CN103603087A - Preparation method of nanofiber for soft tissue regeneration - Google Patents
Preparation method of nanofiber for soft tissue regeneration Download PDFInfo
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- CN103603087A CN103603087A CN201310586082.8A CN201310586082A CN103603087A CN 103603087 A CN103603087 A CN 103603087A CN 201310586082 A CN201310586082 A CN 201310586082A CN 103603087 A CN103603087 A CN 103603087A
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Abstract
The invention relates to a preparation method of an extracellular matrix nanofiber (NF-ECM) for soft tissue regeneration. The preparation method comprises the steps of carrying out electrostatic spinning under different conditions by using collagen, gelatin, polycaprolactone (PCL), poly(l-lactide-co-glycolide) (PLGA) and gelatin polymer solutions with different concentrations, and regulating electrostatic spinning parameters such as the rolling speed of a roller, receiving distance, the distance between an external voltage and an electrode and the like to reach an ideal spinning state. The NF-ECM can be used as an artificial cell ECM skin substitute and can simulate a natural ECM, provide the optimal cell growth environment and even regenerate large-scale wounds or complex organ structures. In addition, industrial needle-free electrostatic spinning equipment is adopted in the preparation method provided by the invention, so that the NF-ECM can be produced in a large scale and has better application prospects on the aspect of large-scale burned skin regeneration.
Description
Technical field
The present invention relates to a kind of preparation method of nanofiber, relate in particular to a kind of preparation method of the nanofiber for soft tissue regeneration.
Background technology
Large-scale soft tissue regeneration remains a challenge in clinical practice, as the guiding regeneration of severe trauma, burn, cancer excision postoperative skin and mucous membrane.Such wound can be cured by dermatoplasty or mucosa transplantation, but any transplanting means all need second operation and suitable donor.
For large-area damage, limited especially for body position and area, engagement technology is by even scope and the area of boring a hole and can expand donor to the method for the stretching of cutify of the surface of a wound, although engagement technology may contribute to reduce the size of graft, but may, due to the contraction after obvious scar and transplanting, cause the skin appearance defect that also can cause function aspects not attractive in appearance.More serious in the situation that, do not have donor to use, in order to prevent the body fluid flow microbial contamination of becoming estranged, interim solution is to use cadaver skin as a mechanical barrier, within a period of time of waiting for epithelium regeneration, may cause fatal complication.Therefore develop a kind of clinical practice, ready-made, artificial extracellular matrix (ECM) Graftskin urgent especially, this artificial extracellular matrix that becomes and can imitate natural ECM, best Growth of Cells environment can be provided, and even large area creation or complicated organ structure can be regenerated.
Natural ECM is complicated because its Fusion of Cells and function of organization cause it to form.Natural ECM normally a kind of three-dimensional of soft connective tissue (3D) nanofiber network structure, comprises collagen, reticular membrane and elastomer.Such structure meets the feature of human skin skin corium.The main component of natural ECM is collagen, and collagen is comprised of the thick collagenous fiber bundle of 1～20mm, and collagenous fiber bundle is comprised of the thick collagenous fibres of 30～10nm, manually produces this collagen fabric and need to adopt nanometer technology.
Electrostatic spinning is being applied as a kind of method of making nanofiber always, but because the productive rate of this technology is extremely low, single spinning head is only 0.005-0.008g/min, becomes a key factor that limits its development.By improving solution fltting speed and polymer solution concentration, can enhance productivity, but this can increase fibre diameter.
Summary of the invention
The present invention relates to a kind of preparation method of the nanofiber for soft tissue regeneration, utilize collagen, gelatin, poly-third to hand over fat caprolactone (PCL), with the gelatin copolymer solution of poly (glycolide-co-lactide) (PLGA) and variable concentrations carries out electrostatic spinning under different condition, by regulating the electrospinning parameters such as distance between rolling speed, receiving range, applied voltage and the electrode of cylinder to reach desirable spinning state.
(1) with 90% (v/v) acetic acid preparation mass fraction, be 10%, 30% and 50% TYPE-A Bos taurus domesticus Gmelin solution; PLGA (the molecule ratio of lactide and glycolide is 50:50), PCL and TYPE-A Bos taurus domesticus Gmelin are dissolved into 2, in 2,2-trifluoroethanol, TYPE-1 bovine collagen is dissolved in 1,1,1,3,3, in 3-hexafluoro-2-propyl alcohol, the concentration of all polymer is 10wt%, and solution prepares rear ultrasonic concussion 30min is dispersed in solvent collagen better, carries out immediately electrostatic spinning after solution prepares.
(2) nanofiber uses the spinning equipment based on line rotation electrode to prepare by the method for electrostatic spinning: an electrode cylinder of being furnished with 10 line electrodes, be immersed in the solution tank that 500mL polymer solution is housed, wherein electrode diameter 100 μ m each other distance be 20mm, electrode diameter of cylinder 50mm, length 300mm, after applying voltage, on the surface of line electrode, can produce spray silk, the metal cylinder that is coated with polypropylene screen of ground connection is as receiving system, the rolling speed that is immersed in the cylinder in solution tank is 10rpm, receiving range is 250mm, spinning voltage is 30～35kV, temperature is 25 ℃.
The present invention relates to a kind of extracellular matrix nanofiber (NF-ECM) for soft tissue regeneration, can be built into the collagenous fiber bundle that 1～20mm is thick, and this collagenous fiber bundle can form collagen, it is the important component part of three-dimensional (3D) nanofiber network Constituent cell epimatrix (ECM).Therefore, product of the present invention can be used as a kind of artificial extracellular matrix (ECM) Graftskin after workman processes, and can imitate natural ECM, and best Growth of Cells environment can be provided, and even large area creation or complicated organ structure can be regenerated.And preparation method of the present invention adopts the equipment of industrial needleless electrospinning, large-scale production extracellular matrix nanofiber (NF-ECM), has reasonable application prospect aspect large-area burns skin regeneration.
The specific embodiment
In order to deepen the understanding of the present invention, below in conjunction with instantiation, the present invention is described in further detail:
(1) with 90% (v/v) acetic acid preparation mass fraction, be 10%, 30% and 50% TYPE-A Bos taurus domesticus Gmelin solution.PLGA (the molecule ratio of lactide and glycolide is 50:50), PCL and TYPE-A Bos taurus domesticus Gmelin are dissolved into 2, in 2,2-trifluoroethanol, TYPE-1 bovine collagen is dissolved in 1,1,1,3,3, in 3-hexafluoro-2-propyl alcohol, the concentration of all polymer is 10wt%, and solution prepares rear ultrasonic concussion 30min is dispersed in solvent collagen better, carries out immediately electrostatic spinning after solution prepares.
(2) nanofiber uses the spinning equipment based on line rotation electrode to prepare by the method for electrostatic spinning: an electrode cylinder of being furnished with 10 line electrodes, be immersed in the solution tank that 500mL polymer solution is housed, wherein electrode diameter 100 μ m each other distance be 20mm, electrode diameter of cylinder 50mm, length 300mm, after applying voltage, on the surface of line electrode, can produce spray silk, the metal cylinder that is coated with polypropylene screen of ground connection is as receiving system, the rolling speed that is immersed in the cylinder in solution tank is 10rpm, receiving range is 250mm, spinning voltage is 30～35kV, temperature is 25 ℃.
Claims (2)
1. for a preparation method for the nanofiber of soft tissue regeneration, its step is as follows:
(1) with 90% (v/v) acetic acid preparation mass fraction, be 10%, 30% and 50% TYPE-A Bos taurus domesticus Gmelin solution; Poly (glycolide-co-lactide) (PLGA), poly-third hands over fat caprolactone (PCL) and TYPE-A Bos taurus domesticus Gmelin to be dissolved into 2, in 2,2-trifluoroethanol, TYPE-1 bovine collagen is dissolved in 1,1,1,3,3, in 3-hexafluoro-2-propyl alcohol, the concentration of all polymer is 10wt%, and solution prepares rear ultrasonic concussion 30min, carries out immediately electrostatic spinning after solution prepares;
(2) nanofiber uses the spinning equipment based on line rotation electrode to prepare by the method for electrostatic spinning:
An electrode cylinder of being furnished with 10 line electrodes, be immersed in the solution tank that 500mL polymer solution is housed, wherein electrode diameter 100 μ m are each other apart from being 20mm, and electrode diameter of cylinder 50mm, length 300mm, after applying voltage, on the surface of line electrode, can produce spray silk, the metal cylinder that is coated with polypropylene screen of ground connection is as receiving system, and the rolling speed that is immersed in the cylinder in solution tank is 10rpm, and receiving range is 250mm, spinning voltage is 30～35kV, and temperature is 25 ℃.
2. the preparation method of nanofiber as claimed in claim 1, is characterized in that: described PLGA is the PLGA that the molecule ratio of lactide and glycolide is 50:50.
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| CN201310586082.8A CN103603087B (en) | 2013-11-15 | 2013-11-15 | Preparation method of nanofiber for soft tissue regeneration |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105420848A (en) * | 2015-11-25 | 2016-03-23 | 中国纺织科学研究院 | Superfine polyglycollide fiber and preparation method and device, application and patch thereof |
| CN106400314A (en) * | 2016-09-10 | 2017-02-15 | 华南理工大学 | Composite nanofiber film of structure bionic skin extracellular matrix and producing method and application thereof |
| CN107059158A (en) * | 2017-05-19 | 2017-08-18 | 中国纺织科学研究院 | Poly (glycolide-lactide) and polycaprolactone composite fibre, its preparation method, purposes and operation suture thread |
| CN109289083A (en) * | 2018-10-25 | 2019-02-01 | 无锡中科光远生物材料有限公司 | A kind of preparation method for the nano fiber scaffold promoting skin wound healing |
| CN112295021A (en) * | 2020-10-19 | 2021-02-02 | 四川大学 | Skin graft with improved topological structure |
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| CN101581010A (en) * | 2009-05-21 | 2009-11-18 | 东华大学 | Preparation of bionic extracellular matrix hyaluronic acid and gelatin compound nanofiber membrane |
| CN102908668A (en) * | 2012-11-09 | 2013-02-06 | 无锡中科光远生物材料有限公司 | Preparation method of induced growth type absorbable patch |
| CN102949751A (en) * | 2012-11-28 | 2013-03-06 | 川北医学院第二临床医学院 | Preparation method for tissue engineering collagen-hyaluronic acid-chondroitin sulfate electrostatic spinning bracket |
| CN103083723A (en) * | 2013-01-18 | 2013-05-08 | 暨南大学 | Collagen/nano-crystalline cellulose skin regenerative material, preparation method and application thereof |
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- 2013-11-15 CN CN201310586082.8A patent/CN103603087B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101581010A (en) * | 2009-05-21 | 2009-11-18 | 东华大学 | Preparation of bionic extracellular matrix hyaluronic acid and gelatin compound nanofiber membrane |
| CN102908668A (en) * | 2012-11-09 | 2013-02-06 | 无锡中科光远生物材料有限公司 | Preparation method of induced growth type absorbable patch |
| CN102949751A (en) * | 2012-11-28 | 2013-03-06 | 川北医学院第二临床医学院 | Preparation method for tissue engineering collagen-hyaluronic acid-chondroitin sulfate electrostatic spinning bracket |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105420848A (en) * | 2015-11-25 | 2016-03-23 | 中国纺织科学研究院 | Superfine polyglycollide fiber and preparation method and device, application and patch thereof |
| CN106400314A (en) * | 2016-09-10 | 2017-02-15 | 华南理工大学 | Composite nanofiber film of structure bionic skin extracellular matrix and producing method and application thereof |
| CN107059158A (en) * | 2017-05-19 | 2017-08-18 | 中国纺织科学研究院 | Poly (glycolide-lactide) and polycaprolactone composite fibre, its preparation method, purposes and operation suture thread |
| CN109289083A (en) * | 2018-10-25 | 2019-02-01 | 无锡中科光远生物材料有限公司 | A kind of preparation method for the nano fiber scaffold promoting skin wound healing |
| CN112295021A (en) * | 2020-10-19 | 2021-02-02 | 四川大学 | Skin graft with improved topological structure |
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| CN103603087B (en) | 2015-07-15 |
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