CN110507853A - A kind of method of biological 3 D-printing interverbebral disc in support liquid medium - Google Patents
A kind of method of biological 3 D-printing interverbebral disc in support liquid medium Download PDFInfo
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- CN110507853A CN110507853A CN201910837486.7A CN201910837486A CN110507853A CN 110507853 A CN110507853 A CN 110507853A CN 201910837486 A CN201910837486 A CN 201910837486A CN 110507853 A CN110507853 A CN 110507853A
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Classifications
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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
- A61L27/36—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
- A61L27/3817—Cartilage-forming cells, e.g. pre-chondrocytes
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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
- A61L27/38—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
- A61L27/3834—Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
- B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
- B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
- B29C64/10—Processes of additive manufacturing
- B29C64/106—Processes of additive manufacturing using only liquids or viscous materials, e.g. depositing a continuous bead of viscous material
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
- B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
- B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
- B29C64/30—Auxiliary operations or equipment
- B29C64/386—Data acquisition or data processing for additive manufacturing
- B29C64/393—Data acquisition or data processing for additive manufacturing for controlling or regulating additive manufacturing processes
-
- 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
- B33Y50/00—Data acquisition or data processing for additive manufacturing
- B33Y50/02—Data acquisition or data processing for additive manufacturing for controlling or regulating additive manufacturing processes
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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
- B33Y70/00—Materials specially adapted for additive manufacturing
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
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- Health & Medical Sciences (AREA)
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- Manufacturing & Machinery (AREA)
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- General Health & Medical Sciences (AREA)
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Abstract
The invention discloses a kind of method of 3 D-printing interverbebral disc biological in support liquid medium, the support liquid medium of biological 3 D-printing interverbebral disc is made of the raw material of following mass fraction in the support liquid medium: chitosan 5-50, alginic acid 5-50, cyclodextrin 5-50, starch 5-50, albumin 5-50, gelatin 5-50, solvent 5-50.The utility model has the advantages that printing path can be accurately controlled, the stability of print structure is greatlyd improve；And use the printing liquid containing seed cell, us can be made to obtain the interverbebral disc biological support for containing a large amount of cells, as needed, printing path parameter is designed, printing obtains artificial bionic interverbebral disc biological support, and this method interverbebral disc biological support obtained has endplate cartilage-fibrous ring-core structure, the precision of structure is high, carrying a large amount of seed cells carries out proliferation reparation simultaneously, conducive to interverbebral disc surrounding microenvironment, reconstruction of intervertebral disc structure and function is quickly established.
Description
Technical field
The present invention relates to three-dimensional printing technology fields, it particularly relates to which biology three-dimensional is beaten in a kind of support liquid medium
Print the method for interverbebral disc.
Background technique
Lumber ertebral disc is made of nucleus pulposus, fibrous ring and cartilage plate three parts, it is transported between upper and lower two centrum in lumbar vertebrae
Play the role of support rotation in dynamic, reduces coefficient of friction.The protrasion of the lumbar intervertebral disci is relatively conventional kinematic system damage,
Disease incidence rises year by year trend.The cause of disease is mainly to induce after a degree of degenerative change occurs for lumber ertebral disc in external force
Lower Anular disruption, nucleus pulposus oppress canalis spinalis inner tissue, cause adjacent spinal nerve root to be stimulated or oppressed, to generate waist
Portion's pain, side lower limb or double lower limb numbness, pain etc..
Non-operative treatment and operative treatment two major classes are divided into the treatment of protrusion of lumber intervertebral disc at present, non-operative treatment is main
Including drug therapy, traction therapeutic, physical therapy, closed treating etc., such method predominantly alleviate symptom, curative effect lasting time compared with
It is short, it can not eradicate.Operative treatment includes conventional open operation, and Minimally Invasive Surgery, percutaneous puncture cuts suction, artificial disc replacement etc..
Wherein, artificial disc replacement can retain the mobility between vertebra section.The artificial intervertebral disk of clinical use is mainly gold at present
Belong to material to be made.Tissue engineering technique provides a kind of side that is ideal, meeting physilogical characteristics for the preparation of clinical artificial intervertebral disk
Method.With tissue engineering technique, i.e., a small amount of seed cell is cultivated in vitro, after amplification and biological support is combined into compound,
After being implanted into diseased region, cell Proliferation, differentiation, secretion matrix finally make the constructor close physilogical characteristics.The compound
It plays a supporting role in organizational project, while providing the place of boarding, growth, differentiation and proliferation for cell.Biological support with
Seed cell jointly guides the regeneration of damaged tissues and controls the structure of regenerating tissues, is the pass of clinical treatment
Key.
For the problems in the relevant technologies, currently no effective solution has been proposed.
Summary of the invention
For the problems in the relevant technologies, the present invention proposes biological 3 D-printing interverbebral disc in a kind of support liquid medium
Method, to overcome above-mentioned technical problem present in existing the relevant technologies.
For this purpose, the specific technical solution that the present invention uses is as follows:
A kind of method of biological 3 D-printing interverbebral disc in support liquid medium, biology is three-dimensional in the support liquid medium beats
The support liquid medium of print interverbebral disc is made of the raw material of following mass fraction:
Chitosan 5-50, alginic acid 5-50, cyclodextrin 5-50, starch 5-50, albumin 5-50, gelatin 5-50, solvent 5-
50。
The method of biological 3 D-printing interverbebral disc in the support liquid medium, comprising the following steps:
The culture of seed cell: being incubated at 37 DEG C, in 5%CO2 incubator with the F12 culture medium containing 10% fetal calf serum, with
α-MEM culture medium containing 10% fetal calf serum, Connective Tissue Growth Factor CTGF 25ng/ml, ascorbic acid 25uM is incubated at 37
DEG C, in 5%CO2 incubator.
It supports the preparation of solvent: measuring suitable chitosan, alginic acid, cyclodextrin, starch, albumin and gelatin, dissolve
In solvent and adjust to required density.
The foundation of artificial intervertebral disk printer model: establishing printer model using 3 D-printing software, and outer layer is multi-layer annular,
Internal layer is multi-layer helical shape, and endplate cartilage is multilayer rectangle, saves, waits next step.
The preparation of artificial intervertebral disk printed material: suitable bioabsorbable polymer material A1, A2, A3, A4 are measured and is sterilized.Amount
It takes suitable natural material B1, B2, B3, B4 and sterilizes, mixed with seed cell, obtain waiting next step containing Cellular gels.
The preparation of artificial intervertebral disk printing device: the technological parameter of adjustment biology 3D printing equipment, print head diameter are
150-400 μm, print temperature is adjusted according to the hot attribute of material, and barrel air pressure is 600-1000KPa in print procedure, is beaten
Print structure is controlled by printing path.
The printing of artificial intervertebral disk: being respectively implanted two barrels for the high-molecular material A 1 of step 2 and B1 containing Cellular gels,
Starting device carries out 3 D-printing, printing alternate high molecular material and Cellular gels in S2 support solvent, supports and contain in solvent
B1 containing Cellular gels is crosslinked using proper method after crosslinking agent, or printing；When prepared by bottom soleplate version cartilage
Material in barrel is changed to A2, B2, printed to nucleus pulposus by Bi Hou；After by the material in barrel be changed to A3,
B3 prints fibrous ring；After the material in barrel is changed to A4, B4, top layer endplate cartilage is printed, often
It is crosslinked after the completion of step printing, method is same as above.
The culture of 3 D-printing artificial intervertebral disk: measuring suitable culture medium, puts after submerging the resulting artificial intervertebral disk of S3
Enter in carbon dioxide incubator and persistently cultivates.
Further, bioabsorbable polymer material A1, A2, A3, A4 is by polylactic acid, polycaprolactone, poly lactic-co-glycolic acid
One or more of copolymer, polylactic acid-caprolactone copolymer, polydioxanone are constituted.
Further, natural material B1, B2, B3, B4 is by gelatin, hyaluronic acid, sodium alginate, chitosan, collagen egg
It is one or more of white to constitute.
Further, seed cell is made of one or more of stem cell, cartilage cell.
Further, crosslinking agent is one or more of ultraviolet light, calcium chloride solution.
Further, after the completion of artificial intervertebral disk prints, seaweed then is removed using the solution containing alginate lyase
Hydrochlorate Supporting Media.
The invention has the benefit that this method uses biological three-dimensional printing technology in support liquid medium, it can be accurate
Control printing path, greatly improve the stability of print structure；And the printing liquid containing seed cell is used, us can be made to obtain
The interverbebral disc biological support containing a large amount of cells is obtained, as needed, designs printing path parameter, printing obtains artificial bionic intervertebral
Disk biological support, this method interverbebral disc biological support obtained have endplate cartilage-fibrous ring-core structure, the essence of structure
Degree is high, while carrying a large amount of seed cells and carrying out proliferation reparation, conducive to interverbebral disc surrounding microenvironment, reconstruction of intervertebral disc is quickly established
Structure and function.
Detailed description of the invention
It in order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, below will be to institute in embodiment
Attached drawing to be used is needed to be briefly described, it should be apparent that, the accompanying drawings in the following description is only some implementations of the invention
Example, for those of ordinary skill in the art, without creative efforts, can also obtain according to these attached drawings
Obtain other attached drawings.
Fig. 1 is the stream of the method for biological 3 D-printing interverbebral disc in a kind of support liquid medium according to an embodiment of the present invention
Cheng Tu；
Fig. 2 is the marrow of the method for biological 3 D-printing interverbebral disc in a kind of support liquid medium according to an embodiment of the present invention
The light absorption value comparative diagram in core area and annular region；
Fig. 3 is the marrow of the method for biological 3 D-printing interverbebral disc in a kind of support liquid medium according to an embodiment of the present invention
The associated products variation diagram in core area and annular region；
Fig. 4 is the marrow of the method for biological 3 D-printing interverbebral disc in a kind of support liquid medium according to an embodiment of the present invention
Core area and annular region mechanical property result figure.
Specific embodiment
To further illustrate that each embodiment, the present invention are provided with attached drawing, these attached drawings are that the invention discloses one of content
Point, mainly to illustrate embodiment, and the associated description of specification can be cooperated to explain the operation principles of embodiment, cooperation ginseng
Examine these contents, those of ordinary skill in the art will be understood that other possible embodiments and advantages of the present invention, in figure
Component be not necessarily to scale, and similar component symbol is conventionally used to indicate similar component.
According to an embodiment of the invention, providing a kind of method of biological 3 D-printing interverbebral disc in support liquid medium.
In the support liquid medium support liquid medium of biological 3 D-printing interverbebral disc by following mass fraction raw material
It is made:
Chitosan 5-50, alginic acid 5-50, cyclodextrin 5-50, starch 5-50, albumin 5-50, gelatin 5-50, solvent 5-
50。
Embodiment one:
The method of biological 3 D-printing interverbebral disc in the support liquid medium, comprising the following steps:
The culture of seed cell: step S101 is incubated at 37 DEG C, 5%CO2 with the F12 culture medium containing 10% fetal calf serum
In incubator, with the α-MEM culture medium containing 10% fetal calf serum, Connective Tissue Growth Factor CTGF25ng/ml, ascorbic acid 25uM
It is incubated at 37 DEG C, in 5%CO2 incubator.
Step S103 supports the preparation of solvent: measure suitable chitosan, alginic acid, cyclodextrin, starch, albumin and
Gelatin is dissolved in solvent and adjusts to required density.
The foundation of artificial intervertebral disk printer model: step S105 establishes printer model using 3 D-printing software, outer layer is
Multi-layer annular, internal layer are multi-layer helical shape, and endplate cartilage is multilayer rectangle, save, wait next step.
The preparation of artificial intervertebral disk printed material: step S107 measures suitable bioabsorbable polymer material A1, A2, A3, A4
And it sterilizes.It measures suitable natural material B1, B2, B3, B4 and sterilizes, mixed with seed cell, obtain waiting containing Cellular gels
Next step.
Step S109, the preparation of artificial intervertebral disk printing device: the technological parameter of adjustment biology 3D printing equipment, printing
Head diameter is 150-400 μm, and print temperature is adjusted according to the hot attribute of material, and barrel air pressure is 600- in print procedure
1000KPa, print structure are controlled by printing path.
The printing of artificial intervertebral disk: step S111 the high-molecular material A 1 of step 2 and B1 containing Cellular gels is set respectively
Enter two barrels, starting device carries out 3 D-printing, printing alternate high molecular material and Cellular gels in S2 support solvent, props up
It supports and B1 containing Cellular gels is crosslinked using proper method after containing crosslinking agent, or printing in solvent；When bottom soleplate version
After prepared by cartilage, the material in barrel is changed to A2, B2, nucleus pulposus is printed；After by the material in barrel
It is changed to A3, B3, fibrous ring is printed；After the material in barrel is changed to A4, B4, to top layer endplate cartilage into
Row printing is crosslinked after the completion of every step printing, and method is same as above.
Step S113, the culture of 3 D-printing artificial intervertebral disk: measuring suitable culture medium, submerges the resulting artificial vertebra of S3
It is put into carbon dioxide incubator after disk and persistently cultivates.
Embodiment two:
Bioabsorbable polymer material A1, A2, A3, A4 are by polylactic acid, polycaprolactone, poly lactide-glycolide acid, gather
One or more of lactic acid-caprol acton copolymer, polydioxanone constitute, further, natural material B1, B2, B3,
B4 is made of one or more of gelatin, hyaluronic acid, sodium alginate, chitosan, collagen, and seed cell is by bone
One or more of marrow stem cell, cartilage cell constitute, crosslinking agent be one or more of ultraviolet light, calcium chloride solution,
After the completion of artificial intervertebral disk printing, alginate Supporting Media then is removed using the solution containing alginate lyase.
Embodiment three:
(1) printer model is established using 3 D-printing software, outer layer is multilayer loop shape, and internal layer is multi-layer helical shape, marrow
Core area, annular area path clearance are different, save, and wait next step；
(2) suitable alginate, xanthan gum, calcium carbonate are measured, D- (+) glucono-delta-lactone and distilled water are dissolved in
In, and adjust to required density；
(3) it measures suitable polycaprolactone and sterilizes.It measures suitable gelatin and sterilizes, mixed with stem cell
It is even, gelatin gel containing stem cell is obtained, next step is waited；
(4) preparation of equipment: the technological parameter of adjustment biology 3D printing equipment: print head diameter is 100 μm, is gathered in oneself
Ester print temperature is 120 DEG C, and barrel air pressure is 600-1000KPa in 18-37 DEG C of print procedure of print temperature containing Cellular gels, is beaten
Print structure is controlled by printing path.Two barrels are respectively implanted by high molecular material polycaprolactone and containing Cellular gels, starting device,
3 D-printing, printing alternate high molecular material and Cellular gels are carried out in support solvent, supports in solvent and has contained crosslinking agent calcium
Ion can be crosslinked to containing Cellular gels simultaneously in printing.After the preparation of nuclear area, modification print head diameter is 200 μ
M prints fibrous ring, obtains artificial bionic interverbebral disc.Then alginic acid is removed using the solution containing alginate lyase
Salt supports liquid.
Example IV:
Artificial bionic interverbebral disc obtained in above-described embodiment 3 is subjected to biocompatibility characterization.By gained artificial bionic
Interverbebral disc is put into incubator and cultivates.With the interverbebral disc printed without Cellular gels, plantation cartilage is control group.It trains respectively
After supporting 1,3,5,7 days, the cell viability of mtt assay detection interverbebral disc is utilized.As indicated with 2, display is artificial with cell for result figure
Cell on bionical interverbebral disc has good proliferation behavior, and cell viability is preferable, and more significant compared with control group proliferation behavior.
Embodiment five:
It will be carried out in above-described embodiment 3 using the artificial bionic interverbebral disc that 3 D-printing biological in support liquid medium obtains
QRT-PCR detection.With the interverbebral disc printed without Cellular gels, plantation cartilage compares for control group.It cultivates 7 days respectively
Afterwards, qRT-PCR detection is carried out.Result figure as indicated at 3, display experimental group cartilage area cartilage associated products (SOX9, Aggrecan,
COL II) expression is obvious more.
Embodiment six:
Artificial intervertebral disk obtained in above-described embodiment 3 is placed in progress mechanics compression verification on omnipotent test machine, respectively
Test nuclear area and annular region mechanical property.It is as shown in Table 1 to measure result, as the result is shown artificial intervertebral disk mechanical property
Preferably.
In conclusion this method is three-dimensional using biology in support liquid medium by means of above-mentioned technical proposal of the invention
Printing technique can be accurately controlled printing path, greatly improve the stability of print structure；And it uses containing seed cell
Liquid is printed, us can be made to obtain the interverbebral disc biological support for containing a large amount of cells, as needed, printing path parameter is designed, beats
Print obtains artificial bionic interverbebral disc biological support, and this method interverbebral disc biological support obtained has endplate cartilage-fibrous ring-
The precision of core structure, structure is high, while carrying a large amount of seed cells and carrying out proliferation reparation, conducive to quickly establishing around interverbebral disc
Microenvironment, reconstruction of intervertebral disc structure and function.
In the present invention unless specifically defined or limited otherwise, term " installation ", " setting ", " connection ", " fixation ",
Terms such as " being screwed on " shall be understood in a broad sense, for example, it may be being fixedly connected, may be a detachable connection, or integral；It can be with
It is mechanical connection, is also possible to be electrically connected；It can be directly connected, two can also be can be indirectly connected through an intermediary
The interaction relationship of connection or two elements inside a element, unless otherwise restricted clearly, for the common of this field
For technical staff, the specific meanings of the above terms in the present invention can be understood according to specific conditions.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Within mind and principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (7)
1. a kind of method of biological 3 D-printing interverbebral disc in support liquid medium, which is characterized in that in the support liquid medium
The support liquid medium of biological 3 D-printing interverbebral disc is made of the raw material of following mass fraction:
Chitosan 5-50, alginic acid 5-50, cyclodextrin 5-50, starch 5-50, albumin 5-50, gelatin 5-50, solvent 5-50.
2. a kind of method of biological 3 D-printing interverbebral disc in support liquid medium, which is characterized in that in the support liquid medium
The method of biological 3 D-printing interverbebral disc, comprising the following steps:
The culture of seed cell: 37 DEG C are incubated at, in 5%CO2 incubator with the F12 culture medium containing 10% fetal calf serum, to contain
10% fetal calf serum, Connective Tissue Growth Factor CTGF 25ng/ml, ascorbic acid 25uM α-MEM culture medium be incubated at 37
DEG C, in 5%CO2 incubator.
It supports the preparation of solvent: measuring suitable chitosan, alginic acid, cyclodextrin, starch, albumin and gelatin, be dissolved in molten
In agent and adjust to required density.
The foundation of artificial intervertebral disk printer model: printer model is established using 3 D-printing software, outer layer is multi-layer annular, internal layer
For multi-layer helical shape, endplate cartilage is multilayer rectangle, saves, waits next step.
The preparation of artificial intervertebral disk printed material: suitable bioabsorbable polymer material A1, A2, A3, A4 are measured and is sterilized.It measures suitable
Natural material B1, B2, B3, B4 of amount simultaneously sterilize, and mix with seed cell, obtain waiting next step containing Cellular gels.
The preparation of artificial intervertebral disk printing device: the technological parameter of adjustment biology 3D printing equipment, print head diameter are 150-
400 μm, print temperature is adjusted according to the hot attribute of material, and barrel air pressure is 600-1000KPa, printing knot in print procedure
Structure is controlled by printing path.
The printing of artificial intervertebral disk: being respectively implanted two barrels for the high-molecular material A 1 of step 2 and B1 containing Cellular gels, starting
Equipment carries out 3 D-printing, printing alternate high molecular material and Cellular gels in S2 support solvent, supports in solvent containing crosslinking
B1 containing Cellular gels is crosslinked using proper method after agent, or printing；After the preparation of bottom soleplate version cartilage,
Material in barrel is changed to A2, B2, nucleus pulposus is printed；After the material in barrel is changed to A3, B3, to fibre
Dimension ring is printed；After the material in barrel is changed to A4, B4, top layer endplate cartilage is printed, every step printing
It is crosslinked after the completion, method is same as above.
The culture of 3 D-printing artificial intervertebral disk: measuring suitable culture medium, is put into two after submerging the resulting artificial intervertebral disk of S3
It is persistently cultivated in carbonoxide incubator.
3. the method for biological 3 D-printing interverbebral disc, feature exist in a kind of support liquid medium according to claim 2
In bioabsorbable polymer material A1, A2, A3, A4 are by polylactic acid, polycaprolactone, poly lactide-glycolide acid, polylactic acid-
One or more of caprolactone copolymer, polydioxanone are constituted.
4. the method for biological 3 D-printing interverbebral disc, feature exist in a kind of support liquid medium according to claim 1
In natural material B1, B2, B3, B4 are by one of gelatin, hyaluronic acid, sodium alginate, chitosan, collagen or several
Kind is constituted.
5. the method for biological 3 D-printing interverbebral disc, feature exist in a kind of support liquid medium according to claim 1
In seed cell is made of one or more of stem cell, cartilage cell.
6. the method for biological 3 D-printing interverbebral disc, feature exist in a kind of support liquid medium according to claim 1
In crosslinking agent is one or more of ultraviolet light, calcium chloride solution.
7. the method for biological 3 D-printing interverbebral disc, feature exist in a kind of support liquid medium according to claim 1
In after the completion of artificial intervertebral disk printing, then using the solution removal alginate Supporting Media containing alginate lyase.
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Cited By (3)
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| CN111346261A (en) * | 2020-02-25 | 2020-06-30 | 浙江大学 | Nucleus pulposus-like scaffold-stem cell combined material subjected to bilateral affinity modification |
| CN113576719A (en) * | 2021-07-14 | 2021-11-02 | 天津市天津医院 | Bionic microchannel integrated intervertebral disc stent and preparation method and application thereof |
| CN115923121A (en) * | 2022-12-05 | 2023-04-07 | 武汉大学 | 3D prints from perception dummy cervical vertebra |
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| US20070003525A1 (en) * | 2003-01-31 | 2007-01-04 | Moehlenbruck Jeffrey W | Hydrogel compositions comprising nucleus pulposus tissue |
| US20080195211A1 (en) * | 2006-10-30 | 2008-08-14 | Chia-Ying Lin | Engineered Scaffolds for Intervertebral Disc Repair and Regeneration and for Articulating Joint Repair and Regeneration |
| CN107139447A (en) * | 2017-05-12 | 2017-09-08 | 安徽大学 | 3D printer and method for 3D printing of sodium alginate hydrogel |
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| US20070003525A1 (en) * | 2003-01-31 | 2007-01-04 | Moehlenbruck Jeffrey W | Hydrogel compositions comprising nucleus pulposus tissue |
| US20080195211A1 (en) * | 2006-10-30 | 2008-08-14 | Chia-Ying Lin | Engineered Scaffolds for Intervertebral Disc Repair and Regeneration and for Articulating Joint Repair and Regeneration |
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| CN111346261A (en) * | 2020-02-25 | 2020-06-30 | 浙江大学 | Nucleus pulposus-like scaffold-stem cell combined material subjected to bilateral affinity modification |
| CN113576719A (en) * | 2021-07-14 | 2021-11-02 | 天津市天津医院 | Bionic microchannel integrated intervertebral disc stent and preparation method and application thereof |
| CN115923121A (en) * | 2022-12-05 | 2023-04-07 | 武汉大学 | 3D prints from perception dummy cervical vertebra |
| CN115923121B (en) * | 2022-12-05 | 2024-02-27 | 武汉大学 | 3D prints from perception dummy cervical vertebra |
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