is in the macroscopic fiber grade, dissolution becomes difficult, which may exist in a swollen form.
Wherein the raw material slurry contains collagen or chitosan at a concentration of 3-10 wt.% and glycosaminoglycan at a concentration of 0.1-2.0 wt.%.
Preferably, the glycosaminoglycan is one or a combination of hyaluronic acid, chondroitin sulfate, dermatan sulfate or keratan sulfate.
Preferably, the drying comprises 5 stages in sequence, respectively:
(1) freezing for 15min at-35-45 ℃;
(2) freeze-drying at-8-10 deg.C and vacuum degree of 0.2bar for 740-920 min;
(3) freeze-drying for 380-460 min at-10-13 deg.C and vacuum degree of 0.2 bar;
(4) freeze-drying at-5-0 deg.C and vacuum degree of 0.2bar for 400-520 min;
(5) drying for 60-100 min at 20-30 ℃ and under the vacuum degree of 0.2 bar.
Based on the same technical concept, the invention further provides a biological scaffold obtained by the forming method. Also, the application of the biological scaffold in repairing nerve, tendon, ligament, skin, meninges and mucosa injury is provided, and the purpose of diagnosing and treating diseases is not provided.
The technical idea of the invention is as follows: structurally, the biological scaffold is a porous scaffold with communicated pores, and in the repair process, the structure is more beneficial to the multi-dimensional growth of nerves, muscles and the like, and the repair effect is better. In order to obtain the structure, raw material slurry is injected into a metal mold in the preparation process, the shape of the metal mold can be selected to be cylindrical or tubular according to needs, the metal mold is vertically lowered to a position 2-20 mm away from the upper surface of a refrigerant (preferably liquid nitrogen, which is low in cost and easy to obtain and can obtain more supercooling degrees as a cold source), the lowering speed does not need to be controlled, the metal mold stays for 15-120 min, and when the tubular mold is adopted, the central column is slowly taken out and pulled out after pre-freezing. It should be emphasized that the distance described in the present invention refers to the distance between the upper surface of the refrigerant and the lower surface of the metal mold, and the overall temperature of the metal mold will eventually be kept consistent due to the conduction effect. After pre-freezing, water in the raw material slurry is condensed into a solid state from a liquid state, ice crystals are formed, the ice crystals are sublimated in the subsequent drying process, and a pore structure in the shape of the ice crystals is left, namely, communicated pores are formed.
Moreover, according to the long-term research and exploration of the inventor, the microstructure and the pore shape of the product are changed along with the change of the distance. Specifically, the method comprises the following steps: when the distance is 2-9 mm, the product has crack defects, and the larger the distance is, the smaller the crack defects are; when the distance is 10-20 mm, the product does not generate crack defects, in the range, the closer the distance is to 10mm, the higher the supercooling degree of the product is, the higher the length-to-axis ratio of finally formed pores is, the closer the shape is to a fusiform shape, and longitudinal pore channels are formed (as shown in fig. 3); the closer the distance is to 20mm, the lower the degree of supercooling the product has, the closer the axial length to diameter ratio of the finally formed pores is to 1, and the closer the shape is to a circle (as shown in fig. 4).
And further it has been found that longitudinal pores are suitable for fibrous isotropic tissue repair, such as nerves and tendons, whereas round pores are suitable for collagen or tissue fibrous anisotropic tissue repair, such as skin, meninges, mucosa and the like.
The invention has the beneficial effects that:
the preparation method of the biological scaffold can achieve the effect of controlling the pre-freezing rate only by adjusting the distance, thereby obtaining the biological scaffold with different pore shapes, meeting different application scenes, being extremely simple and convenient to operate, and reducing the production cost and the production difficulty; and all performance indexes of the obtained product meet the requirements.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a schematic view of a cylindrical metal mold.
Fig. 2 is a schematic view of a circular tubular metal mold.
FIG. 3 is an electron micrograph of a longitudinal pore channel of a biological stent.
FIG. 4 is an electron micrograph of a biological scaffold circular pore channel.
Reference numerals
1-a cylindrical metal mold; 2-a round tubular metal mold; 3-the central column.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be described in detail below. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the examples given herein without any inventive step, are within the scope of the present invention.
Example 1
The embodiment provides a method for molding a biological stent, which comprises the following steps:
(1) mixing acid-soluble collagen dry powder, chitosan dry powder and hyaluronic acid dry powder, and swelling in 0.05mol/L acetic acid aqueous solution to obtain raw material slurry; wherein the total concentration of acid-soluble collagen and chitosan is 3 wt.%, and the concentration of hyaluronic acid is 0.1 wt.%;
(2) pre-freezing the raw material slurry by adopting liquid nitrogen, transferring the raw material slurry into a cylindrical metal mold 1 in an injection mode, and lowering the whole metal mold to a position 2mm away from the upper surface of the liquid nitrogen without controlling the lowering speed, and staying for 15min to keep the whole metal mold at a constant temperature to obtain a pre-frozen biological scaffold;
(3) and drying the pre-frozen biological scaffold, wherein the drying sequentially comprises 5 stages, namely:
(1) freezing at-35 deg.C for 15 min;
(2) freeze drying at-10 deg.C and vacuum degree of 0.2bar for 740 min;
(3) freeze drying at-13 deg.C under vacuum degree of 0.2bar for 380 min;
(4) freeze drying at-5 deg.C under vacuum degree of 0.2bar for 400 min;
(5) drying at 20 deg.C and vacuum degree of 0.2bar for 60 min;
and obtaining the biological scaffold after finishing.
Example 2
The embodiment provides a method for molding a biological stent, which comprises the following steps:
(1) mixing acid-soluble collagen dry powder and chondroitin sulfate dry powder, and swelling in 0.05mol/L acetic acid aqueous solution to obtain raw material slurry; wherein the concentration of acid-soluble collagen is 10 wt.%, and the concentration of chondroitin sulfate is 2 wt.%;
(2) pre-freezing the raw material slurry by adopting liquid nitrogen, transferring the raw material slurry into a tubular metal mold 2 in an injection mode, lowering the whole metal mold to a position 20mm away from the upper surface of the liquid nitrogen without controlling the lowering speed, staying for 120min, keeping the whole metal mold at a constant temperature, taking out and pulling out a central column 3 after completion to obtain a pre-frozen biological scaffold;
(3) and drying the pre-frozen biological scaffold, wherein the drying sequentially comprises 5 stages, namely:
(1) freezing at-35 deg.C for 15 min;
(2) freeze drying at-8 deg.C under vacuum degree of 0.2bar for 920 min;
(3) freeze drying at-10 deg.C and vacuum degree of 0.2bar for 460 min;
(4) freeze drying at 0 deg.C and vacuum degree of 0.2bar for 520 min;
(5) drying at 30 deg.C and vacuum degree of 0.2bar for 100 min;
and obtaining the biological scaffold after finishing.
Example 3
The embodiment provides a method for molding a biological stent, which comprises the following steps:
(1) mixing chitosan dry powder and dermatan sulfate dry powder, and swelling in 0.05mol/L acetic acid aqueous solution to obtain raw material slurry; wherein the chitosan concentration is 7 wt.%, and the dermatan sulfate concentration is 1 wt.%;
(2) pre-freezing the raw material slurry by adopting liquid nitrogen, transferring the raw material slurry into a cylindrical metal mold in an injection mode, and lowering the whole metal mold to a position 10mm above the upper surface of the liquid nitrogen without controlling the lowering speed, and staying for 70min to keep the whole metal mold at a constant temperature to obtain a pre-frozen biological scaffold;
(3) and drying the pre-frozen biological scaffold, wherein the drying sequentially comprises 5 stages, namely:
(1) freezing at-35 deg.C for 15 min;
(2) freeze drying at-9 deg.C under vacuum degree of 0.2bar for 830 min;
(3) freeze drying at-11 deg.C under vacuum degree of 0.2bar for 420 min;
(4) freeze drying at-2 deg.C under vacuum degree of 0.2bar for 460 min;
(5) drying at 25 deg.C and vacuum degree of 0.2bar for 80 min;
and obtaining the biological scaffold after finishing.
Example 4
The embodiment provides a method for molding a biological stent, which comprises the following steps:
(1) swelling the insoluble collagen fiber dry powder in 0.05mol/L acetic acid aqueous solution to obtain raw material slurry; wherein the concentration of the insoluble collagen fibers is 5 wt.%;
(2) pre-freezing the raw material slurry by adopting liquid nitrogen, transferring the raw material slurry into a cylindrical metal mold in an injection mode, and lowering the whole metal mold to a position 10mm above the upper surface of the liquid nitrogen without controlling the lowering speed, and staying for 70min to keep the whole metal mold at a constant temperature to obtain a pre-frozen biological scaffold;
(3) and drying the pre-frozen biological scaffold, wherein the drying sequentially comprises 5 stages, namely:
(1) freezing at-35 deg.C for 15 min;
(2) freeze drying at-9 deg.C under vacuum degree of 0.2bar for 830 min;
(3) freeze drying at-11 deg.C under vacuum degree of 0.2bar for 420 min;
(4) freeze drying at-2 deg.C under vacuum degree of 0.2bar for 460 min;
(5) drying at 25 deg.C and vacuum degree of 0.2bar for 80 min;
and obtaining the biological scaffold after finishing.
In order to show the mechanical property of the biological scaffold, the suture strength test is carried out, and the method comprises the following steps: one end of the sample was fixed to the base of the jig, the other end was passed through with a suture to form a half ring, which was fixed to the jig, and then stretched at a speed of 100mm/min, and the amount of the pulling force by which the suture was pulled out of the sample or the sample was broken was recorded, and the results are shown in table 1.
TABLE 1 test results
The above description is only for the specific embodiments of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention, and all the changes or substitutions should be covered within the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.
Claims (8)
1. A method for forming a biological stent is characterized by comprising the following steps:
(1) mixing the degradable biological material with glycosaminoglycan, and swelling in acetic acid aqueous solution to obtain raw material slurry;
(2) pre-freezing the raw material slurry by using a refrigerant, controlling the refrigeration effect according to the distance between the raw material slurry and the refrigerant, and obtaining a pre-frozen biological scaffold after completion;
(3) and drying the pre-frozen biological scaffold to obtain the biological scaffold.
2. The method for forming a bioscaffold according to claim 1, wherein said degradable biomaterial is collagen or chitosan; the collagen is acid soluble collagen or insoluble collagen fiber.
3. The method for forming a biological scaffold according to claim 1, wherein the glycosaminoglycan is one or a combination of hyaluronic acid, chondroitin sulfate, dermatan sulfate or keratan sulfate.
4. The method of forming a bioscaffold according to claim 1, wherein the refrigerant is liquid nitrogen.
5. The method for forming a bioscaffold according to claim 1, wherein the distance between the raw material slurry and the refrigerant is controlled to 2 to 20 mm.
6. The method for forming a bioscaffold according to claim 1, wherein the drying comprises 5 stages in sequence, respectively:
(1) freezing for 15min at-35-45 ℃;
(2) freeze-drying at-8-10 deg.C and vacuum degree of 0.2bar for 740-920 min;
(3) freeze-drying for 380-460 min at-10-13 deg.C and vacuum degree of 0.2 bar;
(4) freeze-drying at-5-0 deg.C and vacuum degree of 0.2bar for 400-520 min;
(5) drying for 60-100 min at 20-30 ℃ and under the vacuum degree of 0.2 bar.
7. A bioscaffold obtainable by the molding process of any one of claims 1 to 6.
8. Use of the biological scaffold according to claim 7 for repair of nerve, tendon, ligament, skin, meninges, mucosal damage, for non-disease diagnosis and therapeutic purposes.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102078641A (en) * | 2011-01-07 | 2011-06-01 | 北京天新福医疗器材有限公司 | Method for preparing anti-adhesive membrane for tendon ligaments |
| CN104922732A (en) * | 2014-03-19 | 2015-09-23 | 北京天新福医疗器材有限公司 | Preparation method of oral biofilm |
| CN107050519A (en) * | 2016-12-22 | 2017-08-18 | 天新福（北京）医疗器材股份有限公司 | The preparation method of cladding Absorbable membranes |
| WO2019165910A1 (en) * | 2018-02-27 | 2019-09-06 | 上海微创心通医疗科技有限公司 | Dry biological tissue material and preparation method therefor |
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- 2021-12-13 CN CN202111519206.1A patent/CN114177358B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102078641A (en) * | 2011-01-07 | 2011-06-01 | 北京天新福医疗器材有限公司 | Method for preparing anti-adhesive membrane for tendon ligaments |
| CN104922732A (en) * | 2014-03-19 | 2015-09-23 | 北京天新福医疗器材有限公司 | Preparation method of oral biofilm |
| CN107050519A (en) * | 2016-12-22 | 2017-08-18 | 天新福（北京）医疗器材股份有限公司 | The preparation method of cladding Absorbable membranes |
| WO2019165910A1 (en) * | 2018-02-27 | 2019-09-06 | 上海微创心通医疗科技有限公司 | Dry biological tissue material and preparation method therefor |
Non-Patent Citations (1)
| Title |
|---|
| BRENDAN A.HARLEY ET AL.: "Fabricating tubular scaffolds with a radial pore size gradient by a spinning technique", 《BIOMATERIALS》 * |
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