CN106693057A - Cell-biological scaffold complex and 3D printing forming method thereof - Google Patents
Cell-biological scaffold complex and 3D printing forming method thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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
- A61L27/38—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 containing added animal cells
- A61L27/3804—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 containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
- A61L27/3821—Bone-forming cells, e.g. osteoblasts, osteocytes, osteoprogenitor cells
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- 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
- A61L27/38—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 containing added animal cells
- A61L27/3804—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 containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
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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
- A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
- A61L27/42—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having an inorganic matrix
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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
- A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
- A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
- A61L27/48—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix with macromolecular fillers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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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- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y10/00—Processes of additive manufacturing
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Abstract
The invention discloses a cell-biological scaffold complex and a 3D printing forming method thereof. The cell-biological scaffold complex comprises a biological scaffold and a cell-biological gel suspension arranged on the biological scaffold, wherein the biological scaffold comprises a one of or a combination of several of a biological macromolecular material, a natural biological material and an inorganic material, the biological macromolecular material comprises one of or a combination of several of calcium phosphate, polycaprolactone, polytetrafluoroethylene and lactic acid-hydroxyacetic acid copolymer, the natural biological material comprises one of or a combination of two of gelatin and sodium alginate, the inorganic material comprises one of or a combination of several of calcium sulfate, boron silicate and nano hydroxylapatite, the cell-biological gel suspension comprises cell and biological gel, and the cell is a bone cell or a corneal stroma. By adopting the cell-biological scaffold complex and the 3D printing forming method, a complexing efficiency of the cell and the biological gel suspension can be increased, the loss or dropping of a great amount of cells is avoided, the preparation process is simple, and the use is convenient.
Description
Technical field
The present invention relates to biological technical field, more particularly to a kind of cell-biological support complex and its 3D printing shaping
Method.
Background technology
Repair of tissue defect and organ transplant are one of greatest breakthroughs in 20th century medical history, and at present, the whole world has
Nearly 1,000,000 once obtained new life by the patient of dead gloom by organ transplant, but due to cannot get donor organ
The patient that organ transplant cannot be carried out and die be up to several ten million, it is necessary to the patient of Repair of tissue defect be more up to it is more than one hundred million
People, so it is the century of tissue repair and organ transplant that someone says 21 century, the appearance of 3D printing technique makes millions upon millions of to need group
Knit the hope repaired and obtained with the patient of organ transplant and cure and live again.
The patent document of Application No. 201510317506.X discloses a kind of cell-biological support complex and its 3D beats
Transfer molding method, for 3D printing technique to be combined with electrostatic spinning technique, promotes 3D biometric print technologies in organizational project
The application prospect in field, makes the patient for needing tissue repair and organ transplant obtain the hope cured and live again, but, it is biological
Support is low with the combined efficiency of cell, and cell is easily lost or largely comes off, and required incubation time is more long, therefore, Wo Menti
Having gone out a kind of cell-biological support complex and its 3D printing manufacturing process is used to solve the above problems.
The content of the invention
Based on the technical problem that background technology is present, the present invention proposes cell-biological support complex and its 3D printing
Manufacturing process, low with the combined efficiency of cell for solving biological support, cell is easily lost or largely comes off, required culture
Time problem more long.
Cell proposed by the present invention-biological support complex, including biological support and be arranged on biological support cell-
Biogel suspension.
Preferably, one kind of the biological support including bioabsorbable polymer material, natural biologic material and inorganic material or
Various combinations, bioabsorbable polymer material includes calcium phosphate, polycaprolactone, polytetrafluoroethylene (PTFE) and poly lactic coglycolic acid
One or more of combination, natural biologic material includes one or two combination, inorganic material in gelatin and sodium alginate
One or more including calcium sulfate, borosilicic acid and nanometer hydroxyapatite of combination.
Preferably, the cell-biogel suspension includes cell and biogel liquid, and cell is Gegenbaur's cell or angle
Membrane matrix cell, and biogel liquid includes the glacial acetic acid, 20-25 parts of shitosan, 10-15 parts that weight portion is 12-16 parts
NaOH and 8-12 parts of triethanolamine.
The invention allows for the 3D printing manufacturing process of cell-biological support complex, comprise the following steps：
S1：The preparing raw material of biological support is well mixed, will be well mixed with the support printing head of biometric print machine
The preparing raw material of biological support be successively processed as the biological support similar to the actual defect situation of histoorgan；
S2：Shitosan is crushed, 70-80h in glacial acetic acid is then soaked in, fully vibrated once per 5-8h, then
With the centrifugation of 11000-13000rpm/min, supernatant is taken；By in NaOH addition supernatant, adjusting its pH value is
6.5-7.5, then again with the centrifugation of 11000-13000rpm/min, removes supernatant, is subsequently added into glacial acetic acid and three
Monoethanolamine, after being well mixed, is obtained biogel liquid；
S3：Biogel liquid is diluted to required protein concentration with glacial acetic acid, NaOH is subsequently adding, its pH value is adjusted
It is 8-8.7, in the biogel liquid after then weight portion is modulated for 30-35 parts of nutrient solution and the addition of 25-30 parts of serum,
After well mixed, pre- solidifying biogel liquid is obtained, then pre- solidifying biogel liquid and cell are put into centrifuge tube, blown with dropper
Beat and fully mix, cell-biogel suspension is obtained；
S4：The cell printing shower nozzle of cell-biogel suspension biometric print machine is injected on biological support, so
Biological support is inserted in culture dish afterwards, then to nutrient solution needed for being added in culture dish, then culture dish is put into rotary three
Culture in dimension incubator, changes liquid once in two days, after cultivating ten days, cell-biological support complex is obtained.
Preferably, in the S2, shitosan is crushed, is then soaked in 72-78h in glacial acetic acid, it is abundant per 6-7h
Vibration once, then with the centrifugation of 11500-12500rpm/min, takes supernatant；By NaOH add supernatant in,
Its pH value is adjusted for 6.7-7.3, then again with the centrifugation of 11500-12500rpm/min, supernatant is removed, then added
Enter glacial acetic acid and triethanolamine, after being well mixed, biogel liquid is obtained.
Preferably, in the S3, biogel liquid is diluted to required protein concentration with glacial acetic acid, is subsequently adding hydroxide
Sodium, adjusts its pH value for 8.2-8.5, after weight portion then is added into modulation for 31-34 parts of nutrient solution and 26-28 parts of serum
Biogel liquid in, be well mixed after, obtain pre- solidifying biogel liquid, then by pre- solidifying biogel liquid and cell be put into from
In heart pipe, blown and beaten with dropper and fully mixed, cell-biogel suspension is obtained.
In the present invention, the cell-biological support complex and its 3D printing manufacturing process are by using biometric print machine
The preparing raw material of well mixed biological support is successively processed as support printing head the actual defect situation with histoorgan
Similar biological support can precise Printing go out patient needed for biological support, given birth to by by cell-biogel suspension
Cell-biological support that the cell printing shower nozzle of thing printer is injected in needed for patient being accurately obtained on biological support is combined
Body, the combined efficiency of cell and biological support can be improved by cell-biogel suspension, it is to avoid cell loss is a large amount of
Come off, and required incubation time can be reduced, the present invention can improve the combined efficiency of cell and biological support, it is to avoid cell is lost
Lose or largely come off, preparation process is simple is easy to use.
Specific embodiment
The present invention is made with reference to specific embodiment further explain.
Embodiment one
The present embodiment proposes cell-biological support complex, including biological support and is arranged at thin on biological support
Born of the same parents-biogel suspension, the biological support includes one kind of bioabsorbable polymer material, natural biologic material and inorganic material
Or various combinations, bioabsorbable polymer material include calcium phosphate, polycaprolactone, polytetrafluoroethylene (PTFE) and lactic-co-glycolic acid copolymerization
The combination of one or more of thing, natural biologic material includes one or two combination, inorganic material in gelatin and sodium alginate
Material includes one or more of calcium sulfate, borosilicic acid and nanometer hydroxyapatite of combination, the cell-biogel suspension
Including cell and biogel liquid, cell is Gegenbaur's cell or keratocyte, and biogel liquid includes that weight portion is 12
Part glacial acetic acid, 20 parts of shitosan, 10 parts of NaOH and 8 parts of triethanolamine.
The present embodiment also proposed the 3D printing manufacturing process of cell-biological support complex, comprise the following steps：
S1：The preparing raw material of biological support is well mixed, will be well mixed with the support printing head of biometric print machine
The preparing raw material of biological support be successively processed as the biological support similar to the actual defect situation of histoorgan；
S2：Shitosan is crushed, 70h in glacial acetic acid is then soaked in, fully vibrated once per 5h, then with
The centrifugation of 11000rpm/min, takes supernatant；By in NaOH addition supernatant, it is 6.5, Ran Houzai to adjust its pH value
The secondary centrifugation with 11000rpm/min, removes supernatant, is subsequently added into glacial acetic acid and triethanolamine, after being well mixed, system
Obtain biogel liquid；
S3：Biogel liquid is diluted to required protein concentration with glacial acetic acid, NaOH is subsequently adding, its pH value is adjusted
It is 8, during nutrient solution that weight portion is 30 parts and 25 parts of serum then added into the biogel liquid after modulation, is well mixed
Afterwards, pre- solidifying biogel liquid is obtained, then pre- solidifying biogel liquid and cell is put into centrifuge tube, blown and beaten with dropper fully mixed
It is even, cell-biogel suspension is obtained；
S4：The cell printing shower nozzle of cell-biogel suspension biometric print machine is injected on biological support, so
Biological support is inserted in culture dish afterwards, then to nutrient solution needed for being added in culture dish, then culture dish is put into rotary three
Culture in dimension incubator, changes liquid once in two days, after cultivating ten days, cell-biological support complex is obtained.
Embodiment two
The present embodiment proposes cell-biological support complex, including biological support and is arranged at thin on biological support
Born of the same parents-biogel suspension, the biological support includes one kind of bioabsorbable polymer material, natural biologic material and inorganic material
Or various combinations, bioabsorbable polymer material include calcium phosphate, polycaprolactone, polytetrafluoroethylene (PTFE) and lactic-co-glycolic acid copolymerization
The combination of one or more of thing, natural biologic material includes one or two combination, inorganic material in gelatin and sodium alginate
Material includes one or more of calcium sulfate, borosilicic acid and nanometer hydroxyapatite of combination, the cell-biogel suspension
Including cell and biogel liquid, cell is Gegenbaur's cell or keratocyte, and biogel liquid includes that weight portion is 16
Part glacial acetic acid, 25 parts of shitosan, 15 parts of NaOH and 12 parts of triethanolamine.
The present embodiment also proposed the 3D printing manufacturing process of cell-biological support complex, comprise the following steps：
S1：The preparing raw material of biological support is well mixed, will be well mixed with the support printing head of biometric print machine
The preparing raw material of biological support be successively processed as the biological support similar to the actual defect situation of histoorgan；
S2：Shitosan is crushed, 80h in glacial acetic acid is then soaked in, fully vibrated once per 8h, then with
The centrifugation of 13000rpm/min, takes supernatant；By in NaOH addition supernatant, it is 7.5, Ran Houzai to adjust its pH value
The secondary centrifugation with 13000rpm/min, removes supernatant, is subsequently added into glacial acetic acid and triethanolamine, after being well mixed, system
Obtain biogel liquid；
S3：Biogel liquid is diluted to required protein concentration with glacial acetic acid, NaOH is subsequently adding, its pH value is adjusted
It is 8.7, during nutrient solution that weight portion is 35 parts and 30 parts of serum then added into the biogel liquid after modulation, is well mixed
Afterwards, pre- solidifying biogel liquid is obtained, then pre- solidifying biogel liquid and cell is put into centrifuge tube, blown and beaten with dropper fully mixed
It is even, cell-biogel suspension is obtained；
S4：The cell printing shower nozzle of cell-biogel suspension biometric print machine is injected on biological support, so
Biological support is inserted in culture dish afterwards, then to nutrient solution needed for being added in culture dish, then culture dish is put into rotary three
Culture in dimension incubator, changes liquid once in two days, after cultivating ten days, cell-biological support complex is obtained.
Comparative example one, embodiment two and control group, the cell of embodiment two compare with the combined efficiency of biological support
It is 5.3%, the cell and biological support of embodiment one that the cell of embodiment one improves percentage with the combined efficiency of biological support
Combined efficiency compare the cell of control group to improve percentage with the combined efficiency of biological support be 11.4%, therefore, embodiment
Two is most preferred embodiment.
The above, the only present invention preferably specific embodiment, but protection scope of the present invention is not limited thereto,
Any one skilled in the art the invention discloses technical scope in, technology according to the present invention scheme and its
Inventive concept is subject to equivalent or change, should all be included within the scope of the present invention.
Claims (6)
1. cell-biological support complex, it is characterised in that including biological support and the cell-life being arranged on biological support
Thing gel suspension.
2. cell according to claim 1-biological support complex, it is characterised in that the biological support includes biology
The combination of one or more of macromolecular material, natural biologic material and inorganic material, bioabsorbable polymer material include calcium phosphate,
The combination of one or more of polycaprolactone, polytetrafluoroethylene (PTFE) and poly lactic coglycolic acid, natural biologic material includes bright
One or two combination in glue and sodium alginate, inorganic material includes the one of calcium sulfate, borosilicic acid and nanometer hydroxyapatite
Plant or various combinations.
3. cell according to claim 1-biological support complex, it is characterised in that the cell-biogel is suspended
Liquid includes cell and biogel liquid, and cell is Gegenbaur's cell or keratocyte, and biogel liquid is including weight portion
12-16 parts of glacial acetic acid, 20-25 parts of shitosan, 10-15 parts of NaOH and 8-12 parts of triethanolamine.
4. the 3D printing manufacturing process of cell-biological support complex, it is characterised in that comprise the following steps：
S1：The preparing raw material of biological support is well mixed, the life that will be well mixed with the support printing head of biometric print machine
The preparing raw material of thing support is successively processed as the biological support similar to the actual defect situation of histoorgan；
S2：Shitosan is crushed, 70-80h in glacial acetic acid is then soaked in, fully vibrated once per 5-8h, then with
The centrifugation of 11000-13000rpm/min, takes supernatant；By in NaOH addition supernatant, its pH value is adjusted for 6.5-
7.5, then again with the centrifugation of 11000-13000rpm/min, supernatant is removed, it is subsequently added into glacial acetic acid and three ethanol
Amine, after being well mixed, is obtained biogel liquid；
S3：Biogel liquid is diluted to required protein concentration with glacial acetic acid, NaOH is subsequently adding, its pH value is adjusted for 8-
8.7, in the biogel liquid after then weight portion is modulated for 30-35 parts of nutrient solution and the addition of 25-30 parts of serum, mixing
After uniform, pre- solidifying biogel liquid is obtained, then pre- solidifying biogel liquid and cell are put into centrifuge tube, blown and beaten with dropper and filled
Divide and mix, cell-biogel suspension is obtained；
S4：The cell printing shower nozzle of cell-biogel suspension biometric print machine is injected on biological support, then will
Biological support is inserted in culture dish, then to nutrient solution needed for being added in culture dish, then culture dish is put into Rotary three-dimensional training
Culture in device is supported, liquid is changed once within two days, after cultivating ten days, cell-biological support complex is obtained.
5. the 3D printing manufacturing process of cell according to claim 4-biological support complex, it is characterised in that described
In S2, shitosan is crushed, is then soaked in 72-78h in glacial acetic acid, fully vibrated once per 6-7h, then with
The centrifugation of 11500-12500rpm/min, takes supernatant；By in NaOH addition supernatant, its pH value is adjusted for 6.7-
7.3, then again with the centrifugation of 11500-12500rpm/min, supernatant is removed, it is subsequently added into glacial acetic acid and three ethanol
Amine, after being well mixed, is obtained biogel liquid.
6. the 3D printing manufacturing process of cell according to claim 4-biological support complex, it is characterised in that described
In S3, biogel liquid is diluted to required protein concentration with glacial acetic acid, is subsequently adding NaOH, adjust its pH value for 8.2-
8.5, in the biogel liquid after then weight portion is modulated for 31-34 parts of nutrient solution and the addition of 26-28 parts of serum, mixing
After uniform, pre- solidifying biogel liquid is obtained, then pre- solidifying biogel liquid and cell are put into centrifuge tube, blown and beaten with dropper and filled
Divide and mix, cell-biogel suspension is obtained.
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Cited By (7)
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| CN107115561A (en) * | 2017-06-19 | 2017-09-01 | 中山大学附属口腔医院 | A kind of 3D printing timbering material and its preparation method and application |
| CN108144114A (en) * | 2018-01-05 | 2018-06-12 | 浙江省医学科学院 | For the 3D printing material of organizational project and the preparation method of Biodegradable scaffold material |
| CN108297396A (en) * | 2018-02-07 | 2018-07-20 | 华中科技大学鄂州工业技术研究院 | A method of it squeezing out sedimentation type 3D printing and prepares three-dimensional bone tissue engineering scaffold |
| CN108327241A (en) * | 2018-01-28 | 2018-07-27 | 浙江大学 | A kind of manufacturing method of controllable antibacterial trachea bracket |
| CN109821075A (en) * | 2019-01-07 | 2019-05-31 | 华中科技大学 | A kind of biomaterial and preparation method thereof and the application as bone impairment renovation material |
| CN109999226A (en) * | 2018-11-27 | 2019-07-12 | 王翀 | A kind of printed material, the bone tissue engineering scaffold and preparation method for loading stem cell |
| CN111249528A (en) * | 2020-01-20 | 2020-06-09 | 浙江大学 | Tissue engineering bone based on multilayer cell grid and preparation method thereof |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107115561A (en) * | 2017-06-19 | 2017-09-01 | 中山大学附属口腔医院 | A kind of 3D printing timbering material and its preparation method and application |
| CN107115561B (en) * | 2017-06-19 | 2019-11-26 | 中山大学附属口腔医院 | A kind of 3D printing timbering material and its preparation method and application |
| CN108144114A (en) * | 2018-01-05 | 2018-06-12 | 浙江省医学科学院 | For the 3D printing material of organizational project and the preparation method of Biodegradable scaffold material |
| CN108327241A (en) * | 2018-01-28 | 2018-07-27 | 浙江大学 | A kind of manufacturing method of controllable antibacterial trachea bracket |
| CN108327241B (en) * | 2018-01-28 | 2019-09-20 | 浙江大学 | A kind of manufacturing method of controllable antibacterial trachea bracket |
| CN108297396A (en) * | 2018-02-07 | 2018-07-20 | 华中科技大学鄂州工业技术研究院 | A method of it squeezing out sedimentation type 3D printing and prepares three-dimensional bone tissue engineering scaffold |
| CN108297396B (en) * | 2018-02-07 | 2020-01-21 | 华中科技大学鄂州工业技术研究院 | Method for preparing three-dimensional bone tissue engineering scaffold through extrusion deposition type 3D printing |
| CN109999226A (en) * | 2018-11-27 | 2019-07-12 | 王翀 | A kind of printed material, the bone tissue engineering scaffold and preparation method for loading stem cell |
| CN109821075A (en) * | 2019-01-07 | 2019-05-31 | 华中科技大学 | A kind of biomaterial and preparation method thereof and the application as bone impairment renovation material |
| CN111249528A (en) * | 2020-01-20 | 2020-06-09 | 浙江大学 | Tissue engineering bone based on multilayer cell grid and preparation method thereof |
| CN111249528B (en) * | 2020-01-20 | 2021-07-16 | 浙江大学 | Tissue engineering bone based on multilayer cell grid and preparation method thereof |
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