CN102697579A - Three-dimensional bracket preparation method for tissue engineering based on supercritical fluid technology - Google Patents
Three-dimensional bracket preparation method for tissue engineering based on supercritical fluid technology Download PDFInfo
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- CN102697579A CN102697579A CN2012101002467A CN201210100246A CN102697579A CN 102697579 A CN102697579 A CN 102697579A CN 2012101002467 A CN2012101002467 A CN 2012101002467A CN 201210100246 A CN201210100246 A CN 201210100246A CN 102697579 A CN102697579 A CN 102697579A
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Abstract
The invention discloses a three-dimensional bracket preparation method for tissue engineering based on a supercritical fluid technology. The three-dimensional bracket comprises the following steps: dissolving a polymer to be prepared into a porous bracket into a solvent to prepare a solution or a suspension solution with a certain concentration, wherein the solution or the suspension solution is sufficiently contacted with ScCO2; rapidly dissolving the ScCO2 into the solution or the suspension solution to separate the polymer solution or the suspension solution; separating the solution or the suspension solution into a polymer rich phase and a polymer lean phase which can form a metastable balance; meanwhile, rapidly dissolving the solvent in the solution or the suspension solution into the ScCO2; discontinuously carrying out glass transition and curing on the polymer rich phase to form a porous material; forming a crystal nucleus by the polymer lean phase and enabling the crystal nucleus to grow gradually to fully fill gaps in the porous material; and circularly introducing CO2 to wash and dry the porous material and continuously conveying the solvent into an ScCO2 phase to be brought out of the system, so that the single-component or multi-component or composite-activity porous bracket for the tissue engineering which has no solvent residues is obtained.
Description
Technical field
What the present invention relates to is a kind of used in tissue engineering three-dimensional rack method for preparing based on supercritical fluid technology.
Background technology
The core of organizational project is to set up the three dimensions complex that is made up of cell and biomaterial; Its main contents of studying at present concentrate on aspects such as Biodegradable scaffold material, seed cell, condition of in vitro culture, and wherein the structure of ideal three-dimensional rack is the successful precondition of engineered artificial organ research.In recent years, the research tendency of New-support material is one-component to the multicomponent transition, and inert material is to the active material transition.How on the basis of original research, to grind and obtain ideal bioactive bracket, become the core focus and the difficult point of these key subjects of organizational project.
Ideal tissue engineering bracket is except that having excellent biological compatibility, suitable degradability and effectively the surface activity; Also require to have the three-dimensional outside stereochemical structure of suitable volumes, shape; With the reparation of satisfying tissue defect and the needs of filling; And the three-dimensional internal structure of high porosity and internal communication, so that good space and passage is provided for the discharge of seed cell propagation, grow into timbering material and nutrient exchange and metabolite.
The multiple method for preparing three-dimensional porous support for tissue engineering has been proposed so far.More typically have: casting/particle leaching, be separated, fiber connection, three dimensional printing, emulsion lyophilization, melt molding, gas foaming etc.; And some complex methods that on these technological bases, develop, for example be separated/particle leaching, particle leaching/lyophilization, electric spinning/particle leaching/gas foaming method etc.Although these methods respectively have its characteristics and superiority, wherein some problem remains its key in application of restriction.For example, some method relates to high temperature, is unfavorable for the introducing of bioactive substance in the preparation process; Some method relates to organic solvent, and it is removed, and making residual organic solvent reach the acceptable scope of clinical practice just becomes a difficult problem; The porosity that some method forms is low, and it is poor to connect each other between the Kong Yukong; Some method can't realize the control to the aperture, and pore structure and intensity thereof are relatively poor; Some method is complicated; Accomplishing a preparation process at laboratory needs for a long time; For example casting/particle leaching method approximately needed for 1 week; Some method can't be used to prepare multicomponent or composite reactive support, though some method can be used for multicomponent or the preparation of composite reactive support, technical process can be complicated more.
Summary of the invention
Technical problem to be solved by this invention is that the deficiency that is directed against prior art provides a kind of used in tissue engineering three-dimensional rack method for preparing based on supercritical fluid technology.
Technical scheme of the present invention is following:
A kind of used in tissue engineering three-dimensional rack method for preparing based on supercritical fluid technology; May further comprise the steps: the first step; Will process the polymer of porous support; Or the mixture formed of the polymer of two or more different mechanical properties, or polymer with receive micron mixture of active substance composition and be dissolved in and process certain density solution or suspension in the solvent, this solution or suspension and ScCO
2Fully contact is because ScCO
2Be close to the solvability of liquid and the diffusivity of intimate gas, the one, ScCO
2Be dissolved in fast in solution or the suspension and cause that polymer solution or suspension are separated; It is stingy that solution or suspension promptly are separated into the polymer richness phase and the polymer that are in metastable equilibrium; Simultaneously, the solvent in solution or the suspension also can be dissolved in ScCO fast
2In, glass transition constantly takes place and solidifies in polymer richness mutually, forms porous material, and the stingy formation nucleus of polymer is also grown up gradually, is full of the space in the porous material; In second step, circulation feeds CO
2Porous material is cleaned and drying, and solvent constantly gets into ScCO
2Taken out of system, can be obtained the almost one-component or the multicomponent of no solvent residue, or composite reactive used in tissue engineering porous support.
Described method for preparing, the concrete operations step is:
A1, take by weighing the dry polymer of certain mass; Or the mixture of the polymer of two or more different mechanical properties composition; Or the mixture that polymer and micron active substance received are formed dissolves in the solvent, processes solution or the suspension of mass percent concentration 5%-50%, treats that solution or suspension preparation finish; Leave standstill 5-30min, get rid of the bubble in the solution;
A2, solution or suspension that will the row's of leaving standstill bubble be slowly poured in the pouring container that the degree of depth is 1-3cm, and the pouring container that will fill solution or suspension is then put into and risen to 30-50 ℃ autoclave, and sealing autoclave slowly feeds ScCO
2To pressure be 6-30MPa, close plunger displacement pump, close the entrance and exit valve pressurize of autoclave;
A3, the above-mentioned pressure 30min-120min of maintenance, glass transition constantly takes place and solidifies in solution in the pouring container or suspension polymer richness mutually, forms porous material, and the stingy formation nucleus of polymer is also grown up gradually, is full of the space in the porous material;
The ScCO of A4, unlatching autoclave
2Outlet valve is opened inlet valve, and the opening plunger pump makes ScCO
2With the rate of circulating flow of 2-40kg/h flow through autoclave, flash trapping stage still, secondary separating still, mass flowmenter, check valve, until with CO
2The ScCO that steel cylinder comes out
2After mixer mixing, feed the porous material that forms in the dry pouring container of autoclave again, to form exsiccant porous support;
A5, to continue cyclic drying 30-120min fully dry to the porous support;
A6, close plunger displacement pump, close the autoclave terminal valve, slowly to the autoclave pressure release, to pressure be zero, take out pouring container, take out porous support.
Beneficial effect of the present invention is following:
1, the present invention proposes supercritical CO 2 (ScCO2)-cyclic drying process and be used for the preparation of used in tissue engineering porous support; Be Controlled Pore Structure; Porosity is high, and intercommunity is good, and almost the preparation of the used in tissue engineering porous support of no solvent residue provides a new way.
2, among the present invention,, can prepare the porous support of single component, when two kinds of polymer are mixed the multicomponent porous support that can obtain Different Pore Structures and mechanical performance when being dissolved into organic solvent then with certain proportion when polymer when being a kind of.
3,, utilize this process to prepare to have certain bioactive composite tissue engineering and use porous support when polymer and active substance are gone into to form solution or suspension in the organic solvent with the certain proportion mixed dissolution.
4, utilize the pouring container of difformity and volume, can obtain having the porous support of the three-dimensional outside stereochemical structure of different volumes, shape, satisfy the needs of different tissues engineering.
5, ScCO2 not be to can producing vapour-liquid interface in the dry run of support, can dry fast and fully porous material and can not cause caving in of pore structure, and the pore structure form of acquisition is good, porosity is high, intercommunity is strong;
6, near ScCO2 its character critical point is very responsive with the variation of operating parameter, gets final product simple realization to pore Structure Control through the adjustment operating parameter.
7, supercritical fluid is both as being separated derivant, and again as desiccant, pore structure forms with a dry step and accomplishes, and preparation time shortens greatly, and technological operation is simple.
8, a fine setting voltage stabilizing back pressure valve is set between autoclave and flash trapping stage still, it can play pressure stabilization function to high pressure, guarantees pressure reduction certain between two stills, can guarantee the even running of cyclic drying and pressure leak process again, is easy to realize the control to the aperture.
9, behind autoclave, be provided with two groups of separating stills in order to separate ScCO
2Dissolved organic solvent has not contained organic solvent basically through twice isolating supercritical fluid, guarantees CO
2When recycling, can obtain porous support almost completely dry, no solvent residue with organic solvent.
10, the present invention promptly can be used for crystalline polymer, also can be used for the preparation of the porous support of amorphous state polymer.
11, the present invention can carry out at a lower temperature, is easy to realize the introducing of active substance.
Description of drawings
Fig. 1: supercritical fluid inductive phase separation-cyclic drying process flow sketch map;
1, CO
2Steel cylinder; 2, cooling water tank; 3, plunger displacement pump; 4, blender; 5, depurator; 6, preheater; 7, autoclave; 8, pouring container; 9, flash trapping stage still; 10, secondary separating still; 11, effusion meter; 12, check valve; 13, fine setting voltage stabilizing back pressure valve.
The specific embodiment
Below in conjunction with specific embodiment, the present invention is elaborated.
As shown in Figure 1, the present invention proposes a kind of new three-dimensional porous support for tissue engineering method for preparing, i.e. supercritical CO
2(ScCO
2, supercritical CO 2)-the cyclic drying process.: the first step; Will process the polymer of porous support; Or the mixture formed of the polymer of two or more different mechanical properties, or polymer with receive micron mixture of active substance composition and be dissolved in and process certain density solution or suspension in the solvent, this solution or suspension and ScCO
2Fully contact is because ScCO
2Be close to the solvability of liquid and the diffusivity of intimate gas, the one, ScCO
2Be dissolved in fast in solution or the suspension and cause that polymer solution or suspension are separated; It is stingy that solution or suspension promptly are separated into the polymer richness phase and the polymer that are in metastable equilibrium; Simultaneously, the solvent in solution or the suspension also can be dissolved in ScCO fast
2In, glass transition constantly takes place and solidifies in polymer richness mutually, forms porous material, and the stingy formation nucleus of polymer is also grown up gradually, is full of the space in the porous material; In second step, circulation feeds CO
2Porous material is cleaned and drying, and solvent constantly gets into ScCO
2Taken out of system, can be obtained the almost one-component or the multicomponent of no solvent residue, or composite reactive used in tissue engineering porous support.
Its concrete operations step is following:
1, the dry polymer that takes by weighing certain mass is (like degradable polymers such as polylactic acid, polycaprolactone, Polyethylene Glycol, polyglycolic acids; Or it is mixed with certain proportioning according to demands of different; Concrete proportioning can be regulated as required; As when dissolving in solvent with 1: 1 mixed proportion polylactic acid and Polyethylene Glycol; Its mechanical strength of the porous support that obtains is low during than pure polylactic acid, can be used for cartilage tissue engineered etc.) or polymer and active substance (like hydroxyapatite, tricalcium phosphate etc.) incorporate solution or the suspension that (solvent such as acetone, ethanol, chloroform, dichloromethane etc.) in the solvent are processed mass percent concentration 5%-50%.Treat that formulations prepared from solutions finishes, leave standstill 5-30min, get rid of the bubble in the solution.
2, solution or suspension that will the row's of leaving standstill bubble be slowly poured in the pouring container 8 that the degree of depth is 1-3cm, and the pouring container 8 that will fill solution is then put into the autoclave 7 that rises to 30-50 ℃, and sealing autoclave 7 slowly feeds ScCO
2To pressure be 6-30Mpa, close the entrance and exit valve pressurize of autoclave 7.
3, keep above-mentioned pressure 30-120min, glass transition constantly takes place and solidifies in solution in the pouring container or suspension polymer richness mutually, forms porous material, and the stingy formation nucleus of polymer is also grown up gradually, is full of the space in the porous material.
4, open the ScCO of autoclave 7
2Outlet valve is opened inlet valve, and opening plunger pump 3 makes ScCO simultaneously
2With the rate of circulating flow of 2-40kg/h flow through autoclave 7, flash trapping stage still 9, secondary separating still 10, mass flowmenter 11, check valve 12, until with CO
2The ScCO that steel cylinder 1 comes out
2, blender 4 feeds the porous material that forms in the autoclave 7 dry pouring containers 8 again after mixing, to form exsiccant porous support.
5, continuation cyclic drying 30-120min is fully dry to the porous support.
6, closing plunger displacement pump 3, close the autoclave terminal valve, slowly to the autoclave pressure release, is zero to pressure, takes out pouring container 8, takes out porous support.
Whole process time spent 1-4h, porosity reaches more than 85%, and configuration solution can obtain the porous support of different mechanical properties with the kind of polymer difference or mix ingredients difference.
Should be understood that, concerning those of ordinary skills, can improve or conversion, and all these improvement and conversion all should belong to the protection domain of accompanying claims of the present invention according to above-mentioned explanation.
Claims (2)
1. used in tissue engineering three-dimensional rack method for preparing based on supercritical fluid technology; It is characterized in that; May further comprise the steps: the first step, will process the polymer of porous support, or the mixture formed of the polymer of two or more different mechanical properties; Or the mixture formed of polymer and micron active substance received is dissolved in and processes certain density solution or suspension in the solvent, this solution or suspension and ScCO
2Fully contact is because ScCO
2Be close to the solvability of liquid and the diffusivity of intimate gas, the one, ScCO
2Be dissolved in fast in solution or the suspension and cause that polymer solution or suspension are separated; It is stingy that solution or suspension promptly are separated into the polymer richness phase and the polymer that are in metastable equilibrium; Simultaneously, the solvent in solution or the suspension also can be dissolved in ScCO fast
2In, glass transition constantly takes place and solidifies in polymer richness mutually, forms porous material, and the stingy formation nucleus of polymer is also grown up gradually, is full of the space in the porous material; In second step, circulation feeds CO
2Porous material is cleaned and drying, and solvent constantly gets into ScCO
2Taken out of system, can be obtained the almost one-component or the multicomponent of no solvent residue, or composite reactive used in tissue engineering porous support.
2. method for preparing according to claim 1 is characterized in that, the concrete operations step is:
The mixture that the polymer of A1, the dry polymer that takes by weighing certain mass or two or more different mechanical properties is formed; Or the mixture that polymer and micron active substance received are formed dissolves in the solvent; Process solution or the suspension of mass percent concentration 5%-50%; Treat that solution or suspension preparation finish, leave standstill 5-30min, get rid of bubble wherein;
A2, solution or suspension that will the row's of leaving standstill bubble be slowly poured in the pouring container that the degree of depth is 1-3cm, and the pouring container that will fill solution or suspension is then put into and risen to 30-50 ℃ autoclave, and sealing autoclave slowly feeds ScCO
2To pressure be 6-30MPa, close the entrance and exit valve pressurize of autoclave;
A3, the above-mentioned pressure 30-120min of maintenance, glass transition constantly takes place and solidifies in solution in the pouring container or suspension polymer richness mutually, forms porous material, and the stingy formation nucleus of polymer is also grown up gradually, is full of the space in the porous material;
The ScCO of A4, unlatching autoclave
2Outlet valve is opened inlet valve, and the opening plunger pump makes ScCO
2With the rate of circulating flow of 2-40kg/h flow through autoclave, flash trapping stage still, secondary separating still, mass flowmenter, check valve, until with CO
2The ScCO that steel cylinder comes out
2After mixer mixing, feed the porous material that forms in the dry pouring container of autoclave again, to form exsiccant porous support;
A5, to continue cyclic drying 30-120min fully dry to the porous support;
A6, close plunger displacement pump, close the autoclave terminal valve, slowly to the autoclave pressure release, to pressure be zero, take out pouring container, take out porous support.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201210100246.7A CN102697579B (en) | 2012-04-09 | 2012-04-09 | Three-dimensional bracket preparation method for tissue engineering based on supercritical fluid technology |
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| CN201210100246.7A CN102697579B (en) | 2012-04-09 | 2012-04-09 | Three-dimensional bracket preparation method for tissue engineering based on supercritical fluid technology |
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| CN102697579A true CN102697579A (en) | 2012-10-03 |
| CN102697579B CN102697579B (en) | 2015-01-21 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103787509A (en) * | 2014-01-23 | 2014-05-14 | 河北科技大学 | Porous functional carrier preparation device and method for preparing porous macromolecular organic carrier applied to sewage treatment by using porous functional carrier preparation |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002021222A1 (en) * | 2000-09-08 | 2002-03-14 | Ferro Corporation | Orthopedic mixtures prepared by supercritical fluid processing techniques |
| CN1476907A (en) * | 2003-06-30 | 2004-02-25 | 暨南大学 | Bio-active 3-D porous tissue engineering support material and its preparation method |
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2012
- 2012-04-09 CN CN201210100246.7A patent/CN102697579B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002021222A1 (en) * | 2000-09-08 | 2002-03-14 | Ferro Corporation | Orthopedic mixtures prepared by supercritical fluid processing techniques |
| CN1476907A (en) * | 2003-06-30 | 2004-02-25 | 暨南大学 | Bio-active 3-D porous tissue engineering support material and its preparation method |
Non-Patent Citations (2)
| Title |
|---|
| ANA RITA C. DUARTE: "《Perspectives on: Supercritical Fluid Technology for 3D Tissue gineering Scaffold Applications》", 《JOURNAL OF BIOACTIVE AND COMPATIBLE POLYMERS》, vol. 24, 30 July 2009 (2009-07-30) * |
| 桂秋媛: "《超临界流体制备多孔结构的热力学行为研究》", 《中国优秀硕士学位论文全文数据库.工程科学Ⅱ辑》, no. 9, 15 September 2011 (2011-09-15) * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103787509A (en) * | 2014-01-23 | 2014-05-14 | 河北科技大学 | Porous functional carrier preparation device and method for preparing porous macromolecular organic carrier applied to sewage treatment by using porous functional carrier preparation |
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