CN102813562A - Three-dimensional large-aperture nanoscale fibrous scaffold and method for preparing same - Google Patents
Three-dimensional large-aperture nanoscale fibrous scaffold and method for preparing same Download PDFInfo
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Abstract
The invention discloses a three-dimensional large-aperture nanoscale fibrous scaffold and a method for preparing the same. The average diameter of fibers in the scaffold is about 1 nanometer to 900 nanometers, the voidage is about 50%-98%, the scaffold is of a three-dimensional structure, and the thickness of the scaffold is 0.01 micrometer to 10 centimeters. The method includes the following steps of dissolving biodegradable polyester and additives into organic solvent to prepare spinning solution. An electrostatic spinning process is used for spinning the spinning solution to obtain nanoscale/submicron-order fibrofelt. The fibrofelt is soaked in solvent within 48 hours after being spun, is pre-frozen at first, and then is frozen and dried, so that the three-dimensional fibrous scaffold with mutually communicated large holes is obtained. The three-dimensional tissue engineering scaffold material with the communicated large holes is prepared, the structures of the internal holes are uniform, and the three-dimensional large-aperture nanscale fibrous scaffold can be used as a tissue engineering cytoskeleton for bones, cartilages, blood vessels, hearts, nerves and the like, and is suitable for growth of various cells.
Description
Technical field
The present invention relates to three-dimensional large aperture nano-scale fiber support and method for preparing, particularly a kind of static spinning technique preparation is used for the repair deficiency tissue or makes up the method for organ support, belongs to medical material manufacturing approach technical field.
Background technology
How to make up organizational project repair materials with bionical n cell epimatrix 26S Proteasome Structure and Function; Thereby for seed cell provides good growth, increment and functional expression environment; Even induce and influence cell differentiation, be the difficult point and the focus of Tissue Engineering Study always.At present; By the nano-structure engineering rack of electrostatic spinning preparation owing to have great specific surface area, high voidage and high characteristics such as surface energy; Can be from the natural extracellular matrix of nanoscale imitation; Can be used as the porous support of cell growth, promote the migration and the propagation of cell, the research that has been widely used in preparing organizational project repair materials, wound dressing, slow releasing carrier of medication.
Ideal support not only need have high porosity, and needs big aperture, could carry propagation and the differentiation in order to cell of nutrition and oxygen like this.Porosity and average pore size are the important parameters of cell proliferation and formation three-dimensional tissue.The size in aperture depends on cell type, it is generally acknowledged it should is 50～500 μ m, but cell can easily be moved to the zone that the degree of depth is about 100 μ m, for darker zone, is difficult to guarantee the plantation and the infiltration of cell.General nano fiber scaffold has often hindered intercellular contact owing to the aperture is less.Especially myocardial cell is very harsh to the requirement of culture environment and timbering material, and the condition that now can provide is difficult to satisfy the needs of myocardial cell growth and differentiation, and it is good still can not to go out function in external structure, the cardiac muscular tissue of comparative maturity.Though can prepare the tissue engineering bracket with extracellular matrix structure through electrostatic spinning technique, because the aperture of electrospinning fibre is less, the cell of sowing only sticks, grows and breed at fiber surface.When the degree of depth surpassed 100 μ m, cell can't obtain prescribing adequate nutrition and death, therefore needed to combine other technology preparation aperture bigger, the support that fiber is thinner.
Biodegradable polyesters be have purity height, nonhazardous, nonirritant, chemical stability good, degrade, do not cause inflammation, do not have irritated reaction, good biocompatibility, not carcinogenic, content of beary metal is low, pH value is little; Do not cause performances such as haemolysis and blood coagulation, it has more application at biomedical sector.For example; Launched the research about polycaprolactone and polylactic acid based TEHV, porous cultured chondrocytes support, bone tissue engineer composite etc. in recent years both at home and abroad, its tissue engineering bracket has been applied to aspects such as skin, bone, cardiovascular, liver and blood vessel.But conventional biodegradable polyesters fiber is low not enough with cellular affinity than the adhesive force of granular cell, has influenced cell tactophily in its surface.Electrostatic spinning fiber has than minor diameter; And through the fibrous framework of post processing have that bigger hole can not only guarantee that cell is deeply grown and also can the intravital neutrophilic leukocyte of warrantor through repairing the hole of support; Engulf the antibacterial that causes infection easily, the healing at the position that helps being wound and be difficult for causing the infection at position of being wound.
Summary of the invention
The objective of the invention is to propose a kind of combining and prepare the method for macropore high porosity three-dimensional manometer fibrous framework, overcoming the above-mentioned defective that prior art exists, and improve cell compatibility with technologies such as electrostatic spinning process and freeze settled dryings.
The method for preparing macropore high porosity three-dimensional manometer fibrous framework of the present invention comprises the steps:
(1) with biodegradable polyesters, solvent and the additive of different content at 0～80 ℃ of following mixed dissolution; Obtain spinning solution; Wherein: according to the weight meter, 1～40 part of biodegradable polyesters and additive agent mixture, 99～60 parts of solvents; 0～50 part of additive obtains the as-spun fibre felt by dry spinning then.
Said biodegradable polyesters weight average molecular weight is 1～3,000,000, and molecular weight distributing index 2～5, impurity weight content are below 5%.
Said additive is polyvinylpyrrolidone (PVP), polyethylene glycol oxide (PEO), polyacrylamide, polyvinyl alcohol (PVA).Weight average molecular weight is 3000～3,000,000, and the impurity weight content is below 5%.
Described solvent is N; N dimethyl formamide DMF, N; N dimethyl acetylamide DMAc, ethanol, dichloromethane, chloroform, 1, more than one in 2-dichloroethanes, oxolane, acetone, trichloroethylene, ethyl acetate, hexafluoroisopropanol or the trifluoroethanol;
(2) carry out electrostatic spinning with above-mentioned electrostatic spinning liquid, extruded by micro-injection pump, spout connects high-voltage positive electrode; It is voltage 5kv～20kv that receiving device ground connection, the static of control are spun parameter, spout aperture 1 μ m～2mm; Solution flow rate 1 μ l/h～10ml/h; 0～100 ℃ of ambient temperature, receiving range 2～50cm collects nanofiber mats;
This fiber felt can be used for medical field such as medical patch, drug-loading fibre, medicament slow release fabric; Also can be used for being made into civilian environmental protection fields such as oil suction fabric, degradable nonwoven fabrics, degradable fabric, also can its short fiber be used to make many-sided purposes such as cigarette filter.
(3) spin the back and fiber felt is immersed in the solvent in the 48h, between-200 ℃～-4 ℃, carry out precooling,, obtain the stereoscopic three-dimensional support that macropore and hole connect each other again in-40 ℃～4 ℃ lyophilisation.
Said solvent is more than one in ethanol, acetone, the first alcohol and water etc.;
This fibrous framework can be used for the repairing at repairing, cerebrovascular and the carotid artery place of the repairing at cardiac stent, hernia place, various bone transplant operation, brain lobe, also can be used for human bodies such as lip repairing, bridge of the nose finishing everywhere.
The present invention has following advantage:
1. Biodegradable fibers support, not only have nonhazardous, nonirritant, chemical stability good, degrade, do not cause inflammation, do not have irritated reaction, good biocompatibility, not carcinogenic, content of beary metal is low, pH value is little, does not cause performances such as haemolysis and blood coagulation.And have required physics, mechanical property and processing characteristics, and can stand to disinfect.And implanting does not need to take out once more, reduces less patient suffering;
2. through the nanoscale three-dimensional rack of electrostatic spinning process preparation, have that operational approach is simple, Financial cost reduces.But also have the fento effect, and electric property, and mechanical property such as very large specific surface area, flexible and superpower mechanical behavior (like hardness and tensile strength), these excellent characteristics make nanofiber be more suitable for medical and health, fields such as organizational project;
3. nano fiber scaffold not only structurally can effectively imitate extracellular matrix, and has the performance of extracellular matrix on the performance, has functions such as good cell adhesiveness, amplification, migration, differentiation;
4. above-mentioned prepared large hole nano level three-dimensional rack has excellent biological compatibility and tissue regeneration ability; The degradation rate that is complementary with the tissue regeneration of human body corrupted; Good permeability can guarantee the unimpeded of cell; Reach the intensity and the clinical operability of required repair tissue or organ.Bigger hole can the intravital neutrophilic leukocyte of warrantor through repairing the hole of support, engulf the antibacterial that causes infection easily, the healing at the position that helps being wound and be difficult for causing the infection at position of being wound.
Description of drawings
Fig. 1 is the sem photograph of the large hole nano timbering material prepared of the embodiment of the invention 1.
Fig. 2 is the cytocompatibility figure of the large hole nano timbering material prepared of the embodiment of the invention 1.
The specific embodiment
Through embodiment the present invention is carried out concrete description below; It is to be noted; Following examples just are used for the present invention is further specified; Can not be interpreted as the restriction to protection domain of the present invention, the person skilled in the art in this field can make some nonessential improvement and adjustment to the present invention according to the invention described above content.
Embodiment 1
(1) preparation of fiber felt
Static spins parameter control
With weight fraction be 10% through the PCL that purifies and two kinds of polymer dissolution of PVP in organic solvent, obtain transparent homogeneous, contain the organic solution through the PVP of the PCL polymer of purification and 5% weight of 5% weight.It is the static spinning stock solution.
Carrying out static with prepared static spinning stock solution spins.Spinning parameter is controlled at voltage 15kv, spout aperture 0.4mm, solution flow rate 1ml/h, 20 ℃ of ambient temperatures, dash receiver and spout spacing 15cm.
(2) preparation of large hole nano fibrous framework
Spin the back and fiber felt be immersed in the organic solvent ethanol in the 48h, between-200 ℃, carry out precooling 8h, again in-4 ℃ freezing, use CO
2The dry 24h of atmosphere obtains the stereoscopic three-dimensional support that macropore and hole connect each other.
Embodiment 2
(1) preparation of fiber felt
Static spins parameter control
With weight fraction be 40% through the PCL that purifies and two kinds of polymer dissolution of PVP in organic solvent, obtain transparent homogeneous, contain the organic solution through the PVP of the PCL polymer of purification and 36% weight of 4% weight.It is the static spinning stock solution.
Carrying out static with prepared static spinning stock solution spins.Spinning parameter is controlled at voltage 20kv, spout aperture 0.7mm, solution flow rate 2ml/h, 25 ℃ of ambient temperatures, dash receiver and spout spacing 10cm.
(2) preparation of large hole nano fibrous framework
Spin the back and fiber felt be immersed in the organic solvent-acetone in the 48h, carry out precooling 8h in-4 ℃, again in-40 ℃ freezing, use CO
2The dry 12h of atmosphere obtains the stereoscopic three-dimensional support that macropore and hole connect each other.
Implement 3
(1) preparation of fiber felt
Static spins parameter control
With weight fraction be 30% through the PLGA that purifies and two kinds of polymer dissolution of PEO in organic solvent, obtain transparent homogeneous, contain the organic solution through the PVP of the PLGA polymer of purification and 21% weight of 9% weight.It is the static spinning stock solution.Carrying out static with prepared static spinning stock solution spins.Spinning parameter is controlled at voltage 30kv, spout aperture 0.1mm, solution flow rate 0.2ml/h, 40 ℃ of ambient temperatures, dash receiver and spout spacing 2cm.
(2) preparation of large hole nano fibrous framework
Spin the back and fiber felt be immersed in the mixed solvent of organic solvent first alcohol and water in the 48h, carry out precooling 1h in subzero 100 ℃, again in 4 ℃ freezing, use CO
2The dry 48h of atmosphere obtains the stereoscopic three-dimensional support that macropore and hole connect each other.
Implement 4
(1) preparation of fiber felt
Static spins parameter control
With weight fraction be 1% through the PLLA that purifies and two kinds of polymer dissolution of polyacrylamide in organic solvent, obtain transparent homogeneous, contain the organic solution through the PVP of the PLLA polymer of purification and 0.5% weight of 0.5% weight.It is the static spinning stock solution.
Carrying out static with prepared static spinning stock solution spins.Spinning parameter is controlled at voltage 5kv, spout aperture 0.1mm, solution flow rate 0.2ml/h, 80 ℃ of ambient temperatures, dash receiver and spout spacing 2cm.
(2) preparation of large hole nano fibrous framework
Spin the back and fiber felt be immersed in the mixed solvent of organic solvent first alcohol and water in the 48h, carry out precooling 1h in subzero 100 ℃, again in 4 ℃ freezing, use CO
2The dry 48h of atmosphere obtains the stereoscopic three-dimensional support that macropore and hole connect each other.
Implement 5
(1) preparation of fiber felt
Static spins parameter control
With weight fraction be 4% through the PLGA that purifies and two kinds of polymer dissolution of PVA in organic solvent, obtain transparent homogeneous, contain the organic solution through the PVA of the PLGA of purification and 3% weight of 1% weight.It is the static spinning stock solution.
Carrying out static with prepared static spinning stock solution spins.Spinning parameter is controlled at voltage 15kv, spout aperture 0.5mm, solution flow rate 0.5ml/h, 30 ℃ of ambient temperatures, dash receiver and spout spacing 10cm.
(2) preparation of large hole nano fibrous framework
Spin the back and fiber felt be immersed in the mixed solvent of organic solvent first alcohol and water in the 48h, carry out precooling 1h in subzero 100 ℃, again in 4 ℃ freezing, use CO
2The dry 48h of atmosphere obtains the stereoscopic three-dimensional support that macropore and hole connect each other.
Claims (7)
1. three-dimensional large aperture nano-scale fiber support and method for preparing, the average diameter that it is characterized in that fiber in the support is 1nm～900nm; Support is 3-D solid structure; Thickness is 0.01um～10cm; Voidage is about 50%～98%.
2. like claim 1 fibrous framework, it is characterized in that described biodegradable polyesters fiber molecule amount is 10,000 to 3,000,000; Said polyester is polycaprolactone, polylactic acid, gather Acetic acid, hydroxy-, bimol. cyclic ester (PGA), gather hydroxyalkyl acrylate (PHA), a kind of in the polylactide Acetic acid, hydroxy-, bimol. cyclic ester (PLGA).
3. the method for preparing of a three-dimensional large aperture nano-scale fiber support is characterized in that adopting following steps:
(1), degradable polyester, solvent and the additive of different content at 20～80 ℃ of following mixed dissolutions, obtained spinning solution; Wherein: according to the weight meter; 1～40 part of degradable polyester and additive agent mixture, 99～60 parts of solvents obtain the as-spun fibre felt by dry spinning then.
Described degradable polyester such as claim 2
Described additive such as claim 4
Described solvent such as claim 5
(2), carry out electrostatic spinning, extrude by micro-injection pump with above-mentioned electrostatic spinning liquid; Spout connects high-voltage positive electrode, receiving device ground connection, and it is voltage 5kv～20kv that the static of control spins parameter; Spout aperture 1 μ m～2mm, solution flow rate 1 μ l/h～10ml/h, 0～100 ℃ of ambient temperature; Receiving range 2～50cm collects nanofiber mats;
(3), in the 48h fiber felt is immersed in the solvent after spinning,, under uniform temperature, carries out CO again prior to carrying out the precooling drying under the uniform temperature
2Lyophilization obtains the stereoscopic three-dimensional support that macropore and hole connect each other.
Said solvent is more than one in ethanol, acetone, the first alcohol and water etc.
Described precooling temperature such as claim 6
Described lyophilization temperature again such as claim 7
4. method according to claim 3 is characterized in that described additive is more than one in PVP, polyethylene glycol oxide (PEO), polyacrylamide, the polyvinyl alcohol (PVA).
5. method according to claim 3; It is characterized in that described organic solvent is N; N dimethyl formamide DMF, N; N dimethyl acetylamide DMAc, ethanol, dichloromethane, chloroform, 1, one or more mixed solvents in 2-dichloroethanes, oxolane, acetone, trichloroethylene, ethyl acetate, hexafluoroisopropanol or the trifluoroethanol.
6. method according to claim 3 is characterized in that described precooling temperature is-200 ℃～-4 ℃.
7. method according to claim 3 is characterized in that described lyophilization temperature again is-40 ℃～4 ℃.
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103006359A (en) * | 2012-12-24 | 2013-04-03 | 汪泱 | Bionic three-dimensional tissue engineering scaffold and preparation method thereof |
| CN103088452A (en) * | 2013-01-24 | 2013-05-08 | 中山大学 | Preparation device and preparation method of three-dimensional electrospinning fiber support |
| CN103603138A (en) * | 2013-11-15 | 2014-02-26 | 无锡中科光远生物材料有限公司 | Preparation method of PLGA fibrous coat used for corneal tissue transplant |
| CN106491240A (en) * | 2015-09-07 | 2017-03-15 | 先健科技(深圳)有限公司 | absorbable stopper |
| CN107296979A (en) * | 2017-07-02 | 2017-10-27 | 东华大学 | A kind of organizational project nanofiber intravascular stent and preparation method thereof |
| CN108653812A (en) * | 2017-03-28 | 2018-10-16 | 上海微创医疗器械（集团）有限公司 | artificial ligament and preparation method thereof |
| CN108744052A (en) * | 2018-05-08 | 2018-11-06 | 广东职业技术学院 | A kind of compound rest and preparation method thereof can be used for organizational project ligament |
| CN108893872A (en) * | 2018-08-07 | 2018-11-27 | 湖南工业大学 | A kind of preparation method of three-dimensional bulk multi-hole bracket |
| CN109876186A (en) * | 2019-03-21 | 2019-06-14 | 福州大学 | A kind of biological medical degradable double-layer scaffold and preparation method thereof for neural restoration |
| CN109922759A (en) * | 2016-11-10 | 2019-06-21 | 美敦力瓦斯科尔勒公司 | Drug filling bracket and its manufacturing method with the filament for increased surface of internal cavity product |
| CN110747521A (en) * | 2019-11-02 | 2020-02-04 | 东华大学 | Three-dimensional electrostatic spinning micro-fiber scaffold with surface nano-structure and preparation method and application thereof |
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| CN103006359A (en) * | 2012-12-24 | 2013-04-03 | 汪泱 | Bionic three-dimensional tissue engineering scaffold and preparation method thereof |
| CN103088452A (en) * | 2013-01-24 | 2013-05-08 | 中山大学 | Preparation device and preparation method of three-dimensional electrospinning fiber support |
| CN103088452B (en) * | 2013-01-24 | 2015-07-08 | 中山大学 | Preparation device and preparation method of three-dimensional electrospinning fiber support |
| CN103603138A (en) * | 2013-11-15 | 2014-02-26 | 无锡中科光远生物材料有限公司 | Preparation method of PLGA fibrous coat used for corneal tissue transplant |
| CN106491240A (en) * | 2015-09-07 | 2017-03-15 | 先健科技(深圳)有限公司 | absorbable stopper |
| CN106491240B (en) * | 2015-09-07 | 2019-07-05 | 先健科技(深圳)有限公司 | Absorbable plugging device |
| CN109922759A (en) * | 2016-11-10 | 2019-06-21 | 美敦力瓦斯科尔勒公司 | Drug filling bracket and its manufacturing method with the filament for increased surface of internal cavity product |
| CN108653812A (en) * | 2017-03-28 | 2018-10-16 | 上海微创医疗器械（集团）有限公司 | artificial ligament and preparation method thereof |
| CN107296979A (en) * | 2017-07-02 | 2017-10-27 | 东华大学 | A kind of organizational project nanofiber intravascular stent and preparation method thereof |
| CN108744052A (en) * | 2018-05-08 | 2018-11-06 | 广东职业技术学院 | A kind of compound rest and preparation method thereof can be used for organizational project ligament |
| CN108893872A (en) * | 2018-08-07 | 2018-11-27 | 湖南工业大学 | A kind of preparation method of three-dimensional bulk multi-hole bracket |
| CN108893872B (en) * | 2018-08-07 | 2021-11-09 | 湖南工业大学 | Preparation method of three-dimensional fluffy porous scaffold |
| CN109876186A (en) * | 2019-03-21 | 2019-06-14 | 福州大学 | A kind of biological medical degradable double-layer scaffold and preparation method thereof for neural restoration |
| CN109876186B (en) * | 2019-03-21 | 2021-09-28 | 福州大学 | Biomedical degradable double-layer stent for nerve repair and preparation method thereof |
| CN110747521A (en) * | 2019-11-02 | 2020-02-04 | 东华大学 | Three-dimensional electrostatic spinning micro-fiber scaffold with surface nano-structure and preparation method and application thereof |
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Application publication date: 20121212 |