CN109912303A - A kind of calcium phosphate/zirconium oxide gradient porous support and preparation method and application - Google Patents
A kind of calcium phosphate/zirconium oxide gradient porous support and preparation method and application Download PDFInfo
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- CN109912303A CN109912303A CN201910337763.8A CN201910337763A CN109912303A CN 109912303 A CN109912303 A CN 109912303A CN 201910337763 A CN201910337763 A CN 201910337763A CN 109912303 A CN109912303 A CN 109912303A
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- calcium phosphate
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Abstract
The present invention provides a kind of calcium phosphate/zirconium oxide gradient-controllable porous supports and its preparation method and application, this kind of porous support is shaped using calcium phosphate ceramics and Zirconium powder as raw material using photocuring technology, then are made after high temperature is degreasing sintered.Its aperture change of gradient between 10~1000 μm, porosity are 30%~90%, connectivity with higher.The present invention is not required to mold, it can be realized the change of gradient of bracket appearance, the personalized designs of hole and aperture, by controlling the ratio of ceramic powders, the mechanical property and bioactivity of compound rest can be adjusted, to obtain biological property well and there is the porous support of some strength.The complex stephanoporate bracket that the method is prepared has potential application space in bone tissue engineer field.
Description
Technical field
The present invention relates to a kind of calcium phosphate/zirconium oxide gradient-controllable porous support and preparation method and application, belong to biology
Medical material technical field.
Background technique
In Bone Defect Repari engineering, porous support usually has the function of support rib defect, induction New born formation.Cause
This, ideal porous support needs to have good biocompatibility and has some strength to restore Bones morphology and performance,
Meanwhile bracket will have certain solubility in human body environment, gradually osteocyte can be induced to grow into internal stent, realize self
It repairs.
Calcium phosphate ceramics and skeleton ingredient similarity are higher, are a kind of with good bioactive ceramics.From
Bone Defect Repari angle sees that calcium phosphate ceramics have good osteoconductive, and osteoblast is attached to such rack surface, Neng Goushi
It is now proliferated, breaks up, is mature, synthetic bone Collagen, further mineralising forms bone tissue.In addition, most of calcium phosphate ceramics
It can degrade in human body environment, the catabolites such as free calcium ion, phosphate can promote cell Growth and Differentiation, favorably
It is formed in bone tissue.Although calcium phosphate ceramics have superior biological property, there is certain lack in mechanical properties in it
It falls into, such as elasticity modulus is excessive compared with body bone tissue, the problems such as fracture toughness is too small compared with people's bone, various properties all limit
Application of the calcium phosphate ceramics as load-bearing bone renovating material is made.Zirconium oxide is a kind of excellent bio-inert ceramic, mechanics
Function admirable has wearability and chemical stability.However, there is no good cell and tissue affinities for zirconium oxide itself.
Nowadays, ceramic porous bracket can be manufactured by multiple technologies, such as addition pore creating material, foam impregnation, directly hair
Bubble, freeze-drying etc..All there are some problems in the above method, as, there are impurity residual, pore shape, distribution are difficult to after being sintered
Control, needs mold, and cost is higher etc..And the porous bone scaffold of clinical application at present is mostly uniform single aperture, and
Pore structure is more difficult to control.And density is varied everywhere in nature bone, is difficult to after the stenter to implant in single aperture and surrounding bone
The variable density of tissue is consistent, is easy the problems such as causing bone-loss.From the point of view of amechanical angle, the bracket of implantation needs one
Fixed mechanical property supports defect, meets normal activity.Porosity is lower for porous support, and intensity is higher,
But cell grows into bracket and higher porosity is needed to guarantee normal growth metabolism.Therefore, strong mechanical performance, high porosity this to lance
Shield requires to limit the extensive use of ceramic porous bracket.And alleviating this contradictory method is exactly to prepare gradient porous bracket,
This structure is expected to reach mechanical strength requirement and promote to obtain a balance between cell growth.In addition, cell growth,
Aperture needed for metabolism is also had nothing in common with each other, and macropore gauge structure can play nutrition supply, gas exchanges in cell growth process
The effects of with metabolic waste is removed, but will affect cell attachment and Cellular Signaling Transduction Mediated, and aperture gauge structure can provide
State opposite property.Specific aperture requires also related to cell category.In general, aperture size is 15~40 μm, allow to show
Micro-assembly robot is grown into；Aperture is 40~100 μm, and non-mineral class bone tissue is allowed to grow into；Aperture is greater than 100 μm, allows vascular tissue
It grows into (Du J, Zuo Y, Lin L, et al.J Mech Behav Biomed Mater, 2018,88:150-159.).Se
Heang Oh etc. has found that aperture is that 380~405 μm of brace sections are preferable to cartilage cell and Oesteoblast growth under study for action,
Aperture is that 186~200 μm of brace sections are preferable to fibroblastic growth, the brace sections new life bon e formation in 290~310 μm of aperture
Speed be significantly faster than that other apertures brace sections (Se Heang Oh, Il Kyu Park, et al.Biomaterials, 2007,
28:1664-1671.).And the initial stage of cell is adhered to, small-bore bracket is capable of providing higher adhesive surface product, and big
The bracket in aperture provides more sufficient space toward internal stent migration in later period cell.Based on considerations above, gradient porous branch
Frame is more in line with implantation and repairs requirement, can better meet the vegetative activity that cell carries out longer cycle.
It is more difficult that classical production process will produce gradient aperture structure.Optical soliton interaction ceramics printing technique, has
3D printing technique realizes the advantage of personalized customization, can be real while restoring subjects bones' primary morphology to a high degree
The printing of existing some specific structures such as special shape, gradient aperture.It is realized by computer drawing threedimensional model to material
Internal pore shape, size, cellular structure, the accurate control of the indexs such as porosity, thus preferably balancing material intensity and
Relationship between porosity, connected ratio provides greater flexibility and speed for the preparation of porous material.And this method disappears
In addition to mold limits, cost of investment is reduced to a certain extent.
Summary of the invention
For the above-mentioned problems of the prior art and deficiency, the present invention provides a kind of calcium phosphate/zirconium oxide gradient-controllable
Porous support and preparation method and application.The complex stephanoporate bracket using calcium phosphate ceramics and zirconium oxide as porous support matrix,
By computer drawing gradient pore scaffold three-dimensional model, body formation and the degreasing sintered legal system of high temperature are realized using photocuring technology
Obtain bracket finished product.The bracket can realize that gradient aperture customizes design under the conditions of no mold, with certain intensity and good
Good bioactivity, can be applied to bone tissue engineer field, as bone filler, substitute or as cultured cell in vitro branch
Frame.
The technical solution adopted by the invention is as follows:
A kind of preparation method of calcium phosphate/zirconium oxide gradient-controllable porous support, the specific steps of which are as follows:
Step 1, modeling: gradient porous scaffold three-dimensional figure is designed with modeling software, saves as STL formatted file；
Step 2, ball mill mixing: calcium phosphate ceramics powder, Zirconium powder are mixed, ball milling in planetary ball mill is placed in；
Step 3, slurry preparation: photosensitive resin is added in the hybrid ceramic powder that step 2 is obtained and dispersant is uniform；
Step 4, body formation: the model file that step 1 is obtained imports photo solidification machine, and the slurry that step 3 is obtained is used up solid
The method of chemical conversion shape prints porous support green body；
Step 5, ultrasonic cleaning: porous support green body being added to absolute ethanol, sets in supersonic cleaning machine and cleans, and removal surface is not
Cure pastes；
Step 6, drying: the bracket green body that step 5 obtains is put into drying box dry；
Step 7, degreasing sintered: the bracket base substrate degreasing that step 6 is obtained is sintered, and is taken out after furnace cooling to room temperature and is obtained phosphoric acid
Calcium/zirconium oxide gradient porous support.
In the step 2, calcium phosphate ceramics include hydroxyapatite, tricalcium phosphate and biphasic calcium phosphate, fluor-apatite,
At least one of podolite.
In the step 2, selected calcium phosphate ceramics, Zirconium powder partial size be 50nm~100 μm, mass ratio 5
~95: 95~5.
In the step 3, the photosensitive resin of selection includes performed polymer, reactive diluent and photoinitiator, wherein performed polymer
For at least one of epoxy resin, phenolic resin and acrylic resin, reactive diluent is acrylate monomer, and photoinitiator is
At least one of free radical photo-initiation and cation light initiator, dispersing agent are Sodium Polyacrylate, ammonium polyacrylate, poly- second two
At least one of alcohol.
In the step 3, with slurry total weight be 100%, each component content be respectively as follows: hybrid ceramic powder 50%~
80%, photosensitive resin 17%~50% (performed polymer 7%~21%, reactive diluent 8%~24%, photoinitiator 2%~
5%), dispersing agent 0%~3%.
In the step 5, ultrasonic cleaning running parameter be 20~30kHz of frequency, scavenging period 10~30 minutes, frequency mistake
High or overlong time can dissolve rack surface some materials.
In the step 6, Drying Technology Parameter is 20 DEG C~80 DEG C of drying temperature, and drying time is 0.5~2 hour.
In the step 7, degreasing sintered technological parameter is to be warming up between 400 DEG C~700 DEG C to keep the temperature 1~4 hour, is continued
It is warming up between 900 DEG C~1600 DEG C and keeps the temperature 2~5 hours, heating rate is controlled in 0.5~5 DEG C/min.
The present invention provides a kind of calcium phosphate/zirconium oxide gradient-controllable porous support, and the bracket is prepared using the above method
It forms, the graded pore structure with orderly gradual change, pore size is 10~1000 μm, and porosity is 30%~90%.
The present invention also provides a kind of above-mentioned calcium phosphate/zirconium oxide gradient-controllable porous supports to be used as bone in bone tissue engineer
Filler, substitute or the application as cultured cell in vitro bracket.
The beneficial effects of the present invention are:
(1) present invention uses optical soliton interaction technologies, and porous support aperture obtained is at gradient distribution, the ginseng such as shape, hole
Number is controllable, can satisfy personalized customization demand.
(2) bracket of the invention has different pore size porous structure, is conducive to conveying and the longer period skeletonization of nutriment
Cell adheres to, is proliferated and breaks up in rack surface.
(3) calcium phosphate ceramics and zirconium oxide that the present invention uses belong to bioceramic, have good biocompatibility.
Detailed description of the invention
Fig. 1 is calcium phosphate of the present invention/zirconium oxide gradient-controllable porous support preparation flow figure.
Fig. 2 is the gradient porous bracket figure of photocuring printing.
Fig. 3 is MC3T3-E1 preosteoblast and single aperture bracket co-incubation state diagram.
Fig. 4 is different pore size bracket with the absorbance value after MC3T3-E1 preosteoblast co-incubation 1d, 4d, 7d.
Specific embodiment
With reference to the accompanying drawings and detailed description, the invention will be further described.
The preparation of the gradient porous bracket of embodiment 1
As shown in Figure 1, a kind of calcium phosphate/zirconium oxide gradient-controllable porous support preparation method, through the following steps that
It realizes:
Step 1, modeling: gradient porous scaffold three-dimensional figure is designed with modeling software, saves as stl file；
Step 2, ball mill mixing: by partial size be 10 μm hydroxy apatite powder and 300nm Zirconium powder with 90: 10 matter
Amount is placed in planetary ball mill than mixing with 100r/min ball milling 2 hours；
Step 3, slurry preparation: photosensitive resin is added in the hybrid ceramic powder that step 2 is obtained and dispersant is uniform.Slurry
The quality group of every ingredient become: hybrid ceramic powder is 55%, photosensitive resin 43%, dispersing agent 2%；
Step 4, body formation: the model file that step 1 is obtained imports photo solidification machine, and the slurry that step 3 is obtained is used up solid
The method of chemical conversion shape prints porous support green body, as shown in Figure 2；
Step 5, ultrasonic cleaning: porous support green body is added to absolute ethanol, and is set and is carried out cleaning 10 minutes in supersonic cleaning machine；
Step 6, drying: the bracket green body that step 5 obtains is put into 50 DEG C of drying box 1 hour dry；
It is step 7, degreasing sintered: the bracket green body that step 6 obtains is warming up to 500 DEG C from room temperature with 1 DEG C/min and keeps the temperature 2 hours,
It is continuously heating to 1150 DEG C and keeps the temperature 3 hours, taken out after furnace cooling to room temperature and obtain calcium phosphate/porous branch of zirconium oxide gradient-controllable
Frame.
The different aperture bracket cell proliferation experiment of embodiment 2
The difference of this embodiment and embodiment 1 is: step 1 designs single aperture scaffold three-dimensional with modeling software
Figure, remaining place is essentially identical with embodiment 1, no longer repeats one by one herein.
Finally obtain average pore size be 200~300 μm, 300~400 μm, 400~500 μm of porous support.
CCK-8 cell proliferation experiment: it by the bracket of different pore size after autoclave sterilization, is put into 24 orifice plates.Every hole
It is inoculated with the MC3T3-E1 cell suspension that 1ml concentration is 5 × 10^4/ml, 37 DEG C is put into, is trained in the cell incubator of 5%CO2
It supports, as shown in Figure 3.After for 24 hours, bracket is taken out, is placed in 24 new orifice plates, culture solution is added and continues to cultivate, every 2d is changed once
Liquid.Terminate culture after 1d, 4d, 7d, every hole is added 100 μ L of CCK-8 solution, 37 DEG C be protected from light be incubated for 2h after be sucked out respectively
100 μ l liquid are added in 96 new orifice plates, detect each hole absorbance value in microplate reader 450nm.As a result as shown in Figure 4.
As a result: it is good with the cell growth state of bracket co-incubation in Fig. 3, and branch frame peripheral and hole inner cell
It is evenly distributed, without apparent blank space, shows that the bracket has good biocompatibility.Fig. 4 shows in three kinds of aperture branch
In the presence of frame, cell can keep good proliferation.Specifically, average pore size is 200~300 μm after culture 1d
Bracket group cell viability be higher than other two groups, average pore size be 400~500 μm bracket group cell viability it is minimum, it is this
Situation is still obvious after 4d.It may be cell culture initial stage, rack surface product in small-bore is bigger, is conducive to cell on its surface
Adherency growth.After cultivating 7d, group of cells all keeps good proliferative conditions, and average pore size is that 300~400 μm of bracket group is thin
Born of the same parents' vigor is slightly above the bracket group that average pore size is 200~300 μm.It traces it to its cause, it may be possible to which the continuous proliferation apoptosis of cell needs
Biggish interstitial space guarantees normal growth metabolism.Therefore, small-bore is conducive to cell and is adhered to rack surface, large aperture early period
Be conducive to migrate for later period cell Proliferation and sufficient space is provided.The bracket of gradient porous structure is had ready conditions for the cell of different phase
It grows migratory movement and place is provided.
3 composite porous material Mechanics Performance Testing of embodiment
The difference of this embodiment and embodiment 1 is: step 1 designs single aperture scaffold three-dimensional with modeling software
The mass ratio of figure, step 2 hydroxyapatite and zirconium oxide is 100/0,90/10,80/20,70/30,0/100.Remaining place with
Embodiment 1 is essentially identical, no longer repeats one by one herein.
Intensity test is carried out to complex stephanoporate bracket obtained, test result is as follows shown in table.The result shows that with
The increase of hydroxy apatite-base vivo oxidation zirconium incorporation, compression strength step up.The porous compound support frame of different ratio can
To be applied to filling or displacement at different parts bone defect.
| Hydroxyapatite/zirconium oxide mass ratio | Compression strength/MPa |
| 100/0 | 15.25 |
| 90/10 | 21.34 |
| 80/20 | 27.27 |
| 70/30 | 31.41 |
| 0/100 | 61.23 |
In conjunction with attached drawing, the embodiment of the present invention is explained in detail above, but the present invention is not limited to above-mentioned
Embodiment within the knowledge of a person skilled in the art can also be before not departing from present inventive concept
It puts and makes a variety of changes.
Claims (8)
1. a kind of calcium phosphate/zirconium oxide gradient-controllable porous support preparation method, it is characterised in that specific step is as follows:
Step 1, modeling: gradient porous scaffold three-dimensional illustraton of model is designed with modeling software, saves as STL formatted file；
Step 2, ball mill mixing: calcium phosphate ceramics powder, Zirconium powder are mixed, ball milling in planetary ball mill is placed in；
Step 3, slurry preparation: photosensitive resin is added in the hybrid ceramic powder that step 2 is obtained and dispersant is uniform；
Step 4, body formation: the model file that step 1 is obtained imports photo solidification machine, and the slurry that step 3 is obtained is used up solid
The method of chemical conversion shape prints gradient porous bracket green body；
Step 5, ultrasonic cleaning: porous support green body being added to absolute ethanol, sets in supersonic cleaning machine and cleans, and removal surface is not
Cure pastes；
Step 6, drying: the bracket green body that step 5 obtains is put into drying box dry；
Step 7, degreasing sintered: the bracket base substrate degreasing that step 6 is obtained is sintered, and is taken out after furnace cooling to room temperature and is obtained phosphoric acid
Calcium/zirconium oxide gradient porous support.
2. calcium phosphate according to claim 1/zirconium oxide gradient-controllable porous support preparation method, it is characterised in that:
In the step 2, calcium phosphate ceramics include hydroxyapatite, tricalcium phosphate and biphasic calcium phosphate, fluor-apatite, carbonic acid phosphorus ash
At least one of stone.
3. calcium phosphate according to claim 2/zirconium oxide gradient-controllable porous support preparation method, it is characterised in that:
Selected calcium phosphate ceramics, Zirconium oxide powder partial size be 50nm~100 μm, mass ratio be 5~95: 95~5.
4. calcium phosphate according to claim 1/zirconium oxide gradient-controllable porous support preparation method, it is characterised in that:
In the step 3, the photosensitive resin of selection includes performed polymer, reactive diluent and photoinitiator, and wherein performed polymer is asphalt mixtures modified by epoxy resin
At least one of rouge, phenolic resin and acrylic resin, reactive diluent are acrylate monomer, and photoinitiator is free radical light
At least one of initiator and cation light initiator, dispersing agent be Sodium Polyacrylate, ammonium polyacrylate, in polyethylene glycol at least
It is a kind of.
5. calcium phosphate according to claim 4/zirconium oxide gradient-controllable porous support preparation method, it is characterised in that:
With slurry total weight for 100%, each component content is respectively as follows: hybrid ceramic powder 50%~80%, and photosensitive resin 17%~
50% (performed polymer 7%~21%, reactive diluent 8%~24%, photoinitiator 2%~5%), dispersing agent 0%~3%.
6. calcium phosphate according to claim 1/zirconium oxide gradient-controllable porous support preparation method, it is characterised in that:
In the step 7, degreasing sintered technological parameter is to be warming up between 400 DEG C~700 DEG C to keep the temperature 1~4 hour, is continuously heating to
2~5 hours are kept the temperature between 900 DEG C~1600 DEG C, heating rate is controlled in 0.5~5 DEG C/min.
7. a kind of calcium phosphate/zirconium oxide gradient-controllable porous support, it is characterised in that use above-mentioned any one claim institute
The method stated is prepared, and the bracket has the graded pore structure of orderly gradual change, and pore size is 10~1000 μm, porosity
It is 30%~90%.
8. a kind of calcium phosphate/zirconium oxide gradient-controllable porous support of method as described in any one of claims 1 to 6 preparation or
Calcium phosphate/zirconium oxide gradient-controllable porous support as claimed in claim 7 is in bone tissue engineer as bone filler, displacement
Object or application as cultured cell in vitro bracket.
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| CN115385721B (en) * | 2022-08-29 | 2023-08-08 | 广东工业大学 | Porous bionic human bone with controllable pore structure based on photo-curing molding and preparation method thereof |
| CN115634311A (en) * | 2022-09-27 | 2023-01-24 | 润原生物科技(苏州)有限公司 | Multi-structure cartilage repair implant and preparation method thereof |
| CN116120051A (en) * | 2023-02-13 | 2023-05-16 | 武汉理工大学 | Preparation method of gradient biological composite ceramic skeleton based on photo-curing |
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