Aligned and electrospun piezoelectric polymer fiber assembly and scaffold

• US 9,005,604 B2
• Filed: 12/15/2010
• Issued: 04/14/2015
• Est. Priority Date: 12/15/2009
• Status: Active Grant

• Current Assignee: United States Of America As Represented By The Administrator Of The National Aeronautics And Space Administration
• Original Assignee: United States Of America As Represented By The Administrator Of The National Aeronautics And Space Administration
• Inventors: Scott-Carnell, Lisa A., Siochi, Emilie J., Holloway, Nancy M., Leong, Kam W., Kulangara, Karina
• Primary Examiner(s): Lankford, Blaine
• Assistant Examiner(s): BERKE-SCHLESSEL, DAVID W
• Application Number:
  US12/969,076
• Publication Number:
  US 20110142806A1
• Time in Patent Office: 1,581 Days
• Field of Search: None
• US Class Current: 424/93.7
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61L 27/16 obtained by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: A61L 27/3834 Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: C08L 27/16 Homopolymers or copolymers or vinylidene fluoride
• CPC Class Codes: C12N 11/082 obtained by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• CPC Class Codes: C12N 2529/00 Culture process characterised by the use of electromagnetic stimulation
• CPC Class Codes: C12N 2533/30 Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06

Abstract

A scaffold assembly and related methods of manufacturing and/or using the scaffold for stem cell culture and tissue engineering applications are disclosed which at least partially mimic a native biological environment by providing biochemical, topographical, mechanical and electrical cues by using an electroactive material. The assembly includes at least one layer of substantially aligned, electrospun polymer fiber having an operative connection for individual voltage application. A method of cell tissue engineering and/or stem cell differentiation uses the assembly seeded with a sample of cells suspended in cell culture media, incubates and applies voltage to one or more layers, and thus produces cells and/or a tissue construct. In another aspect, the invention provides a method of manufacturing the assembly including the steps of providing a first pre-electroded substrate surface; electrospinning a first substantially aligned polymer fiber layer onto the first surface; providing a second pre-electroded substrate surface; electrospinning a second substantially aligned polymer fiber layer onto the second surface; and, retaining together the layered surfaces with a clamp and/or an adhesive compound.

Claims

1. An assembly configured to utilize stimuli through bioactive fibers comprising:

a first substrate film;

a first electrode and a second electrode disposed on said first substrate film; and

a layer that comprises a plurality of aligned, electrospun-piezoelectric polymer fibers,

such that the layer of aligned, electrospun piezoelectric polymer fibers forms;

a first outer periphery region configured to be connected to said first electrode;

a second outer periphery region configured to be connected to the second electrode; and

an inner region that is suspended over a bottom surface between the first electrode and second electrode.

2. The assembly of claim 1, wherein the layer is a first layer and the assembly comprises a second layer having a plurality of aligned, electrospun piezoelectric polymer fibers,

such that the second layer forms:

a first outer periphery region configured to be connected to a third electrode on a second substrate film;

a second outer periphery region configured to be connected to a fourth electrode on the second substrate film; and

an inner region that is suspended over a bottom surface located between the third electrode and the fourth electrode; and

the said second substrate film is layered on the first substrate film in a configuration that permits electrical stimulation of the first layer independent of the second layer.

3. The assembly of claim 2, wherein the first layer and the second layer are independently connected to individual voltage sources.

4. The assembly of claim 1, wherein the plurality of aligned, electrospun piezoelectric polymer fibers of the layer comprises polyvinylidene fluoride (PVDF).

5. The assembly of claim 1, further comprising human mesenchymal stem cells and effective cell culture media.

6. The assembly of claim 1, further comprising induced pluripotent stem cells and effective cell culture media.

7. The assembly of claim 1, further comprising embryonic stem cells and effective cell culture media.

8. The assembly of claim 1, wherein the plurality of aligned, electrospun piezoelectric polymer fibers of layer are substantially parallel to one another.

9. The assembly of claim 2, wherein the plurality of aligned, electrospun piezoelectric polymer fibers of the first layer are substantially parallel to one another, and wherein the plurality of aligned, electrospun piezoelectric polymer fibers of the second layer are substantially parallel to one another.

10. The assembly of claim 1, wherein an electrical stimuli is applied to the comprising the plurality of aligned, electrospun piezoelectric polymer fibers.

11. The assembly of claim 1, wherein said first substrate film is a pre-electroded substrate film, and wherein said first and second electrodes are configured to be deposited onto said first substrate film prior to attachment of said layer comprising the aligned, electrospun piezoelectric polymer fibers to said pre-electroded substrate film.

12. The assembly of claim 1, wherein said fibers are connected to said first and second electrodes with a silicon adhesive.

13. The assembly of claim 1, wherein said first and second electrodes are comprised of gold.

14. The assembly of claim 1, wherein said layer is porous.

15. The assembly of claim 14, wherein the second layer is porous.

16. A three dimensional scaffolding system configured to transmit stimuli through fibers comprising:

a first layer of aligned electrospun piezoelectric polymer fibers provided on a first substrate film;

a second layer of aligned electrospun piezoelectric polymer fibers provided on a second substrate film; and

an electrical stimulation system configured to selectively apply electrical stimulation to the first layer of fibers independently of the second layer of fibers, comprising;

a first electrode and a second electrode disposed on the first substrate film configured to independently apply electrical stimulation to the first layer by enabling the conversion of the applied electrical stimulation to a mechanical strain on the fibers of the first layer on a molecular level without eliciting a mechanical response at a macroscale level; and

a third electrode and a fourth electrode disposed on the second substrate film configured to independently apply electrical stimulation to the second layer by enabling the conversion of the applied electrical stimulation to a mechanical strain on the fibers of the second layer on a molecular level without eliciting a mechanical response at a macroscale level.

17. The three dimensional scaffolding system of claim 16, wherein the electrical stimulation system is configured to provide electrical stimulation at about 9 mV/cm.

18. The three dimensional scaffolding system of claim 16, wherein the electrical stimulation system is configured to provide electrical stimulation below 10 mV/cm.

19. An assembly configured to utilize stimuli through bioactive fibers comprising:

a first substrate film;

a first electrode and a second electrode disposed on said first substrate film; and

a non-woven layer of aligned, electrospun piezoelectric polymer fibers having;

a first outer periphery region configured to be connected to said first electrode;

a second outer periphery region configured to be connected to the second electrode; and

an inner region that is suspended over a bottom surface between the first electrode and second electrode.