Electroactive Scaffold

• US 20110142806A1
• Filed: 12/15/2010
• Published: 06/16/2011
• 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: The United States of America As Represented By The Secretary of Agriculture
• Inventors: Leong, Kam W., Siochi, Emilie J., Holloway, Nancy M., Kulangara, Karina, Scott-Carnell, Lisa A.
• Granted Patent: US 9,005,604 B2
• Time in Patent Office: Days
• 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
• 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 for tissue engineering and/or stem cell differentiation using electrical and/or mechanical stimuli through bioactive fibers comprising at least one layer of substantially aligned, electrospun polymer fiber having an operative connection for individual voltage application.

2. The assembly of claim 1, further comprising at least one pre-electroded substrate.

3. The assembly of claim 2, wherein the layer of polymer fiber is secured to the substrate with an adhesive compound.

4. The assembly of claim 1, wherein the assembly comprises at least two or more layers of polymer fibers.

5. The assembly of claim 4, wherein the layers are semi-woven and/or interwoven.

6. The assembly of claim 4, wherein the layers are independently connected to individual voltage sources.

7. The assembly of claim 4, wherein the layers are independently connected to individual mechanical force generators.

8. The assembly of claim 4, wherein the layers are dependently connected to a voltage source.

9. The assembly of claim 4, wherein the layers are dependently connected to a mechanical force generator.

10. The assembly of claim 1, wherein the polymer fiber comprises an electroactive polymer.

11. The assembly of claim 10, wherein the electroactive polymer comprises polyvinylidene fluoride (PVDF).

12. The assembly of claim 1, further comprising a clamp to hold the assembly together.

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

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

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

16. A method of manufacturing an assembly for tissue engineering and/or stem cell differentiation comprising:

(a) providing a first pre-electroded substrate surface;

(b) electrospinning a first substantially aligned polymer fiber layer onto the first surface;

(c) providing a second pre-electroded substrate surface;

(d) electrospinning a second substantially aligned polymer fiber layer onto the second surface; and

,

(e) retaining together the layered surfaces with a clamp and/or an adhesive compound.

17. The method of claim 16, wherein the pre-electroded substrate surface is prepared by fabricating a substrate from a polymer sheet and evaporating metal onto the substrate.

18. The method of claim 17, wherein the polymer sheet comprises polyimide and the metal comprises gold.

19. The method of claim 16, wherein the electrospinning step(s) comprise using an auxiliary electrode to create a dipole field.

20. The method of claim 19, wherein the polymer fiber comprises an electroactive polymer.

21. The method of claim 20, wherein the electroactive polymer comprises polyvinylidene fluoride (PVDF).

22. A method of cell tissue engineering and/or stem cell differentiation, said method comprising:

(a) providing an assembly having at least two layers of substantially aligned, electrospun polymer fiber having an operative connection for individual voltage application to each layer;

(b) seeding the assembly with a sample of cells suspended in cell culture media;

(c) incubating for an effective time period;

(d) applying an effective voltage to one or more layers; and

(e) recovering cells and/or a tissue construct.

23. The method of claim 22, wherein the polymer fiber comprises an electroactive polymer.

24. The method of claim 23, wherein the electroactive polymer comprises polyvinylidene fluoride (PVDF).

25. The method of claim 22, further comprising the step of echanically stimulating one or more layers.

26. The method of claim 22, further comprising the step of using the recovered cells and/or the tissue construct to treat a human disability or disease condition.