CN102501348B - Preparation method of high polymer material scaffolds for tissue engineering - Google Patents
Preparation method of high polymer material scaffolds for tissue engineering Download PDFInfo
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- CN102501348B CN102501348B CN201110301671.8A CN201110301671A CN102501348B CN 102501348 B CN102501348 B CN 102501348B CN 201110301671 A CN201110301671 A CN 201110301671A CN 102501348 B CN102501348 B CN 102501348B
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Abstract
The invention discloses a method for preparing high polymer material scaffolds for the tissue engineering by collaboratively adopting mixing flow field and supercritical fluid. The method comprises the following steps: (1) two or more types of high polymer material are proportionally added into an extruder or an injection molding machine for melting; (2) the blend forms a continuous phase structure under the effect of mixing flow in a machine barrel (or under the combined action of the flow field and supercritical fluid which is injected); (3) the prepared blend with the continuous phase structure is foamed through supercritical fluid; and (4) the multi-component phase in foamed blend samples is removed, and less-component phase is remained as biological scaffolds. The method of the invention collaboratively adopts chaotic mixing and supercritical fluid to prepare the blend with the continuous phase structure in the condition that the less-component phase content is low, and enables the blend to be applied to the preparation of biological scaffolds for the tissue engineering. Moreover, the method has fewer processes, can realize batch production, and is easy to promote during the industrial production.
Description
Technical field
The present invention relates to prepare the technology of high polymer material scaffolds for tissue engineering, particularly a kind of mixing flow field and supercritical fluid coordinate system method for high polymer material scaffolds for tissue engineering that adopts.
Background technology
The basic principle of organizational engineering and method are that the functioning cell of in vitro culture is planted on support good in a kind of biocompatibility and that can be absorbed by body, form compound, then by this compound implanting to human body tissue and lesions position, to reach the object of reconstruction.In organizational project, support plays key effect, and it not only provides support structure for specific cell, but also plays template action, guide tissue regeneration and control institutional framework.Therefore, selecting the good material of biocompatibility except needs prepares support, the most important thing is to make support to there is higher voidage, to provide enough spaces to make Growth of Cells, and must there is good connectedness between space, to the required nutriment of Growth of Cells is flowed into, cell excretion refuse is flowed out.Therefore, how to prepare high porosity, high connectivity support just seems particularly important.
Employing is extruded or injection molding technology is prepared high polymer material scaffolds for tissue engineering, is conducive to realize the specific three-dimensional profile of support to obtain required tissue or organ shape, and can be mass, and is easy to promote in industrial production.But, how, by extruding or injection molding technology is prepared high porosity, high connectivity, the better support of mechanical property, how further reducing operation, reduce costs, these difficult problems all need to be solved by the new method of preparing support.
The present invention relates to a kind of mixing flow field and supercritical fluid coordinate system method for high polymer material scaffolds for tissue engineering that adopts, wherein relate to and adopt mixing flow field (the particularly mixing flow field of chaos) preparation to there is the method for macromolecular material blend of common continuous phase structure, and can in this process, introduce supercritical fluid further promote blend in the generation of continuous phase structure altogether.
The generation that is total to continuous phase structure in blend is mainly subject to the impact of the factors such as content ratio between each component, viscosity ratio, elasticity ratio and interfacial tension.After macromolecular material is determined, the viscosity of material itself and elasticity just determined, thus between each component content than and interfacial tension become formation and be total to the key of continuous phase structure.
Adopt chaos mixing in the process of preparing blend, can content between fixing each component than in the situation that, original position forms multiple phase morphology, such as drop, lamella structure, filamentary structure and common continuous phase structure etc., thus make blend material there is different characteristics.And mixing can the realization in the alap situation of few component phase content of chaos makes macromolecular material blend form common continuous phase structure.In addition, adopt that chaos is mixing while preparing macromolecular material blend, can overcome the strong shortcomings such as the inhomogeneous of mixing middle mixability and molecular chain rupture that strong shear action causes of shearing of tradition.
In macromolecular material Blending Processes, due to the difference of the molecular composition between each component and chemical constitution, make their thermodynamics incompatible, in the situation that few component phase content is lower, form continuous phase structure altogether thereby cannot realize.Add compatilizer can effectively reduce the interfacial tension between each component in co-mixing system, be conducive to form common continuous phase structure.But compatilizer mostly is oligomer, can reduce to a certain extent the mechanical property of blend.Supercritical fluid has stronger mass transfer ability, easily enter between macromolecular material strand, increase its free volume, reduce its vitrification point, reduce the entanglement of strand, make its strand have larger activity space, thereby can effectively reduce the interfacial tension between viscosity and the macromolecular material of melt.Therefore, supercritical fluid can play the effect that is similar to compatilizer in melt blended.And different from compatilizer, supercritical fluid is just present in preparation process, and is not present in final blend.Therefore, supercritical fluid has higher practical value aspect reduction interfacial tension.
The patent No. is that the patent of WO 2007/115367Al discloses a kind of method of porous polymer materials blend structure as biological support of preparing, whole process relates to two or more macromolecular material is dissolved in to solution, then through overcuring, etch away wherein phase or a two-phase, form loose structure, as stent applications in organizational project.The process of the method is comparatively loaded down with trivial details, wherein relates to and adopts solution to dissolve macromolecular material, so application surface is narrower, and is difficult to batch production, is unfavorable for promoting in industrial production.
The patent No. is that the patent of WO 2003/4103925A1 discloses the method that macromolecular material blend that a kind of employing has common continuous phase structure is prepared used in tissue engineering biological support.This patent relates to by regulating and controlling viscosity ratio and Volume fraction between each component makes macromolecular material blend form the method for common continuous phase structure.The generation of continuous phase structure requires the volume content of few component phase higher than 40% altogether.For the higher material of cost, this will increase cost undoubtedly; If reduce its content, the continuation degree of continuous phase structure will reduce altogether, thereby the integrity degree of whole support and mechanical property will can not be guaranteed.
Summary of the invention
The object of the invention is to overcome the existing deficiency of preparing high polymer material scaffolds for tissue engineering technology, provide a kind of simple to operate, work flow is reasonable, operation is few, reliable and stable and can be used for the preparation method of industrial high polymer material scaffolds for tissue engineering.
Object of the present invention is achieved through the following technical solutions: a kind of preparation method of high polymer material scaffolds for tissue engineering, comprises the steps:
(1) two or more macromolecular material mixed and add in the former such as extruder or injection machine and carry out melting, various high polymer material plasticizings are melt;
(2) blend forms continuous phase structure altogether under the effect of mixing flow field (shearing flow field, elongation flow field and/or chaos flow field);
(3) the blend sample of preparation is put into autoclave;
(4) adopt supercritical fluid formation and induction system control gas in supercriticality, and be injected in autoclave, with intermittent mode, sample is foamed;
(5) prepared foaming blend sample is put into particular solution, remove multicomponent phase, retain few component as biological support.
Also above-mentioned steps (2), (3) and (4) can be merged into a step, that is: supercritical fluid forms with induction system control gas in supercriticality, and be injected in extruder or injection machine machine barrel, the supercritical fluid injecting and macromolecular material melt carry out mixing in extruder or injection machine.Blend forms continuous phase structure altogether under mixing flow field (shearing flow field, elongation flow field and/or chaos flow field) acts synergistically with supercritical gas, and inject molding mold cavity or pump into extruder head, by quick release pressure, make blend foaming.
The preparation process of above-mentioned two kinds of modes as shown in Figure 2.
In step (1), described macromolecular material can be this class Biodegradable material of PLA, polyvinyl alcohol, polycaprolactone, starch, polyethylene glycol oxide, polyglycolic acid and polyhydroxyalkanoate, also can be this class common high molecular materials of polypropylene, high density polyethylene (HDPE), low density polyethylene (LDPE), nylon, polymethyl methacrylate and polystyrene.
In step (1), described extruder comprises single screw extrusion machine and double screw extruder.
In step (2), described mixing flow field comprises shears flow field, elongation flow field and chaos flow field.
In step (4), described supercritical fluid comprises supercritical carbon dioxide and overcritical nitrogen.
In step (4), described supercritical fluid makes blend foaming comprise extrusion foaming and injection foaming.
In step (5), described solution comprises distilled water, salt solution and oxolane.
The inventive method more preferably can comprise following preparation process and parameter:
(i) take 25～50 parts of PLAs, 75～50 parts of polyethylene glycol oxides and evenly mix by quality and obtain blend.
(II) blend obtaining in step (I) is added to continuity supercritical carbon dioxide auxiliary polyalcohol contour machining equipment (patent No. is the Chinese invention patent of ZL 200720061588.7), and adopt multi-side thread-groove convective screw (patent No. is the Chinese invention patent of ZL 92114886.0), under supercritical carbon dioxide content 0.1～10%, the processing temperature of 170～210 ℃, carry out blend foaming, extrudate is the polyethylene glycol oxide/polylactic acid blend after foaming.This blend is put into distilled water and carry out leaching removal polyethylene glycol oxide phase, obtain polylactic acid bracket.
This method may be used on the equipment including single screw extrusion machine, double screw extruder and injection machine, realizes the preparation of biological support.And this method can effectively be prepared the porous polymer materials with high porosity, except as scaffold for tissue engineering, also can be applicable to the aspects such as sound-absorbing, filtration, buffering.
Principle of the present invention is: in the melt blended process of macromolecular material, by the effect in flow field, macromolecular material melt is produced shunting, convection current, supercharging-decompression, Surface Renewal, continuously changes the effects such as VELOCITY DISTRIBUTION, make melt constantly be stretched and fold.Simultaneously, the thermodynamics causing for the difference of the molecular composition between each component and chemical constitution in macromolecular material blend is incompatible, add supercritical fluid, its stronger mass transfer ability can make it enter rapidly between macromolecular material strand, increase the free volume of material, reduce molecular entanglement, reduce the interfacial tension between each component, further the few component phase of refinement.And mixing flow field stretches continuously and folding effect can make supercritical fluid be dissolved in more fully in macromolecular material, and few component is further stretched mutually and folded.Under the synergy of mixing flow field and supercritical fluid, make blend in the situation that few component phase content is lower, form the common continuous phase structure that continuation degree is higher.Utilize supercritical fluid to foam to blend sample, improve its porosity and degree of communication, then remove the few component phase of multicomponent phase, reservation in blend, be prepared into the biological support of used in tissue engineering.
The present invention has following advantage and effect with respect to existing technology and preparation method:
(1) this method operation is less, and the equipment adopting is continuous type process equipment (extruder or injection machine) comparatively general in industrial production, therefore guaranteed the continuity of producing, can realize batch production, be easy to promote in industrial production, there is wide prospects for commercial application.
(2) in this method, the introducing of supercritical fluid can improve the mobility of macromolecular material melt, reduces processing temperature, thereby can make to extrude or the power consumption of polymer processing of injection moulding reduces by 25% left and right.
(3) in this method, the few component in blend is as final reserve part, and its content is lower, thereby has reduced the cost of support.
(4) in this method, remove multicomponent mutually after, because few component phase content is higher compared with the low porosity of whole support that makes, then adopt supercritical fluid to foam mutually to few component, can further improve the porosity (30% left and right) of support.
(5), in this method, in the macromolecular material blend of preparation, formed the higher common continuous phase structure of continuation degree, therefore, remove multicomponent mutually after, can guarantee structural intergrity and the mechanical property of support.
Accompanying drawing explanation
Fig. 1 is the schematic flow sheet of preparing macromolecular material support that the inventive method proposes.
Fig. 2 is the schematic diagram of preparing each stage macromolecular material blend morphology in support process.
Fig. 3 is polystyrene/polymethyl methacrylate blend (a) and the electron scanning micrograph of employing supercritical fluid to its foaming rear (b) that the mass ratio of the mixing preparation of employing chaos is 50/50.
Fig. 4 is the electron scanning micrograph that adopts PLA/low density polyethylene blends that the mass ratio of chaos mixing preparation is 70/30.
Fig. 5 is the electron scanning micrograph that adopts acetic starch/polylactic acid blend that the mass ratio of chaos mixing preparation is 65/35.
Fig. 6 is that employing supercritical fluid is the electron scanning micrograph after 70/30 PLA/sodium chloride mixture foams to the mass ratio by shearing mixing preparation.
Fig. 7 is that employing supercritical fluid is the electron scanning micrograph after 70/30 PLA/acetic starch blend foams to the mass ratio by the mixing preparation of chaos.
In above-mentioned each figure, symbol description is as follows: A-extruder or injection machine; 1-feeding hopper; 2-macromolecular material; 3-screw rod; 4-macromolecular material blend melt; 5-extruder or injection machine machine barrel; 6-supercritical fluid forms and induction system; 7-autoclave; 8-injection mold; 9-extruder head; 10-leaching equipment; 11-scaffold for tissue engineering; The few component phase of 12-; 13-multicomponent phase; 14-abscess; 15-low density polyethylene (LDPE) phase; 16-PLA phase; 17-acetic starch phase
The specific embodiment
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited to this.
Embodiment 1
Fig. 1 shows the specific implementation process of the inventive method.As shown in Figure 1, the implementation process equipment needed thereby of the inventive method comprises extruder or injection machine A, supercritical fluid forms and induction system 6, autoclave 7 and leaching equipment 10.Supercritical fluid forms and is connected with extruder or injection machine machine barrel 5 with induction system 6; Injection machine or extruder barrel 5 are connected with mould 8 or head 9; Autoclave 7 forms and is connected with induction system 6 with supercritical fluid, and the structure of screw rod 3 can change, and machine barrel 5 outer walls are equipped with heating collar (not shown).
This mixing flow field and supercritical fluid coordinate system for the whole process of high polymer material scaffolds for tissue engineering are: start extruder or injection machine A, two or more macromolecular material 2 is mixed by a certain percentage, join in the feeding hopper 1 of extruder or injection machine, external heat and screw rod 3 through machine barrel 5 rotate the shear heat producing, and melt polymer material forms melt 4.The phase morphology schematic diagram of macromolecular material blend is as shown in Fig. 2 (a).Subsequently, the inventive method can be divided into one-step method and two-step method is carried out, and wherein the process of one-step method is: open supercritical fluid and form and induction system 6, supercritical fluid is injected in machine barrel; The macromolecular material melt that contains supercritical fluid injected in the die cavity of mould 7 or extrude from head 8, through the macromolecular material blend of release pressure preparation foaming fast.Two-step method is: by macromolecular material melt cooling, put into autoclave 9; Start supercritical fluid and form and induction system 6, supercritical fluid is injected to autoclave, macromolecular material blend sample is carried out after certain hour saturated, make its foaming through release pressure fast.After foaming, the form schematic diagram of macromolecular material blend is as shown in Fig. 2 (b).Macromolecular material blend after one-step method or two-step method foaming is placed in to leaching equipment 10 leachings, removes multicomponent phase 13, thereby obtain support 11, its form schematic diagram is as shown in Fig. 2 (c).
Fig. 3 shows implementation result of the present invention from the microcosmic point of material.Polystyrene/polymethyl methacrylate blend (a) that the mass ratio that Figure 3 shows that the mixing preparation of employing chaos is 50/50 and the electron scanning micrograph of employing supercritical fluid to its foaming rear (b).Observe for convenient, PMMA is carried out to etching mutually, therefore, in Fig. 3 (a), black cavity is PMMA phase.From figure, can obviously find out, PS/PMMA blend has formed common continuous structure.From Fig. 3 (b), can find, adopt supercritical carbon dioxide (CO
2) foam after, blend sample has formed that abscess is tiny, densification and the high foam structure of porosity.
Embodiment 2
(1) take 70 parts of PLAs (PLA) and 30 parts of low density polyethylene (LDPE)s (HDPE) by quality, and mix.
(2) blend of step (1) is added to single screw extrusion machine, prepare PLA/HDPE blend by the mixing mode of chaos.
Figure 4 shows that the electron scanning micrograph of the PLA/HDPE blend of preparation.As can be seen from the figure, be 30% at the content of HDPE, blend has formed good common continuous phase structure.
(1) take 65 portions of acetic starches (AS) and two kinds of Biodegradable materials of 35 parts of PLA by quality, and mix.
(2) blend of step (1) is added in extruder, prepare AS/PLA blend by the mixing mode of chaos.
Figure 5 shows that the electron scanning micrograph of the AS/PLA blend of preparation.As can be seen from the figure,, in the situation that PLA content is 35%, blend has formed common continuous phase structure.
(1) take 60 parts of PLA and 40 parts of sodium chloride (NaCl) particle by quality, and mix.
(2) mixture of step (1) is added in extruder, extrude preparation PLA/NaCl mixture.
(3) the PLA/NaCl mixture of being prepared by step (2) is put into autoclave, under 170 ℃, the condition of 18MPa, passes into supercritical CO
2after saturated 7h, carry out release of pressure foaming.
Figure 6 shows that the electron scanning micrograph of the PLA/NaCl mixture after foaming.As can be seen from the figure, in the mixture after foaming, formed open-celled structure, the percent opening of sample is about 35%, and this significantly increases the connectedness of sample.
Embodiment 5
(1) take 70 parts of PLA and 30 parts of AS by quality, and mix.
(2) blend of step (1) is added in extruder, extrude preparation PLA/AS blend by the mixing mode of chaos.
(3) the PLA/AS blend of being prepared by step (2) is put into autoclave, under 150 ℃, the condition of 16MPa, passes into supercritical CO
2, after saturated 1h, carry out release of pressure foaming.
Figure 7 shows that the electron scanning micrograph of the PLA/AS blend after foaming.As can be seen from the figure,, after foaming, blend material inside has formed the loose structure of high degree of communication.
Above-mentioned example is part embodiment of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other are any does not deviate from change, the modification done under Spirit Essence of the present invention and principle, substitute, combination, simplify; all should be equivalent substitute mode, within being included in protection scope of the present invention.
Claims (1)
1. the preparation method of a high polymer material scaffolds for tissue engineering, it is characterized in that comprising the steps: that (1) mixes in proportion two or more macromolecular material, and add in former and carry out melting, various high polymer material plasticizings are melt;
(2) blend forms continuous phase structure altogether under the effect in mixing flow field;
(3) the blend sample of preparation is put into autoclave;
(4) supercritical fluid forms with induction system control gas in supercriticality, and is injected in autoclave, with intermittent mode, sample is foamed;
(5) prepared foaming blend sample is put into particular solution, remove the multicomponent phase in blend, retain few component as biological support;
Specifically comprise following preparation process and parameter:
(I) by quality take 25～35 parts of PLAs of raw material, 65～75 parts of polyethylene glycol oxides mix, and obtain blend;
(II) blend step (I) being obtained adds continuity supercritical carbon dioxide assistant high molecular material contour machining equipment, and employing multi-side thread-groove convective screw, under supercritical carbon dioxide content 0.1～10%, the processing temperature of 170～210 ℃, carry out blend foaming, extrudate is the polyethylene glycol oxide/polylactic acid blend after foaming, this blend is put into distilled water and carry out leaching removal polyethylene glycol oxide phase, obtain polylactic acid bracket.
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| CN103350510B (en) * | 2013-07-04 | 2015-07-22 | 浙江海虹控股集团有限公司 | Equipment and method for preparing polymer supercritical foaming material |
| CN103520770B (en) * | 2013-09-27 | 2015-06-17 | 郑州大学 | Porous material for tissue engineering stent |
| CN104312113B (en) * | 2014-08-21 | 2016-10-05 | 华南理工大学 | Polylactic acid base ultra-toughness intermingling material/high-strength blended nano composite material of ultra-toughness with shape memory effect and preparation method thereof |
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| CN108014372A (en) * | 2017-10-27 | 2018-05-11 | 郑州大学 | A kind of method that supercritical gas prepares small-caliber vascular stent |
| CN108310458B (en) * | 2018-03-21 | 2021-05-11 | 奚桢浩 | Method for preparing tissue engineering scaffold material by adopting degradable synthetic polymer material |
| CN108559236B (en) * | 2018-03-21 | 2020-11-17 | 奚桢浩 | Tissue engineering scaffold material with micro-nano structure and preparation method thereof |
| CN110755672A (en) * | 2019-10-31 | 2020-02-07 | 慧生医学科技(徐州)有限公司 | Antibacterial hemostatic sponge and preparation method thereof |
| CN113877004B (en) * | 2021-10-09 | 2023-02-03 | 南京聚隆科技股份有限公司 | Biological bone nail material and preparation method thereof |
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