CN107412855A - Has cated 3D printing support and its preparation method and application - Google Patents

Has cated 3D printing support and its preparation method and application Download PDF

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Publication number
CN107412855A
CN107412855A CN201710657006.XA CN201710657006A CN107412855A CN 107412855 A CN107412855 A CN 107412855A CN 201710657006 A CN201710657006 A CN 201710657006A CN 107412855 A CN107412855 A CN 107412855A
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magnesium silicate
mesoporous magnesium
copper
support
silicate microballoon
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魏杰
袁朝
魏武
吴涵
唐亮琛
胡兴龙
苏佳灿
吴钊英
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East China University of Science and Technology
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East China University of Science and Technology
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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
    • A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
    • 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
    • A61L27/28—Materials for coating prostheses
    • 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
    • A61L27/28—Materials for coating prostheses
    • A61L27/30—Inorganic materials
    • A61L27/306—Other specific inorganic materials not covered by A61L27/303 - A61L27/32
    • 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
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J7/00—Chemical treatment or coating of shaped articles made of macromolecular substances
    • C08J7/04—Coating
    • C08J7/06—Coating with compositions not containing macromolecular substances
    • 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
    • A61L2300/10—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing inorganic materials
    • A61L2300/102—Metals or metal compounds, e.g. salts such as bicarbonates, carbonates, oxides, zeolites, silicates
    • 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
    • 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
    • A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
    • A61L2300/62—Encapsulated active agents, e.g. emulsified droplets
    • A61L2300/622—Microcapsules
    • 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
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2367/00—Characterised by the use of polyesters obtained by reactions forming a carboxylic ester link in the main chain; Derivatives of such polymers
    • C08J2367/02—Polyesters derived from dicarboxylic acids and dihydroxy compounds

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  • Polymers & Plastics (AREA)
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Abstract

The invention discloses cated 3D printing support of tool and its preparation method and application.The support includes poly butylene succinate rest body, dopamine film and mesoporous magnesium silicate microballoon coating or mixes the mesoporous magnesium silicate microballoon coating of copper, described mesoporous magnesium silicate microballoon coating is mixed in the mesoporous magnesium silicate microballoon coating of copper, microspherulite diameter is 180 220nm, microballoon aperture is 3 5nm, and dopamine film thickness is 50 100nm.The results show, the present invention with mesoporous magnesium silicate microballoon or mix the 3D printing support of the mesoporous magnesium silicate microballoon coating of copper with good histocompatbility and external activity, cell adherence, propagation and differentiation can be promoted, also there is excellent promotion new bone and angiogenesis in vivo, there is preferable application prospect preparing bone impairment renovation material field.

Description

Has cated 3D printing support and its preparation method and application
Technical field
The invention belongs to biological medicine engineering field, in particular it relates to have cated 3D printing support and its preparation side Method and application.
Background technology
Bone is human body important component, has certain mechanical performance, plays body support's effect.But due to the external world Wound, bone disease (bone tumour, osteomyelitis etc.) and Cranial defect caused by infection many reasons, it is clinically very common and be difficult to Repair completely, the defect repair research medium vessels problem of especially bone is the significant challenge faced at present.Recent studies suggest that During bone tissue regeneration, the formation and extension of capilary, oxygen and nutriment are provided for the growth and differentiation of bone tissue, It is ultimately formed the major influence factors of calcified tissue.And in currently used bone impairment renovation material, 3D printing poly-succinic Butanediol ester (PBSu) support has the pore structure of height rule and excellent hole connectivity, and possessing bone repairing support sample will The pore structure and mechanical strength asked, but there is no bioactivity.Therefore, one kind is badly in need of in this area has bioactivity and prepares letter Single easy-operating PBSu supports.
The content of the invention
Present invention aim to address bone defect healing field to lack the poly butylene succinate with bioactivity (PBSu) the problem of support, and then cated 3D printing support of a tool and its preparation method and application is provided.The present invention's Support coats mesoporous magnesium silicate microballoon (NMS) on its surface or mixes copper on the basis of dopamine is modified 3D printing PBSu supports Mesoporous magnesium silicate microballoon (CuNMS), assigns its Bioactivity.The results show, of the invention has mesoporous magnesium silicate Microballoon or the 3D printing support for mixing the mesoporous magnesium silicate microballoon coating of copper have good histocompatbility and external activity, also have The excellent promotion newly effect of bone and angiogenesis in vivo, before preparing bone impairment renovation material field and there is preferably application Scape.
The present invention is that solve above-mentioned technical problem by the following technical programs.
First aspect present invention provides the cated poly butylene succinate of a tool (PBSu) support, wherein, it is described Support includes poly butylene succinate rest body, dopamine film and mesoporous magnesium silicate microballoon (NMS) coating or to mix copper mesoporous Magnesium silicate microballoon (CuNMS) coating；
Described mesoporous magnesium silicate microballoon coating mixes the outer layer that the mesoporous magnesium silicate microballoon coating of copper is coated in dopamine film；
Described dopamine film is located at the surface of poly butylene succinate rest body；
Described mesoporous magnesium silicate microballoon coating is mixed in the mesoporous magnesium silicate microballoon coating of copper, microspherulite diameter 180- 220nm, the aperture of microballoon is 3-5nm；
The thickness of the dopamine film is 50-100nm.
Described support, wherein,
The poly butylene succinate is commonly used in the art, and number-average molecular weight is preferably 4 × 104-7×104, weight Average molecular weight/number-average molecular weight is preferably 1.2-2.4.
The thickness of the dopamine film is preferably 60-80nm.
The specific surface area of described mesoporous magnesium silicate microballoon is preferably 500-570m2/ g, more preferably 538.1 m2/g。
The described specific surface area for mixing the mesoporous magnesium silicate microballoon of copper is preferably 450-550m2/ g, more preferably 490.4m2/g。
Described mixes in the mesoporous magnesium silicate microballoon of copper, and the content of copper is preferably 3-7wt%, more preferably 5wt%.
Described mesoporous magnesium silicate microballoon or the aperture for mixing the mesoporous magnesium silicate microballoon of copper are preferably 4nm.
Described mesoporous magnesium silicate microballoon or the particle diameter for mixing the mesoporous magnesium silicate microballoon of copper are preferably 200nm.
Described mesoporous magnesium silicate microballoon and to mix the mesoporous magnesium silicate microballoon of copper be commonly used in the art, its preparation method Also it is the conventional preparation method in this area, is typically prepared using template and self-assembly method.
In the preferred embodiment of the present invention, mesoporous magnesium silicate microballoon is prepared using the method comprised the following steps：
At S1,35-40 DEG C, cetyl trimethylammonium bromide (CTAB) is dissolved in the water, after CTAB is completely dissolved, Sequentially add ammoniacal liquor, tetraethyl orthosilicate (TEOS) and magnesium nitrate hexahydrate (Mg (NO3)2·6H2O reaction) is stood；
S2, the reaction product obtained by S1 is centrifuged and washs gained white depositions, drying；
S3, drying product obtained by S2 is placed in Muffle furnace sintered, remove CTAB, obtained white powder is mesoporous silicon Sour magnesium microballoon (NMS).
Wherein, in step S1, the water can be deionized water.In a preferred embodiment of the present invention, step S1 is according to such as Lower operation is carried out：At 35-40 DEG C, 0.6-0.8g cetyl trimethylammonium bromides (CTAB) are dissolved in 30-35mL deionizations In water, after CTAB is completely dissolved, 6.5-7.5 mL 1M ammoniacal liquor, 3.2-3.8 tetraethyl orthosilicates (TEOS) and 2.3- are sequentially added 2.5g magnesium nitrate hexahydrate.
Wherein, in step S1, the standing reaction time is 3-6 hours, preferably 4 hours.
Wherein, in step S2, the washing white depositions include being washed with ethanol and being once washed with deionized water twice The step of washing.
Wherein, in step S2, the operation and condition of the operation of the drying and condition for the conventional drying in this area, preferably Ground is carried out in 60 DEG C of electric drying oven with forced convection.
Wherein, in step S3, the temperature of the sintering is preferably 500-700 DEG C, is more preferably 600 DEG C, the sintering The time of insulation is preferably 2-4 hours, is more preferably 3 hours.
In the preferred embodiment of the present invention, the mesoporous magnesium silicate microballoon of copper is mixed using the method system comprised the following steps It is standby：
M1, at 35-40 DEG C, cetyl trimethylammonium bromide (CTAB) is dissolved in the water, CTAB is completely dissolved Afterwards, magnesium nitrate hexahydrate (Mg (NO are added3)2·6H2) and Gerhardite (Cu (NO O3)2·3H2O), reaction is stood；
M2, the reaction product obtained by M1 is centrifuged and washs gained light green sediment, drying；
M3, drying product obtained by M2 is placed in Muffle furnace sintered, remove CTAB, obtained pale green powder is to mix copper Mesoporous magnesium silicate microballoon (CuNMS).
Wherein, in step M1, the water can be deionized water.In a preferred embodiment of the present invention, step M1 is according to such as Lower operation is carried out：At 35-40 DEG C, 0.6-0.8g cetyl trimethylammonium bromides (CTAB) are dissolved in 30-35mL deionizations In water, after CTAB is completely dissolved, 1.6-1.8g magnesium nitrate hexahydrates and 0.5-0.6g Gerhardites, reaction is stood.
Wherein, in step M1, the standing reaction time is 3-6 hours, preferably 4 hours.
Wherein, in step M2, the washing light green sediment includes being washed with ethanol twice and once being used deionized water The step of washing.
Wherein, in step M2, the operation and condition of the operation of the drying and condition for the conventional drying in this area, preferably Ground is carried out in 60 DEG C of electric drying oven with forced convection.
Wherein, in step M3, the temperature of the sintering is preferably 500-700 DEG C, is more preferably 600 DEG C, the sintering The time of insulation is preferably 2-4 hours, is more preferably 3 hours.
Second aspect of the present invention provides a kind of cated poly-succinic of the tool prepared as described in the first aspect of the invention The method of butanediol ester support, the described method comprises the following steps：
(1) 3D printing prepares poly butylene succinate rest body；
(2) dopamine film is formed in poly butylene succinate rest body surface prepared by step (1), obtains dopamine Surface modified butanediol ester poly succinic acid support (DP)；
(3) coat the mesoporous magnesium silicate microballoon on DP prepared by step (2) or mix the mesoporous magnesium silicate microballoon of copper, produce To the cated poly butylene succinate support of tool of the present invention.
In step (1), the method that 3D printing prepares poly butylene succinate rest body is beaten for the conventional 3D in this area Impression method.In the preferred embodiment of the present invention, poly butylene succinate rest body is using the side comprised the following steps It is prepared by method：
Poly butylene succinate (PBSu) is added in the barrel of 3D printing equipment, heating cylinder temperature to 110- 115 DEG C, 3D printing is carried out, printing finishes, and cools down, produces PBSu rest bodies.
Wherein, the 3D printing equipment is that this area is conventional, is preferably carried by Shanghai Fuqifan Electromechanical Science ＆ Technology Co., Ltd. For unit type HTS-300.
Wherein, the technical parameter of the 3D printing includes as follows：
The discharge velocity of 3D printing equipment is preferably 100-200g/min, is more preferably 150g/min；
The mechanical arm rate travel of 3D printing equipment is preferably 0.1-1m/s, is more preferably 0.5m/s；
The cooling is preferably natural cooling.
In step (2), the method that dopamine film is formed on poly butylene succinate rest body surface is conventional for field, In the preferred embodiment of the present invention, carried out using the method comprised the following steps：
PBSu rest bodies are cleaned by ultrasonic, is then immersed in dopamine solution and stirs；Cleaned, dried with deionized water, So as to obtain support sample DP.
Wherein, the ultrasonic cleaning of the PBSu rest bodies is preferably carried out in deionized water, and ultrasonic time is preferably It it is more preferably 10 minutes for 5-20 minutes.
Wherein, described dopamine solution is preferably the Tris-HCl cushioning liquid of dopamine, the dopamine solution Concentration be preferably 1-3mg/ml, more preferably 2mg/ml.
In a preferred embodiment, the preparation of the dopamine solution is carried out according to following operation：
180-220mg dopamine powder is weighed, is dissolved in 100mL 10mM Tris-HCl cushioning liquid (pH=8.5) In, so as to obtain the dopamine solution.
Wherein, mixing speed of the PBSu rest bodies in dopamine solution is preferably 100-300 revs/min, More preferably it is 150-200 revs/min；Mixing time is preferably 18-36 hours, is more preferably 24 hours.
Wherein, the operation of the drying and condition are operation and the condition of the conventional drying in this area, preferably 60 DEG C electric drying oven with forced convection in carry out.
In step (3), the coating mesoporous magnesium silicate microballoon or the method for mixing the mesoporous magnesium silicate microballoon of copper can be that field is normal Rule, in the preferred embodiment of the present invention, carried out using the method comprised the following steps：
The support sample DP that step (2) obtains is soaked in mesoporous magnesium silicate microsphere suspension liquid or to mix the mesoporous magnesium silicate of copper micro- In ball suspension and stir；Cleaned, dried with deionized water, so as to obtain the described cated poly butylene succinate of tool Support.
Wherein, the support sample DP in mesoporous magnesium silicate microsphere suspension liquid or mixes the mesoporous magnesium silicate microsphere suspension liquid of copper Mixing speed be preferably 100-300 revs/min, be more preferably 150-200 revs/min；Mixing time is preferably 18-36 Hour, it is more preferably 24 hours.
Wherein, the mesoporous magnesium silicate microsphere suspension liquid or the compound method of the mesoporous magnesium silicate microsphere suspension liquid of copper is mixed for this The conventional compound method in field, in the preferred embodiment of the present invention, the compound method is carried out according to following operation：Take 0.9-1.1 grams of mesoporous magnesium silicate microballoon mixes the mesoporous magnesium silicate microballoon of copper, is distributed in 100ml deionized waters, produces respectively.
Wherein, the operation of the drying and condition are operation and the condition of the conventional drying in this area, preferably 60 DEG C electric drying oven with forced convection in carry out.
It is cated poly- that third aspect present invention provides tool prepared by a kind of method as described in second aspect of the present invention Succinic acid-butanediol ester (PBSu) support.
It is cated poly- that fourth aspect present invention provides a kind of tool as described in first aspect present invention or the third aspect Application of succinic acid-butanediol ester (PBSu) support in bone impairment renovation material field is prepared.
It on the basis of common sense in the field is met, above-mentioned each optimum condition, can be combined, it is each preferably real to produce the present invention Example.
Agents useful for same and raw material of the present invention are commercially available.
The positive effect of the present invention is：
The technical scheme is that dopamine film is formed on 3D printing poly butylene succinate rest body surface, and Coated with mesoporous magnesium silicate microballoon or the mesoporous magnesium silicate microballoon coating of copper is mixed on dopamine film surface.The support has good Histocompatbility and external activity, cell adherence, propagation and differentiation can be promoted, also with excellent promotion new bone and blood vessel in vivo The effect of generation, there is higher application value in bone impairment renovation material field.
Brief description of the drawings
Fig. 1 is mesoporous magnesium silicate microballoon (NMS) powder of the present invention and mixes mesoporous magnesium silicate microballoon (CuNMS) powder of copper Digital photo figure.Wherein (a) is mesoporous magnesium silicate microsphere powder, and (b) is to mix the mesoporous magnesium silicate microsphere powder of copper.
Fig. 2 is mesoporous magnesium silicate microballoon (NMS) powder of the present invention and mixes mesoporous magnesium silicate microballoon (CuNMS) powder of copper Transmission electron microscope picture.Wherein, (a) and (b) is NMS powder, and (c) and (d) is CuNMS powder.
Fig. 3 is the mesoporous magnesium silicate microballoon (NMS) of the present invention and the N2 adsorption for mixing the mesoporous magnesium silicate microballoon (CuNMS) of copper/take off Attached isollaothermic chart (figure A) and pore size distribution curve figure (figure B).
Fig. 4 be the mesoporous magnesium silicate microballoon (NMS) of the present invention and mix the XPS of the mesoporous magnesium silicate microballoon (CuNMS) of copper scheme and Enlarged drawing at 923-966eV, wherein (A) and (C) is NMS, (B) and (D) is CuNMS.
Fig. 5 is the PBSu supports of comparative example 1 of the present invention, and the dopamine surface of comparative example 2 is modified PBSu supports (DP), real The PBSu supports (NDP) and embodiment 2 for applying the mesoporous magnesium silicate microballoon coating of example 1 mix what the mesoporous magnesium silicate microballoon of copper coated The digital photo figure of PBSu supports (CNDP), wherein (a) is PBSu, (b) is DP, and (c) is NDP, and (d) is CNDP.
Fig. 6 is the PBSu supports of comparative example 1 of the present invention, and the dopamine surface of comparative example 2 is modified PBSu supports (DP), real The PBSu supports (NDP) and embodiment 2 for applying the mesoporous magnesium silicate microballoon coating of example 1 mix what the mesoporous magnesium silicate microballoon of copper coated The XRD spectrum (figure A) and FTIR collection of illustrative plates (figure B) of PBSu supports (CNDP), wherein (a) is PBSu, (b) is DP, and (c) is NDP, (d) it is CNDP.
The PBSu supports of Fig. 7 comparative examples 1 of the present invention, the dopamine surface of comparative example 2 are modified PBSu supports (DP), implemented The PBSu supports (NDP) of the mesoporous magnesium silicate microballoon coating of example 1 and the PBSu for mixing the mesoporous magnesium silicate microballoon coating of copper of embodiment 2 The scanning electron microscope (SEM) photograph of support (CNDP), wherein (a) is PBSu, (b) is DP, and (c) is NDP, and (d) is CNDP.
Fig. 8 be comparative example 1 of the present invention PBSu supports, embodiment 1 mesoporous magnesium silicate microballoon coating PBSu supports (NDP) and the PBSu supports (CNDP) for mixing the coating of copper mesoporous magnesium silicate microballoon of embodiment 2 are immersed in simulated body fluid (SIMULATED BODY FLUID, SBF) the 4 days and surface SEM photograph after 7 days.Wherein (a) and (d) be PBSu, (b) and (c) For NDP, (c) and (f) is CNDP；And (a), (b), (c) be immersion 4 days after SEM photograph, (d), (e), (f) for immersion 7 days SEM photograph afterwards.
In Fig. 9, figure (A) is the PBSu supports of comparative example 1 of the present invention, the mesoporous magnesium silicate microballoon coating of embodiment 1 PBSu supports (NDP) and the PBSu supports (CNDP) for mixing the mesoporous magnesium silicate microballoon coating of copper of embodiment 2 soak in SBF solution EDS spectrogram of the bubble after 7 days；Figure (B) is the change that CNDP soaks the ion concentration after different time in SBF solution.
Scheme in (A) and figure (B), (a) is PBSu, and (b) is NDP, and (c) is CNDP.
Figure 10 is the PBSu supports that BMSCs cells are adhered to comparative example 1 respectively, and the mesoporous magnesium silicate microballoon of embodiment 1 applies PBSu supports (CNDP) upper 12 hour for mixing the mesoporous magnesium silicate microballoon coating of copper of the PBSu supports (NDP) that cover and embodiment 2 SEM figures and the adhesion rate figure of 3,6 and 12 hours (figure d), wherein (a) is PBSu, (b) is NDP, and (c) is CNDP.
In Figure 11, figure (A) is that BMSCs cells are micro- in the PBSu supports of comparative example 1, the mesoporous magnesium silicate of embodiment 1 respectively The PBSu supports (NDP) of ball coating and PBSu supports (CNDP) surface training for mixing the mesoporous magnesium silicate microballoon coating of copper of embodiment 2 Cell proliferation rate figure after supporting 1,3,5 day；Figure (B) is BMSCs cells respectively in PBSu/NDP/CNDP support sample surfaces cultures 7th, the ALP activity figures after 10 and 14 days.
Scheme in (A) and figure (B), (a) is PBSu, and (b) is NDP, and (c) is CNDP.
Figure 12 be HUVECs cells respectively in the PBSu supports of comparative example 1, the mesoporous magnesium silicate microballoon coating of embodiment 1 Cell propagation feelings on PBSu supports (NDP) and the PBSu supports (CNDP) for mixing the mesoporous magnesium silicate microballoon coating of copper of embodiment 2 Condition figure, wherein (a) is PBSu, (b) is NDP, and (c) is CNDP.
Figure 13 is the HUVECs cells PBSu supports with comparative example 1 respectively, the mesoporous magnesium silicate microballoon coating of embodiment 1 After PBSu supports (NDP) and the PBSu supports (CNDP) for mixing the mesoporous magnesium silicate microballoon coating of copper of embodiment 2 co-culture 7 days, The laser confocal microscope figure of 1 hour is cultivated on matrigel, wherein (a) and (d) be PBSu, (b) and (e) is DP, (c) with (f) it is NDP, (d) is CNDP；And (a), (b), the nucleus photo that (c) is DAPI dyeing, (d), (e), (f) they are FITC The F-actin photos of dyeing.
Figure 14 femoral defects in rabbits repairing model and surgical procedure figure.
Figure 15 be comparative example 1 PBSu supports, embodiment 1 mesoporous magnesium silicate microballoon coating PBSu supports (NDP) and The PBSu supports (CNDP) for mixing the coating of copper mesoporous magnesium silicate microballoon of embodiment 2 were implanted into rabbit femoral 1,2 and after 3 months respectively Digital photograph, wherein (a) is PBSu, (b) is NDP, and (c) is CNDP.
Embodiment
The present invention is further illustrated below by the mode of embodiment, but does not therefore limit the present invention to described reality Apply among a scope.The experimental method of unreceipted actual conditions in the following example, conventionally and condition, or according to business Product specification selects.
In the embodiment of the present invention, magnesium nitrate hexahydrate (Mg (NO3)2·6H2O), purchased from traditional Chinese medicines chemical reagent Co., Ltd, Purity is AR；Gerhardite (Cu (NO3)2·3H2O), purchased from traditional Chinese medicines chemical reagent Co., Ltd；Tetraethyl orthosilicate (TEOS), purchased from Shanghai Ling Feng chemical reagent Co., Ltd, purity AR；Poly butylene succinate (PBSu) purchased from Anqing and Xinghua work Co., Ltd, purity AR.
Electric drying oven with forced convection is DHG-9070A, purchased from one permanent Scientific Instruments Corporation of Shanghai；Muffle furnace is SGM 2892A, Purchased from Shanghai sigma high-temperature electric resistance furnace company；3D printer is FFS-MDJ (The Freeform Fabrication System With Micro-Droplet Jetting), purchased from Shanghai Fuqifan Electromechanical Science ＆ Technology Co., Ltd., model HTS-300.
Other raw materials and reagent are all commercially available.
The preparation of the poly butylene succinate of comparative example 1 (PBSu) support
Poly butylene succinate (PBSu) support of this comparative example is prepared using following methods：
30 grams of poly butylene succinates (PBSu) are added in the stainless steel barrel of 3D printer, heating cylinder temperature 112 DEG C are increased to, carries out 3D printing, so as to which PBSu supports be made.
The preparation of the dopamine modified butanediol ester poly succinic acid support (DP) of comparative example 2
Dopamine modified butanediol ester poly succinic acid (PBSu) support of this comparative example is prepared using following methods：
200mg dopamine powder accurately is weighed, is dissolved into 100mL 10mM Tris-HCl cushioning liquid (pH=8.5) In, prepare solution (2mg/mL).Obtained PBSu supports immersion in comparative example 1 in deionized water, is cleaned by ultrasonic 10 minutes, Immersed after drying in above-mentioned dopamine solution.After stirring 24 hours, support is taken out, is cleaned with deionized water, in 50 degrees Celsius of bakings Case is dried.Now, rack surface has formd the dopamine film of black, and gained support sample is DP.
The preparation of the poly butylene succinate support (NDP) of the mesoporous magnesium silicate microballoon coating of embodiment 1
(1) preparation of mesoporous magnesium silicate microballoon (NMS) suspension
1g NMS powder is weighed, is distributed in 100mL deionized waters, produces mesoporous magnesium silicate microsphere suspension liquid.
(2) preparation of the poly butylene succinate support (NDP) of mesoporous magnesium silicate microballoon coating
Support sample DP is soaked in NMS suspension and stirred, speed is 200 revs/min, after stirring 24 hours, is taken out Support, cleaned with deionized water, dried in 60 DEG C of electric drying oven with forced convection, produce NDP.
Embodiment 2 mixes the preparation of the poly butylene succinate support (CNDP) of the mesoporous magnesium silicate microballoon coating of copper
(1) preparation of mesoporous magnesium silicate microballoon (CNMS) suspension of copper is mixed
1g CNMS powder is weighed, is distributed in 100mL deionized waters, produces and mixes the mesoporous magnesium silicate microsphere suspension liquid of copper.
(2) preparation of the poly butylene succinate support (CNDP) of the mesoporous magnesium silicate microballoon coating of copper is mixed
Support sample DP is soaked in CNMS suspension and stirred, speed is 200 revs/min, after stirring 24 hours, is taken out Support, cleaned with deionized water, dried in 60 DEG C of electric drying oven with forced convection, produce CNDP.
The preparation of the poly butylene succinate support (NDP-1) of the mesoporous magnesium silicate microballoon coating of embodiment 3
(1) preparation of mesoporous magnesium silicate microballoon (NMS) suspension
1.1g NMS powder is weighed, is distributed in 100mL deionized waters, produces mesoporous magnesium silicate microsphere suspension liquid.
(2) preparation of the poly butylene succinate support (NDP-1) of mesoporous magnesium silicate microballoon coating
Support sample DP is soaked in NMS suspension and stirred, speed is 200 revs/min, after stirring 24 hours, is taken out Support, cleaned with deionized water, dried in 60 DEG C of electric drying oven with forced convection, produce NDP-1.
Embodiment 4 mixes the preparation of the poly butylene succinate support (CNDP-1) of the mesoporous magnesium silicate microballoon coating of copper
(1) preparation of mesoporous magnesium silicate microballoon (CNMS) suspension of copper is mixed
1.1g CNMS powder is weighed, is distributed in 100mL deionized waters, produces and mixes the mesoporous magnesium silicate microsphere suspension liquid of copper.
(2) preparation of the poly butylene succinate support (CNDP-1) of the mesoporous magnesium silicate microballoon coating of copper is mixed
Support sample DP is soaked in CNMS suspension and stirred, speed is 200 revs/min, after stirring 24 hours, is taken out Support, cleaned with deionized water, dried in 60 DEG C of electric drying oven with forced convection, produce CNDP-1.
Effect example 1NMS and CuNMS sign
1. digital photo figure
Mesoporous magnesium silicate microballoon (NMS) powder and mix mesoporous magnesium silicate microballoon (CuNMS) powder of copper that the present invention uses Digital photo figure is as shown in Figure 1.
As seen from the figure, NMS powder is white, and CuNMS powder is in light green.
2. transmission electron microscope (TEM) is observed
Using projection electron microscope (TEM, JEM-1400, Jeol Ltd.) respectively to mesoporous magnesium silicate microballoon (NMS) powder and mix mesoporous magnesium silicate microballoon (CuNMS) powder of copper and observed.Observation result is shown in Fig. 2.
As seen from the figure, NMS and CuNMS particle size distribution is more uniform, about 200nm；And there is obvious aperture, About 4nm.Illustrate before and after adulterating copper, the particle diameter of particle and aperture are without significant difference.
3. BET is analyzed
Using nitrogen adsorption desorption instrument (Tristar-3000, Merck ＆ Co., Inc of the U.S.), mesoporous magnesium silicate microballoon is determined respectively (NMS) and adsorption/desorption curve of the mesoporous magnesium silicate microballoon (CuNMS) of copper to nitrogen is mixed, using BJH models (Barrett- Joyer-Hanlenda) mesopore orbit is analyzed (specific surface area, pore volume and aperture).As a result Fig. 3 is seen.
From adsorption/desorption isotherms (Fig. 3 A), two kinds of materials are respectively provided with H1 types and return stagnant ring, illustrate that the material is ordered into Mesoporous material.NMS and CuNMS specific surface area is respectively 538.1m2/ g and 490.4m2/g.NMS and CuNMS average pore size Respectively 3.4nm and 3.2nm.As a result show, after adulterating copper, its specific surface area and aperture slightly reduce, without obvious shadow Ring.
4. XPS analysis
With x-ray photoelectron spectroscopy (ESCALAB 250Xi, Thermo Fisher, US) to mesoporous before and after copper doped The component of silicic acid magnesium material is measured, and obtains the Momentum profiles figure of the total spectrograms of XPS and contained element.As a result Fig. 4 is seen.
As seen from the figure, after copper doped, Mg1s peak intensities are declined slightly in spectrogram.Expansion between 923 and 966 eV, It can be clearly seen that NMS does not have peak appearance, and substantially there are Cu2p peaks in CuNMS.As a result show, except Mg, Si and O elements Outside, Cu elements are contained in CuNMS, it is consistent with pale green powder result in digital photo.
Effect example 2 carries out the sign of structure, pattern, element etc. to the support of the present invention
1. the digital photo figure of support
The PBSu supports of the present invention, dopamine surface are modified PBSu supports (DP), the PBSu of mesoporous magnesium silicate microballoon coating Support (NDP) and mix the digital photo figure of PBSu supports (CNDP) of copper mesoporous magnesium silicate microballoon coating and see Fig. 5.
As seen from the figure, PBSu supports are white, and DP is in brownish black, NDP and CNDP colors are slightly shallow, are still brown.In addition, Support NDP-1 and CNDP-1 observing effect are suitable with NDP and CNDP effect respectively.
2. XRD analysis
Using Fourier transformation infrared spectrometer (type of IR, Nicolet 5700, Nicolet companies of the U.S.), respectively to branch Frame carries out molecular structure and chemical composition analysis, test scope use mid-infrared light (about 4000-400cm-1)。
Using X-ray diffractometer (XRD, Rigaku D/max 2550/PC 18KW, Rigaku motor rigaku), divide Do not tested in wide-angle (10-80 °), test result is as shown in Figure 6.
From scheming B, there is obvious diffraction maximum at 19.3 °, 22.2 ° and 29 °, this is PBSu characteristic peak.Surface changes After property, do not change significantly.On the one hand it is because NMS and CuNMS is indefiniteness material；On the other hand be because coating very It is thin, so coating does not influence on XRD.
3. SEM is observed
The present invention four kinds of supports PBSu, DP, NDP and CNDP have been carried out with the observation of microstructure scanning electron microscope sem.As a result see Fig. 7.
As seen from the figure, PBSu rack surfaces are smooth.And it is modified through dopamine, DP surfaces are still more smooth, slightly recessed It is convex.As can be seen that rack surface has all been paved with one layer of evengranular spheroidal particle from figure c and figure d, particle is NMS respectively And CuNMS.Spheroidal particle is more uniform in surface distributed, is signal layer coating.As a result show, porous branch modified using dopamine One layer of spheroidal particle can be adhered in frame surface, and particle is evenly distributed.In addition, support NDP-1 and CNDP-1 observing effect difference It is suitable with NDP and CNDP effect.
The Bioactivity measure of 3 support of the present invention of effect example
Support sample (PBSu, NDP and CNDP) is dipped in SBF solution, SEM observation branch was used at the 4th day and the 7th day Frame modification of surface morphology, and its surface-element is analyzed by EDS, determine the ion concentration change (ICP) in solution.As a result figure is seen 8 and Fig. 9.
As shown in Figure 8, after being immersed in SBF solution 4 days, PBSu surfaces are smooth, without significant change.NDP surfaces have on a small quantity Irregular particle, CNDP surfaces then form the mineral crystal of regular shape.After immersion 7 days, except PBSu surfaces are not obvious Outside change, the mineral quality of NDP and CNDP surfaces deposition is significantly increased, and CNDP surface spherical material aggregations are connected to Together, material surface is substantially completely covered.As a result show, after surface coating NMS and CuNMS, support sample possesses good Good Bioactivity, class bone hydroxyapatite deposition can be induced in surface, contribute in skeletonization in vivo with surrounding bone Tissue produces chemical bonding, so as to form freshman bone tissue.
From Fig. 9 A, PBSu surfaces only have C and O elements, and the peak that there are obvious calcium and P elements in NDP and CNDP surfaces goes out It is existing.Stoichiometry Ca/P ≈ 1.67.
From Fig. 9 B, during whole soak cycle, the concentration of Ca and P ion is always maintained at downward trend, and Si, Mg and Cu ions show ascendant trend, and this result is mainly due to caused by CNDP degraded.
In addition, support NDP-1 and CNDP-1 observing effect are suitable with NDP and CNDP effect respectively.
To sum up result shows, in PBSu stent surface coated NMS and CuNMS coatings, improves the external life of support sample Thing activity.
The cytoactive detection of 4 support of the present invention of effect example
1. the separation and culture of mesenchymal stem cells MSCs (BMSCs)
The SD rats of 4 week old are taken, block femur both ends after execution, bone marrow extraction 2mL, are injected into containing heparin (200U/ ML in PBS), 1500r/min is centrifuged 10 minutes, abandoning supernatant, cell suspension is made with PBS, then be carefully added to 2 times of bodies Long-pending lymphocyte separation medium upper strata, centrifuged 25 minutes with 2500r/min at room temperature.Liquid level intersection cloud and mist is drawn with suction pipe Shape liquid, it is placed in the centrifuge tube of the mL containing PBS 5, piping and druming mixes, supernatant discarding after 1000r/min is centrifuged 5 minutes at room temperature Liquid.After the precipitation in pipe is cleaned into one time with PBS, it is suspended in the α-MEM nutrient solutions containing 20%FCS, through cell counting count board After counting, by density (1 × 104/cm2) be inoculated into 100mL blake bottles, it is placed in incubator (5%CO2, 37 DEG C) in routinely train Support.Culture changes liquid after 24 hours.When observation cell is bred to 80%~90% degree of converging, Secondary Culture.
When primary BMSCs carries out Secondary Culture, 1min is digested in 37 DEG C using 0.25% trypsase, then adds and contains 20%FCS α-MEM nutrient solutions terminate digestion, after centrifugation, using containing 20%FCS, 100U/mL penicillin, 100 μ g/mL strepto-s α-the MEM of element are resuspended into BMSCs suspensions, are fully counted after piping and druming, the cell density as required for subsequent experimental is inoculated with.Choosing With 2-5 subsequent experimental is carried out for BMSCs.
2. cell adherence and morphology observation
Cell is characterized in the adhesion situation of rack surface by observing its cellular morphology in the present embodiment, is specially existed At the 12nd hour, 3 days and 5 days of culture, fixed with PBS sample, then with 2.5% glutaraldehyde solution, for SEM and CLSM is observed.
SEM is observed：When cultivating the 12nd hour, 3 days, 5 days, with PBS sample 2-3 times, glutaraldehyde fixer is washed Only, then it is with 1mL 10%, 30%, 50%, 70%, 85% and 90% ethanol solution dewater treatment, each time successively 15min, finally it is dehydrated twice with 100% absolute ethyl alcohol again.Sample is dried at 37 DEG C.Using SEM to support (PBSu, NDP And CNDP) carry out surface cellular morphology observation.Testing result is shown in Figure 10.
As shown in Figure 10, cell has good adhesion form and upgrowth situation on three kinds of support samples, shows three Kind support no cytotoxicity.The shape of cell mainly shows strip and fusiform.NDP (figure b) and CNDP (figure c) surface The cell number of adhesion is significantly more than PBSu (figure a), and cell is gradually sprawled and stretches out pseudopodium, shows polygon.As a result table Bright, NDP and CNDP surface adhering cells numbers are more, have more preferable cell compatibility.In addition, support NDP-1 and CNDP-1 Testing result it is suitable with NDP and CNDP result respectively.
3. cell is bred and alkaline phosphatase (ALP) activity
When cell rack surface attachment, sprawl after, it is necessary to express certain proliferative as early as possible, can just produce near support The microenvironment of raw enriched cell, so that the secretion of extracellular matrix and the synthesis of each albuminoid, promote material and week Enclose tissue tight's combination.
This effect example determines the proliferative of BMSCs cells using mtt assay and carries out statistical analysis.In cell culture 1st, 3 and 5 days when, per hole add 100 μ L 5%MTT liquid (Sigma Aldrich Chemistry), 37 DEG C culture 4h, Ran Houjia Enter 200 μ L dimethyl sulfoxide (DMSO)s (DMSO), finally with ELIASA at 570nm, survey absorbance (O.D.).5 groups are at least selected to put down Row sample is tested, p<0.05 represents there is statistical significant difference, and all quantitative datas are used using average value Origin pro8.2 are analyzed.Testing result is shown in Figure 11 (a).
Alkaline phosphatase (ALP) activity determination method is：Cell is inoculated on support sample, and tissue culturing plate (TCP) makees For control.After culture 4 days and 7 days, culture medium is discarded, sample is flushed three times with PBS.500 μ L 1%Nonidet is added per hole P-40 (NP-40) solution, cell lysis 1 hour, obtains cell pyrolysis liquid at room temperature.50 μ L1mg/mL nitro is added per hole Benzenephosphonic acid disodium salt (p-nitrophenylphosphate；Songon, China) solution (glycine containing 0.1mol/L and 1mmol/ L magnesium chloride hexahydrates, pH=9), it is placed in 37 DEG C of incubators and is incubated 1 hour.After end, 100 μ L 0.1mol/L is added per hole NaOH solution color development stopping is reacted.Finally existed using ELIASA (384SPECTRAmax, Molecular Devices, the U.S.) At 405nm, O.D. values are determined.The content of total protein uses bovine serum albumin(BSA) to be tried for standard protein using BCA in lysate Agent box (green skies biotechnology, China) is measured.ALP activity is expressed as O.D. values/total protein content at 405nm.Detection As a result Figure 11 (a) is seen.
From Figure 11 (a), when cultivating the 1st day, the O.D. values of three groups of cells are all relatively low, no significant difference.At the 3rd day During with the 5th day, the O.D. values of cell are significantly lifted on NDP and CNDP, hence it is evident that higher than PBSu.This is due to Jie of coating surface Hole magnesium silicate microballoon (NMS) and mix the mesoporous magnesium silicate microballoon (CuNMS) of copper and can form weak base around material and Cellular interfaces Property microenvironment, be adapted to cell adhesion and growth.In addition, NMS can be sustained out Mg and Si ions in nutrient solution, the two is equal The propagation of cell can be promoted.
From Figure 11 (a), at 7 days, the cell ALP activity no significant differences on three kinds of support samples.Over time Passage, the ALP activity of three kinds of support sample surfaces cells all gradually steps up.The ALP activity of the cell on NDP and CNDP surfaces All apparently higher than PBSu, there is significant difference.As a result show, NDP and CNDP have aobvious to the ALP activity expressions of BMSCs cells The facilitation of work.In addition, support NDP-1 and CNDP-1 testing result are suitable with NDP and CNDP result respectively.
4. the propagation of HUVECs cells
Proliferation rate of the HUVECs cells when three kinds of support sample P BSu/NDP/CNDP surfaces are cultivated 1,3 and 5 day is shown in figure 12.As seen from the figure, when cultivating the 1st day, the O.D. values of three kinds of support sample surfaces HUVECs cells are essentially identical.At 3 days, The obvious increase of the O.D. values of cell, and CNDP O.D. values are significantly higher than PBSu (P<0.05).At 5 days, NDP and CNDP's O.D. value is all remarkably higher than PBSu (P<0.05, P<0.01).In addition, support NDP-1 and CNDP-1 testing result respectively with NDP It is suitable with CNDP result.
As a result show, CNDP can be obviously promoted HUVECs cells propagation.
5. HUVECs external sprout occurs
After HUVECs cells and three kinds of support sample P BSu/NDP/CNDP are co-cultured 7 days, it is transferred on matrigel and cultivates 1 Hour, the situation that then its external sprout occurs with confocal laser scanning microscope, as a result see Figure 13.As seen from the figure, After HUVECs co-cultures with PBSu supports, obvious change does not occur；Cell is in spot distribution, and compared to other two groups, Cell number is less (figure a, b).After HUVECs co-cultures with NDP supports, cell starts mutually to draw close with iuntercellular, and exchange increases By force, pseudopodium is connected between part cell and cell, and the blank for having had sprout produces (figure c, d).HUVECs and CNDP supports are total to After culture, it is observed that cell is connected with iuntercellular filopodia, sprout a situation arises highly significant (figure e, f).In addition, branch Frame NDP-1 and CNDP-1 testing result are suitable with NDP and CNDP result respectively.
The zoopery of 5 support of the present invention of effect example and effect
The optimum growing environment of cell is that in vivo, whole animal body is the entirety that can not be split, and works as body Some position receive the stimulation in the external world and damage occur and during defect, its own, which can be produced, promotes the growth of damage location peripheral cell Environment.Due to Cell culture invitro environment can not the real state that grows in vivo of analog cell, so animal is real in vivo Test just particularly important.Autoimmune response and body fluid microenvironment in vivo be present, in vivo the influence to implantation material or ring The promoting factor in border is still not clear, and is observed so only material is implanted into animal body, could really judge the skeletonization of the material Property and whether can be with repairing bone defect.
Support PBSu/NDP/CNDP is implanted into and lacked as animal model by femur defect on the right side of this effect example structure rabbit Position is damaged, femur defect region is taken out in the postoperative 1st, 2 and 3 months.Using synchrotron radiation micro-CT two and three dimensions be imaged into Row detection and analysis, assess freshman bone tissue's growing state and mature bone is organized into bone amount analysis.
1. rabbit femoral model implantation experiment
The support sample specification that zoopery uses is 6 × 6mm of Φ.First support is sealed, then entered using ultraviolet radioactive Row sterilizing.
The body of three kinds of support samples (PBSu/NDP/CNDP) is investigated in this experiment using femur end defect model on the right side of rabbit Interior skeletonization effect.In national tissue engineering technique research center, (Shanghai, China) is carried out for all surgical procedures and animal feeding, Ratified by local Ethics Committee.With NZw, (5 months ages, average weight are for the experiment of 27 male and female half and half of selection Zoopery 3kg) is carried out, stochastic averagina is divided into three groups, and every group 9 is only implanted into a kind of porous support sample respectively.
Zoopery is carried out in aseptic operating room.Anesthetic dosage by animal body re-computation (25mg/kg it is free from worries and 0.15mL/kg 2%Rompon), anaesthetized sb. generally after intramuscular injection.New zealand white rabbit right hind shaving, sterilization, are pressed Prone position is placed on operating table.Right side femur condyle position takes about long 1cm otch, successively separates skin, muscle, periosteum etc. Tissue, appears rabbit femoral ectocondyle.From orthopedic drill, 6mm × 6mm cylinders are manufactured along the femur longitudinal axis and sagittal axis vertical direction Spinal joints hole defect, above-mentioned three kinds of supports distinguish implantation in rabbit Cranial defect model, successively tight sewing-up cut, and iodine is smeared in wound surface Volt carries out wound disinfection.Postoperative single cage raising, diet, intramuscular injection penicillin is with anti-inflammatory for three days on end.Surgical procedure is shown in figure 14。
Animal was put to death by injecting overdose of sodium pentobarbital respectively at postoperative 4,8 and 12 weeks, takes out defect repair position, stripping Except acquisition bone sample sample after musculature.Digital photograph is shot, gross examination of skeletal muscle is carried out to Cranial defect situation.Then use in 10% Property formalin buffer carrys out fixed preparation, carries out various detections.
2. gross examination of skeletal muscle
Digital photograph in support sample P BSu, NDP and CNDP implantation rabbit femoral after 1,2 and 3 month is shown in Figure 15.Can by figure See, for stenter to implant after 1 month, PBSu is still exposed to outside, can be clearly apparent support, surface Cranial defect is more apparent, does not obtain Significantly repair.NDP and CNDP implant site is with the presence of obvious pit, but support has been wrapped in organization internal in itself. After stenter to implant 2 months, three pack support defects have obvious healing trend.After stenter to implant 3 months, PBSu implantation Position, obvious Cranial defect still be present and fail to repair；NDP implant sites, defect and the obscurity boundary of support sample, only The hole of remaining very little is not yet repaired；CNDP implant sites, defect and the border of support sample disappear, and can not can be visually seen The boundary of host bone and implantation material.As a result show, CNDP has compatibility and osteoinductive inside preferably, can be effective Promote the regeneration of freshman bone tissue.In addition, support NDP-1 and CNDP-1 testing result the result phase with NDP and CNDP respectively When.

Claims (10)

  1. A 1. cated poly butylene succinate support of tool, it is characterised in that the support includes poly-succinic acid-butanediol Ester rest body, dopamine film and mesoporous magnesium silicate microballoon coating mix the mesoporous magnesium silicate microballoon coating of copper；
    Described mesoporous magnesium silicate microballoon coating mixes the outer layer that the mesoporous magnesium silicate microballoon coating of copper is coated in dopamine film；
    Described dopamine film is located at the surface of poly butylene succinate rest body；
    Described mesoporous magnesium silicate microballoon coating is mixed in the mesoporous magnesium silicate microballoon coating of copper, microspherulite diameter 180-220nm, micro- The aperture of ball is 3-5nm；
    The thickness of described dopamine film is 50-100nm.
  2. 2. support as claimed in claim 1, it is characterised in that
    The thickness of the dopamine film is 60-80nm；
    And/or the mesoporous magnesium silicate microballoon or to mix the aperture of the mesoporous magnesium silicate microballoon of copper be 4nm；
    And/or the mesoporous magnesium silicate microballoon or to mix the particle diameter of the mesoporous magnesium silicate microballoon of copper be 200nm；
    And/or the specific surface area of the mesoporous magnesium silicate microballoon is 500-570m2/ g, preferably 538.1m2/g；
    And/or the specific surface area for mixing the mesoporous magnesium silicate microballoon of copper is 450-550m2/ g, preferably 490.4m2/g；
    And/or it is described mix in the mesoporous magnesium silicate microballoon of copper, the content of copper is 3-7wt%, preferably 5wt%；
    And/or the number-average molecular weight of the poly butylene succinate is 4 × 104-7×104, the equal molecule of weight average molecular weight/number Measure as 1.2-2.4；
    And/or the mesoporous magnesium silicate microballoon is made by the method comprised the following steps：
    S1, cetyl trimethylammonium bromide is dissolved completely in water, adds ammoniacal liquor, tetraethyl orthosilicate and six nitric hydrates Magnesium, stand reaction；
    S2, the reaction product obtained by S1 is centrifuged and washs gained white depositions, drying；
    S3, drying product obtained by S2 is placed in Muffle furnace sintered, remove cetyl trimethylammonium bromide, gained white powder As mesoporous magnesium silicate microballoon；
    And/or the mesoporous magnesium silicate microballoon of copper of mixing is made by the method comprised the following steps：
    M1, cetyl trimethylammonium bromide is dissolved completely in water, adds magnesium nitrate hexahydrate and Gerhardite, it is quiet Put reaction；
    M2, the reaction product obtained by M1 is centrifuged and washs gained light green sediment, drying；
    M3, drying product obtained by M2 is placed in Muffle furnace sintered, remove cetyl trimethylammonium bromide, gained pale green toner End is to mix the mesoporous magnesium silicate microballoon of copper.
  3. 3. support as claimed in claim 2, it is characterised in that
    Water in step S1 is deionized water；
    And/or in step S1, the concentration of the ammoniacal liquor is 1-1.5M, preferably 1M；
    And/or in step S1, the time for standing reaction is 3-6 hours, preferably 4 hours；
    And/or step S1 is carried out according to following operation：At 35-40 DEG C, the dissolving of 0.6-0.8g cetyl trimethylammonium bromides In 30-35mL deionized waters, 6.5-7.5mL 1M ammoniacal liquor, 3.2-3.8mL tetraethyl orthosilicates and 2.3-2.5g six are sequentially added Nitric hydrate magnesium, stand reaction；
    And/or in step S2, the washing is washed using deionized water and absolute ethyl alcohol；
    And/or in step S2, the temperature of the drying is 50-70 DEG C, preferably 60 DEG C；
    And/or in step S3, the temperature of the sintering is 500-700 DEG C, preferably 600 DEG C；The soaking time of the sintering is 2- 4 hours, preferably 3 hours；
    And/or in step M1, the concentration of the ammoniacal liquor is 1-1.5M, preferably 1M；
    And/or in step M1, the water is deionized water；
    And/or in step M1, the time for standing reaction is 3-6 hours, preferably 4 hours；
    And/or step M1 is carried out according to following operation：At 35-40 DEG C, the dissolving of 0.6-0.8g cetyl trimethylammonium bromides In 30-35mL deionized waters, 1.6-1.8g magnesium nitrate hexahydrates and 0.5-0.6g Gerhardites are added, stands reaction；
    And/or in step M2, the washing is washed using deionized water and absolute ethyl alcohol；
    And/or in step M2, the temperature of the drying is 50-70 DEG C, preferably 60 DEG C；
    And/or in step M3, the temperature of the sintering is 500-700 DEG C, and preferably 600 DEG C, the soaking time of the sintering is 2- 4 hours, preferably 3 hours.
  4. 4. a kind of method of the cated poly butylene succinate support of tool prepared as described in claim any one of 1-3, Characterized in that, it the described method comprises the following steps：
    (1) 3D printing prepares poly butylene succinate rest body；The technical parameter of the 3D printing is preferably as follows：3D printing The barrel temperature of equipment is 105-120 DEG C, preferably 112 DEG C；The discharge velocity of 3D printing equipment is 100-200g/min；3D printing The mechanical arm rate travel of equipment is 0.1-1m/s；
    (2) dopamine film is formed in poly butylene succinate rest body surface prepared by step (1), obtains dopamine surface Modified butanediol ester poly succinic acid support；
    (3) mesoporous magnesium silicate microballoon is coated on the dopamine surface modified butanediol ester poly succinic acid support that step (2) obtains Or the mesoporous magnesium silicate microballoon of copper is mixed, so as to which the described cated poly butylene succinate support of tool be made.
  5. 5. method as claimed in claim 4, it is characterised in that step (2) is carried out using the method for including following operation：
    Poly butylene succinate rest body is cleaned by ultrasonic, immerses in dopamine solution and stirs；Cleaned, dried with deionized water It is dry, obtain dopamine surface modified butanediol ester poly succinic acid support.
  6. 6. method as claimed in claim 5, it is characterised in that
    The ultrasonic cleaning of the rest body is carried out in deionized water, and ultrasonic time is 5-20 minutes, preferably 10 minutes；
    And/or the Tris-HCl cushioning liquid that the dopamine solution is dopamine；
    And/or the concentration of the dopamine solution is 1-3mg/ml, preferably 2mg/ml；
    And/or mixing speed of the rest body in dopamine solution be 100-300 revs/min, preferably 150-200 turn/ Minute；Mixing time is 18-36 hours, preferably 24 hours；
    And/or the temperature of the drying is 50-70 DEG C, preferably 60 DEG C.
  7. 7. method as claimed in claim 4, it is characterised in that step (3) is carried out using the method for including following operation：
    The dopamine surface modified butanediol ester poly succinic acid support that step (2) obtains is soaked in into mesoporous magnesium silicate microballoon to suspend Liquid is mixed in the mesoporous magnesium silicate microsphere suspension liquid of copper and stirred；Cleaned, dried with deionized water, applied so as to which described having be made The poly butylene succinate support of layer.
  8. 8. method as claimed in claim 7, it is characterised in that
    The dopamine surface modified butanediol ester poly succinic acid support is in mesoporous magnesium silicate microsphere suspension liquid or mixes copper mesoporous silicon Mixing speed in sour magnesium microsphere suspension liquid is 100-300 revs/min, preferably 150-200 revs/min；Mixing time is 18-36 Hour, preferably 24 hours；
    And/or the mesoporous magnesium silicate microsphere suspension liquid or mix in the mesoporous magnesium silicate microsphere suspension liquid of copper, microballoon and dispersing liquid Mass volume ratio be 0.5-2g/100ml, preferably 1g/100ml；
    And/or the temperature of the drying is 50-70 DEG C, preferably 60 DEG C.
  9. A kind of 9. cated poly butylene succinate support of tool prepared by method as described in claim any one of 4-8.
  10. 10. a kind of cated poly butylene succinate support of tool as described in claim 1-3 or 9 any one is preparing bone Application in impairment renovation material field.
CN201710657006.XA 2017-08-03 2017-08-03 Has cated 3D printing support and its preparation method and application Pending CN107412855A (en)

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CN113041403B (en) * 2021-03-25 2022-04-08 四川大学 Bone repair n-HA/CS porous scaffold, preparation method and application

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