CN103893817A - Coaxial electrostatic spinning fibrous scaffold and preparation method thereof - Google Patents
Coaxial electrostatic spinning fibrous scaffold and preparation method thereof Download PDFInfo
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- CN103893817A CN103893817A CN201410105971.2A CN201410105971A CN103893817A CN 103893817 A CN103893817 A CN 103893817A CN 201410105971 A CN201410105971 A CN 201410105971A CN 103893817 A CN103893817 A CN 103893817A
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Abstract
The invention discloses a coaxial electrostatic spinning fibrous scaffold and a preparation method thereof, belonging to the field of biological tissue engineering. The scaffold comprises an aliphatic polyester shell and a polyvinylpyrrolidone core, not only is safe and non-toxic and has excellent biocompatibility but also has the functions of loading and slowly releasing drugs and/or bioactive factors, and can be widely applied to the field of biological tissue engineering. The invention also provides the preparation method of the coaxial electrostatic spinning fibrous scaffold. The preparation method comprises the step of carrying out coaxial electrostatic spinning by using an aliphatic polyester shell solution and a polyvinylpyrrolidone core solution in the windless environment to prepare a nanoscale electrostatic spinning fibrous scaffold. The nanoscale electrostatic spinning fibrous scaffold is safe and non-toxic, has a relatively high specific surface area and porosity, and has a good slow release function as a carrier of the drugs and/or bioactive factors. The method is simple, is easy to control, and has relatively strong practicability.
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 n cell epimatrix, construction features that can bionic extracellular matrix, has the effects such as the Growth of Cells of support, guide tissue regeneration, control organizational structure and the delivery of biologically active factor.Because material and the structure of fibrous framework are the principal elements that affects its function, so very important to the preparation of fibrous framework.Utilize the prepared nano-scale fiber of coaxial electrostatic spinning technology not only to there is 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 is used the chloroformic solution of aliphatic polyester as shell solution, the chloroformic solution of protein-Polyethylene Glycol (PEG) is as sandwich layer solution, by adopting coaxial electrostatic spinning technology to prepare the coaxial electrostatic spinning fibrous framework with the nanoscale structures close with n cell epimatrix.
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 not only easily cause anaphylaxis when PEG local application, as urticaria or retardance anaphylaxis, also easily cause the disease such as acidosis and renal hypofunction of burn patient generation hypertonicity, metabolite.Visible, PEG cannot be applied in some specific occasion, and its application has certain limitation, thereby makes the application of prepared coaxial electrostatic spinning fibrous framework have certain limitation.
Summary of the invention
For the application that solves prior art coaxial electrostatic spinning fibrous framework has circumscribed problem, the embodiment of the present invention provides a kind of coaxial electrostatic spinning fibrous framework.Described technical scheme is as follows:
On the one hand, the embodiment of the present invention provides a kind of coaxial electrostatic spinning fibrous framework, and 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 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 also provides a kind of preparation method of coaxial electrostatic spinning fibrous framework, and described method comprises:
Prepare aliphatic polyester shell solution: aliphatic polyester is dissolved in organic solvent, is stirred to completely and dissolves, obtain described aliphatic polyester shell solution;
Prepare polyvinylpyrrolidone sandwich layer solution: polyvinylpyrrolidone is dissolved in organic solvent of the same race, is stirred to completely and dissolves, obtain described polyvinylpyrrolidone sandwich layer solution;
Without under wind environment, respectively described aliphatic polyester shell solution and described polyvinylpyrrolidone sandwich layer solution are injected to 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: while preparing polyvinylpyrrolidone sandwich layer solution, in described polyvinylpyrrolidone sandwich layer solution, add medicine and/or bioactie agent described.
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:
The embodiment of the present invention provides a kind of coaxial electrostatic spinning fibrous framework that comprises aliphatic polyester shell and polyvinylpyrrolidone sandwich layer.Because polyvinylpyrrolidone is a kind of water-soluble pharmaceutical intermediate and pharmaceutical adjuvant of safety non-toxic, can dissolve each other with many kinds of substance or compound, make coaxial electrostatic spinning fibrous framework safety non-toxic and there is excellent biocompatibility; Because the N-H of polyvinylpyrrolidone or O-H bond energy and multi-medicament and/or bioactie agent form intermolecular association, and by release time and the action intensity of this association control medicine and/or bioactie agent, extend this medicine and/or the bioactie agent emission and absorption time in vivo, the function of giving the load of coaxial electrostatic spinning fibrous framework 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 also provides a kind of preparation method of coaxial electrostatic spinning fibrous framework, by to aliphatic polyester shell solution and polyvinylpyrrolidone sandwich layer solution without carrying out coaxial electrostatic spinning under wind environment, can prepare nanoscale electrospun fibers support, this nanoscale electrospun fibers support safety non-toxic, and there is higher specific surface area and porosity, make its carrier as medicine and/or bioactie agent there is good slow-release function.The inventive method is simple, easy to control, and 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 of required use during embodiment is described is briefly described, apparently, accompanying drawing in the following describes is only some embodiments of the present invention, for those of ordinary skills, do not paying under the prerequisite of creative work, can also obtain according to these accompanying drawings other accompanying drawing.
Fig. 1 is the preparation flow figure of the coaxial electrostatic spinning fibrous framework that provides of the embodiment of the present invention;
Fig. 2 is the preparation flow figure of the coaxial electrostatic spinning fibrous framework that provides of further embodiment of this invention;
Fig. 3 is the preparation process schematic diagram of the coaxial electrostatic spinning fibrous framework that provides of further embodiment of this invention;
Fig. 4 is the transmission electron microscope picture of the coaxial electrostatic spinning fibrous framework that provides of further embodiment of this invention;
Fig. 5 is the scanning electron microscope (SEM) photograph of the coaxial electrostatic spinning fibrous framework that provides of further embodiment of this invention;
Fig. 6 is the diameter Distribution figure of the coaxial electrostatic spinning fibrous framework that provides of further embodiment of this invention;
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.
The specific embodiment
For making the object, technical solutions and advantages of the present invention clearer, below in conjunction with accompanying drawing, embodiment of the present invention is described further in detail.
The embodiment of the present invention provides a kind of coaxial electrostatic spinning fibrous framework 5, and this coaxial electrostatic spinning fibrous framework 5 comprises: aliphatic polyester shell 51 and polyvinylpyrrolidone sandwich layer 52.
The embodiment of the present invention provides a kind of coaxial electrostatic spinning fibrous framework 5 that comprises aliphatic polyester shell 51 and polyvinylpyrrolidone sandwich layer 52.Because polyvinylpyrrolidone is a kind of water-soluble pharmaceutical intermediate and pharmaceutical adjuvant of safety non-toxic, can dissolve each other with many kinds of substance or compound, make coaxial electrostatic spinning fibrous framework 5 safety non-toxics and there is excellent biocompatibility; Because the N-H of polyvinylpyrrolidone or O-H bond energy and multi-medicament and/or bioactie agent form intermolecular association, and by release time and the action intensity of this association control medicine and/or bioactie agent, extend this medicine and/or the bioactie agent emission and absorption time in vivo, the function of giving 5 loads of coaxial electrostatic spinning fibrous framework 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.
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 add medicine and/or bioactie agent in polyvinylpyrrolidone sandwich layer 52, make prepared coaxial electrostatic spinning fibrous framework 5 there is medicinal function, patient is carried out to orientation and cure.
Particularly, this aliphatic polyester is selected from least one in polycaprolactone, polylactic acid, Poly(D,L-lactide-co-glycolide, poly-(lactic acid-hexanol) copolymer, preferably polycaprolactone.
The requirement of Biodegradable, mechanical performance and processing characteristics based on to shell, the Shell Materials of embodiment of the present invention fibrous framework is selected at least one in polycaprolactone, polylactic acid, Poly(D,L-lactide-co-glycolide, poly-(lactic acid-hexanol) copolymer.And polycaprolactone due to have good biocompatibility and biological degradability, with the good advantage such as the compatibility, good solvent solubility of other high molecular polymer, the preferred polycaprolactone of the embodiment of the present invention is as Shell Materials, thereby gives the performance of prepared coaxial electrostatic spinning fibrous framework 5 excellences.
N-H based on polyvinylpyrrolidone or O-H bond energy and multi-medicament and/or bioactie agent form intermolecular association, and by release time and the action intensity of this association control medicine and/or bioactie agent, extend this medicine and/or the bioactie agent emission and absorption time in vivo.Visible, polyvinylpyrrolidone sandwich layer 52 not only can be served as the carrier of medicine and/or bioactie agent, and the medicine of effectively 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 preferably the structure of prepared coaxial electrostatic spinning fibrous framework 5, make it have higher specific surface area and porosity, make the load capacity of medicine and/or bioactie agent and slow-release capability in equilibrium point preferably, the mass ratio of polyvinylpyrrolidone and medicine and/or bioactie agent in aliphatic polyester, polyvinylpyrrolidone sandwich layer in aliphatic polyester shell is controlled 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.
As shown in Figure 1, the embodiment of the present invention provides a kind of preparation method of coaxial electrostatic spinning fibrous framework, comprising:
Step 101: prepare aliphatic polyester shell solution: aliphatic polyester is dissolved in organic solvent, is stirred to completely and dissolves, obtain aliphatic polyester shell solution.
Step 102: prepare polyvinylpyrrolidone sandwich layer solution: polyvinylpyrrolidone is dissolved in organic solvent, is stirred to completely and dissolves, obtain polyvinylpyrrolidone sandwich layer solution.
Step 103: without under wind environment, respectively aliphatic polyester shell solution and polyvinylpyrrolidone sandwich layer solution are injected to shell solution injector 1 and sandwich layer solution injector 2, carry 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 to aliphatic polyester shell solution and polyvinylpyrrolidone sandwich layer solution without carrying out coaxial electrostatic spinning under wind environment, can prepare nanoscale electrospun fibers support, this nanoscale electrospun fibers support safety non-toxic, and there is higher specific surface area and porosity, make its carrier as medicine and/or bioactie agent there is good slow-release function.The inventive method is simple, easy to control, and practicality is stronger.
As shown in Figure 2, the embodiment of the present invention provides a kind of preparation method of coaxial electrostatic spinning fibrous framework, comprising:
Step 201: aliphatic polyester is dissolved in organic solvent, is stirred to completely and dissolves, obtain aliphatic polyester shell solution, the concentration of controlling this aliphatic polyester shell solution is 0.14g/ml-0.16g/ml.
Coaxial electrostatic spinning crosses range request shell and sandwich layer solution has suitable concentration, is understandable that, concentration and viscosity are directly proportional.In order to guarantee 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 guarantee that prepared fibrous framework has the structure of stable uniform, the embodiment of the present invention is can spinning in the situation that, the concentration of controlling this aliphatic polyester shell solution is 0.14g/ml-0.16g/ml, preferably 0.15g/ml.
Step 202: polyvinylpyrrolidone and medicine and/or bioactie agent are dissolved in organic solvent, be stirred to completely and dissolve, obtain polyvinylpyrrolidone sandwich layer solution, the concentration of 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, in the embodiment of the present invention, preparing aliphatic polyester shell solution organic solvent used and preparing organic solvent used in polyvinylpyrrolidone sandwich layer solution is same.
Because the concentration of sandwich layer solution can not too greatly again can not be too little, because in the situation that other conditions are constant, the concentration of sandwich layer solution is excessive, what shell solution produced at interface place can not overcome self viscoelastic power to the viscous friction of sandwich layer solution, and then good drawing-off formation composite injection thread; The viscosity of sandwich layer solution is too small, can not form continuous stable injection thread, causes the unstability of this coaxial electrostatic spinning process to increase.So the concentration of this polyvinylpyrrolidone sandwich layer solution of embodiment of the present invention control is 0.065g/ml-0.075g/ml, preferably 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 that contains medicine and/or bioactie agent, the concentration of controlling this polyvinylpyrrolidone sandwich layer solution Chinese medicine that contains medicine and/or bioactie agent and/or bioactie agent is 0.0015g/ml-0.0025g/ml.
There is in order to prepare the coaxial electrostatic spinning fibrous framework 5 that discharges medicine and/or bioactie agent function, the embodiment of the present invention is dissolved in polyvinylpyrrolidone and medicine and/or bioactie agent in organic solvent simultaneously, prepares the polyvinylpyrrolidone sandwich layer solution that contains medicine and/or bioactie agent.In order to guarantee drug loading and the slow-release capability of prepared fibrous framework the best, the concentration of this polyvinylpyrrolidone sandwich layer solution that contains medicine and/or bioactie agent of embodiment of the present invention control is 0.0015g/ml-0.0025g/ml, preferably 0.002g/ml.
Step 204: without under wind environment, respectively aliphatic polyester shell solution and the polyvinylpyrrolidone sandwich layer solution that contains medicine and/or bioactie agent are injected to 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, at same spinning head, 3 places form fiber jet, 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 be subject to the impact of extraneous factor, and in order to guarantee structure and the length stable homogeneous of obtained fibrous framework, the embodiment of the present invention is again without carrying out coaxial electrostatic spinning operation under wind environment.
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, make shell solution drive sandwich layer solution to carry out better drawing-off, form good composite injection thread; And guarantee that fibrous framework center core layer-shell border of obtaining is clearly demarcated, structure is more perfect, the embodiment of the present invention selects at least one in trifluoroethanol, formic acid, hexafluoroisopropanol, chloroform, ethanol as electrostatic spinning organic solvent, preferably 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 of controlling sandwich layer and shell solution is very crucial for obtaining the fibrous framework of the good core-shell structure of form.In order to prevent that sandwich layer solution flow rate is too fast to such an extent as to break through the parcel of shell solution, to form stable composite injection thread; Carry out independent electrostatic spinning for fear of shell solution, obtain the fibrous framework of the continuous core-shell structure of the good even thickness of form, the fltting speed of polycaprolactone shell solution is controlled 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 to 0.13-0.15 ml/hour, preferably 0.14 ml/hour.
For spinning voltage, in the time that added voltage is different, for breaking surface tension and electric field equilibrium of forces, the drop on capillary tube top will produce different surface configurations, the liquid droplets that impact produces subsequently and distribution situation, fibre morphology and its size of current of conducting of thread size.In the time that applied voltage is lower, initial injection point is dashed forward in the outside of shower nozzle, sprays thread and also will produce in drop is most advanced and sophisticated, and now the diameter of drop is greater than the aperture of spraying syringe needle, and the nanofiber of gained is thinner, and knot pearl is less; After voltage increases, liquid droplets sprays retraction in syringe needle, makes to spray thread and is produced by needle tip thereupon, and the line density of gained nanofiber and knot pearl density also have corresponding increase; Voltage continues to be increased to critical, and thread will be gone out by the direct sharp spray of syringe needle inwall, no longer forms and sprays thread, and only form liquid droplets, makes to connect pearl density and sharply rises.Based on more than, spinning voltage is controlled at 10.5-10.8Kv by the embodiment of the present invention, preferably 10.6Kv, thereby guarantee that the fibrous framework obtaining is nanoscale structures, and smooth without knot pearl.
Distance for spinning head 3 to collecting board 4, on the one hand, in order to prevent that prepared fibrous framework from too disperseing, collecting board 4 is collected certain thickness fibrous framework required time process; On the other hand, in order to prevent that 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, preferably 20cm.
Further, in order to make the core-shell structure of prepared fibrous framework clear and even, the syringe needle internal diameter of shell solution injector 1 is controlled 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 to 0.40-0.42 millimeter, preferably 0.41 millimeter.
0.75 gram of polycaprolactone is dissolved in 5 milliliters of trifluoroethanols, and 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, without under wind environment, above-mentioned polycaprolactone shell solution and bovine serum albumin 6-polyvinylpyrrolidone sandwich layer solution are added respectively in shell solution injector 1 and sandwich layer solution injector 2, carry 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 prepared fibrous framework of transmission electron microscope observing, as shown in Figure 4, fibrous framework prepared by the embodiment of the present invention " shell-core " double-decker significantly clearly as seen, and the structural integrity homogeneous of aliphatic polyester shell 51 and polyvinylpyrrolidone sandwich layer 52, illustrates 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 prepared fibrous framework of scanning electron microscopic observation, as shown in Figure 5, the fibre morphology homogeneous of fibrous framework prepared by the embodiment of the present invention, smooth, form without knot pearl.And the diameter of fibrous framework prepared by the embodiment of the present invention is mainly distributed in 600-700 nanometer left and right, 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, reached the application requirements of bioengineered tissue.
Bovine serum albumin in this fibrous framework 6 is carried out to fluorescent labeling, and in the fibrous framework of preparing by the confocal microscopy embodiment of the present invention, bovine serum albumin 6 evenly and continuous distribution.
In order to verify fibrous framework prepared by the embodiment of the present invention load factor to bovine serum albumin 6, the embodiment of the present invention is measured bovine serum albumin 6 load factors wherein.Detailed process is as follows:
Cut-off footpath is 2 centimetres of big or small circular supports (weight is 30 milligrams), with 1 milliliter of trifluoroethanol dissolving, obtains mixed solution.Then get the above-mentioned mixed solution of 200 microlitre join 96 orifice plates in, (use 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 factors is:
Albumen load-carry duty (%)=100 × Mn/M0
Mn represents the quality of the bovine serum albumin 6 carrying in this fibrous framework, and M0 is illustrated in the coaxial electrostatic spinning preparation process of this fibrous framework, the consumption of bovine serum albumin 6.Result demonstration, 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.
Utilize the dispose procedure of the bovine serum albumin 6 of fibrous framework center core layer prepared by the scanning electron microscopic observation embodiment of the present invention, as shown in accompanying drawing 7a and accompanying drawing 7b, bovine serum albumin 6 release of succeeding 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 can effectively discharge it as the carrier of bovine serum albumin 6.
The embodiment of the present invention is tested the effect of prepared fibrous framework slow release bovine serum albumin 6, and detailed process is: cut-off footpath is 2 centimetres of big or small circular supports (weight is 30 milligrams), with 1.5 milliliters of phosphate buffers immersions, and is placed in 37 ℃.Every day, soak 500 microlitres were got in timing, as sample, then with fresh phosphoric buffer, soak were increased to 1.5 milliliters.Repeat said process 21 days, with multichannel spectrophotometer (use exciting light: 575 nanometers; Utilizing emitted light: 620 nanometers) detect the content of bovine serum albumin 6 in collected sample.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, when Mt is illustrated in time point t, the quality of bovine serum albumin that this fibrous framework discharges 6, Mn represents the quality of the bovine serum albumin 6 carrying 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 release periods of bovine serum albumin 6.Visible this fibrous framework has successfully carried out slow release to Ox blood serum, and slow release effect is stable, and slow-release time is long.
0.7 gram of polycaprolactone is dissolved in 5 milliliters of hexafluoro b propanols, and 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, without under wind environment, above-mentioned polycaprolactone shell solution and bovine serum albumin 6-polyvinylpyrrolidone sandwich layer solution are added respectively in shell solution injector 1 and sandwich layer solution injector 2, carry 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.
Not only safety non-toxic and there is excellent biocompatibility of 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, also there is the function of load slow releasing pharmaceutical and/or bioactie agent, can be widely used in bioengineered tissue field.
Embodiment 7
0.80 gram of polycaprolactone is dissolved in 5 milliliters of trifluoroethanols, and 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, without under wind environment, above-mentioned polycaprolactone shell solution and bovine serum albumin 6-polyvinylpyrrolidone sandwich layer solution are added respectively in shell solution injector 1 and sandwich layer solution injector 2, carry 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.
Not only safety non-toxic and there is excellent biocompatibility of 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, also there is the function of load slow releasing pharmaceutical and/or bioactie agent, can be widely used in bioengineered tissue field.
The foregoing is only preferred embodiment of the present invention, in order to limit the present invention, within the spirit and principles in the present invention not all, any modification of doing, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (10)
1. a coaxial electrostatic spinning fibrous framework, is characterized in that, described coaxial electrostatic spinning fibrous framework comprises: aliphatic polyester shell and polyvinylpyrrolidone sandwich layer.
2. coaxial electrostatic spinning fibrous framework according to claim 1, it is characterized in that, in described aliphatic polyester shell, aliphatic polyester is selected from least one in polycaprolactone, polylactic acid, Poly(D,L-lactide-co-glycolide, poly-(lactic acid-hexanol) copolymer.
3. coaxial electrostatic spinning fibrous framework according to claim 1, it is characterized in that, described coaxial electrostatic spinning fibrous framework also comprises medicine and/or bioactie agent, and described medicine and/or bioactie agent load in described polyvinylpyrrolidone sandwich layer.
4. coaxial electrostatic spinning fibrous framework according to claim 3, is characterized in that, described medicine is bovine serum albumin.
5. coaxial electrostatic spinning fibrous framework according to claim 3, it is characterized in that, 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.
6. a preparation method for the coaxial electrostatic spinning fibrous framework described in claim 1-5 any one, is characterized in that, described method comprises:
Prepare aliphatic polyester shell solution: aliphatic polyester is dissolved in organic solvent, is stirred to completely and dissolves, obtain described aliphatic polyester shell solution;
Prepare polyvinylpyrrolidone sandwich layer solution: polyvinylpyrrolidone is dissolved in organic solvent, is stirred to completely and dissolves, obtain described polyvinylpyrrolidone sandwich layer solution;
Without under wind environment, respectively described aliphatic polyester shell solution and described polyvinylpyrrolidone sandwich layer solution are injected to 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.
7. the preparation method of coaxial electrostatic spinning fibrous framework according to claim 6, it is characterized in that, described method also comprises: while preparing polyvinylpyrrolidone sandwich layer solution, in described polyvinylpyrrolidone sandwich layer solution, add medicine and/or bioactie agent described.
8. the preparation method of coaxial electrostatic spinning fibrous framework according to claim 6, 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.
9. the preparation method of coaxial electrostatic spinning fibrous framework according to claim 6, is characterized in that, described organic solvent is selected from least one in trifluoroethanol, formic acid, hexafluoroisopropanol, chloroform, ethanol.
10. according to the preparation method of the coaxial electrostatic spinning fibrous framework described in claim 6-9 any one, it is characterized in that, 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.
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