CN112807491A - Elastic tissue engineering scaffold with communicated macroporous structure and preparation method thereof - Google Patents
Elastic tissue engineering scaffold with communicated macroporous structure and preparation method thereof Download PDFInfo
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- CN112807491A CN112807491A CN202011619646.XA CN202011619646A CN112807491A CN 112807491 A CN112807491 A CN 112807491A CN 202011619646 A CN202011619646 A CN 202011619646A CN 112807491 A CN112807491 A CN 112807491A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/56—Porous materials, e.g. foams or sponges
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61L2400/16—Materials with shape-memory or superelastic properties
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/34—Materials or treatment for tissue regeneration for soft tissue reconstruction
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Abstract
The invention relates to an elastic tissue engineering scaffold with a communicated macroporous structure and a preparation method thereof. The scaffold comprises PLLA and PPS. The method comprises the following steps: PLLA and PPS were added to an organic solvent, stirred, and the resulting polymer solution was cooled to cause phase separation and to displace the organic solvent. The scaffold has good elasticity, biocompatibility and biodegradability, and has a macroporous microstructure which is mutually communicated.
Description
Technical Field
The invention belongs to the field of tissue repair and regeneration, and particularly relates to an elastic tissue engineering scaffold with a communicated macroporous structure and a preparation method thereof.
Background
Tissue repair and regeneration are always a great problem in the field of biomedicine, and the appearance and development of tissue engineering technology provides a new idea for the field and greatly promotes the development of the field. The tissue engineering comprises three elements: cells, scaffolds and growth factors. Scaffolds are the core part of tissue engineering, where they are intended to serve as a substitute for the native extracellular matrix (ECM), providing temporary support for cells. Tissue engineering scaffolds need to have good biocompatibility, biodegradability, and also need to have mechanical properties similar to those of natural tissues and an interconnected pore structure that facilitates material exchange. At present, the tissue engineering scaffolds applied in more applications mainly comprise: biodegradable high molecular porous foam scaffold, tissue acellular matrix scaffold, hydrogel and the like. The tissue acellular matrix scaffold and the hydrogel have good biocompatibility, biodegradability and a micropore structure, but the strength is low, and the tissue acellular matrix scaffold cannot meet the actual requirement for many clinical applications. The porous foam scaffold prepared from the biodegradable high polymer material is a thermoplastic material with high crystallinity, high modulus, poor elasticity and slow degradation, so that the scaffold has poor elasticity and degradability and is not suitable for application in soft tissue engineering. Therefore, the development of the tissue engineering scaffold with good elasticity, biodegradability, interconnected microporous structure and certain mechanical strength has important significance for the development of the field of tissue engineering, particularly soft tissue engineering.
Poly L-lactic acid (PLLA) is a biodegradable polymer material commonly used in biomedicine, has good mechanical properties and processability, and a polymer solution thereof can form a tissue engineering scaffold with a nanofiber structure through Thermally Induced Phase Separation (TIPS), thereby having a highly interconnected pore structure (Journal of biological Materials Research,1999,46(1): 60-72). However, PLLA itself has high crystallinity and modulus, so that its scaffold has poor elasticity and is not suitable for soft tissue engineering, and its nanofiber structure has too small pores to facilitate cell ingrowth, so that its application is limited. Therefore, there have been attempts to introduce a porogen (sugar spheres, salt particles, etc.) during the scaffold molding process so that the scaffold has a macroporous structure (Journal of biological Materials Research,2000,52(2): 430-. However, this method makes the preparation process of the scaffold more complicated, and the pore size of the obtained macroporous structure is too large, which is in the range of 100-. In addition, researchers have tried TIPS by combining various materials with PLLA, respectively, to obtain scaffolds with a certain macroporous structure (Acta biomaterials, 2018,79: 168-. However, the macropores of the obtained scaffold exist independently, the connectivity among the macropores is poor, the cell infiltration is not facilitated, and the introduced materials such as PLCL, PCL, PS and the like have poor elasticity, so that the obtained composite scaffold still has poor elasticity and is not suitable for soft tissue engineering application. Poly (sebacic acid-polyethylene glycol 200) ester (PPS) is a thermoplastic high-elasticity biodegradable polyester material disclosed in Chinese patent CN111892703A, and has good biocompatibility and biodegradability, ultrahigh elasticity and good processability. Therefore, the PPS and the PLLA are used in a composite way, and the tissue engineering scaffold with better comprehensive performances such as mechanical property, microstructure and the like is expected to be constructed.
Disclosure of Invention
The invention aims to solve the technical problem of providing an elastic tissue engineering scaffold with a communicated macroporous structure and a preparation method thereof, so as to overcome the defects of poor elasticity, biocompatibility, biodegradability, microstructure and the like of the tissue engineering scaffold in the prior art.
The invention provides an elastic tissue engineering scaffold with a communicated macroporous structure, which comprises the following components in percentage by mass of 0.05-19: 1 PLLA and PPS, a poly (sebacic acid-polyethylene glycol 200) ester.
Preferably, in the stent, the molecular weight of the PLLA is 5 to 50 ten thousand, and PDI is 1.01 to 5.0. More preferably, the poly-L-lactic acid PLLA has a molecular weight of 7-20 ten thousand, 1.01< PDI < 2.0.
Preferably, in the stent, the molecular weight of the poly (sebacic acid-polyethylene glycol 200) ester PPS is 5-50 ten thousand, and the PDI is 1.01-5.0. More preferably, the molecular weight of the PPS is 20-40 ten thousand, 1.01< PDI < 3.5.
Preferably, in the above stent, the mass ratio of PLLA to PPS is 0.1 to 5.0: 1.
preferably, in the above scaffold, the scaffold has, on a microscopic scale, interconnected macropores having an average pore diameter of 5 to 100 μm.
The invention also provides a preparation method of the elastic tissue engineering scaffold with the communicated macroporous structure, the scaffold is prepared from PLLA/PPS mixed polymer solution by a thermally induced phase separation method, and the preparation method comprises the following steps:
mixing poly (L-lactic acid) PLLA and poly (sebacic acid-polyethylene glycol 200) ester PPS in a mass ratio of 0.05-19: 1, adding the mixture into an organic solvent, stirring, carrying out cold treatment on the obtained polymer solution to carry out phase separation, and replacing the organic solvent to obtain the elastic tissue engineering scaffold with a communicated macroporous structure.
Preferably, in the above method, the preparation method of the poly (sebacic acid-polyethylene glycol 200) ester PPS comprises: mixing sebacic acid and polyethylene glycol 200 in a molar ratio of 0.99:1-1.10:1, stirring, carrying out esterification reaction under a vacuum condition, adding a catalyst, carrying out pre-condensation reaction under a vacuum condition, continuously reacting the obtained prepolymer, and purifying to obtain the modified polyester prepolymer.
More preferably, in the above method, the stirring is: stirring the mixture at the temperature of 160 ℃ under the protection of nitrogen until the sebacic acid is completely dissolved.
More preferably, in the above method, the process parameters of the esterification reaction are: the reaction temperature is 120 ℃ and 180 ℃, the pressure is 0.1-4000Pa, and the reaction time is 0.5-8 hours.
More preferably, in the above method, the catalyst is one of stannous octoate, stannous chloride, stannous octoate/p-toluenesulfonic acid, and stannous chloride/p-toluenesulfonic acid, the molar amount of p-toluenesulfonic acid is equal to that of stannous octoate or stannous chloride, and the molar amount of stannous octoate or stannous chloride is 0.0005-0.02 times of that of hydroxyl or carboxyl in the reaction system.
More preferably, in the above method, the process parameters of the precondensation reaction are: the reaction temperature is 120-180 ℃, the pressure is 0.1-3000Pa, and the reaction time is 2-20 hours.
More preferably, in the above method, the process parameters of the continuous reaction are: the reaction temperature is 180 ℃ and 280 ℃, the pressure is lower than 3000Pa, and the reaction time is 1-20 hours.
Preferably, in the above method, the organic solvent comprises one or more of tetrahydrofuran, N-dimethylformamide, N-dimethylacetamide, 1, 4-dioxane, hexafluoroisopropanol, trifluoroacetic acid and trifluoroethanol. More preferably, the organic solvent is tetrahydrofuran.
Preferably, in the above method, the mass concentration of the polymer solution is 1% to 40% g/mL. More preferably, the polymer solution has a mass/volume concentration of 4% to 20% g/mL.
Preferably, in the above method, the stirring temperature is 20 to 80 ℃. More preferably, the stirring temperature is 40-70 ℃.
Preferably, in the above method, the cold treatment temperature is-200-0 ℃, and the cold treatment time is not less than 10 minutes. More preferably, the cold treatment time is 2 to 48 hours.
Preferably, in the above method, the process parameters of the replacement of the organic solvent are as follows: the temperature is-100-50 ℃, the displacer comprises one or more of deionized water, methanol, ethanol, propanol, isopropanol, cyclohexane and normal hexane, the liquid changing frequency of the deionized water is at least 2 times, and the displacement time is 2 hours to 2 weeks.
The invention also provides application of the elastic tissue engineering scaffold with the communicated macroporous structure in soft tissue engineering. For example for soft tissue engineering.
Advantageous effects
(1) Because the adopted stent materials PLLA and PPS are both biocompatible and biodegradable, the composite stent also has good biocompatibility and biodegradability.
(2) Because the high-elasticity PPS material is introduced into the PLLA stent, the composite stent has better elasticity than a pure PLLA stent, and is more favorable for practical application, particularly application in the field of soft tissue engineering.
(3) Due to the introduction of PPS, the PLLA/PPS composite scaffold microscopically forms a uniform and interconnected macroporous structure, the aperture of the macroporous structure is obviously larger than that of a pure PLLA scaffold, the connectivity among pores is better, and the PLLA/PPS composite scaffold is more favorable for the growth of cells and the regeneration of new tissues.
Drawings
FIG. 1 is a photograph of the scaffold obtained in example 1 in a wet (aqueous) state and showing elastic recovery.
FIG. 2 is a cross-sectional Scanning Electron Microscope (SEM) image of the stent obtained in example 1.
FIG. 3 is a cross-sectional Scanning Electron Microscope (SEM) image of the stent obtained in comparative example 1.
Detailed Description
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should be understood that various changes or modifications of the present invention may be made by those skilled in the art after reading the teaching of the present invention, and such equivalents may fall within the scope of the present invention as defined in the appended claims.
The preparation method of the PPS comprises the following steps:
(1) adding 0.05mol of sebacic acid and 0.05mol of polyethylene glycol-200 (Vocko) into a 250ml two-neck flask, stirring for 0.5h at the temperature of 140 ℃ under the protection of nitrogen, and forming a clear and transparent liquid after the sebacic acid is completely dissolved;
(2) reducing the pressure of a reaction system to 3000Pa at 140 ℃, stirring and reacting for 0.5h, then adding 0.0005mol of p-toluenesulfonic acid, keeping the pressure to 3000Pa, and reacting for 4 h;
(3) adding 0.0005mol of stannous octoate into a reaction system, keeping the pressure at 3000Pa, and slowly raising the temperature from 140 ℃ to 260 ℃ within 8 h;
(4) when the temperature is increased to 260 ℃, the pressure is reduced to 100Pa, the reaction is continued for 4.5h, and a primary product of the polyester elastic material with high molecular weight is obtained;
(5) dissolving the obtained polyester primary product in tetrahydrofuran to obtain 10% (w/v) polymer solution, dropwise adding the solution into methanol, wherein the volume ratio of the tetrahydrofuran to the methanol is 1:4, and generating precipitate; and repeating the purification for 3 times, and drying the precipitate obtained in the last time in vacuum for 2 days to obtain the purified polyester elastic material.
Example 1
(1) Adding 0.1g of PLLA (Mw is 10 ten thousand, PDI is 1.5) and 0.4g of PPS (molecular weight is 30 ten thousand, PDI is 2.5) into a reaction bottle, adding 5ml of tetrahydrofuran, heating in a water bath at 60 ℃, stirring, and forming a clear and transparent liquid after the PLLA and the PPS are completely dissolved;
(2) pouring the obtained polymer solution into a polytetrafluoroethylene mold, and quickly placing the mold at-80 ℃ for 12 hours;
(3) taking out the mold from-80 ℃, placing the mold in 300ml of deionized water at 4 ℃, replacing the primary liquid for 12h, and keeping for 2 days;
(4) and taking the PLLA/PPS composite scaffold out of deionized water at 4 ℃, freezing and freezing at-20 ℃, and then carrying out freeze drying to obtain the dry PLLA/PPS composite scaffold.
FIG. 1 shows that: the PLLA/PPS (mass ratio of 20:80) composite stent obtained in example 1 has good elasticity, can deform when compressed, and can completely recover after external force is removed; and the support has good water absorption and retention effects, water contained in the support can be extruded out during compression, and the water is absorbed by the support again after external force is removed. These effects are mainly due to the high elasticity of PPS, and the microscopic porous structure of the PLLA/PPS composite scaffold, which is similar to a sponge.
FIG. 2 shows that: the Scanning Electron Microscope (SEM) picture of the cross section of the PLLA/PPS composite scaffold shows that the PLLA/PPS composite scaffold obtained in example 1 has a microscopically uniform macroporous structure, and the connectivity among pores is good, the pore diameter is about 20 μm, and the size of the PLLA/PPS composite scaffold is equivalent to that of human cells.
Example 2
A dry PLLA/PPS composite scaffold was obtained in the same manner as in example 1 except that the mass of PLLA and the mass of PPS were changed to 0.25g and 0.25g, respectively, according to example 1.
Comparative example 1
A dry PLLA stent was obtained in the same manner as in example 1 except that the mass of PLLA was changed to 0.5g without adding PPS according to example 1.
FIG. 3 shows: SEM pictures of the cross-section of a pure PLLA scaffold show that the PLLA scaffold obtained from comparative example 1 has a micro-porous structure, but the pore size is small.
Claims (10)
1. An elastic tissue engineering scaffold with an interconnected macroporous structure, which is characterized by comprising two materials of poly (L-lactic acid) PLLA and poly (sebacic acid-polyethylene glycol 200) ester PPS, wherein the mass ratio of the PLLA to the PPS is 0.05-19: 1.
2. the stent of claim 1, wherein the poly-L-lactic acid PLLA has a molecular weight of 5 to 50 ten thousand and a PDI of 1.01 to 5.0.
3. The stent of claim 1, wherein the poly (sebacic acid-polyethylene glycol 200) ester PPS has a molecular weight of 5-50 ten thousand and a PDI of 1.01-5.0.
4. The scaffold according to claim 1, wherein the scaffold has an average pore size of interconnected macropores on a microscopic scale of 5 to 100 μm.
5. A preparation method of an elastic tissue engineering scaffold with a communicated macroporous structure comprises the following steps:
mixing poly (L-lactic acid) PLLA and poly (sebacic acid-polyethylene glycol 200) ester PPS in a mass ratio of 0.05-19: 1, adding the mixture into an organic solvent, stirring, carrying out cold treatment on the obtained polymer solution to carry out phase separation, and replacing the organic solvent to obtain the elastic tissue engineering scaffold with a communicated macroporous structure.
6. The method as claimed in claim 5, wherein the preparation method of the poly (sebacic acid-polyethylene glycol 200) ester PPS comprises the following steps: mixing sebacic acid and polyethylene glycol 200 in a molar ratio of 0.99:1-1.10:1, stirring, carrying out esterification reaction under a vacuum condition, adding a catalyst, carrying out pre-condensation reaction under a vacuum condition, continuously reacting the obtained prepolymer, and purifying to obtain the modified polyester prepolymer.
7. The method according to claim 5, wherein the organic solvent comprises one or more of tetrahydrofuran, N-dimethylformamide, N-dimethylacetamide, 1, 4-dioxane, hexafluoroisopropanol, trifluoroacetic acid and trifluoroethanol; the mass/volume concentration of the polymer solution is 1-40% g/mL.
8. The method of claim 5, wherein the stirring temperature is 20-80 ℃; the cold treatment temperature is-200-0 deg.C, and the cold treatment time is not less than 10 min.
9. The method according to claim 5, wherein the process parameters for replacing the organic solvent are as follows: the temperature is-100-50 ℃, the displacer comprises one or more of deionized water, methanol, ethanol, propanol, isopropanol, cyclohexane and normal hexane, the liquid replacement frequency of the displacer is at least 2 times, and the replacement time is 2 hours to 2 weeks.
10. Use of the scaffold of claim 1 in tissue engineering.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102908208A (en) * | 2012-10-11 | 2013-02-06 | 东华大学 | Preparation method of porous nano-fiber tubular scaffold |
| CN103394125A (en) * | 2013-07-11 | 2013-11-20 | 东华大学 | Tissue engineering double-layered tubular support and preparation method thereof |
| CN104629026A (en) * | 2015-02-15 | 2015-05-20 | 东华大学 | Biomedical polybasic copolymerized crosslinked polyester elastomer material and preparation method thereof |
| US20170035932A1 (en) * | 2015-08-03 | 2017-02-09 | University Of Pittsburgh - Of The Commonwealth System Of Higher Education | Biodegradable Poly(Ester Amide) Elastomers and Uses Therefor |
| CN106581748A (en) * | 2016-12-09 | 2017-04-26 | 东华大学 | Production method of three-dimensional structured poly(glycerol-sebacate)-based macro-porous scaffold |
| CN111892703A (en) * | 2020-06-28 | 2020-11-06 | 东华大学 | Biodegradable thermoplastic polyester elastic material and preparation method thereof |
-
2020
- 2020-12-31 CN CN202011619646.XA patent/CN112807491A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102908208A (en) * | 2012-10-11 | 2013-02-06 | 东华大学 | Preparation method of porous nano-fiber tubular scaffold |
| CN103394125A (en) * | 2013-07-11 | 2013-11-20 | 东华大学 | Tissue engineering double-layered tubular support and preparation method thereof |
| CN104629026A (en) * | 2015-02-15 | 2015-05-20 | 东华大学 | Biomedical polybasic copolymerized crosslinked polyester elastomer material and preparation method thereof |
| US20170035932A1 (en) * | 2015-08-03 | 2017-02-09 | University Of Pittsburgh - Of The Commonwealth System Of Higher Education | Biodegradable Poly(Ester Amide) Elastomers and Uses Therefor |
| CN106581748A (en) * | 2016-12-09 | 2017-04-26 | 东华大学 | Production method of three-dimensional structured poly(glycerol-sebacate)-based macro-porous scaffold |
| CN111892703A (en) * | 2020-06-28 | 2020-11-06 | 东华大学 | Biodegradable thermoplastic polyester elastic material and preparation method thereof |
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