ELECTRIFIED COMPOSITE MEMBRANE WITH EXTRACELLULAR MATRIX ELECTRICAL TOPOLOGY CHARACTERISTICS, AND PREPARATION METHOD THEREOF

Abstract

The invention involves a kind of electrified composite membrane with extracellular matrix electrical topology characteristics and its preparation method, which resolves the technical problems of poor matching of electric characteristics and natural extracellular matrix characteristics in the existing materials and limited restoration effect of materials. The invention provides a kind of electrified composite membrane with extracellular matrix electrical topology characteristics mainly composed of ferroelectric polymer matrix and piezoelectric active fiber fillings. By regulating the draw ratio, content and of piezoelectric active fiber and thickness of composite film, the invention can realized the flexibility of film material and electrical topological features of bionic extracellular matrix, with proper tissue adhesion and good electric adaptability and high clinical operability.

Claims

1. A kind of electrified composite membrane with extracellular matrix electrical topology characteristics, wherein said electrified composite membrane is composed of high polymer and piezoelectric fiber filling.

2. The electrified composite membrane of claim 1 wherein said high polymer is a kind of or composite of ferroelectric polymer polyvinylidene fluoride or polyvinylidene fluoride-trifluroethylene.

3. The electrified composite membrane of claim 2 wherein said piezoelectric fiber filling is piezoelectric active fiber material and the said piezoelectric active fiber material is a kind of or multi-composite of barium titanate fiber, zinc oxide fiber, KNN fiber and so on.

4. The electrified composite membrane of claim 1 wherein said electrified composite membrane is thin-film material and the thickness of electrified composite membrane is 10 m500 m.

5. The electrified composite membrane of claim 3 wherein said electrified composite membrane with extracellular matrix electrical topology characteristics is a kind of thin-film material with a thickness of 30-300 m and the volume fraction of barium titanate fiber in the electrified composite membrane is 5-15 vol %.

6. The preparation method for the electrified composite membrane of claim 1, wherein said electrified composite membrane is prepared by dispersing, solving and casting with organic solvent, in the following steps: (1) Take glacial acetic acid and acetylacetone for mixing evenly, stirring for 5 min, prepare the mixed solvent of glacial acetic acid and acetylacetone, add barium acetate into the mixed solvent of glacial acetic acid and acetylacetone, stir for 1 h, add tetrabutyl titanate with the same molar weight as barium acetate, stir for 15 min, then add in polyvinylpyrrolidone, with the mass ratio of 9:32 with as barium acetate, and stir for 3 h; (2) Inject into the injector the mixed solvent obtained in Step (1), and obtain barium titanate nanofiber with the diameter of 50 nm500 nm, with the static electrospinning technology under a voltage of 10 kV16 kV; (3) Add the barium titanate nanofiber obtained in Step (2) into 0.010.1 mol/L of dopamine aqueous solution to form the solution with a concentration range of 0.010.1 g/mL stir for 6 h12 h at a temperature of 40 C.80 C., then apply ultrasonic concussion for 1 min15 min, centrifugal washing for 35 times, and then ultrasonic concussion for 110 min with a power of 180 W, to obtain barium titanate nanofiber fillings with a draw ratio of 820; (4) Take barium titanate nanofiber fillings obtained in Step (3), add into the organic solvent, NN-dimethylformamide, apply ultrasonic concussion and stir for 1 h3 h to obtain the fiber filling dispersion liquid; (5) Weigh and take ferroelectric high polymers, add into the organic solvent DMF, stir for 3 h6 h till completely solved to obtain the polymer solution. The concentration of solution obtained is 0.14 g/ml. The said ferroelectric high polymer is polyvinylidene fluoride or polyvinylidene fluoride-trifluoroethylene; (6) Add the dispersion liquid obtained in Step (4) into the polymer solution obtained in Step (5) so that the volume fraction range of barium titanate fiber fillings in polymer matrix id 120 vol %, stir for 6 h12 h so that the fiber fillings are evenly dispersed in polymer matrix to obtain the polymer mixed liquid containing ceramic fiber filling; (7) Take mixed liquid obtained in Step (6) for casting in the casting machine, place the cast film obtained under a temperature of 40 C.100 C. for drying to obtain a kind of composite film material with a thickness of 10 m500 m; (8) Polarize the film material obtained in Step (7), as per parameters: 1 kV30 kV (voltage), 0 mm50 mm (distance), 25 C.150 C. (temperature) and 1 min60 min (time), to obtain a kind of electrified composite membrane with extracellular matrix electrical topology characteristics.

7. The preparation method of electrified composite membrane of claim 6 is featured for that in the mixed solvent of glacial acetic acid and acetylacetone as mentioned in in Step (1), the volume ratio of glacial acetic acid and acetylacetone is 9:1.34 ml/g.

8. The preparation method of electrified composite membrane of claim 6 wherein feeding volume ratio of barium acetate and glacial acetic acid in Step (1) is 1.703:9 g/ml.

9. The preparation method of electrified composite membrane of claim 6 wherein mass percentage of fiber fillings obtained in Step (4) in the organic solvent is 1.5%30%.

Description

DESCRIPTIONS OF FIGURES

[0030] FIG. 1 is a photo of electrified composite membrane with extracellular matrix electrical topology characteristics prescribed in Example 1.

[0031] FIG. 2 is a surface SEM photo of electrified composite membrane with extracellular matrix electrical topology characteristics as prescribed in Example 1.

[0032] FIG. 3 is a section SEM photo of electrified composite membrane with extracellular matrix electrical topology characteristics as prescribed in Example 1.

[0033] FIG. 4 is the electrical stability result of electrified composite membrane with extracellular matrix electrical topology characteristics as prescribed in Example 1.

[0034] FIG. 5 is the analytical result of surface potential distribution simulation of electrified composite membrane with extracellular matrix electrical topology characteristics as prescribed in Example 1.

[0035] FIG. 6 is a LSCM photo of adhesion and spreading out 12 hours after the electrified composite membrane with extracellular matrix electrical topology characteristics acts on Rat BM MSC as prescribed in Example 1.

[0036] FIG. 7 is a Micro-CT photo of the test set and reference set of the experiment for the electrified composite membrane to restore rat skull defect as prescribed in Example 1.

PARTICULAR IMPLEMENTING MODE

[0037] With the following example, the invention can be better understood. However, the technicians of the field may understand easily that the content prescribed in the example is used to explain the invention only and neither shall nor will limit the invention as prescribed in the Claims.

Example 1

[0038] (1) Take 9 ml of glacial acetic acid and 1.340 g of acetylacetone, mix and stir for 5 min before adding 1.703 g of barium acetate, stirring for 1 h, adding 2.266 g of tetrabutyl titanate, stirring for 15 min, then adding 0.45 g of polyvinylpyrrolidone (PVP), stirring for 3 h;

[0039] (2) Inject into the injector the mixed solvent obtained in Step (1), and obtain barium titanate nanofiber with the diameter of 200 nm, with the static electrospinning technology under a voltage of 14 kV;

[0040] (3) Add the barium titanate nanofiber obtained in Step (2) into 0.01 mol/L of dopamine aqueous solution for bathing in water of 60 degrees, heating and stirring for 12 h, then apply ultrasonic concussion for 5 min, centrifugal drying to obtain barium titanate nanofiber fillings with a draw ratio of 12;

[0041] (4) Take an appropriate amount of barium titanate nanofiber fillings obtained in Step (3) for ultrasonic oscillation dispersion in 3 mL of organic solvent DMF, apply ultrasonic oscillation and stir for 1.5 h to obtain the fiber filling, dispersion liquid;

[0042] (5) Weigh and take 1 g of P (VDF-TrFE), add in 7 mL of organic solvent DMF, and stir for 5 h till completed solved to obtain P (VDF-TrFE) solution;

[0043] (6) Add the dispersion liquid obtained in Step (4) in P(VDF-TrFE) solution obtained in Step (5) so that the volume fraction of barium titanate fiber fillings in polymer is 7 vol %, stir for 10 h so that barium titanate fiber fillings are evenly dispersed in P (VDF-TrFE) matrix to obtain the mixed liquid;

[0044] (7) Take the mixed liquid obtained in Step (6) for casting in the casting machine, place the cast film obtained under a temperature of 55 C. for drying so that the solvent is completely evaporated to obtain a kind of composite film material with a thickness of 30 m;

[0045] (8) Polarize the film material obtained in Step (7), as per parameters: 20 kV (voltage), 20 mm (distance), 25 C. (temperature) and 30 min (time), to obtain a kind of electrified composite membrane with extracellular matrix electrical topology characteristics Its main components are P (VDF-TrFE) and barium titanate nanofiber, as the thin-film material with a thickness of 30 m. The volume fraction of barium titanate fiber fillings in electrified composite membrane is 5 vol %. For the photo the product in the example, see FIG. 1. For the surface SEM photo, see FIG. 2. For the section SEM photo, see FIG. 3.

[0046] Product Performance Testing:

[0047] (1) With COMSOL software, simulate and analyze the potential distribution features of the film material surface. For the results of simulation and analysis, see FIG. 4.

[0048] (2) Place the film material obtained in Step (8) in serum-free medium, for incubation for 0, 1, 3, 7, 14 and 21 days respectively under the condition of 37 C. At each time point, take out the material for piezoelectric constant testing. For the test result, see FIG. 5.

[0049] (3) Sterilize the film material obtained in (8) with Cobalt-60, then vaccinate, on its top, RMSC with a density of 510.sup.4, for incubation for 12 hours in total before immunostaining fixed and adherent plagues and then observing with LSCM. For the test result, see FIG. 6.

[0050] (4) Upon sterilizing with Cobalt-60, cut the film material obtained in Step (8) into a circular diaphragm with a diameter of 8 mm, for covering the rat skull defect with a diameter of 5 mm, while the defect not covered with any material is taken as the reference set. 4 weeks after the surgery, kill the animal and separate the film material for micro-CT scanning and observation. For the test result, see FIG. 7.

Example 2

[0051] (1) Take 9 ml of glacial acetic acid and 1.340 g of acetylacetone, mix and stir for 5 min before adding 1.703 g of barium acetate, stirring for 1 h, adding 2.266 g of tetrabutyl titanate, stirring for 15 min, then adding 0.45 g of polyvinylpyrrolidone (PVP), stirring for 3 h;

[0052] (2) Inject into the injector the mixed solvent obtained in Step (1), and obtain barium titanate nanofiber with the diameter of 300 nm, with the static electrospinning technology under a voltage of 16 kV;

[0053] (3) Add the barium titanate nanofiber obtained in Step (2) into 0.05 mol/L of dopamine aqueous solution for bathing in water of 60 degrees, heating and stirring for 12 h, then apply ultrasonic concussion for 10 min, centrifugal drying to obtain barium titanate nanofiber fillings with a draw ratio of 8;

[0054] (4) Take an appropriate amount of barium titanate nanofiber fillings obtained in Step (3) for ultrasonic oscillation dispersion in 3 mL of organic solvent DMF, apply ultrasonic oscillation and stir for 1.5 h to obtain the fiber filling, dispersion liquid;

[0055] (5) Weigh and take 1 g of P (VDF-TrFE), add in 7 mL of organic solvent DMF, and stir for 3 h till completed solved to obtain P (VDF-TrFE) solution;

[0056] (6) Add the dispersion liquid obtained in Step (4) in P(VDF-TrFE) solution obtained in Step (5) so that the volume fraction of barium titanate fiber fillings in polymer is 7 vol %, stir for 10 h so that barium titanate fiber fillings are evenly dispersed in P (VDF-TrFE) matrix to obtain the mixed liquid;

[0057] (7) Take the mixed liquid obtained in Step (6) for casting in the casting machine, place the cast film obtained under a temperature of 45 C. for drying so that the solvent is completely evaporated to obtain a kind of composite film material with a thickness of 50 m;

[0058] (8) Polarize the film material obtained in Step (7), as per parameters: 15 kV (voltage), 20 mm (distance), 50 C. (temperature) and 60 min (time), to obtain a kind of electrified composite membrane with extracellular matrix electrical topology characteristics Its main components are P (VDF-TrFE) and barium titanate nanofiber, as the thin-film material with a thickness of 30 m. The volume fraction of barium titanate fiber fillings in electrified composite membrane is 7 vol %.

[0059] Product Performance Testing:

[0060] (1) With COMSOL software, simulate and analyze the potential distribution features of the film material surface.

[0061] (2) Place the film material obtained in Step (8) in serum-free medium, for incubation for 0, 1, 3, 7, 14 and 21 days respectively under the condition of 37 C. At each time point, take out the material for piezoelectric constant testing.

[0062] (3) Sterilize the film material obtained in (8) with Cobalt-60, then vaccinate, on its top, RMSC with a density of 510.sup.4, for incubation for 12 hours in total before immunostaining fixed and adherent plagues and then observing with LSCM.

[0063] (4) Upon sterilizing with Cobalt-60, cut the film material obtained in Step (8) into a circular diaphragm with a diameter of 8 mm, for covering the rat skull defect with a diameter of 5 mm, while the defect not covered with any material is taken as the reference set. 4 weeks after the surgery, kill the animal and separate the film material for micro-CT scanning and observation.

Example 3

[0064] (1) Take 9 ml of glacial acetic acid and 1.340 g of acetylacetone, mix and stir for 5 min before adding 1.703 g of barium acetate, stirring for 1 h, adding 2.266 g of tetrabutyl titanate, stirring for 15 min, then adding 0.45 g of polyvinylpyrrolidone (PVP), stirring for 3 h;

[0065] (2) Inject into the injector the mixed solvent obtained in Step (1), and obtain barium titanate nanofiber with the diameter of 400 nm, with the static electrospinning technology under a voltage of 12 kV;

[0066] (3) Add the barium titanate nanofiber obtained in Step (2) into 0.08 mol/L of dopamine aqueous solution for bathing in water of 60 degrees, heating and stirring for 12 h, then apply ultrasonic concussion for 10 min, centrifugal drying to obtain barium titanate nanofiber fillings with a draw ratio of 20;

[0067] (4) Take an appropriate amount of barium titanate nanofiber fillings obtained in Step (3) for ultrasonic oscillation dispersion in 3 mL of organic solvent DMF, apply ultrasonic oscillation and stir for 1.5 h to obtain the fiber filling, dispersion liquid;

[0068] (5) Weigh and take 1 g of P (VDF-TrFE), add in 7 mL of organic solvent DMF, and stir for 5 h till completed solved to obtain P (VDF-TrFE) solution;

[0069] (6) Add the dispersion liquid obtained in Step (4) in P(VDF-TrFE) solution obtained in Step (5) so that the volume fraction of barium titanate fiber fillings in polymer is 10 vol %, stir for 10 h so that barium titanate fiber fillings are evenly dispersed in P (VDF-TrFE) matrix to obtain the mixed liquid;

[0070] (7) Take the mixed liquid obtained in Step (6) for casting in the casting machine, place the cast film obtained under a temperature of 80 C. for drying so that the solvent is completely evaporated to obtain a kind of composite film material with a thickness of 100 m;

[0071] (8) Polarize the film material obtained in Step (7), as per parameters: 10 kV (voltage), 15 mm (distance), 100 C. (temperature) and 40 min (time), to obtain a kind of electrified composite membrane with extracellular matrix electrical topology characteristics Its main components are P (VDF-TrFE) and barium titanate nanofiber, as the thin-film material with a thickness of 100 m. The volume fraction of barium titanate fiber fillings in electrified composite membrane is 10 vol %.

[0072] Product Performance Testing:

[0073] (1) With COMSOL software, simulate and analyze the potential distribution features of the film material surface.

[0074] (2) Place the film material obtained in Step (8) in serum-free medium, for incubation for 0, 1, 3, 7, 14 and 21 days respectively under the condition of 37 C. At each time point, take out the material for piezoelectric constant testing.

[0075] (3) Sterilize the film material obtained in (8) with Cobalt-60, then vaccinate, on its top, RMSC with a density of 510.sup.4, for incubation for 12 hours in total before immunostaining fixed and adherent plagues and then observing with LSCM.

[0076] (4) Upon sterilizing with Cobalt-60, cut the film material obtained in Step (8) into a circular diaphragm with a diameter of 8 mm, for covering the rat skull defect with a diameter of 5 mm, while the defect not covered with any material is taken as the reference set. 4 weeks after the surgery, kill the animal and separate the film material for micro-CT scanning and observation.

Example 4

[0077] (1) Take 9 ml of glacial acetic acid and 1.340 g of acetylacetone, mix and stir for 5 min before adding 1.703 g of barium acetate, stirring for 1 h, adding 2.266 g of tetrabutyl titanate, stirring for 15 min, then adding 0.45 g of polyvinylpyrrolidone (PVP), stirring for 3 h;

[0078] (2) Inject into the injector the mixed solvent obtained in Step (1), and obtain barium titanate nanofiber with the diameter of 400 nm, with the static electrospinning technology under a voltage of 12 kV;

[0079] (3) Add the barium titanate nanofiber obtained in Step (2) into 0.09 mol/L of dopamine aqueous solution for bathing in water of 60 degrees, heating and stirring for 12 h, then apply ultrasonic concussion for 10 min, centrifugal drying to obtain barium titanate nanofiber fillings with a draw ratio of 20;

[0080] (4) Take an appropriate amount of barium titanate nanofiber fillings obtained in Step (3) for ultrasonic oscillation dispersion in 3 mL of organic solvent DMF, apply ultrasonic oscillation and stir for 1.5 h to obtain the fiber filling, dispersion liquid;

[0081] (5) Weigh and take 1 g of P (VDF-TrFE), add in 7 mL of organic solvent DMF, and stir for 5 h till completed solved to obtain P (VDF-TrFE) solution;

[0082] (6) Add the dispersion liquid obtained in Step (4) in P(VDF-TrFE) solution obtained in Step (5) so that the volume fraction of barium titanate fiber fillings in polymer is 10 vol %, stir for 10 h so that barium titanate fiber fillings are evenly dispersed in P (VDF-TrFE) matrix to obtain the mixed liquid;

[0083] (7) Take the mixed liquid obtained in Step (6) for casting in the casting machine, place the cast film obtained under a temperature of 80 C. for drying so that the solvent is completely evaporated to obtain a kind of composite film material with a thickness of 100 m;

[0084] (8) Polarize the film material obtained in Step (7), as per parameters: 10 kV (voltage), 15 mm (distance), 100 C. (temperature) and 40 min (time), to obtain a kind of electrified composite membrane with extracellular matrix electrical topology characteristics Its main components are P (VDF-TrFE) and barium titanate nanofiber, as the thin-film material with a thickness of 120 m. The volume fraction of barium titanate fiber fillings in electrified composite membrane is 10 vol %.

[0085] Product Performance Testing:

[0086] (1) With COMSOL software, simulate and analyze the potential distribution features of the film material surface.

[0087] (2) Place the film material obtained in Step (8) in serum-free medium, for incubation for 0, 1, 3, 7, 14 and 21 days respectively under the condition of 37 C. At each time point, take out the material for piezoelectric constant testing.

[0088] (3) Sterilize the film material obtained in (8) with Cobalt-60, then vaccinate, on its top, RMSC with a density of 510.sup.4, for incubation for 12 hours in total before immunostaining fixed and adherent plagues and then observing with LSCM.

[0089] (4) Upon sterilizing with Cobalt-60, cut the film material obtained in Step (8) into a circular diaphragm with a diameter of 8 mm, for covering the rat skull defect with a diameter of 5 mm, while the defect not covered with any material is taken as the reference set. 4 weeks after the surgery, kill the animal and separate the film material for micro-CT scanning and observation.

Example 5

[0090] (1) Take 9 ml of glacial acetic acid and 1.340 g of acetylacetone, mix and stir for 5 min before adding 1.703 g of barium acetate, stirring for 1 h, adding 2.266 g of tetrabutyl titanate, stirring for 15 min, then adding 0.45 g of polyvinylpyrrolidone (PVP), stirring for 3 h;

[0091] (2) Inject into the injector the mixed solvent obtained in Step (1), and obtain barium titanate nanofiber with the diameter of 500 nm, with the static electrospinning technology under a voltage of 10 kV;

[0092] (3) Add the barium titanate nanofiber obtained in Step (2) into 0.1 mol/L of dopamine aqueous solution for bathing in water of 60 degrees, heating and stirring for 12 h, then apply ultrasonic concussion for 15 min, centrifugal drying to obtain barium titanate nanofiber fillings with a draw ratio of 8;

[0093] (4) Take an appropriate amount of barium titanate nanofiber fillings obtained in Step (3) for ultrasonic oscillation dispersion in 3 mL of organic solvent DMF, apply ultrasonic oscillation and stir for 1.5 h to obtain the fiber filling, dispersion liquid;

[0094] (5) Weigh and take 1 g of P (VDF-TrFE), add in 7 mL of organic solvent DMF, and stir for 5 h till completed solved to obtain P (VDF-TrFE) solution;

[0095] (6) Add the dispersion liquid obtained in Step (4) in P(VDF-TrFE) solution obtained in Step (5) so that the volume fraction of barium titanate fiber fillings in polymer is 15 vol %, stir for 10 h so that barium titanate fiber fillings are evenly dispersed in P (VDF-TrFE) matrix to obtain the mixed liquid;

[0096] (7) Take the mixed liquid obtained in Step (6) for casting in the casting machine, place the cast film obtained under a temperature of 100 C. for drying so that the solvent is completely evaporated to obtain a kind of composite film material with a thickness of 300 m;

[0097] (8) Polarize the film material obtained in Step (7), as per parameters: 30 kV (voltage), 50 mm (distance), 150 C. (temperature) and 10 min (time), to obtain a kind of electrified composite membrane with extracellular matrix electrical topology characteristics Its main components are P (VDF-TrFE) and barium titanate nanofiber, as the thin-film material with a thickness of 300 m. The volume fraction of barium titanate fiber fillings in electrified composite membrane is 15 vol %.

[0098] Product Performance Testing:

[0099] (1) With COMSOL software, simulate and analyze the potential distribution features of the film material surface.

[0100] (2) Place the film material obtained in Step (8) in serum-free medium, for incubation for 0, 1, 3, 7, 14 and 21 days respectively under the condition of 37 C. At each time point, take out the material for piezoelectric constant testing.

[0101] (3) Sterilize the film material obtained in (8) with Cobalt-60, then vaccinate, on its top, RMSC with a density of 510.sup.4, for incubation for 12 hours in total before immunostaining fixed and adherent plagues and then observing with LSCM.

[0102] (4) Upon sterilizing with Cobalt-60, cut the film material obtained in Step (8) into a circular diaphragm with a diameter of 8 mm, for covering the rat skull defect with a diameter of 5 mm, while the defect not covered with any material is taken as the reference set. 4 weeks after the surgery, kill the animal and separate the film material for micro-CT scanning and observation. For the product performance test results of Examples 25, refer to Example 1.