CN103893817B - A kind of coaxial electrostatic spinning fibrous framework and preparation method thereof - Google Patents
A kind of coaxial electrostatic spinning fibrous framework and preparation method thereof Download PDFInfo
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- CN103893817B CN103893817B CN201410105971.2A CN201410105971A CN103893817B CN 103893817 B CN103893817 B CN 103893817B CN 201410105971 A CN201410105971 A CN 201410105971A CN 103893817 B CN103893817 B CN 103893817B
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Abstract
The invention discloses a kind of coaxial electrostatic spinning fibrous framework and preparation method thereof, belong to bioengineered tissue field.This support comprises aliphatic polyester shell and polyvinylpyrrolidone sandwich layer, not only safety non-toxic and biocompatibility excellence is good, also has load and the function of slow releasing pharmaceutical and/or bioactie agent, can be widely used in bioengineered tissue field.Present invention also offers the preparation method of this coaxial electrostatic spinning fibrous framework, by carrying out coaxial electrostatic spinning to aliphatic polyester shell solution and polyvinylpyrrolidone sandwich layer solution under no-wind environment, prepare nanoscale electrospun fibrous scaffolds, this nanoscale electrospun fibrous scaffolds safety non-toxic, and there is higher specific surface area and porosity, make its carrier as medicine and/or bioactie agent be provided with good slow-release function.The inventive method is simple, and easy to control, practicality is stronger.
Description
Technical field
The present invention relates to bioengineered tissue field, particularly a kind of coaxial electrostatic spinning fibrous framework and preparation method thereof.
Background technology
Fibrous framework is a kind of bioengineered tissue support, because it has the nanoscale structures close with natural extracellular matrix, can the construction features of bionic extracellular matrix, there is effects such as supporting Growth of Cells, guide tissue regeneration, control organizational structure and the delivery of biologically active factor.Materials and structures due to fibrous framework is the principal element affecting its function, so very important to the preparation of fibrous framework.The nano-scale fiber prepared by coaxial electrostatic spinning technology is utilized not only to have higher specific surface area and porosity, also there is the structure similar with extracellular matrix, so conventional coaxial electrostatic spinning technology preparation has " shell-core " double-deck coaxial electrostatic spinning fibrous framework.The shell that biocompatibility is good supports Growth of Cells as supporting structure, and protects medicine and/or the bioactie agent of the load of sandwich layer institute.
Prior art uses the chloroformic solution of aliphatic polyester as shell solution, the chloroformic solution of protein-Polyethylene Glycol (PEG), as sandwich layer solution, prepares the coaxial electrostatic spinning fibrous framework with the nanoscale structures close with natural extracellular matrix by adopting coaxial electrostatic spinning technology.
Realizing in process of the present invention, inventor finds that prior art at least exists following problem:
Prior art coaxial electrostatic spinning fibrous framework uses PEG as core material, and during PEG local application, not only easily cause anaphylaxis, as urticaria or retardance anaphylaxis, and the disease such as the acidosis of easy initiation burn patient generation hypertonicity, metabolite and renal hypofunction.Visible, PEG cannot be applied in some specific occasion, and its application has certain limitation, thus makes the application of prepared coaxial electrostatic spinning fibrous framework have certain limitation.
Summary of the invention
In order to the application solving prior art coaxial electrostatic spinning fibrous framework has circumscribed problem, embodiments provide a kind of coaxial electrostatic spinning fibrous framework.Described technical scheme is as follows:
On the one hand, embodiments provide a kind of coaxial electrostatic spinning fibrous framework, described coaxial electrostatic spinning fibrous framework comprises: aliphatic polyester shell and polyvinylpyrrolidone sandwich layer.
Particularly, described aliphatic polyester is selected from least one in polycaprolactone, polylactic acid, Poly(D,L-lactide-co-glycolide, poly-(lactic acid-hexanol) copolymer.
As preferably, described coaxial electrostatic spinning fibrous framework also comprises medicine and/or bioactie agent, and described medicine and/or bioactie agent load are in described polyvinylpyrrolidone sandwich layer.
As preferably, described medicine is bovine serum albumin.
As preferably, in aliphatic polyester shell, in aliphatic polyester, polyvinylpyrrolidone sandwich layer, the mass ratio of polyvinylpyrrolidone and described medicine and/or bioactie agent is 1-1.5:0.35-0.55:0.01-0.015.
On the other hand, the embodiment of the present invention additionally provides a kind of preparation method of coaxial electrostatic spinning fibrous framework, and described method comprises:
Prepare aliphatic polyester shell solution: be dissolved in by aliphatic polyester in organic solvent, be stirred to and dissolve completely, obtain described aliphatic polyester shell solution;
Prepare polyvinylpyrrolidone sandwich layer solution: be dissolved in organic solvent of the same race by polyvinylpyrrolidone, be stirred to and dissolve completely, obtain described polyvinylpyrrolidone sandwich layer solution;
Under no-wind environment, respectively described aliphatic polyester shell solution and described polyvinylpyrrolidone sandwich layer solution are injected shell solution injector and sandwich layer solution injector, carry out coaxial electrostatic spinning, prepare described coaxial electrostatic spinning fibrous framework;
Preparing aliphatic polyester shell solution organic solvent used and preparing organic solvent used in polyvinylpyrrolidone sandwich layer solution is same.
As preferably, described method also comprises: described prepare polyvinylpyrrolidone sandwich layer solution time, in described polyvinylpyrrolidone sandwich layer solution, add medicine and/or bioactie agent.
As preferably, the concentration of described aliphatic polyester shell solution is 0.14g/ml-0.16g/ml, and the concentration of described polyvinylpyrrolidone sandwich layer solution is 0.065g/ml-0.075g/ml.
Particularly, described organic solvent is selected from least one in trifluoroethanol, formic acid, hexafluoroisopropanol, chloroform, ethanol.
As preferably, the operating parameter of described coaxial electrostatic spinning is: spinning voltage is 10.5-10.8Kv; Spinning head is 18-20cm to the distance of collecting board; The fltting speed of aliphatic polyester shell solution is 0.20-0.22 ml/hour; The fltting speed of polyvinylpyrrolidone sandwich layer solution is 0.13-0.15 ml/hour; The syringe needle internal diameter of shell solution injector is 0.89-0.91 millimeter; The syringe needle internal diameter of sandwich layer solution injector is 0.40-0.42 millimeter.
The beneficial effect that the technical scheme that the embodiment of the present invention provides is brought is:
Embodiments provide a kind of coaxial electrostatic spinning fibrous framework comprising aliphatic polyester shell and polyvinylpyrrolidone sandwich layer.Due to water-soluble pharmaceutical intermediate and pharmaceutical adjuvant that polyvinylpyrrolidone is a kind of safety non-toxic, can dissolve each other or compound with many kinds of substance, make coaxial electrostatic spinning fibrous framework safety non-toxic and there is excellent biocompatibility; Because N-H or the O-H bond energy of polyvinylpyrrolidone and multi-medicament and/or bioactie agent form intermolecular association, and release time and the action intensity of medicine and/or bioactie agent is controlled by this association, extend this medicine and/or bioactie agent emission and absorption time in vivo, give coaxial electrostatic spinning fibrous framework load and the function of slow releasing pharmaceutical and/or bioactie agent.Visible, the coaxial electrostatic spinning fibrous framework of the safety non-toxic that the embodiment of the present invention provides can be widely used in bioengineered tissue field.
The embodiment of the present invention additionally provides a kind of preparation method of coaxial electrostatic spinning fibrous framework, by carrying out coaxial electrostatic spinning to aliphatic polyester shell solution and polyvinylpyrrolidone sandwich layer solution under no-wind environment, nanoscale electrospun fibrous scaffolds can be prepared, this nanoscale electrospun fibrous scaffolds safety non-toxic, and there is higher specific surface area and porosity, make its carrier as medicine and/or bioactie agent be provided with good slow-release function.The inventive method is simple, and easy to control, practicality is stronger.
Accompanying drawing explanation
In order to be illustrated more clearly in the technical scheme in the embodiment of the present invention, below the accompanying drawing used required in describing embodiment is briefly described, apparently, accompanying drawing in the following describes is only some embodiments of the present invention, for those of ordinary skill in the art, under the prerequisite not paying creative work, other accompanying drawing can also be obtained according to these accompanying drawings.
Fig. 1 is the preparation flow figure of the coaxial electrostatic spinning fibrous framework that the embodiment of the present invention provides;
Fig. 2 is the preparation flow figure of the coaxial electrostatic spinning fibrous framework that further embodiment of this invention provides;
Fig. 3 is the preparation process schematic diagram of the coaxial electrostatic spinning fibrous framework that further embodiment of this invention provides;
Fig. 4 is the transmission electron microscope picture of the coaxial electrostatic spinning fibrous framework that further embodiment of this invention provides;
Fig. 5 is the scanning electron microscope (SEM) photograph of the coaxial electrostatic spinning fibrous framework that further embodiment of this invention provides;
Fig. 6 is the diameter distribution profile of the coaxial electrostatic spinning fibrous framework that further embodiment of this invention provides;
Fig. 7 a is the scanning electron microscope (SEM) photograph before the slow release of bovine serum albumin in the coaxial electrostatic spinning fibrous framework that provides of further embodiment of this invention;
Fig. 7 b is the scanning electron microscope (SEM) photograph after the slow release of bovine serum albumin in the coaxial electrostatic spinning fibrous framework that provides of further embodiment of this invention;
Fig. 8 is the slow release effect schematic diagram of bovine serum albumin in the coaxial electrostatic spinning fibrous framework that provides of further embodiment of this invention.
Wherein, 1 shell solution injector,
2 sandwich layer solution injectors,
3 spinning heads,
4 collecting boaries,
5 coaxial electrostatic spinning fibrous frameworks,
51 aliphatic polyester shells,
52 polyvinylpyrrolidone sandwich layers,
6 bovine serum albumins,
7 hollow form coaxial electrically spun silk fiber supports.
Detailed description of the invention
For making the object, technical solutions and advantages of the present invention clearly, below in conjunction with accompanying drawing, embodiment of the present invention is described further in detail.
Embodiment 1
Embodiments provide a kind of coaxial electrostatic spinning fibrous framework 5, this coaxial electrostatic spinning fibrous framework 5 comprises: aliphatic polyester shell 51 and polyvinylpyrrolidone sandwich layer 52.
Embodiments provide a kind of coaxial electrostatic spinning fibrous framework 5 comprising aliphatic polyester shell 51 and polyvinylpyrrolidone sandwich layer 52.Due to water-soluble pharmaceutical intermediate and pharmaceutical adjuvant that polyvinylpyrrolidone is a kind of safety non-toxic, can dissolve each other or compound with many kinds of substance, make coaxial electrostatic spinning fibrous framework 5 safety non-toxic and there is excellent biocompatibility; Because N-H or the O-H bond energy of polyvinylpyrrolidone and multi-medicament and/or bioactie agent form intermolecular association, and release time and the action intensity of medicine and/or bioactie agent is controlled by this association, extend this medicine and/or bioactie agent emission and absorption time in vivo, give coaxial electrostatic spinning fibrous framework 5 load and the function of slow releasing pharmaceutical and/or bioactie agent.Visible, the coaxial electrostatic spinning fibrous framework 5 of the safety non-toxic that the embodiment of the present invention provides can be widely used in bioengineered tissue field.
Embodiment 2
The invention provides a kind of coaxial electrostatic spinning fibrous framework 5, this coaxial electrostatic spinning fibrous framework 5 comprises: aliphatic polyester shell 51 and load have the polyvinylpyrrolidone sandwich layer 52 of medicine and/or bioactie agent.
By adding medicine and/or bioactie agent in polyvinylpyrrolidone sandwich layer 52, making prepared coaxial electrostatic spinning fibrous framework 5 be provided with medicinal function, orientation being carried out to patient and cures.
Particularly, this aliphatic polyester is selected from least one in polycaprolactone, polylactic acid, Poly(D,L-lactide-co-glycolide, poly-(lactic acid-hexanol) copolymer, preferred polycaprolactone.
Based on the requirement of the Biodegradable to shell, mechanical performance and processing characteristics, the Shell Materials of embodiment of the present invention fibrous framework selects at least one in polycaprolactone, polylactic acid, Poly(D,L-lactide-co-glycolide, poly-(lactic acid-hexanol) copolymer.And polycaprolactone is owing to having good biocompatibility and biological degradability, the advantage such as the compatibility, good solvent solubility good with other high molecular polymer, the preferred polycaprolactone of the embodiment of the present invention is as Shell Materials, thus the performance of coaxial electrostatic spinning fibrous framework 5 excellence prepared by giving.
Intermolecular association is formed based on N-H or the O-H bond energy of polyvinylpyrrolidone and multi-medicament and/or bioactie agent, and release time and the action intensity of medicine and/or bioactie agent is controlled by this association, extend this medicine and/or bioactie agent emission and absorption time in vivo.Visible, polyvinylpyrrolidone sandwich layer 52 can not only as the carrier of medicine and/or bioactie agent, and can the effectively medicine of slow release load and/or bioactie agent, is widely used in bioengineered tissue field.
As preferably, this medicine is bovine serum albumin 6.
Because bovine serum albumin 6 is widely used carriers in a kind of biochemical test, the preferred bovine serum albumin 6 of this medicine in the embodiment of the present invention.Be understandable that, this medicine also can be selected from the medicines such as other antibiotic, vitamin, protein and nucleic acid.
Particularly, in aliphatic polyester shell, in aliphatic polyester, polyvinylpyrrolidone sandwich layer, the mass ratio of polyvinylpyrrolidone and medicine and/or bioactie agent is 1-1.5:0.35-0.55:0.01-0.015.
In order to control the structure of prepared coaxial electrostatic spinning fibrous framework 5 preferably, make it have higher specific surface area and porosity, make the load capacity of medicine and/or bioactie agent and slow-release capability be in preferably equilibrium point, the mass ratio of polyvinylpyrrolidone and medicine and/or bioactie agent in aliphatic polyester, polyvinylpyrrolidone sandwich layer in aliphatic polyester shell controls at 1-1.5:0.35-0.55:0.01-0.015 by the embodiment of the present invention simultaneously.As preferably, in aliphatic polyester shell, in aliphatic polyester, polyvinylpyrrolidone sandwich layer, the mass ratio of polyvinylpyrrolidone and medicine and/or bioactie agent is 1:0.5:0.013.
Embodiment 3
As shown in Figure 1, embodiments provide a kind of preparation method of coaxial electrostatic spinning fibrous framework, comprising:
Step 101: prepare aliphatic polyester shell solution: be dissolved in by aliphatic polyester in organic solvent, be stirred to and dissolve completely, obtains aliphatic polyester shell solution.
Step 102: prepare polyvinylpyrrolidone sandwich layer solution: be dissolved in by polyvinylpyrrolidone in organic solvent, be stirred to and dissolve completely, obtains polyvinylpyrrolidone sandwich layer solution.
Step 103: under no-wind environment, injects shell solution injector 1 and sandwich layer solution injector 2 by aliphatic polyester shell solution and polyvinylpyrrolidone sandwich layer solution respectively, carries out coaxial electrostatic spinning, prepare coaxial electrostatic spinning fibrous framework 5.
The preparation method of the coaxial electrostatic spinning fibrous framework 5 that the embodiment of the present invention provides, by carrying out coaxial electrostatic spinning to aliphatic polyester shell solution and polyvinylpyrrolidone sandwich layer solution under no-wind environment, nanoscale electrospun fibrous scaffolds can be prepared, this nanoscale electrospun fibrous scaffolds safety non-toxic, and there is higher specific surface area and porosity, make its carrier as medicine and/or bioactie agent be provided with good slow-release function.The inventive method is simple, and easy to control, practicality is stronger.
Embodiment 4
As shown in Figure 2, embodiments provide a kind of preparation method of coaxial electrostatic spinning fibrous framework, comprising:
Step 201: be dissolved in by aliphatic polyester in organic solvent, be stirred to and dissolve completely, obtains aliphatic polyester shell solution, and the concentration controlling this aliphatic polyester shell solution is 0.14g/ml-0.16g/ml.
Coaxial electrostatic spinning process entails shell and sandwich layer solution have suitable concentration, are understandable that, concentration and viscosity are directly proportional.In order to ensure that aliphatic polyester shell solution has enough large surface tension and electric field force to balance each other, and form stable taylor cone by the spinning nozzle place that acts on of this equilibrant, and then the fibrous framework prepared by ensureing has the structure of stable uniform, the embodiment of the present invention when can spinning, the concentration controlling this aliphatic polyester shell solution is 0.14g/ml-0.16g/ml, preferred 0.15g/ml.
Step 202: polyvinylpyrrolidone and medicine and/or bioactie agent are dissolved in organic solvent, be stirred to and dissolve completely, obtain polyvinylpyrrolidone sandwich layer solution, the concentration controlling this polyvinylpyrrolidone sandwich layer solution is 0.065g/ml-0.075g/ml.
In order to improve the compatibility of shell and sandwich layer in prepared fibrous framework, preparing aliphatic polyester shell solution organic solvent used in the embodiment of the present invention and preparing organic solvent used in polyvinylpyrrolidone sandwich layer solution is same.
Due to sandwich layer solution concentration can not too large again can not be too little, because when other conditions constant, the concentration of sandwich layer solution is excessive, the viscous friction to sandwich layer solution that then shell solution produces at interface place can not overcome the viscoelastic power of self, and then well drawing-off can not form composite injection thread; The viscosity of sandwich layer solution is too small, then can not form the injection thread that continuous print is stable, causes the unstability of this coaxial electrostatic spinning process to increase.So the concentration that the embodiment of the present invention controls this polyvinylpyrrolidone sandwich layer solution is 0.065g/ml-0.075g/ml, preferred 0.07g/ml.
Step 203: medicine and/or bioactie agent are added in polyvinylpyrrolidone sandwich layer solution, be stirred to mix homogeneously, obtain the polyvinylpyrrolidone sandwich layer solution containing medicine and/or bioactie agent, the concentration controlling the polyvinylpyrrolidone sandwich layer drug in solution and/or bioactie agent that this contains medicine and/or bioactie agent is 0.0015g/ml-0.0025g/ml.
In order to prepare the coaxial electrostatic spinning fibrous framework 5 with release medicine and/or bioactie agent function, polyvinylpyrrolidone and medicine and/or bioactie agent are dissolved in organic solvent by the embodiment of the present invention simultaneously, prepare the polyvinylpyrrolidone sandwich layer solution containing medicine and/or bioactie agent.In order to ensure drug loading and the slow-release capability of prepared fibrous framework the best, it is 0.0015g/ml-0.0025g/ml that the embodiment of the present invention controls this concentration containing the polyvinylpyrrolidone sandwich layer solution of medicine and/or bioactie agent, preferred 0.002g/ml.
Step 204: under no-wind environment, respectively aliphatic polyester shell solution and the polyvinylpyrrolidone sandwich layer solution containing medicine and/or bioactie agent are injected shell solution injector 1 and sandwich layer solution injector 2, carry out coaxial electrostatic spinning, prepare coaxial electrostatic spinning fibrous framework 5.
As shown in Figure 3, in coaxial electrostatic spinning process, shell solution and sandwich layer solution are injected through shell solution injector 1 and sandwich layer solution injector 2 respectively, fiber jet is formed at same spinning head 3 place, and by the preferred aluminium foil of collecting board 4() collect, obtain coaxial electrostatic spinning fibrous framework 5.This fiber jet is before uncured, and its structure is easy to the impact being subject to extraneous factor, and in order to ensure structure and the length stable homogeneous of obtained fibrous framework, the embodiment of the present invention carries out coaxial electrostatic spinning operation under no-wind environment again.
Particularly, this organic solvent is selected from least one in trifluoroethanol, formic acid, hexafluoroisopropanol, chloroform, ethanol.
In order to effectively reduce the interfacial tension between shell and sandwich layer solution, making shell solution drive sandwich layer solution to carry out better drawing-off, forming good composite injection thread; And ensureing that fibrous framework center core layer-shell border of obtaining is clearly demarcated, structure is more perfect, and the embodiment of the present invention selects at least one in trifluoroethanol, formic acid, hexafluoroisopropanol, chloroform, ethanol as electrostatic spinning organic solvent, preferred trifluoroethanol.
As preferably, the operating parameter of this coaxial electrostatic spinning process is: spinning voltage is 10.5-10.8Kv; Spinning head 3 is 18-20cm to the distance of collecting board 4; The fltting speed of aliphatic polyester shell solution is 0.20-0.22 ml/hour; The fltting speed of polyvinylpyrrolidone sandwich layer solution is 0.13-0.15 ml/hour; The syringe needle internal diameter of shell solution injector 1 is 0.89-0.91 millimeter; The syringe needle internal diameter of sandwich layer solution injector 2 is 0.40-0.42 millimeter.
In coaxial electrostatic spinning process, very crucial to the control of its operating parameter.Particularly, the flow velocity controlling sandwich layer and shell solution is for very crucial the fibrous framework obtaining the good core-shell structure of form.In order to prevent sandwich layer solution flow rate too fast to such an extent as to the parcel of breakthrough shell solution, to form stable composite injection thread; In order to avoid shell solution carries out independent electrostatic spinning, obtain the fibrous framework of the continuous print core-shell structure of the good even thickness of form, the fltting speed of polycaprolactone shell solution controls at 0.20-0.22 ml/hour by the embodiment of the present invention, preferably 0.21 ml/hour; The fltting speed of polyvinylpyrrolidone sandwich layer solution is controlled at 0.13-0.15 ml/hour, preferably 0.14 ml/hour.
For spinning voltage, when added voltage is different, for breaking the balance of surface tension and electric field force, the drop of capillary tip will produce different surface configurations, affects the distribution situation of liquid droplets and the thread size produced subsequently, fibre morphology and its size of current of conducting.When applied voltage is lower, initial injection point is dashed forward in the outside of shower nozzle, and spray thread and also will produce in drop tip, now the diameter of drop is greater than the aperture of spraying syringe needle, and the nanofiber of gained is comparatively thin, and knot pearl is less; After voltage increases, retraction is sprayed in syringe needle by liquid droplets, and injection thread is produced by needle tip thereupon, and line density and the knot pearl density of gained nanofiber also have corresponding increase; Voltage continues to be increased to critical, then thread will be gone out by the direct sharp spray of inner needle wall, is no longer formed and sprays thread, and only form liquid droplets, make to connect pearl density and sharply rise.Based on more than, spinning voltage controls at 10.5-10.8Kv by the embodiment of the present invention, preferred 10.6Kv, thus ensures that the fibrous framework that obtains is nanoscale structures, and smooth without knot pearl.
For the distance of spinning head 3 to collecting board 4, on the one hand, in order to prevent prepared fibrous framework from too disperseing, collecting board 4 collects certain thickness fibrous framework required time process; On the other hand, in order to prevent obtained fibrous framework from forming band shape or beading structure, embodiment of the present invention control spinning head 3 is 18-20cm to the distance of collecting board 4, preferred 20cm.
Further, in order to make the core-shell structure of prepared fibrous framework clear and evenly, the syringe needle internal diameter of shell solution injector 1 controls at 0.89-0.91 millimeter by the embodiment of the present invention, preferably 0.90 millimeter; The syringe needle internal diameter of sandwich layer solution injector 2 is controlled at 0.40-0.42 millimeter, preferably 0.41 millimeter.
Embodiment 5
Be dissolved in by 0.75 gram of polycaprolactone in 5 milliliters of trifluoroethanols, stirring at low speed 2 hours, dissolves completely to polycaprolactone, obtains the polycaprolactone shell solution of 0.15g/ml; 0.35 gram of polyvinylpyrrolidone is dissolved in 5 milliliters of trifluoroethanols, stirring at low speed 4 hours, dissolve completely to polyvinylpyrrolidone, obtain the polyvinylpyrrolidone sandwich layer solution of 0.07g/ml, then using 10 milligrams of bovine serum albumins 6 as model protein, join in this polyvinylpyrrolidone sandwich layer solution, be made into the bovine serum albumin 6-polyvinylpyrrolidone sandwich layer solution of 0.002g/ml.
As shown in Figure 3, under no-wind environment, above-mentioned polycaprolactone shell solution and bovine serum albumin 6-polyvinylpyrrolidone sandwich layer solution are added in shell solution injector 1 and sandwich layer solution injector 2 respectively, carries out coaxial electrostatic spinning, prepare coaxial electrostatic spinning fibrous framework 5.In this coaxial electrostatic spinning process, control spinning voltage is 10.6Kv; Spinning head 3 is 20cm to the distance of collecting board 4; The fltting speed of polycaprolactone shell solution is 0.21 ml/hour; The fltting speed of polyvinylpyrrolidone sandwich layer solution is 0.14 ml/hour; The syringe needle internal diameter of shell solution injector 1 is 0.90 millimeter; The syringe needle internal diameter of sandwich layer solution injector 2 is 0.41 millimeter.
Utilize the fibrous framework prepared by transmission electron microscope observing, as shown in Figure 4, visible obvious " shell-core " double-decker clearly of fibrous framework prepared by the embodiment of the present invention, and the structural integrity of aliphatic polyester shell 51 and polyvinylpyrrolidone sandwich layer 52 is homogeneous, illustrate that load has the polyvinylpyrrolidone sandwich layer 52 of medicine and/or bioactie agent successfully to be wrapped up by aliphatic polyester shell 51.
Utilize the fibrous framework prepared by scanning electron microscopic observation, as shown in Figure 5, the fibre morphology of fibrous framework prepared by the embodiment of the present invention is homogeneous, smooth, is formed without knot pearl.And the diameter of fibrous framework prepared by the embodiment of the present invention is mainly distributed in 600-700 ran, belongs to nanoscale structures, with reference to Fig. 6.Visible, coaxial electrostatic spinning fibrous framework 5 structural parameters that the embodiment of the present invention provides are all better, reach the application requirement of bioengineered tissue.
Bovine serum albumin 6 in this fibrous framework is carried out fluorescent labeling, and in the fibrous framework prepared by the confocal microscopy embodiment of the present invention, bovine serum albumin 6 evenly and continuous distribution.
In order to verify that fibrous framework prepared by the embodiment of the present invention is to the load factor of bovine serum albumin 6, the embodiment of the present invention measures bovine serum albumin 6 load factor wherein.Detailed process is as follows:
Cut-off footpath is the circular support (weight is 30 milligrams) of 2 centimetres of sizes, dissolves, obtain mixed solution with 1 milliliter of trifluoroethanol.Then getting the above-mentioned mixed solution of 200 microlitre joins in 96 orifice plates, (uses exciting light: 575 nanometers with multichannel spectrophotometer; Utilizing emitted light: 620 nanometers) detect the content of bovine serum albumin 6, getting sample size is 6.In this fibrous framework, the computing formula of bovine serum albumin 6 load factor is:
Albumen load-carry duty (%)=100 × Mn/M0
Mn represents the quality of the bovine serum albumin 6 carried in this fibrous framework, and M0 represents in the coaxial electrostatic spinning preparation process of this fibrous framework, the consumption of bovine serum albumin 6.Result shows, and fibrous framework prepared by the embodiment of the present invention is 80.7 ± 4.5% to the load factor of bovine serum albumin 6, has stronger load capacity.
The dispose procedure of the bovine serum albumin 6 of the fibrous framework center core layer utilizing the scanning electron microscopic observation embodiment of the present invention to prepare, as shown in accompanying drawing 7a and accompanying drawing 7b, bovine serum albumin 6 is succeeded release in this fibrous framework, visible, after bovine serum albumin 6 discharges, this coaxial electrically spun silk fiber support becomes hollow form coaxial electrically spun silk fiber support 7.Visible, the fibrous framework that the embodiment of the present invention provides effectively can discharge it as the carrier of bovine serum albumin 6.
The effect of the embodiment of the present invention to prepared fibrous framework slow release bovine serum albumin 6 is tested, and detailed process is: cut-off footpath is the circular support (weight is 30 milligrams) of 2 centimetres of sizes, soaks, and be placed in 37 DEG C with 1.5 milliliters of phosphate buffers.Every day, soak 500 microlitre was got in timing, as sample, then with fresh phosphoric buffer, soak was increased to 1.5 milliliters.Repeat said process 21 days, (use exciting light: 575 nanometers with multichannel spectrophotometer; Utilizing emitted light: 620 nanometers) detect collected by sample in the content of bovine serum albumin 6.Getting sample size is 6, and calculates cumulative release amount according to this, reacts the slow release situation of bovine serum albumin 6 in this fibrous framework.Wherein, the computing formula of cumulative release amount is:
Cumulative release amount (%)=100 × Mt/Mn
Wherein, Mt represents when time point t, this fibrous framework discharge the quality of bovine serum albumin 6, Mn represents the quality of the bovine serum albumin 6 carried in support.
As shown in Figure 8, the dispose procedure of this fibrous framework can be divided into two stages, within 1-7 days, is release periods gradually of bovine serum albumin 6, within 8-21 days, is slow releasing phases of bovine serum albumin 6.This fibrous framework visible has successfully carried out slow release to Ox blood serum, and slow release effect is stablized, and slow-release time is long.
Embodiment 6
Be dissolved in by 0.7 gram of polycaprolactone in 5 milliliters of hexafluoro b propanols, stirring at low speed 2 hours, dissolves completely to polycaprolactone, obtains polycaprolactone shell solution; 0.325 gram of polyvinylpyrrolidone is dissolved in 5 milliliters of hexafluoro b propanols, stirring at low speed 4 hours, dissolve completely to polyvinylpyrrolidone, obtain polyvinylpyrrolidone sandwich layer solution, then using 9 milligrams of bovine serum albumins 6 as model protein, join in this polyvinylpyrrolidone sandwich layer solution, be made into bovine serum albumin 6-polyvinylpyrrolidone sandwich layer solution.
As shown in Figure 3, under no-wind environment, above-mentioned polycaprolactone shell solution and bovine serum albumin 6-polyvinylpyrrolidone sandwich layer solution are added in shell solution injector 1 and sandwich layer solution injector 2 respectively, carries out coaxial electrostatic spinning, prepare coaxial electrostatic spinning fibrous framework 5.In this coaxial electrostatic spinning process, control spinning voltage is 10.5Kv; Spinning head 3 is 18cm to the distance of collecting board 4; The fltting speed of polycaprolactone shell solution is 0.20 ml/hour; The fltting speed of polyvinylpyrrolidone sandwich layer solution is 0.13 ml/hour; The syringe needle internal diameter of shell solution injector 1 is 0.89 millimeter; The syringe needle internal diameter of sandwich layer solution injector 2 is 0.40 millimeter.
The coaxial electrostatic spinning fibrous framework 5 of what the embodiment of the present invention provided comprise aliphatic polyester shell 51 and polyvinylpyrrolidone sandwich layer 52 not only safety non-toxic and there is excellent biocompatibility, also there is load and the function of slow releasing pharmaceutical and/or bioactie agent, bioengineered tissue field can be widely used in.
Embodiment 7
Be dissolved in by 0.80 gram of polycaprolactone in 5 milliliters of trifluoroethanols, stirring at low speed 2 hours, dissolves completely to polycaprolactone, obtains polycaprolactone shell solution; 0.375 gram of polyvinylpyrrolidone is dissolved in 5 milliliters of trifluoroethanols, stirring at low speed 4 hours, dissolve completely to polyvinylpyrrolidone, obtain polyvinylpyrrolidone sandwich layer solution, then using 10.5 milligrams of bovine serum albumins 6 as model protein, join in this polyvinylpyrrolidone sandwich layer solution, be made into bovine serum albumin 6-polyvinylpyrrolidone sandwich layer solution.
As shown in Figure 3, under no-wind environment, above-mentioned polycaprolactone shell solution and bovine serum albumin 6-polyvinylpyrrolidone sandwich layer solution are added in shell solution injector 1 and sandwich layer solution injector 2 respectively, carries out coaxial electrostatic spinning, prepare coaxial electrostatic spinning fibrous framework 5.In this coaxial electrostatic spinning process, control spinning voltage is 10.8Kv; Spinning head 3 is 19cm to the distance of collecting board 4; The fltting speed of polycaprolactone shell solution is 0.22 ml/hour; The fltting speed of polyvinylpyrrolidone sandwich layer solution is 0.15 ml/hour; The syringe needle internal diameter of shell solution injector 1 is 0.91 millimeter; The syringe needle internal diameter of sandwich layer solution injector 2 is 0.42 millimeter.
The coaxial electrostatic spinning fibrous framework 5 of what the embodiment of the present invention provided comprise aliphatic polyester shell 51 and polyvinylpyrrolidone sandwich layer 52 not only safety non-toxic and there is excellent biocompatibility, also there is load and the function of slow releasing pharmaceutical and/or bioactie agent, bioengineered tissue field can be widely used in.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (6)
1. a preparation method for coaxial electrostatic spinning fibrous framework, is characterized in that, described method comprises:
Prepare aliphatic polyester shell solution: be dissolved in by aliphatic polyester in organic solvent, be stirred to and dissolve completely, obtain described aliphatic polyester shell solution;
Prepare polyvinylpyrrolidone sandwich layer solution: be dissolved in by polyvinylpyrrolidone in organic solvent, be stirred to and dissolve completely, obtain described polyvinylpyrrolidone sandwich layer solution;
Under no-wind environment, respectively described aliphatic polyester shell solution and described polyvinylpyrrolidone sandwich layer solution are injected shell solution injector and sandwich layer solution injector, carry out coaxial electrostatic spinning, prepare described coaxial electrostatic spinning fibrous framework;
Preparing aliphatic polyester shell solution organic solvent used and preparing organic solvent used in polyvinylpyrrolidone sandwich layer solution is same;
Described organic solvent is trifluoroethanol;
The operating parameter of described coaxial electrostatic spinning is: spinning voltage is 10.5-10.8Kv; Spinning head is 18-20cm to the distance of collecting board; The fltting speed of aliphatic polyester shell solution is 0.20-0.22 ml/hour; The fltting speed of polyvinylpyrrolidone sandwich layer solution is 0.13-0.15 ml/hour; The syringe needle internal diameter of shell solution injector is 0.89-0.91 millimeter; The syringe needle internal diameter of sandwich layer solution injector is 0.40-0.42 millimeter.
2. the preparation method of coaxial electrostatic spinning fibrous framework according to claim 1, it is characterized in that, described method also comprises: described prepare polyvinylpyrrolidone sandwich layer solution time, in described polyvinylpyrrolidone sandwich layer solution, add medicine and/or bioactie agent.
3. the preparation method of coaxial electrostatic spinning fibrous framework according to claim 1, it is characterized in that, the concentration of described aliphatic polyester shell solution is 0.14g/ml-0.16g/ml, and the concentration of described polyvinylpyrrolidone sandwich layer solution is 0.065g/ml-0.075g/ml.
4. the preparation method of coaxial electrostatic spinning fibrous framework according to claim 1, it is characterized in that, described aliphatic polyester is selected from least one in polycaprolactone, polylactic acid, Poly(D,L-lactide-co-glycolide, poly-(lactic acid-hexanol) copolymer.
5. the preparation method of coaxial electrostatic spinning fibrous framework according to claim 2, it is characterized in that, the mass ratio of described aliphatic polyester, polyvinylpyrrolidone and described medicine and/or bioactie agent is 1-1.5:0.35-0.55:0.01-0.015.
6. the preparation method of coaxial electrostatic spinning fibrous framework according to claim 2, is characterized in that, described medicine is bovine serum albumin.
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