CN107412879A

CN107412879A - 3D printing compound rest and its preparation method and application

Info

Publication number: CN107412879A
Application number: CN201710657054.9A
Authority: CN
Inventors: 苏佳灿; 苏奕铭; 魏杰
Original assignee: Shanghai State Of Bio Technology Co Ltd
Current assignee: Shanghai State Of Bio Technology Co Ltd
Priority date: 2017-08-03
Filing date: 2017-08-03
Publication date: 2017-12-01

Abstract

The invention discloses 3D printing compound rest and its preparation method and application.Described 3D printing compound rest is mesoporous magnesium silicate/poly butylene succinate binary compound rest and mesoporous magnesium silicate/poly butylene succinate/gliadin tri compound support.It is 1 that described binary compound rest, which includes mesoporous magnesium silicate and the ratio between poly butylene succinate, its content,:1‑1:5, the aperture of support is 300 500 μm；It is 1 that described tri compound support, which includes mesoporous magnesium silicate/poly butylene succinate/the ratio between gliadin, its content,:1:1‑5:5:1, the aperture of support is 300 500 μm.The compound rest of the present invention has preferable Bioactivity, improves the hydrophilicity of material, effectively raises the degradation rate of support；And show preferably to promote cell propagation and Osteoblast Differentiation ability, there is preferable application prospect preparing bone impairment renovation material field.

Description

3D printing compound rest and its preparation method and application

Technical field

The invention belongs to biological medicine engineering field, in particular it relates to 3D printing compound rest and preparation method thereof and should With.

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, therefore, the research for Bone Defect Repari and alternative materials is significant.Preferable bone renovating material should have Bionical physical arrangement, chemical composition and degradability.

In recent years, mesoporous magnesium silicate is of great interest for the research of bone defect healing.Mesoporous material is a kind of New nanostructured functional material, there is preferable bioactivity, cell can be promoted to breed and break up, and effectively activate bone Related gene expression；Magnesium ion can adjust the affinity of cell and material surface, and promote the propagation of Gegenbaur's cell.Therefore it is situated between Hole magnesium silicate is preferable bone tissue regeneration repair materials.In addition, aliphatic polyester has been widely used for bio-medical Material Field, because it has good biological degradability.Such material can by multiple-microorganism or in animal body enzyme degraded, Metabolism, and it is finally decomposed to CO₂And H₂O, and then excrete, among these with poly butylene succinate (poly (butylene Succinate, PBSu) it is representative.However, PBSu is hydrophobic material there is also shortcoming, such as PBSu, PBSu surfaces can be influenceed Cell adhesion behavior；PBSu or bio-inert material, the combination of itself and host bone can be hindered, influence its bone formation performance, lacked Effect is obviously promoted to bone tissue regeneration.

Although above-mentioned single repair materials have the advantages of certain, support shortcoming is also bright made of single material Aobvious, therefore, a kind of this area material hole of urgent need is uniform, mechanical strength is high, making is simple, the suitable composite support of degradation rate Frame.

The content of the invention

Present invention aim to address lacking in this area, a kind of material hole is uniform, mechanical strength is high, making is simple, drop Solve the suitable compound rest of speed, and then provide a kind of mesoporous magnesium silicate/poly butylene succinate binary compound rest and A kind of mesoporous magnesium silicate/poly butylene succinate/gliadin tri compound support.The compound rest of the present invention uses 3D Prepared by Method of printing, have preferable Bioactivity, improve the hydrophilicity of material, effectively raise the drop of support Solve speed；And show preferably to promote cell propagation and Osteoblast Differentiation ability, preparing bone impairment renovation material field tool There is preferable application prospect.

The present invention solves above-mentioned technical problem by following technical proposals.

First aspect present invention provides a kind of mesoporous magnesium silicate/poly butylene succinate binary compound rest, wherein, The binary compound rest includes following components：Mesoporous magnesium silicate and poly-succinic acid-butanediol；The mesoporous magnesium silicate compares surface Product is 445-455m²/ g, pore volume 0.3-0.5cm³/g；

The mass ratio of the mesoporous magnesium silicate and poly butylene succinate is 1:1-1:5；

The aperture of the binary compound rest is 300-500 μm.

Described binary compound rest,

Wherein, the mass ratio of the mesoporous magnesium silicate and poly butylene succinate is preferably 1:1-1:3, more preferably for 1:1-1:1.5。

Wherein, the binary compound rest specification is preferably (Φ 6-12) × (1-8) mm, be more preferably 12 × 2mm of Φ, 6 × 6mm of Φ 10 × 2mm and Φ.

Wherein, the aperture of the binary compound rest is preferably 400 μm.

Wherein, the compressive strength of the binary compound rest is preferably 60-80Mpa, more preferably 65-75Mpa.

Wherein, the porosity of the binary compound rest is preferably 40-60%, more preferably 42-52%.

Wherein, the mesoporous magnesium silicate is commonly used in the art that macroscopic view exists with powdered form.It is it is preferred that described mesoporous The particle diameter of magnesium silicate is 800-1500nm, average pore size 6-7nm.

Wherein, the mesoporous magnesium silicate can be prepared using this area conventionally used for preparing the method for mesoporous magnesium silicate, Typically use sol-gal process.In a preferred embodiment of the present invention, the mesoporous magnesium silicate is using the side comprised the following steps It is prepared by method：

S1, at 45-55 DEG C, template and water and mixed in hydrochloric acid, are clarified at stirring to solution；

S2, at 45-55 DEG C, by magnesium nitrate hexahydrate (Mg (NO₃)₂·6H₂O), tetraethyl orthosilicate (TEOS) adds above-mentioned molten In liquid, stir to stand after 4-6h and filter, dry afterwards white powder；

S3, gained white powder is calcined to removing template at 500-700 DEG C.

Wherein, in step S1, the template is template commonly used in the art, generally use PEO- PPOX-PEO triblock copolymer, such as the average weight-average molecular weight that Sigma Aldrich are provided is 5800 P123 triblock copolymers.The water can be deionized water.The concentration of the hydrochloric acid is preferably 1.7-2.5mol/L.At this Invent in a preferred embodiment, step S1 is carried out according to following operation：By 5.0-7.0g P123 block copolymers and 20-40mL Water and the mixed in hydrochloric acid that 110-130mL concentration is 1.7-2.5mol/L, stirring, are clarified to solution.

Wherein, in step S2, add magnesium nitrate hexahydrate and tetraethyl orthosilicate substep can be used to add into above-mentioned solution, first add Enter magnesium nitrate hexahydrate, add tetraethyl orthosilicate.In a preferred embodiment of the present invention, step S2 is carried out according to following operation： At 45-55 DEG C, 4.6-5.0g magnesium nitrate hexahydrates and 8.3-8.7g tetraethyl orthosilicates are added in above-mentioned solution successively, stir 4- Stand and filter after 6h.

Wherein, in step S2, the suction filtration can be carried out repeatedly, generally 2~5 times, preferably 3 times.Knot is filtered every time Cleaned using ionized water as Shu Houyi, filtered again afterwards.

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 operation and condition of the operation of the calcining and condition for the conventional calcining in this area, preferably Ground is carried out in 600 DEG C of Muffle furnace.

Second aspect of the present invention provides a kind of 3D printing and prepares mesoporous magnesium silicate as described in the first aspect of the invention/poly- The method of succinic acid-butanediol ester binary compound rest, wherein, it the described method comprises the following steps：

A1, the mesoporous magnesium silicate and the poly butylene succinate mixed, obtain binary mix material；

A2, the binary mix material obtained to step A1 using 3D printing equipment carry out 3D printing, produce described mesoporous Magnesium silicate/poly butylene succinate binary compound rest.

In step A1, the preparation of binary mix material can be carried out in device commonly used in the art, such as in quark banburying Carried out in machine.The process prepared using quark banbury includes entering the mesoporous magnesium silicate and poly-succinic acid-butanediol of addition The step of row stirring；Wherein, the speed of the stirring is preferably 100-500rpm, such as 300rpm；The time of the stirring is excellent Elect 2-5 hours, such as 3 hours as.

In step A2, the 3D printing equipment is the conventional 3D printer in this area, preferably electromechanical by Shanghai Fu Qifan Science and Technology Ltd. provides, model HTS-300.

Wherein, the method for the 3D printing and operation are the conventional Method of printing in this area and operation, it is preferred that the 3D The technical parameter of printing includes as follows：The injection rate of the mixed material is 20-30g/min, such as 25g/min；The 3D The barrel temperature of printing device is 100-125 DEG C, preferably 110-115 DEG C, such as 112 DEG C.

Third aspect present invention provides mesoporous magnesium silicate prepared by a kind of method as described in second aspect of the present invention/poly- Succinic acid-butanediol ester binary compound rest.

Fourth aspect present invention provides a kind of mesoporous magnesium silicate/poly butylene succinate/gliadin ternary and answered Support is closed, wherein, the tri compound support includes following components：Mesoporous magnesium silicate, poly butylene succinate and wheat alcohol are molten Albumen；The specific surface area of the mesoporous magnesium silicate is 445-455m²/ g, pore volume 0.3-0.5cm³/g；

The mass ratio of the mesoporous magnesium silicate, poly butylene succinate and gliadin is 1:1:1-5:5:1；

The aperture of the tri compound support is 300-500 μm.

Described tri compound support,

Wherein, the mass ratio of the mesoporous magnesium silicate, poly butylene succinate and gliadin is preferably 1:1:1- 3:3:1；More preferably 2:2:1.

Wherein, the specification of the tri compound support be preferably (Φ 6-12) × (1-8) mm, more preferably for Φ 12 × 6 × 6mm of 2mm, Φ 10 × 2mm and Φ.

Wherein, the compressive strength of the tri compound support is preferably 60-80Mpa, more preferably 65-75Mpa.

Wherein, the porosity of the tri compound support is preferably 40-60%, more preferably 42-52%.

Wherein, the aperture of the tri compound support is preferably 400 μm.

Wherein, the mesoporous magnesium silicate is commonly used in the art that macroscopic view exists with powdered form, it is preferred that described mesoporous The particle diameter of magnesium silicate is 800-1500nm, average pore size 6-7nm.The mesoporous magnesium silicate can be as described in first aspect present invention The method for preparing mesoporous magnesium silicate be made.

Fifth aspect present invention provides a kind of 3D printing and prepares mesoporous magnesium silicate as described in fourth aspect present invention/gather The method of succinic acid-butanediol ester/gliadin tri compound support, wherein, it the described method comprises the following steps：

B1, by the mesoporous magnesium silicate, the poly-succinic acid-butanediol and the gliadin mix, obtain ternary mixing Material；

B2, the ternary mixture material obtained to step B1 using 3D printing equipment carry out 3D printing, produce described mesoporous Magnesium silicate/poly butylene succinate/gliadin tri compound support.

In step B1, the preparation of ternary mixture material can be carried out in device commonly used in the art, such as in quark banburying Carried out in machine.The process prepared using quark banbury include to the mesoporous magnesium silicate of addition, poly-succinic acid-butanediol and The step of gliadin is stirred；Wherein, the speed of the stirring is preferably 100-500rpm, such as 300rpm, described The time of stirring is preferably 2-5 hours, such as 3 hours.

In step B2, the 3D printing equipment is the conventional 3D printer in this area, preferably electromechanical by Shanghai Fu Qifan Science and Technology Ltd. provides, model HTS-300.

Wherein, the method for the 3D printing and operation are the conventional Method of printing in this area and operation, it is preferred that the 3D The technical parameter of printing is preferably as follows：The injection rate of mixed material is 20-30g/min, such as 25g/min；Barrel temperature is 100-125 DEG C, preferably 110-115 DEG C, such as 112 DEG C.

In the present invention, the poly-succinic acid-butanediol and gliadin are material commonly used in the art, are It is commercially available.

In the preferred embodiment of the present invention, step A2 or step B2 are carried out according to following operation：By obtained mixing Material is added in the stainless steel barrel of 3D printing equipment, then heating cylinder temperature carries out 3D printing to 110-115 DEG C, from And mesoporous magnesium silicate/poly butylene succinate binary compound rest described in first aspect present invention or the present invention the 3rd is made Mesoporous magnesium silicate/poly butylene succinate/gliadin tri compound support described in aspect.

Sixth aspect present invention provides mesoporous magnesium silicate prepared by a kind of method as described in fifth aspect present invention/poly- Succinic acid-butanediol ester/gliadin tri compound support.

Seventh aspect present invention provides a kind of mesoporous magnesium silicate as described in first aspect present invention or the third aspect/poly- Mesoporous magnesium silicate/polybutadiene described in succinic acid-butanediol ester binary compound rest or fourth aspect present invention or the 6th aspect Sour butanediol ester/application of the gliadin tri compound support in bone renovating 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：

(1) present invention uses 3D printing technique, is prepared for mesoporous magnesium silicate/poly butylene succinate (m-MS/PBSu) Binary compound rest and mesoporous magnesium silicate-poly-succinic acid-butanediol-gliadin (m-MS/PBSu/Gliadin) tri compound Support.Described compound rest forms a large amount of microcellular structures on its surface, so as to form the multi-stage porous knot of macropore-micropore-mesopore Structure, there is more preferable Bioactivity, improve the hydrophilicity of material, effectively raise the degradation rate of support；

(2) mesoporous magnesium silicate/poly butylene succinate (m-MS/PBSu) binary compound rest and mesoporous silicon of the invention Sour magnesium-poly-succinic acid-butanediol-gliadin tri compound support shows preferably to promote cell propagation and Osteoblast Differentiation Ability, there is higher application value in Bone Defect Repari and osteogenic materials field.

Brief description of the drawings

The 3D printing support S2 and implementation that Fig. 1 is the 3D printing support S1 of comparative example 1 of the present invention preparation, prepared by embodiment 1 3D printing support S3 prepared by example 2 digital photo figure, wherein (a) is S1, (b) is S2, and (c) is S3.

In Fig. 2, the 3D that (A) and (B) is the 3D printing support S1 that respectively prepared by comparative example 1 of the present invention, prepared by embodiment 1 is beaten Print support S2 and the 3D printing support S3 of the preparation of embodiment 2 wide-angle x-ray diffraction collection of illustrative plates and FTIR spectrum figure；Its In (a) be S1, (b) is S2, and (c) is S3, and (d) is gliadin and (e) is mesoporous magnesium silicate (m-MS).

The 3D printing support S2 and implementation that Fig. 3 is the 3D printing support S1 of comparative example 1 of the present invention preparation, prepared by embodiment 1 3D printing support S3 prepared by example 2 microstructure scanning electron microscope (SEM) photograph：Wherein (a) is S1, and (b) is S2, and (c) is S3.

The 3D printing support S2 and implementation that Fig. 4 is the 3D printing support S1 of comparative example 1 of the present invention preparation, prepared by embodiment 1 3D printing support S3 prepared by example 2 section microstructure scanning electron microscope (SEM) photograph, wherein (a) and (d) is S1, (b) and (e) is S2, (c) and (f) is S3.

The 3D printing support S2 and implementation that Fig. 5 is the 3D printing support S1 of comparative example 1 of the present invention preparation, prepared by embodiment 1 3D printing support S3 prepared by example 2 SR μ CT electronics section photo and Three-dimensional Gravity composition, wherein (a) and (d) be S1, (b) with (e) it is S2, (c) and (f) is S3.

The 3D printing support S2 and implementation that Fig. 6 is the 3D printing support S1 of comparative example 1 of the present invention preparation, prepared by embodiment 1 Microstructures of the 3D printing support S3 under laser co-focusing 3D microscopes prepared by example 2, wherein (a) and (d) be S1, (b) with (e) it is S2, (c) and (f) is S3.

The 3D printing support S2 and implementation that Fig. 7 is the 3D printing support S1 of comparative example 1 of the present invention preparation, prepared by embodiment 1 Scanning after 3D printing support S3 prepared by example 2 soaks 5 days in simulated body fluid (SIMULATED BODY FLUID, SBF) is electric Mirror photo, wherein (a) and (d) is S1, (b) and (e) is S2, and (c) and (f) is S3.

Fig. 8 (A) is 3D printing support S1, the 3D printing support S2 and reality of the preparation of embodiment 1 prepared by comparative example 1 of the present invention Apply the EDS spectrums after 3D printing support S3 prepared by example 2 soaks 5 days in simulated body fluid (Simulated Body Fluid, SBF) Figure, wherein (a) is S1, (b) is S2, and (c) is S3；Figure (B) is that S3 soaks the ion concentration in solution after different time in SBF Change.

Fig. 9 (A) is 3D printing support S1, the 3D printing support S2 and reality of the preparation of embodiment 1 prepared by comparative example 1 of the present invention Apply the adsorption protein content after 3D printing support S3 prepared by example 2 soaks 24 hours in bovine serum albumen solution (BSA)； (B) weight-loss ratio after different time is soaked in Tris-HCl for S1, S2 and S3, wherein (a) is S1, (b) is S2, and (c) is S3.

In Figure 10, (a), (b) and (c) be BMSCs cell adherences prepared in comparative example 1 of the present invention 3D printing support S1, SEM figures on 3D printing support S3 samples prepared by 3D printing support S2 and embodiment 2 prepared by embodiment 1 after 12 hours；Figure (d) it is its 3,6 and 12 hours adhesion rate, wherein (a) is S1, (b) is S2, and (c) is S3.

Figure 11 (A) be comparative example 1 of the present invention prepare 3D printing support S1, embodiment 1 prepare 3D printing support S2 and 3D printing support S3 prepared by embodiment 2 is it 7,10 in the influence to MC3T3-E1 cell proliferative conditions in 1,3 and 7 day, (B) With the influence to BMSCs cells ALP activity in 14 days；Wherein (a) is S1, and (b) is S2, and (c) is S3.

The 3D printing support S2 and implementation that Figure 12 is the 3D printing support S1 of comparative example 1 of the present invention preparation, prepared by embodiment 1 3D printing support S3 prepared by example 2 is implanted into the digital photograph of femur after 4,8 and 12 weeks in animal body respectively, wherein (a) is S1, (b) it is S2, (c) is S3, and circle indicates implant site.

The 3D printing support S2 and implementation that Figure 13 is the 3D printing support S1 of comparative example 1 of the present invention preparation, prepared by embodiment 1 MicroCT three-dimensionalreconstruction photos in 3D printing support S3 implantation animal bodies prepared by example 2 after 4,8 and 12 weeks, wherein (a) is S1, (b) are S2, and (c) is S3.

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 following embodiments, P123 (EO₂₀PO₇₀EO₂₀, molecular weight 5800), purchased from Sigma Aldriches,

Magnesium nitrate hexahydrate (Mg (NO₃)₂·6H₂O), purchased from traditional Chinese medicines chemical reagent Co., Ltd, purity AR,

Tetraethyl orthosilicate (TEOS), purchased from Shanghai Ling Feng chemical reagent Co., Ltd, purity AR,

Poly butylene succinate (PBSu) is purchased from Anqing and Xinghua work Co., Ltd, purity AR,

Gliadin (Gliadin from Wheat) is pure purchased from uncommon love (Shanghai) the chemical conversion industry Development Co., Ltd of ladder Spend for AR；

Quark banbury is purchased from Bao Lun precision detecting instruments company,

Electric drying oven with forced convection is DHG-9070A, purchased from one permanent Scientific Instruments Corporation of Shanghai,

Tablet press machine is YP-15t, purchased from Tianjin Jin Fu Lun Science ＆ technology Co., Ltd,

Muffle furnace is SGM 2892A, purchased from Shanghai sigma high-temperature electric resistance furnace company,

3D printing equipment is FFS-MDJ (The Freeform Fabrication System with Micro- Droplet Jetting), purchased from Shanghai Fuqifan Electromechanical Science ＆ Technology Co., Ltd..

Other raw materials and reagent are all commercially available.

The preparation of the mesoporous magnesium silicate of embodiment 1/poly butylene succinate binary compound rest (S2)

(1) preparation of mesoporous magnesium silicate (m-MS)

The m-MS of the present embodiment is prepared with the following method：

30mL deionized waters and the mixed solution of 120mL 2.0M watery hydrochloric acid are placed in 50 DEG C of water-baths.Then, it is accurate to claim 4.0g P123 are taken, are added into above-mentioned mixed solution, stirring is extremely clarified for 30 minutes.Then, 4.8g six is weighed successively is hydrated nitre Sour magnesium and 8.5g tetraethyl orthosilicate is added in above-mentioned settled solution, is stirred 5 hours.Filtered after standing, deionized water cleaning, then Filter, after being repeated 3 times, be placed in 60 DEG C of electric drying oven with forced convections and obtain white powder.Powder is put into stainless steel mould, adopted Wafer sample (12 × 2mm of Φ) is made under 2MPa pressure with tablet press machine.Above-mentioned powder and wafer sample are finally placed in Muffle In stove, (heating rate is sintered under the conditions of 600 DEG C：1 DEG C/min), so as to obtain m-MS powder and wafer sample (Φ 12 ×2mm)。

(2) preparation of mesoporous magnesium silicate/poly butylene succinate binary compound rest (S2)

The present embodiment prepares mixed material using quark banbury, and wherein in mixed material, mesoporous magnesium silicate dosage is 20 Gram, poly butylene succinate dosage is 30 grams, then gained mixed material is added to the stainless steel barrel of 3D printing equipment Interior, heating cylinder temperature prepares binary compound rest S2 to 112 DEG C.The present invention is prepared for the support material of three kinds of different sizes altogether Material, wherein 12 × 2mm of Φ support is used for material characterization and cell experiment, and 10mm × 10mm × 2mm support is used for upper sea light Source SR μ CT are tested, and 6 × 6mm of Φ support is used for zoopery.

The preparation of the mesoporous magnesium silicate/poly butylene succinate of embodiment 2/gliadin tri compound support (S3)

(1) preparation of mesoporous magnesium silicate (m-MS)

The m-MS of the present embodiment is prepared with the following method：

(2) preparation of mesoporous magnesium silicate/poly butylene succinate/gliadin tri compound support (S3)

The present embodiment prepares mixed material using quark banbury, and wherein in mixed material, mesoporous magnesium silicate dosage is 20 Gram, poly butylene succinate dosage is 20 grams, and gliadin dosage is 10 grams, and gained mixed material then is added into 3D In the stainless steel barrel of printing device, heating cylinder temperature prepares tri compound support S3 to 112 DEG C.The present invention is prepared for altogether The timbering material of three kinds of different sizes, wherein 12 × 2mm of Φ support are used for material characterization and cell experiment, and 10mm × 10mm × 2mm support is tested for SSRF SR μ CT, and 6 × 6mm of Φ support is used for zoopery.

The preparation of the mesoporous magnesium silicate of embodiment 3/poly butylene succinate binary compound rest (S4)

(1) preparation of mesoporous magnesium silicate (m-MS)

The m-MS of the present embodiment is prepared with the following method：

(2) preparation of mesoporous magnesium silicate/poly butylene succinate binary compound rest (S4)

The present embodiment prepares mixed material using quark banbury, and wherein in mixed material, mesoporous magnesium silicate dosage is 25 Gram, poly butylene succinate dosage is 25 grams, then gained mixed material is added to the stainless steel barrel of 3D printing equipment Interior, heating cylinder temperature prepares binary compound rest S4 to 112 DEG C.

The preparation of the mesoporous magnesium silicate/poly butylene succinate of embodiment 4/gliadin tri compound support (S5)

(1) preparation of mesoporous magnesium silicate (m-MS)

The m-MS of the present embodiment is prepared with the following method：

(2) preparation of mesoporous magnesium silicate/poly butylene succinate/gliadin tri compound support (S5)

The present embodiment prepares mixed material using quark banbury, and wherein in mixed material, mesoporous magnesium silicate dosage is 35 Gram, poly butylene succinate dosage is 35 grams, and gliadin dosage is 30 grams, and gained mixed material then is added into 3D In the stainless steel barrel of printing device, heating cylinder temperature prepares tri compound support S5 to 112 DEG C.

The preparation of the poly butylene succinate of comparative example 1 (PBSu) support (S1)

In this comparative example, only 3D printing support, the 3D printing system are prepared using poly butylene succinate (PBSu) Preparation Method is identical with the method described in Examples 1 and 2.

Three kinds of 3D printing support S1/S2/S3, respective formula composition and physicochemical constant are as shown in table 1：

Table 1

Support S1/S2/S3 digital photograph is shown in Fig. 1.

Structure, pattern, the sign of element composition of the support of effect example 1 (S1/S2/S3)

1. IR, XRD analysis

Using Fourier transformation infrared spectrometer (type of IR, Nicolet 5700, Nicolet companies of the U.S.), to support (S1/S2/S3) carry 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), at wide-angle (10-80 °) Tested, test result is as shown in Figure 2.

From Fig. 2 (A), in the XRD spectral lines (d) of gliadin (Gliadin), nearby there is one in 2 θ=20 ° Wider diffraction maximum, it is amorphous amorphous materials to illustrate Gliadin；In m-MS XRD spectral lines (e), 2 θ=25 ° are nearby present One wider diffraction maximum, it is its characteristic peak.In S1 XRD spectral lines (a), can be observed at 19.3 °, 22.2 °, 25.9 ° and There is obvious diffraction maximum at 29 °, this is PBSu characteristic peak.In S2 and S3 XRD spectral lines (b, c), it can be observed PBSu's Characteristic peak, but the intensity at peak substantially reduces.As a result show, m-MS and Gliadin addition so that the crystallinity of compound rest It is obvious to reduce.

From Fig. 2 (B), in S1 spectrogram (a), 1730cm^-1Locate the strong absworption peak occurred, corresponding is C=O in PBSu The symmetrical stretching vibration peak of key.In Gliadin spectrogram (d), 1650cm^-1And 1550cm^-1The absworption peak at place corresponds to C=O respectively Stretching vibration and C-N stretching vibrations, show amido link in Gliadin be present.In m-MS spectrograms (e), 1082cm^-1, 799cm^-1With 457cm^-1Locate the absorption band occurred, correspond to Si-O-Si antisymmetric stretching vibration respectively, symmetrical stretching vibration and bending are shaken It is dynamic.In S3 spectrogram (c), while the characteristic peak of PBSu, m-MS and Gliadin three are found that, show successfully to be prepared for Mesoporous magnesium silicate-poly-succinic acid-butanediol-gliadin tri compound support.In addition, support S4 and S5 observation result point It is unsuitable with S2 and S3.

2. sem analysis

Support sample microscopic appearance is observed using electron scanning Electronic Speculum (S-3400N, Hitachi, Japan) and fracture is microcosmic Pattern.Observe result as shown in Figure 3 and Figure 4.

From the figure 3, it may be seen that being uniform-distribution with about 400 × 400 μm of BODY WITH SQUARE APERTURE by support made from 3D printing, and hang down Nogata is to insertion.The microfilament of S1 and S2 final moldings is linear, and diameter is about 500 μm；And the microfilament of S3 final moldings is upper and lower Layer junction is slightly bent, and diameter is about 550 μm.In addition, S1 surfaces are smooth, and S2 is compared with S1, except surface slightly have it is coarse in addition to, There is no significant difference；And S3 surface irregularities, form several microns of microcellular structure.This is probably high temperature to the molten egg of wheat alcohol Caused by white structure impacts.

As shown in Figure 4, S1 fractures are more regular, and it is brittle fracture to illustrate it.After adding m-MS, the fracture on S2 surfaces becomes It is coarse, not only have it is raised caused by inorganic particle, but also have inorganic particle come off caused by pit.M-MS is evenly distributed on branch Inside frame, particle diameter is about 5 μm, is reunited without apparent inorganic phase.After adding gliadin, the fracture on S3 surfaces becomes more Overstriking is rough, and has obvious dispersed phase (Gliadin) to exist.As a result show, in S3, each component is uniformly dispersed.In addition, support S4 It is suitable with S2 and S3 respectively with S5 observation result.

3. synchrotron radiation SR μ CT are imaged

Using the X-ray imaging of Shanghai synchrotron radiation light source (SSRF) and biomedical applications line station (BL13W), to branch Frame (S1/S2/S3) sample carries out x-ray scanning, obtains Two-dimensional electron section picture and three-dimensionalreconstruction image, and observed.See It is as shown in Figure 5 to examine result.

As shown in Figure 5, this experiment can be according to the model print carriage being pre-designed, and microfilament thickness is equal in gained sample Even, the pore size of formation is homogeneous；Internal stent structure is intact, no fracture of wire, tomography, cave in, channel blockage phenomena such as, repeat Property it is high.Wire vent Swelling in print procedure be present, after adding gliadin, swelling effect is more obvious, so as to which aperture is omited Less than other two groups.In addition, support S4 and S5 observation result are suitable with S2 and S3 respectively.

4. laser co-focusing 3D microscopic analyses

Using laser confocal microscope (offer of Keyence companies), under 40 times and 200 times of two different multiples, see The surface topography of analysis support (S1/S2/S3) is examined, and tests its roughness.It is as shown in Figure 6 to observe result.

It will be appreciated from fig. 6 that S1 surfaces are smooth, difference in height is that normal cylindrical radian causes；There is obvious projection on S2 surfaces, may It is the m-MS of addition；S3 surface irregularities, several microns of height fall be present.In addition, the microfilament diameter of S1 and S2 supports About 500 μm, the microfilament diameter of S3 supports is about 550 μm.This result is consistent with SEM and SR μ CT results.In addition, S1 is measured, S2 and S3 surface roughness (profile arithmetic average error, Ra) is respectively 0.72 μm, 1.39 μm and 2.29 μm.This explanation, m- The addition of MS and gliadin improves the roughness of the timbering material.In addition, support S4 and S5 observation result respectively with S2 and S3 are suitable.

Bioactivity and the physicochemical property test of the support of effect example 2 (S1/S2/S3)

1. Bioactivity and plasma diffusing W,Mo capability analysis

Support (S1/S2/S3) (12 × 2mm of Φ) is immersed in simulated body fluid (Simulated Body Fluid, SBF) 5 After it, the surface topography of material is characterized using SEM and EDS respectively and element forms, and determines the change of the ion concentration in SBF solution Change.Test result is shown in Fig. 7 and Fig. 8.

As shown in Figure 7, the S1 surfaces after soaking 5 days are still smooth, and there is substantial amounts of spherical apatite particle covering on S2 surfaces In microfilament surface, the spherical apatite particle of S3 Surface Creations is more more than S2, connects and is superimposed between apatite particle, Substantially completely cover microfilament surface.The result shows m-MS addition, substantially increase the external of 3D printing support sample Bioactivity；And the addition of gliadin, its Bioactivity is further increased, it is more suitable as Bone Defect Repari material Material.

From Fig. 8 (A), after being soaked 5 days in SBF solution, S1 surface-elements do not change, and S2 and S3 surfaces calcium and The content of P elements all increased, and the content of S3 surfaces calcium and P elements is far more than S2.This explanation S3 surface has more Apatite generation, this result and SEM results (Fig. 7) are consistent.

From Fig. 8 (B), in immersion process, Ca and P concentration continuous decrease in SBF solution, and Si concentration persistently increases Add.This shows that the Si ions in S3 materials are gradually released in SBF solution, and Ca and P ion gradually deposit in SBF solution S3 material surfaces.The concentration of Mg ions first raises the reason for declining afterwards and is probably in SBF solution, the Mg ions in initial stage S3 materials It is promptly released into SBF solution；After 72 hours, Mg ions reach dynamic with the Mg ions discharged in S3 materials in SBF solution Balance, and be held essentially constant.This result is consistent with SEM (Fig. 7) and EDS (Fig. 8 A) result.In addition, support S4 and S5 Test result is suitable with S2 and S3 respectively.

2. the porosity test of support (S1/S2/S3)

The present invention determines the porosity of support (S1/S2/S3) by Archimedes principle.From test result, S1 holes Gap rate (%) is that 48.60 ± 5.94, S2 porositys (%) are that 47.93 ± 4.20, S3 porositys (%) are 45.35 ± 1.32.This Outside, support S4 and S5 test result is suitable with S2 and S3 respectively.

3. surface protein adsorption capacity is tested

This experiment is model protein to determine support (S1/S2/ using bovine serum albumin(BSA) (Bovine albumin, BSA) S3) the ability of adsorption protein.Support sample is placed in 10 μ g/mL BSA-PBS solution, be placed in cell culture incubator After 4 hours, 2 samples are rinsed with PBS to remove unadsorbed albumen.Determined using ELISA kit in BSA-PBS solution BSA concentration, the BSA adsorbances of rack surface are obtained by the concentration difference of BSA in solution before and after calculating absorption, according to unit matter The adsorbable BSA mass (mg protein/g scaffold) of support sample institute of amount characterizes end product.As a result figure is seen 9A。

From Fig. 9 A, the quality of S2 and S3 adsorptions bovine serum albumin(BSA) (Bovine albumin, BSA) is respectively 0.16mg, 0.43mg and 1.37mg.This may be a kind of hydrophobic polymer material due to PBSu, so as to cause its adsorption Protein content is seldom, and m-MS and gliadin are hydroaropic substance, so as to be advantageous to improve timbering material to hydrophily egg The absorption of white matter.In addition, the addition of gliadin so that timbering material surface forms microcellular structure, adds it and compares surface Product, the contact with solution is increased, and then another step improves the adsorption protein content of timbering material；And BSA is that the later stage is thin The important composition composition of culture medium, will directly influence cell in the adhesion on timbering material surface and sprawls situation in born of the same parents' experiment. In addition, support S4 and S5 test result are suitable with S2 and S3 respectively.

4. external degradation performance test

Support (S1/S2/S3) (12 × 2mm of Φ) is immersed in Tris-HCl solution, measures and calculates its weight loss Rate, external degradation performance is characterized with this.Measurement result is shown in Fig. 9 B.

From Fig. 9 B, increase over time, S1, S2 and S3 weight-loss ratio gradually step up.Wherein, S1 degradation speeds It is most slow, also only degrade 8.5% after 84 days.After adding m-MS, the degradation rate of support significantly improves；And add gliadin Afterwards, the degradation rate of support further improves.After 84 days, S2 and S3 weight-loss ratios have respectively reached 32.3% and 53.1%.As a result Show, the addition of m-MS and gliadin, substantially increase the degradation rate of 3D printing support.

The cell experiment of the support of effect example 3 (S1/S2/S3)

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 volumes Lymphocyte separation medium upper strata, at room temperature with 2500r/min centrifuge 25 minutes.Liquid level intersection cloud liquid is drawn with suction pipe Body, it is placed in the centrifuge tube of the 5mL containing PBS, piping and druming mixes, abandoning supernatant after 1000r/min is centrifuged 5 minutes at room temperature.With After PBS cleans the precipitation in pipe one time, it is suspended in the α-MEM nutrient solutions containing 20%FCS, is counted through cell counting count board Afterwards, by density (1 × 10⁴/cm²) be inoculated into 100mL blake bottles, it is placed in incubator (5%CO₂, 37 DEG C) in cellar culture, training Liquid is changed after supporting 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 adhesion rate test

Cell is observed in support (S1/S2/S3) surface adhesion situation (12 hours) by SEM, examined by flow cytometer Cell adherence rate (3,6 and 12 hours) is surveyed, testing result is shown in Figure 10.

As seen from the figure, BMSCs cell adherences are in three kinds of rack surfaces, and sprawl in good condition.Wherein cell is sprawled on S3 Area is maximum, has more pseudopodium.Adhesion rate test result shows that S3 cell adherence rate is far above S1；12 hours, S2's Cell adherence rate also has significant difference to S1.This is probably because S2 and S3 surfaces can adsorb the albumen in more training bases (Fig. 9), beneficial to the adhesion of BMSCs cells.On the other hand, Mg and Si ions can be discharged in S2 and S3, can promotes BMSCs The adhesion of cell.In addition, support S4 and S5 test result are suitable with S2 and S3 respectively.

3. cell is bred and ALP activity

Using the proliferative conditions (1,3 and 7 day) of CCK-8 methods detection support (S1/S2/S3) superficial cell, ALP reagents are used Box detector ALP activity (7,10 and 14 days).Testing result is shown in Figure 11.

From Figure 11 (A), cell quantity increases with the increase of incubation time on support.At 3 days and 7 days, S2 and S3 O.D. values all be significantly larger than S1 O.D. values, show that S2 and S3 have obvious facilitation to the propagation of BMSCs cells.

From Figure 11 (B), during whole culture, the ALP activity of S2 and the upper cells of S3 is all apparently higher than S1 ALP work Property, and there is no significant difference between S2 and S3.This mainly due to, contain m-MS in S2 and S3, can be gone out with sustained release Mg and Si ions, there is facilitation to the propagation and Osteoblast Differentiation of BMSCs cells.In addition, support S4 and S5 test result difference It is suitable with S2 and S3.

The zoopery of the support of effect example 4 (S1/S2/S3)

This experiment carries out vivo biodistribution phase using new zealand white rabbit femur end defect model to support (S1/S2/S3) Capacitive and bone formation performance evaluation.

1. gross examination of skeletal muscle

Support (S1/S2/S3) puts to death animal after being implanted 4,8,12 weeks, is drawn materials from implant site, at once with neutral Fu Er Malin's liquid is fixed.Cranial defect position is taken pictures using digital camera, observes its reparation situation.Observation result is shown in Figure 12.

As seen from the figure, postoperative 4 weeks, S1 implant sites had obviously bone defect, and S2 and S3 implant sites are hidden About visible Cranial defect position.Elapse over time, the Cranial defect position of each group is all gradually recovered.At 12 weeks, S1 implant sites are still With the presence of obvious pit, and S2 and S3 implant sites can't see the area with bone tissue from directly perceived on surface Not.In addition, support S4 and S5 test result are suitable with S2 and S3 respectively.

2. MicroCT observation analysis

This experiment is using GE companies of U.S. eXplore Locus SP types Micro-CT to new zealand white rabbit femur end Defect sample is scanned analysis.The anglec of rotation is 360 °, and rotation angle increment is 0.4 °, voltage 80kV, and electric current is 80μA.Sample is positioned in flying-spot tube during test and is scanned, obtains continuous level image, scanning resolution is 14 μm.Adopt Image data acquiring and three-dimensional reconstruction are carried out with GE Microview ABA softwares, and calculates New Bone Quantity.CT values between It is set as bone tissue between 1300-3500.Observation result is shown in Figure 13.

As shown in Figure 13,4 weeks when, S1 implant sites only have fragmentary white point to exist, and represent a small amount of freshman bone tissue； S2 and S3 implant sites and host bone boundary reduction of area are small, but centre generates still without bone tissue.At 8 weeks, S1 implant sites There is diminution trend at interface between host bone, but still with the presence of larger black hole；Interface between S2 and S3 implant sites and host bone Further reduce, only in centre still with the presence of smaller black hole；And freshman bone tissue is in latticed presence.12 weeks When, it is close when interface was with 8 weeks between S1 implant sites and host bone, and indistinction, it may be possible to because material can not degrade, cause new Osteogenic tissue can not grow into；The latticed freshman bone tissue of S2 implant sites further grows to centre, but still remains one It is individual compared to 8 weeks when less black hole；S3 implant sites are covered by latticed freshman bone tissue substantially.As a result show, S3 can be obviously promoted internal bone tissue regeneration.In addition, support S4 and S5 test result are suitable with S2 and S3 respectively.

Claims

A kind of 1. mesoporous magnesium silicate/poly butylene succinate binary compound rest, it is characterised in that the binary compound rest Including following components：Mesoporous magnesium silicate and poly butylene succinate；The specific surface area of the mesoporous magnesium silicate is 445- 455m²/ g, pore volume 0.3-0.5cm³/g；

The mass ratio of the mesoporous magnesium silicate and poly butylene succinate is 1:1-1:5；

The aperture of the binary compound rest is 300-500 μm.
2. binary compound rest as claimed in claim 1, it is characterised in that

The mass ratio of the mesoporous magnesium silicate and poly butylene succinate is 1:1-1:3, preferably 1:1-1:1.5；

And/or the specification of the binary compound rest be (Φ 6-12) × (1-8) mm, preferably 12 × 2mm of Φ, 10 × 2mm of Φ or Φ6×6mm；

And/or the aperture of the binary compound rest is 400 μm；

And/or the compressive strength of the binary compound rest is 60-80Mpa, preferably 65-75Mpa；

And/or the porosity of the binary compound rest is 40-60%, preferably 42-52%；

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 particle diameter of the mesoporous magnesium silicate is 800-1500nm, average pore size 6-7nm；

And/or the mesoporous magnesium silicate is made using sol-gal process, is preferably comprised following steps：

S1, at 45-55 DEG C, template, water and mixed in hydrochloric acid, are clarified at stirring to solution；

S2, at 45-55 DEG C, magnesium nitrate hexahydrate, tetraethyl orthosilicate are added in the solution obtained by step S1, stirred quiet after 4-6h Suction filtration is put, is dried, obtains white powder；

S3, white powder obtained by step S2 is calcined to removing template at 500-700 DEG C；

Wherein, it is preferred that

In step S1, the template is PEO-PPOX-PEO triblock copolymer, preferably The P123 triblock copolymers for being 5800 for average weight-average molecular weight；

In step S1, the water is deionized water；

In step S1, the concentration of the hydrochloric acid is 1.7-2.5mol/L；

In step S1, the amount ratio of the template, water and hydrochloric acid is (5.0-7.0g)：(20-40mL):(110-130mL)；

In step S2, add magnesium nitrate hexahydrate and tetraethyl orthosilicate to be added using substep into step S1 resulting solutions, first add six Water magnesium nitrate, adds tetraethyl orthosilicate；

Step S2 is carried out according to following operation：At 45-55 DEG C, successively by 4.6-5.0g magnesium nitrate hexahydrates and the positive silicon of 8.3-8.7g Acetoacetic ester is added in above-mentioned solution, and suction filtration is stood after stirring 4-6h；

In step S2, it is described filter carry out 2-5 time, preferably 3 times, every time filter terminate it is latter as using deionized water progress Cleaning, is filtered again afterwards；

In step S2, the drying is carried out in 50-70 DEG C of electric drying oven with forced convection, and the preferably electric heating air blast at 60 DEG C is done Carried out in dry case；

In step S3, the temperature of the calcining is preferably 600 DEG C.
3. a kind of 3D printing prepares mesoporous magnesium silicate/poly butylene succinate binary composite support as claimed in claim 1 or 2 The method of frame, it is characterised in that the described method comprises the following steps：

A1, the mesoporous magnesium silicate and the poly butylene succinate mixed, obtain binary mix material；The mixing is preferable Ground is to be stirred；The preferred 100-500rpm of speed of the stirring；The time preferred 2-5 hours of the stirring；

A2, the binary mix material obtained using 3D printing equipment to step A1 carry out 3D printing, produce described mesoporous silicic acid Magnesium/poly butylene succinate binary compound rest；The technical parameter of the 3D printing is preferably as follows：The spray of the mixed material Firing rate rate is 20-30g/min；The barrel temperature of the 3D printing equipment is 100-125 DEG C.
A kind of 4. mesoporous magnesium silicate/poly butylene succinate binary compound rest prepared by method as described in claim 3.
5. a kind of mesoporous magnesium silicate/poly butylene succinate/gliadin tri compound support, it is characterised in that described Tri compound support includes following components：Mesoporous magnesium silicate, poly butylene succinate and gliadin；

The specific surface area of the mesoporous magnesium silicate is 445-455m²/ g, pore volume 0.3-0.5cm³/g；

The mass ratio of the mesoporous magnesium silicate, poly butylene succinate and gliadin is 1:1:1-5:5:1；

The aperture of the tri compound support is 300-500 μm.
6. tri compound support as claimed in claim 5, it is characterised in that

The mass ratio of the mesoporous magnesium silicate, poly butylene succinate and gliadin is 1:1:1-5:5:1；

And/or the specification of the tri compound support be (Φ 6-12) × (1-8) mm, preferably 12 × 2mm of Φ, 10 × 2mm of Φ or Φ6×6mm；

And/or the compressive strength 60-80Mpa of the tri compound support, preferably 65-75Mpa；

And/or the porosity of the tri compound support is 40-60%, preferably 42-52%；

And/or the aperture of the tri compound support is 400 μm；

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 particle diameter of the mesoporous magnesium silicate is 800-1500nm, average pore size 6-7nm；

And/or the mesoporous magnesium silicate is made using sol-gal process, is preferably comprised following steps：

M1, at 45-55 DEG C, template, water and mixed in hydrochloric acid, are clarified at stirring to solution；

M2, at 45-55 DEG C, magnesium nitrate hexahydrate, tetraethyl orthosilicate are added in the solution obtained by step M1, stirred quiet after 4-6h Suction filtration is put, dries, obtains white powder；

M3, white powder obtained by step M2 is calcined to removing template at 500-700 DEG C；

Wherein, it is preferred that

In step M1, the template is PEO-PPOX-PEO triblock copolymer, preferably The P123 triblock copolymers for being 5800 for average weight-average molecular weight；

In step M1, the water is deionized water；

In step M1, the concentration of the hydrochloric acid is 1.7-2.5mol/L；

In step M1, the amount ratio of the template, water and hydrochloric acid is (5.0-7.0g)：(20-40mL):(110-130mL)；

In step M2, add magnesium nitrate hexahydrate and tetraethyl orthosilicate to be added using substep into step M1 resulting solutions, first add six Water magnesium nitrate, adds tetraethyl orthosilicate；

Step M2 is carried out according to following operation：At 45-55 DEG C, successively by 4.6-5.0g magnesium nitrate hexahydrates and the positive silicon of 8.3-8.7g Acetoacetic ester is added in above-mentioned solution, and suction filtration is stood after stirring 4-6h；

In step M2, it is described filter carry out 2-5 time, preferably 3 times, every time filter terminate it is latter as using deionized water progress Cleaning, is filtered again afterwards；

In step M2, the drying is carried out in 50-70 DEG C of electric drying oven with forced convection, and the preferably electric heating air blast at 60 DEG C is done Carried out in dry case；

In step M3, the temperature of the calcining is preferably 600 DEG C.
7. a kind of 3D printing prepares mesoporous magnesium silicate/poly butylene succinate/molten egg of wheat alcohol as described in claim 5 or 6 The method of white tri compound support, it is characterised in that the described method comprises the following steps：

B1, by the mesoporous magnesium silicate, the poly-succinic acid-butanediol and the gliadin mix, obtain ternary mixture Material；The mixing is preferably stirred；The preferred 100-500rpm of speed of the stirring；The time preferred 2- of the stirring 5 hours；

B2, the ternary mixture material obtained using 3D printing equipment to step B1 carry out 3D printing, produce described mesoporous silicic acid Magnesium/poly butylene succinate/gliadin tri compound support；The technical parameter of the 3D printing is preferably following：It is described The injection rate of mixed material is 20-30g/min；The barrel temperature of the 3D printing equipment is 110-120 DEG C.
A kind of 8. mesoporous magnesium silicate/poly butylene succinate/gliadin three prepared by method as described in claim 7 First compound rest.
A kind of 9. mesoporous magnesium silicate/poly butylene succinate binary compound rest as described in any one of claim 1,2,4 Or mesoporous magnesium silicate/poly butylene succinate/gliadin tri compound as described in any one of claim 5,6,8 Application of the support in bone renovating material field is prepared.