CN106729988A - A kind of 3D printing bone repairing support with anti-microbial property and preparation method thereof - Google Patents
A kind of 3D printing bone repairing support with anti-microbial property and preparation method thereof Download PDFInfo
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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
- A61L27/54—Biologically active materials, e.g. therapeutic substances
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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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- 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
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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
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
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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
- A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
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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
- A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
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Abstract
The invention belongs to biomedical engineering technology field, particularly skin wound dressing material is for technical field, disclose a kind of 3D printing bone repairing support with anti-microbial property and preparation method thereof, the described 3D printing bone repairing support with anti-microbial property is that multilayer column structure has with good three-dimensional pore space structure, is made up of polycaprolactone, poly-dopamine and antibacterial peptide LL37.This kind of bone repairing support has good biocompatibility and antibacterial ability, and has good osteogenic ability and osteoconductive potential, plays the role of to promote new bone tissue growth to Cranial defect position.
Description
Technical field
It is open the invention belongs to biomedical engineering technology field, particularly osseous tissue renovating material preparing technical field
A kind of 3D printing bone repairing support with anti-microbial property and preparation method thereof.
Background technology
Due to the caused bulk Cranial defect such as wound, tumor resection, operation, bone grafting reparation is often needed.Conventional bone graft method
Including two kinds of autologous bone transplanting and allogenic bone transplantation.Autologous bone transplanting typically uses patient itself Iliac Bone Grafts, although immune anti-
Should be relatively low, but limited source, and to causing new wound for bone area, some complication are produced therewith；Allogenic bone transplantation is not received
Size shape etc. is limited, but can cause stronger immune response, and healing time is also relatively slow, and discovered in recent years its cross-infection
And later infections rate is greatly increased, originate also relatively limited.
Tissue engineering bracket is a kind of important replacement therapy measure, is expected to finally solve this problem.Traditional is porous
Support manufacturing technology has the three-dimensional friendship of particulate leaching method, melt molding method, Emulsion freeze-drying, high pressure gas expansion method, fiber
Weave, phase separation method etc.；Its key deficiency is that inadequate brace aperture insertion degree, porosity and pore size distribution controllability are poor,
Both support performance had been influenceed, had also been unfavorable for cell growth and tissue vascularization.Bone tissue engineer is related to support, Gegenbaur's cell, bone to lure
The multiple factors such as the stimulation of the ostosis factor and suitable biomechanical environment are led and promote, to mechanical strength, porosity, hole connection
Logical degree has requirement very high.3D printing technique can be largely fulfilled porosity, aperture, pore volume, the sky of support
Between arrangement and other surfaces characteristic controllability, it is thus possible to realize the preparation of excellent bone tissue engineering scaffold.But it is logical at present
Cross 3D printing prepare obtained by bone repairing support function it is more single, do not possess anti-microbial property, often lead to it in the application
Easily a series of inflammation and complication is caused to be occurred by bacterium infection.
Antibacterial peptide can be to multiple types pathogen, with broad spectrum antibiotic activity, to most of gram-positive bacterias, feminine gender
Bacterium, also there is very strong inhibitory action to fungi, the bacterium that significantly antibacterial peptide is tolerated to drug have significantly kill
Go out to act on and include the staphylococcus aureus that first chlorine Asia Linne is received and the enterococcus that vancomycin is tolerated.Antibacterial peptide is by bacterium
Cell membrane accumulation and occurred conformation change so that destroy membrane structure integrality, cause cellular content to outflow, bacterium is thin
Born of the same parents are ruptured, it is also possible to which, while in the presence of some intracellular antibacterial mechanisms, such as suppressing nucleic acid synthesis, interferencing protein is closed with cell membrane is suppressed
Into etc..And the antibacterial process of antibacterial peptide is rapid, can just occur contacting in the microorganism several seconds, this is that conventional antibiotic cannot phase
Analogy.Additionally, it is all to carry positive charge that antibacterial peptide is most, and the cell membrane of bacterium is rich in phosphatidyl glycerol or serine phosphorus
The phosphatide of fat etc and it is carried negative electrical charge, so easily there is charge effect between the two, and then produce antibacterial peptide to kill
Go out the biological effect of bacterium.And the cell membrane of mammal is mainly made up of amphion, do not exist sterol on cell membrane
Quasi-molecule makes antibacterial peptide be difficult to produce mammalian cell injury, and the high specific of antibacterial peptide also determines that its clinic quotes tool
There is security higher.
Wherein antimicrobial peptide LL-37 broad-spectrum antiseptic, has inhibitory action to bacterium, fungi, virus, mycoplasma.Especially may be used
The infection bacteria species common to suppress wound, such as Strep A (Group A Streptococcus), MRSE
(Staphylococcus epidermidis), staphylococcus aureus (Staphylococcus aureus), Pseudomonas aeruginosa
(Pseudomonas aeruginosa), Escherichia coli (Escherichia coli), Klebsiella Pneumoniae (Klebsiella
Pneumoniae) etc..Additionally, antimicrobial peptide LL-37 can participate in the formation of blood vessel, contribute to the vascularization at Cranial defect position.
The content of the invention
It is an object of the invention to the bone repairing support hole insertion journey being directed to obtained by existing Bone Defect Repari technology of preparing
Spend not high, porosity and pore size distribution controllability is poor, and the function list for passing through the bone repairing support prepared by 3D printing technique
One deficiency for not possessing anti-microbial property and vascularization promoting ability, there is provided a kind of 3D printing bone repairing support with anti-microbial property and
Its preparation method, this kind of bone repairing support has a good biocompatibility and anti-microbial property, and with good osteogenic ability,
Osteoconductive potential and vascularization ability.The present invention by with polycaprolactone be host material system using 3D printing technique for go out it is poly- oneself
Lactone bone repaiies support, then biologic antibiotic peptide LL-37 is fixed on rack surface by the method for surface modification, entirely prepares
Journey takes short, and method is simple, easily realizes industrialization, and the size and profile of support can go customization according to damaged part.
To achieve the above object, the technical scheme is that a kind of 3D printing bone repairing support with anti-microbial property by
It is made up of the polycaprolactone bone repairing support obtained by 3D printing and modification antibacterial peptide LL37 in its surface.
Further, the described 3D printing bone repairing support with anti-microbial property is counted by polycaprolactone 10 by weight
Part, poly-dopamine 0.1-0.5 parts and antimicrobial peptide LL-37 0.01-0.05 parts of composition.
Preferably, the molecular weight of described polycaprolactone is 1 × 106, purchased from Sigma Reagent Companies, antibacterial peptide LL37 purchases
From BeadTech companies of the U.S..
To achieve the above object, another technical scheme of the invention is：A kind of 3D printing Bone Defect Repari with anti-microbial property
The preparation method of support, described preparation method is realized by following steps：The printing of polycaprolactone bone repairing support and support
The poly- bar in surface is amine-modified；The fixation of antimicrobial peptide LL-37.
Further, the printing of the polycaprolactone bone repairing support and poly- bar of rack surface is amine-modified concretely comprises the following steps：
（1）Print the preparation of collecting platform：Dopamine hydrochloride is dissolved in trishydroxymethylaminomethane-hydrochloride buffer that pH is 8.5 molten
In liquid, the Dopamine hydrochloride solution that concentration is 10mg/mL is configured to, Dopamine hydrochloride solution is then placed in batch cultur ware
In, the batch cultur ware that then will fill Dopamine hydrochloride solution is placed on the receiving platform of 3D printer, and has adhesive tape to consolidate
It is fixed；（2）The printing of bone repairing support：Polycaprolactone coiled material is put into the charging aperture of 3D printer, setting print temperature is
75 DEG C, print speed be 2-3 minute every layer, for 4-8 layers, top of form is set to the middle of collecting platform to the printing number of plies；
（3）After the completion of printing, the bone repairing support of gained is removed into 3D printer in the lump together with batch cultur ware, by batch cultur
Dopamine hydrochloride solution in ware is all discarded, and Dopamine hydrochloride solution is then added again to bone repairing support is submerged, in aseptic
Placed in super-clean bench 3-4 hours, support is finally removed into batch cultur ware, it is standby with distilled water flushing.
Further, the fixation of the antimicrobial peptide LL-37 is specific as follows：（1）Antibacterial peptide LL37 is dissolved in into pH is
In 7.4 PBS, gentle agitation 1 hour is configured to the antibacterial peptide LL37 solution that concentration is 10 ~ 50 μ g/mL, so
After be positioned over it is standby under gnotobasis；（2）The bone repairing support that will be processed through poly-dopamine surface modification, is soaked in antibacterial peptide
It is placed in LL37 solution and is incubated in 37 DEG C of CO2gas incubators 12-18 hours, then with the deionized water rinsing of sterilizing, is put
The freeze-drying in -80 DEG C of vacuum freeze driers.Obtain final product a kind of 3D printing bone repairing support with anti-microbial property.
Further, the 3D printer in the printing of described polycaprolactone bone repairing support be using fusion sediment into
Type（FDM）The 3D printer of operation principle, purchased from 3D systems companies, model 3D systems Cubepro.
Beneficial effects of the present invention are：
（1）Bone repairing support obtained by the present invention has stronger suppression to Gram-negative bacteria, gram-positive bacteria and fungi
Bacterium acts on, and rapid-action（24 hours after contact bacterium, bacteriostasis rate can be up to more than 95%）, can effectively reduce bone repairing support and exist
By bacterium or the risk of fungal infection during implantation；
（2）The present invention prepares polycaprolactone bone repairing support by 3D printing, then carries out surface to it by poly-dopamine and repair
Decorations, because poly-dopamine has substantial amounts of active amino and hydroxyl, making the surface-active site of support increases, so as to improve support
To cell and the adhesion of protein, nutriment or growth factor is gathered on the surface of support, be conducive to damaged part
The regeneration of bone tissue；
（3）It with polycaprolactone is host material by obtained by 3D printing technique that bone repairing support obtained by the present invention is, warp
After poly-dopamine surface modification, then antimicrobial peptide LL-37 is fixed on its surface, compared to the antibacterial bone repairing support of support type,
The antibacterial timeliness of this kind of bone repairing support is more long, the problem for being discharged in the absence of antimicrobial component and causing anti-microbial property to decline.In addition
The method fixed by surface, can effectively be solved antimicrobial peptide LL-37 and cause poison to host cell because local concentration is too high
Property effect, occur haemolysis.
（4）The present invention is repaiied on support by the way that antimicrobial peptide LL-37 is fixed on into bone, bone repairing support is had good resisting
Outside bacterium performance, also assigning support has good vascularization ability, and angiogenesis is complicated multi-step process, is related to thin
Interaction between born of the same parents, soluble factor, extracellular components.In processes of wound repair, angiogenesis deficiency can be directly resulted in
The necrosis of local microcirculation obstacle, tissue ischemia and anoxic.Therefore, vascularization is the important step in wound repair, and LL-
37 are proved to take part in vascularization process.LL-37 is mediated by formyl peptides sample acceptor -1 (FPRL1) can stimulate people's navel quiet
The propagation of arteries and veins endothelial cell (HUVECs), and degree of cell proliferation is proportionate with the concentration of LL-37.
Brief description of the drawings
Fig. 1 is the antibacterial ability evaluation experimental comparative result figure of embodiment 1~3 and comparative example；
Fig. 2 is the Cytotoxic evaluation experimental result comparison diagram of embodiment 1~3 and comparative example；
Fig. 3 is the protein adsorption merit rating comparative result figure of embodiment 1~3 and comparative example；
Fig. 4 is the alkaline phosphatase testing result comparison diagram of embodiment 1~3 and comparative example.
Specific embodiment
Technical scheme is described further with reference to embodiment.
Embodiment 1
The technical scheme is that a kind of 3D printing bone repairing support with anti-microbial property is as obtained by by 3D printing
Polycaprolactone bone repairing support and modification antibacterial peptide LL37 compositions in its surface；A kind of described 3D with anti-microbial property
Printing bone repairing support is counted by 10 parts, poly-dopamine 0.1-0.5 parts of polycaprolactone and antimicrobial peptide LL-37 by weight
0.01-0.05 parts of composition.Further, the molecular weight of described polycaprolactone is 1 × 106, purchased from Sigma Reagent Companies, resist
Bacterium peptide LL37 is purchased from BeadTech companies of the U.S..
Embodiment 2
The technical scheme is that a kind of 3D printing bone repairing support with anti-microbial property is multilayer column structure, by passing through
Polycaprolactone bone repairing support obtained by 3D printing is constituted with the antibacterial peptide LL37 for modifying in its surface；A kind of described tool
The 3D printing bone repairing support for having anti-microbial property is counted by 10 parts of polycaprolactone, 0.3 part of poly-dopamine and antibacterial peptide by weight
0.03 part of composition of LL-37.Further, the molecular weight of described polycaprolactone is 1 × 106, purchased from Sigma Reagent Companies,
Antibacterial peptide LL37 is purchased from BeadTech companies of the U.S..
Embodiment 3
The technical scheme is that a kind of 3D printing bone repairing support with anti-microbial property is multilayer column structure, by passing through
Polycaprolactone bone repairing support obtained by 3D printing is constituted with the antibacterial peptide LL37 for modifying in its surface；A kind of described tool
The 3D printing bone repairing support for having anti-microbial property is counted by 10 parts of polycaprolactone, 0.5 part of poly-dopamine and antibacterial peptide by weight
0.05 part of composition of LL-37.Further, the molecular weight of described polycaprolactone is 1 × 106, purchased from Sigma Reagent Companies,
Antibacterial peptide LL37 is purchased from BeadTech companies of the U.S..
Embodiment 4
A kind of 3D printing bone repairing support with anti-microbial property of any example of 1~embodiment of embodiment 3, its preparation method it is specific
Step is as follows：
1. the printing of the polycaprolactone bone repairing support and poly- bar of rack surface is amine-modified concretely comprises the following steps：
（1）Print the preparation of collecting platform：Dopamine hydrochloride is dissolved in into trishydroxymethylaminomethane-hydrochloric acid that pH is 8.5 to delay
Rush in solution, be configured to the Dopamine hydrochloride solution that concentration is 10mg/mL, Dopamine hydrochloride solution is then placed in disposable training
Support in ware, the batch cultur ware that then will fill Dopamine hydrochloride solution is placed on the receiving platform of 3D printer, and has glue
Band is fixed；
（2）The printing of bone repairing support：Polycaprolactone coiled material is put into the charging aperture of 3D printer, setting print temperature is
75 DEG C, print speed be 2-3 minute every layer, for 4-8 layers, top of form is set to the middle of collecting platform to the printing number of plies；
（3）After the completion of printing, the bone repairing support of gained is removed into 3D printer in the lump together with batch cultur ware, will be disposable
Dopamine hydrochloride solution in culture dish is all discarded, and Dopamine hydrochloride solution is then added again to submerging bone repairing support, in
Placed in aseptic super-clean bench 3-4 hours, support is finally removed into batch cultur ware, it is standby with distilled water flushing.
2. the fixation of antimicrobial peptide LL-37：
（1）Antibacterial peptide LL37 is dissolved in the PBS that pH is 7.4, gentle agitation 1 hour, being configured to concentration is
The antibacterial peptide LL37 solution of 10 ~ 50 μ g/mL, is then placed into standby under gnotobasis；
（2）The bone repairing support that will be processed through poly-dopamine surface modification, is soaked in antibacterial peptide LL37 solution and is placed in 37 DEG C two
It is incubated in carbonoxide incubator 12-18 hours, then with the deionized water rinsing of sterilizing, is placed in -80 DEG C of vacuum freeze driers
Middle freeze-drying.Obtain final product a kind of 3D printing bone repairing support with anti-microbial property.
Embodiment 5
Comparative example：A kind of polycaprolactone support obtained by 3D printing（Application reference number is disclosed in CN201510188649.5
A kind of human body of FDM technology 3D printing can absorb enhanced bone fixation structural material and preparation method thereof disclosed preparation side
Obtained by method）.
Experimental group 1 ~ 3：It is a kind of 3D printing bone repairing support with anti-microbial property of the gained of embodiment 1~3, using reality
The method for applying example 4 is prepared from.
The 3D printing bone repairing support with anti-microbial property prepared by above-described embodiment 1~3 is resisted with comparative example
Bacterium merit rating is tested, and comparative example 1 ~ 3 and comparative example are to Escherichia coli, Jinhua staphylococcus and Candida albicans contact 24
Fungistatic effect after hour.Experimental result such as Fig. 1 shows.
From the visible embodiment 1 ~ 3 of result after contacting 24 hours with Escherichia coli, Jinhua staphylococcus and Candida albicans
Bacteriostasis rate can reach more than 95%, and comparative example is respectively less than 20%, it was demonstrated that embodiment 1 ~ 3 is respectively provided with potent, and action time is fast
Broad spectrum antibacterial performance.
Embodiment 6
Comparative example：A kind of polycaprolactone support obtained by 3D printing（Application reference number is disclosed in CN201510188649.5
A kind of human body of FDM technology 3D printing can absorb enhanced bone fixation structural material and preparation method thereof disclosed preparation side
Obtained by method）.
Experimental group 1 ~ 3：It is a kind of 3D printing bone repairing support with anti-microbial property of the gained of embodiment 1~3, using reality
The method for applying example 4 is prepared from.
The 3D printing bone repairing support with anti-microbial property prepared by above-described embodiment 1~3 is carried out carefully with comparative example
Cellular toxicity evaluation experimental（Tested by GB GB/T 16886.5-2003）, comparative example 1 ~ 3 and comparative example pair.Experiment
Result such as Fig. 2 shows.
Cytotoxicity testing result shows embodiment 1 ~ 3, and its is corresponding after being co-cultured 3 days and 7 days with Gegenbaur's cell MG-63
Cell with respect to proliferation rate more than 90%, cytotoxicity is rated 0 grade, it was demonstrated that it has good describing property of cell, and right
Ratio and HF co-culture 24 hours and 48 hours after the relative proliferation rate of its corresponding cell in 70% or so, cell
Toxicity is rated 3 grades, with serious cytotoxicity.Additionally, the extension embodiment 1 ~ 3 of the time of co-cultivation is relative compared to propagation
Rate is significantly improved, and its cell is high with respect to more negative group of proliferation rate after 7 days for embodiment 2 and 3（It is above 100%）, card
The bright bone repairing support using obtained by preparation method disclosed in this invention can promote the growth of Gegenbaur's cell MG-63, favorably
In the growth of Cranial defect position new bone tissue.
Embodiment 7
Comparative example：A kind of polycaprolactone support obtained by 3D printing（Application reference number is disclosed in CN201510188649.5
A kind of human body of FDM technology 3D printing can absorb enhanced bone fixation structural material and preparation method thereof disclosed preparation side
Obtained by method）.
Experimental group 1 ~ 3：It is a kind of 3D printing bone repairing support with anti-microbial property of the gained of embodiment 1~3, using reality
The method for applying example 4 is prepared from.
With BSA as model protein, the tool prepared by embodiment 1~3 is evaluated using BCA protein adsorption aptitude tests method and is induced
The repair of cartilage hydrogel of repair and the protein adsorption ability of comparative example.Result of the test such as Fig. 3 shows.
Protein adsorption capability result display embodiment 1 ~ 3 protein adsorption ability compared with comparative example will height, this with beneficial to life
The factor long and nutriment stick thereon, are that Gegenbaur's cell grows one good microenvironment of offer in the above, are conducive to.
Embodiment 8
Comparative example：A kind of polycaprolactone support obtained by 3D printing（Application reference number is disclosed in CN201510188649.5
A kind of human body of FDM technology 3D printing can absorb enhanced bone fixation structural material and preparation method thereof disclosed preparation side
Obtained by method）.
Experimental group 1 ~ 3：It is a kind of 3D printing bone repairing support with anti-microbial property of the gained of embodiment 1~3, using reality
The method for applying example 4 is prepared from.
By repair of cartilage hydrogel and the comparative example of the tool induction repair prepared by above-described embodiment 1~3 respectively with
MG-63（OS-732 cells）Detect its alkaline phosphatase enzyme values after co-culturing 7 days, contrast experiment's group and contrast
The self-bone grafting ability of example.Experimental result is as shown in Figure 4：
Alkaline phosphatase（alkaline phosphate ALP）It is the mark for breaking up Gegenbaur's cell, the ore deposit of bone matrix can be promoted
Change.Understand from the graph, compared with comparative example, the ALP values of embodiment 1 ~ 3 substantially will height compared with comparative example.As can be seen here, by this hair
A kind of bright disclosed 3D printing bone repairing support with anti-microbial property has self-bone grafting ability and osteoconductive potential higher.
Obviously, the above embodiment of the present invention is only intended to clearly illustrate example of the present invention, and is not right
The restriction of embodiments of the present invention；For those of ordinary skill in the field, may be used also on the basis of the above description
To make other changes in different forms, there is no need and unable to be exhaustive to all of implementation method；It is all this
Any modification, equivalent and improvement made within the spirit and principle of invention etc., should be included in the claims in the present invention
Protection domain within.
Claims (7)
1. a kind of 3D printing bone repairing support with anti-microbial property, it is characterised in that the described 3D with anti-microbial property beats
Polycaprolactone bone repairing support and modification of the print bone repairing support as obtained by by 3D printing antibacterial peptide in its surface
LL37 is constituted.
2. the 3D printing bone repairing support with anti-microbial property according to claim 1, it is characterised in that described has
The 3D printing bone repairing support of anti-microbial property is counted by 10 parts, poly-dopamine 0.1-0.5 parts of polycaprolactone and antibacterial by weight
0.01-0.05 parts of composition of peptide LL-37.
3. the 3D printing bone repairing support with anti-microbial property according to claim 2, it is characterised in that described gathers oneself
The molecular weight of lactone is 1 × 106。
4. the preparation method of the 3D printing bone repairing support with anti-microbial property described in any one of claim 1 ~ 3, its feature
It is that described preparation method is realized by following steps：The printing of polycaprolactone bone repairing support and the poly- bar amine of rack surface
Modification, the fixation of antimicrobial peptide LL-37.
5. the preparation method of the 3D printing bone repairing support with anti-microbial property according to claim 4, it is characterised in that
The printing of the polycaprolactone bone repairing support and poly- bar of rack surface is amine-modified concretely comprises the following steps：
（1）Print the preparation of collecting platform：Dopamine hydrochloride is dissolved in into trishydroxymethylaminomethane-hydrochloric acid that pH is 8.5 to delay
Rush in solution, be configured to the Dopamine hydrochloride solution that concentration is 10mg/mL, Dopamine hydrochloride solution is then placed in disposable training
In supporting ware, and the batch cultur ware of Dopamine hydrochloride solution will be filled it is placed on the receiving platform of 3D printer, it is solid with adhesive tape
It is fixed；
（2）The printing of bone repairing support：Polycaprolactone coiled material is put into the charging aperture of 3D printer, setting print temperature is
75 DEG C, print speed be 2-3 minute every layer, for 4-8 layers, top of form is set to the middle of collecting platform to the printing number of plies；
（3）After the completion of printing, the bone repairing support of gained is removed into 3D printer in the lump together with batch cultur ware, will be disposable
Dopamine hydrochloride solution in culture dish is all discarded, and Dopamine hydrochloride solution is then added again to submerging bone repairing support, in
Placed in aseptic super-clean bench 3-4 hours, support is finally removed into batch cultur ware, it is standby with distilled water flushing.
6. the preparation method of the preparation method of the 3D printing bone repairing support with anti-microbial property according to claim 4,
Characterized in that, the fixation of antimicrobial peptide LL-37 is concretely comprised the following steps：
（1）Antibacterial peptide LL37 is dissolved in the PBS that pH is 7.4, gentle agitation 1 hour, being configured to concentration is
The antibacterial peptide LL37 solution of 10 ~ 50 μ g/mL, is then placed into standby under gnotobasis；
（2）The bone repairing support that will be processed through poly-dopamine surface modification, is soaked in antibacterial peptide LL37 solution and is placed in 37 DEG C two
It is incubated in carbonoxide incubator 12-18 hours, then with the deionized water rinsing of sterilizing, is placed in -80 DEG C of vacuum freeze driers
Middle freeze-drying, obtains final product the 3D printing bone repairing support with anti-microbial property.
7. the preparation method of the 3D printing bone repairing support with anti-microbial property according to claim 5, it is characterised in that
Described 3D printer is the 3D printer using fused glass pellet operation principle.
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