CN103599563A - Preparation method of nanofiber scaffold for heart tissue engineering - Google Patents
Preparation method of nanofiber scaffold for heart tissue engineering Download PDFInfo
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Abstract
The invention relates to a preparation method of a nanofiber scaffold for heart tissue engineering. The preparation method comprises the steps of preparing the nanofiber scaffold from poly(glyceroi sebacate) (PGS)/a gelatin nanofiber scaffold material with different components through electrostatic spinning, and crosslinking the nanofiber scaffold in a mixed solution of hydrochlorinated N,N-(3-dimethylaminopropyl)-N'-ethyl-carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to obtain the nanofiber scaffold. The nanofiber scaffold related by the invention can be used for promoting the formation of sarcomere and has wide prospects on the aspect of being used as a material for cardiac muscle cell regeneration and repair.
Description
Technical field
The present invention relates to cardiac muscular tissue and repair and regeneration field, relate in particular to a kind of preparation of the nano fiber scaffold for heart tissue engineering.
Background technology
A very important challenge in heart tissue engineering field is the development that can potentially promote the bionical graft technology of cardiac muscular tissue's reparation and regeneration.A lot of research all concentrates on the anisotropic structure of simulating myocardial cell by building the method for a series of biophysics environment.For example, electrostatic spinning technique has been widely used as a kind of simple method still with property versatile and flexible, prepares and has nano fiber scaffold internal crosslinking pore structure, high voidage, and be used for simulating myocardium structure.Such nano fiber scaffold can the arrangement of inducing cell and the cell transfer of enhancing iuntercellular junction the structure of final induction of vascular tissue.Certain methods is simulated myocardial tissue structure with engineering rack and is is accurately regulated and controled the function of myocardial cell.But current technology is chemistry, mechanics and the structural behaviour of comprehensive extracellular Matrix simultaneously.
Gelatin has been widely used for the timbering material for the preparation of heart transplantation as a kind of protein of good mechanical properties.Gelatin is easy to degraded, and the slower synthetic polymer of degradation rate of other kind, as poly-adipic acid (PGA), polylactic acid (PLA) and polyurethane (PU) are usually used to prepare myocardium transplantation material.But these materials are easy to deform and cause degenerating of structure after the stretching through long-term.
Poly-decanedioic acid glyceride (PGS) is a kind of virose biodegradable elastomer (hardness :～0.056-1.5MPa), be widely used in some field of tissue engineering technology, as nerve, heart and vascular system that do not have.For example, PGS can be used to build the favous micro structure of a kind of anisotropic class and carrys out synchronously beating of supporting core myocyte.Yet this timbering material usually preparation technology is very complicated, can not accurately control the performance of support, cavernous structure is random formation, the size limited (micro-meter scale) of fiber, and be easy to degraded.
Summary of the invention
The present invention relates to a kind of preparation method of the nano fiber scaffold for heart tissue engineering.By elastomer polymer (as PGS) with there is relatively high-intensity biomaterial (as crosslinked gelatin) and combine and just there is extraordinary application prospect.Be the biomaterial that a kind of chemistry and mechanical property can regulate and control, can be used as the dimensional culture that suitable microenvironment is carried out heart cell (as Cardiac Fibroblasts and myocardial cell).
Concrete preparation method is as follows:
(1) poly-decanedioic acid glyceride (PGS) preparation (Mw=10000): decanedioic acid and glycerol are according to mol ratio 2:1～1:2 150～180 ℃ of synthetic PGS of reaction 30～48h in the atmosphere of nitrogen and fine vacuum (<50mTorr).
(2) there is the method preparation that the different PGS/ gelatine nano fiber timbering materials that form utilize electrostatic spinning: PGS and gelatin are according to different mass ratio (2:1,1:2 and 0:1) in the acetic acid solution of 80% (v/v), be made into the spinning liquid of 15～20% (w/v), with syringe pump, spinning liquid is injected in the syringe of 1～5mL and carries out electrostatic spinning, spinning speed is 2～4mL/h, spinning distance 10～15cm, spinning voltage 16～20kV.Prepare ordered nano-fibers time standby aluminium foil parallel pole as receiving device, preparation random alignment nanofiber time standby sheet glass as collection material, the nano fiber scaffold material of preparation is dry 24～48h in vacuum drying oven.
(3) crosslinked: the nano fiber scaffold of electrostatic spinning is immersed to hydrochlorinate N, in the mixed solution of N-(3-dimethyl aminopropyl)-N`-ethyl-carbodiimide (EDC) and N-hydroxy-succinamide (NHS), at 1～4 ℃, react 24～36h, when gelatin is cross-linked, EDC concentration is 85mmol/L, the mol ratio of EDC and NHS is 4:1, then timbering material is washed to three times to remove unnecessary cross-linking agent with phosphate buffer.
The present invention has built a kind of elastomer PGS/ gelatine nano fiber support and for cardiac muscle tissue engineering field.The arrangement of cell and be also subject to the impact of its structure (in order and random) when being organized in the intensity effect that is subject to timbering material, the contraction of myocardial cell, tissue and the ripe impact that is subject to structure, composition and the intensity of timbering material.
On ordered nano-fibers timbering material, myocardial cell shows the sarcomere structure of anisotropy tissue, and no matter is the timbering material of ordered structure and random structure, and PGS adds the formation that can both improve sarcomere.The present invention shows that PGS/ gelatine nano fiber support is the material that can be used in myocardial cell regeneration and repair.
The specific embodiment
In order to deepen the understanding of the present invention, below in conjunction with embodiment, the invention will be further described.
The preparation method that is used for the nano fiber scaffold material with adjustable mechanics and structural behaviour in cardiac muscle tissue engineering field, its step is as follows:
(1) preparation of PGS (Mw=10000): decanedioic acid and glycerol are according to the ratio of mol ratio 1:1 160 ℃ of synthetic PGS of reaction 36h in the atmosphere of nitrogen and fine vacuum (<50mTorr).
(2) there is the method preparation that the different PGS/ gelatine nano fiber timbering materials that form utilize electrostatic spinning: PGS and gelatin are made into the spinning liquid of 20% (w/v) in the acetic acid solution of 80% (v/v) according to different mass ratio (2:1,1:2 and 0:1).With syringe pump, spinning liquid is injected in the syringe of 1mL and carries out electrostatic spinning, spinning speed is 2ml/h, and spinning is apart from 10cm, spinning voltage 18kV.Prepare ordered nano-fibers time standby aluminium foil parallel pole as receiving device, preparation random alignment nanofiber time standby sheet glass as collection material, nano fiber scaffold material orderly and random alignment is distinguished called after A (2PGS:Gelatin), A (PGS:2Gelatin), A (Gelatin) and R (2PGS:Gelatin), R (PGS:2Gelatin), R (Gelatin), the nano fiber scaffold material of preparation is dry 24h in vacuum drying oven.
(3) crosslinked: electrostatic spinning nano fiber support is immersed to hydrochlorinate N, in the mixed solution of N-(3-dimethyl aminopropyl)-N`-ethyl-carbodiimide (EDC) and N-hydroxy-succinamide (NHS), at 4 ℃, react 24h, when gelatin is cross-linked, EDC concentration is 85mmol/L, the mol ratio of EDC and NHS is 4: 1, then timbering material is washed to three times to remove unnecessary cross-linking agent with phosphate buffer.
Claims (4)
1. for a preparation method for the nano fiber scaffold of heart tissue engineering, tool step is as follows:
(1) poly-decanedioic acid glyceride (PGS) preparation (Mw=10000):
Decanedioic acid and glycerol are less than 150～180 ℃ of synthetic PGS of reaction 30～48h in the atmosphere of 50mTorr according to mol ratio 2:1～1:2 in nitrogen and vacuum;
(2) by the method for electrostatic spinning, prepare PGS/ gelatine nano fiber timbering material:
PGS and gelatin are according to different mass ratio (2:1,1:2 and 0:1) in the acetic acid solution of 80% (v/v), be made into the spinning liquid of 15～20% (w/v), with syringe pump, spinning liquid is injected in the syringe of 1～5mL and carries out electrostatic spinning, spinning speed is 2～4mL/h, spinning distance 10～15cm, spinning voltage 16～20kV; Prepare ordered nano-fibers time standby aluminium foil parallel pole as receiving device, preparation random alignment nanofiber time standby sheet glass as collection material, the nano fiber scaffold material of preparation is dry 24～48h in vacuum drying oven;
(3) crosslinked:
The nano fiber scaffold of electrostatic spinning is immersed to hydrochlorinate N, in the mixed solution of N-(3-dimethyl aminopropyl)-N`-ethyl-carbodiimide (EDC) and N-hydroxy-succinamide (NHS), at 1～4 ℃, react 24～36h; When gelatin is cross-linked, EDC concentration is 85mmol/L, and the mol ratio of EDC and NHS is 4:1, then timbering material is washed to three times to remove unnecessary cross-linking agent with phosphate buffer.
2. the preparation method of nano fiber scaffold as claimed in claim 1, is characterized in that: the reaction temperature in step (1) is 170 ℃.
3. the preparation method of nano fiber scaffold as claimed in claim 1, is characterized in that: in step (2), spinning speed is 4mL/h.
4. the preparation method of nano fiber scaffold as claimed in claim 1, is characterized in that: in step (3), mixed solution reaction temperature is 4 ℃.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104815351A (en) * | 2015-05-21 | 2015-08-05 | 东南大学 | Myocardial bionic scaffold made from composite conducting material and preparation method thereof |
| CN106367820A (en) * | 2016-12-02 | 2017-02-01 | 苏州大学 | Air bubble electrostatic spinning device |
| 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 |
| WO2018058874A1 (en) * | 2016-09-29 | 2018-04-05 | 国家纳米科学中心 | Gel fiber composite scaffold material formed through one step and preparation method and use thereof |
| CN109097910A (en) * | 2018-07-26 | 2018-12-28 | 北京化工大学常州先进材料研究院 | The preparation and application of PGS/ polylactic acid nuclear shell structure nano tunica fibrosa |
| CN110409059A (en) * | 2019-07-30 | 2019-11-05 | 北京化工大学常州先进材料研究院 | The preparation method of the acrylated PGS nano fibrous membrane of dimethylaminoethyl methacrylate enhancing |
| CN111714704A (en) * | 2020-06-09 | 2020-09-29 | 南通纺织丝绸产业技术研究院 | PGS/SF electrospun artificial blood vessel and preparation method thereof |
| CN111719243A (en) * | 2020-06-09 | 2020-09-29 | 紫罗兰家纺科技股份有限公司 | PGS/SF electrospun membrane and preparation method thereof |
-
2013
- 2013-11-15 CN CN201310586350.6A patent/CN103599563A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| MAHSHID KHARAZIHA ET AL.: "PGS: Gelatin nanofibrous scaffolds with tunable mechanical and structural properties for engineering cardiac tissues", 《BIOMATERIALS》 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104815351A (en) * | 2015-05-21 | 2015-08-05 | 东南大学 | Myocardial bionic scaffold made from composite conducting material and preparation method thereof |
| WO2018058874A1 (en) * | 2016-09-29 | 2018-04-05 | 国家纳米科学中心 | Gel fiber composite scaffold material formed through one step and preparation method and use thereof |
| CN106367820A (en) * | 2016-12-02 | 2017-02-01 | 苏州大学 | Air bubble electrostatic spinning device |
| 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 |
| CN106581748B (en) * | 2016-12-09 | 2019-06-04 | 东华大学 | A kind of preparation method of three-dimensional structure sebacic acid and propyl tri-alcohol ester sill macropore bracket |
| CN109097910A (en) * | 2018-07-26 | 2018-12-28 | 北京化工大学常州先进材料研究院 | The preparation and application of PGS/ polylactic acid nuclear shell structure nano tunica fibrosa |
| CN110409059A (en) * | 2019-07-30 | 2019-11-05 | 北京化工大学常州先进材料研究院 | The preparation method of the acrylated PGS nano fibrous membrane of dimethylaminoethyl methacrylate enhancing |
| CN111714704A (en) * | 2020-06-09 | 2020-09-29 | 南通纺织丝绸产业技术研究院 | PGS/SF electrospun artificial blood vessel and preparation method thereof |
| CN111719243A (en) * | 2020-06-09 | 2020-09-29 | 紫罗兰家纺科技股份有限公司 | PGS/SF electrospun membrane and preparation method thereof |
| CN111719243B (en) * | 2020-06-09 | 2022-03-01 | 紫罗兰家纺科技股份有限公司 | PGS/SF electrospun membrane and preparation method thereof |
| CN111714704B (en) * | 2020-06-09 | 2022-03-29 | 南通纺织丝绸产业技术研究院 | PGS/SF electrospun artificial blood vessel and preparation method thereof |
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Application publication date: 20140226 |