Films for controlled cell growth and adhesion

• US 20050287111A1
• Filed: 05/17/2005
• Published: 12/29/2005
• Est. Priority Date: 05/17/2004
• Status: Active Grant

• Current Assignee: Florida State University Research Foundation, Inc. (State University System of Florida)
• Original Assignee: Florida State University Research Foundation, Inc. (State University System of Florida)
• Inventors: Olenych, Scott G., Keller, Thomas C. III, Salloum, David S., Schlenoff, Joseph B.
• Granted Patent: US 9,056,125 B2
• Time in Patent Office: Days
• US Class Current: 424/78.3
• CPC Class Codes: A61K 31/16 Amides, e.g. hydroxamic acids
• CPC Class Codes: A61K 31/785 Polymers containing nitrogen
• CPC Class Codes: A61L 2300/424 Anti-adhesion agents
• CPC Class Codes: A61L 2400/18 Modification of implant surfaces in order to improve biocompatibility, cell growth, fixation of biomolecules, e.g. plasma treatment
• CPC Class Codes: A61L 2420/06 Coatings containing a mixture of two or more compounds
• CPC Class Codes: A61L 2430/00 Materials or treatment for tissue regeneration
• CPC Class Codes: A61L 2430/12 for dental implants or prostheses
• CPC Class Codes: A61L 2430/16 for reconstruction of eye parts, e.g. intraocular lens, cornea
• CPC Class Codes: A61L 27/26 Mixtures of macromolecular compounds
• CPC Class Codes: A61L 27/34 Macromolecular materials
• CPC Class Codes: A61L 27/54 Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: A61L 29/085 Macromolecular materials
• CPC Class Codes: A61L 29/16 Biologically active materials, e.g. therapeutic substances A61L29/048 takes precedence
• CPC Class Codes: A61L 31/10 Macromolecular materials
• CPC Class Codes: A61L 31/16 Biologically active materials, e.g. therapeutic substances A61L31/047 takes precedence
• CPC Class Codes: C12N 2537/00 Supports and/or coatings for cell culture characterised by physical or chemical treatment
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06

Abstract

An article for controlling the attachment and growth of cells on a surface of the article, and a method for the use of the article is provided. The article comprises a substratum having a surface and a film on the surface, the film comprising a network of a net positively charged composition and a net negatively charged composition, wherein the net positively charged composition comprises a net positively charged polyelectrolyte or the net negatively charged composition comprises a net negatively charged polyelectrolyte, and the net positively charged polyelectrolyte or the net negatively charged polyelectrolyte contain (i) a polymer repeat unit having at least two fluorine atoms, or (ii) a polymer repeat unit having a zwitterion group. The method comprises contacting the article with living tissue, living organisms, or with water in an aqueous system comprising living organisms.

Claims

1. An article adapted for use in combination with living tissue or in a marine environment, the article comprising:

a substratum having a surface;

a polyelectrolyte film on the surface, the polyelectrolyte film comprising a network of a net positively charged composition and a net negatively charged composition, wherein the net positively charged composition comprises a net positively charged polyelectrolyte or the net negatively charged composition comprises a net negatively charged polyelectrolyte, and the net positively charged polyelectrolyte or the net negatively charged polyelectrolyte contain (i) a polymer repeat unit having at least two fluorine atoms, or (ii) a polymer repeat unit having a zwitterion group; and

the substratum having a composition and shape adapting the article for use in combination with living tissue or in a marine environment.

2. The article of claim 1 wherein the polyelectrolyte film comprises a combination selected from the group consisting of the following combinations:

(1) a net positively charged polyelectrolyte containing a polymer repeat unit having at least two fluorine atoms and a net negatively charged polyelectrolyte containing a polymer repeat unit having at least two fluorine atoms;

(2) a net positively charged polyelectrolyte containing a polymer repeat unit having at least two fluorine atoms and a net negatively charged polyelectrolyte containing polymer repeat units having (i) fewer than two fluorine atoms and (ii) no zwitterion groups;

(3) a net positively charged polyelectrolyte containing a polymer repeat unit having at least two fluorine atoms and a net negatively charged polyelectrolyte containing a polymer repeat unit having a zwitterion group;

(4) a net positively charged polyelectrolyte containing polymer repeat units having (i) fewer than two fluorine atoms and (ii) no zwitterion groups and a net negatively charged polyelectrolyte containing a polymer repeat unit having at least two fluorine atoms;

(5) a net positively charged polyelectrolyte containing a polymer repeat unit having a zwitterion group and a net negatively charged polyelectrolyte containing a polymer repeat unit having at least two fluorine atoms;

(6) a net positively charged polyelectrolyte containing polymer repeat units having (i) fewer than two fluorine atoms and (ii) no zwitterion groups and a net negatively charged polyelectrolyte containing a polymer repeat unit having a zwitterion group;

(7) a net positively charged polyelectrolyte containing a polymer repeat unit having a zwitterion group and a net negatively charged polyelectrolyte containing polymer repeat units having (i) fewer than two fluorine atoms and (ii) no zwitterion groups; and

(8) a net positively charged polyelectrolyte containing a polymer repeat unit having a zwitterion group and a net negatively charged polyelectrolyte containing a polymer repeat unit having a zwitterion group.

3. The article of claim 1 wherein the polyelectrolyte film comprises a combination selected from the group consisting of the following combinations:

(1) a net positively charged particle and a net negatively charged polyelectrolyte containing a polymer repeat unit having at least two fluorine atoms, (2) a net positively charged polyelectrolyte containing a polymer repeat unit having at least two fluorine atoms and a net negatively charged particle, (3) a net positively charged fluorinated particle and a net negatively charged polyelectrolyte and (4) a net positively charged polyelectrolyte and a net negatively charged fluorinated particle.

4. The article of claim 1 wherein the substratum is selected from the group consisting of stents, catheters, vascular grafts, vascular prostheses, contact lenses, intraocular implants, artificial valves for in vivo use, artificial organs, dental implants, metal implants into bone, corneal implants, Petri dishes, roller bottles, microcarriers, porous structural supports for three dimensional cell growth, synthetic elastomeric polymers, and metal objects adapted for use in aqueous systems containing living organisms.

5. The article of claim 1 wherein the polyelectrolyte film further comprises agents selected from the group consisting of growth factors, cell nutrients, and combinations thereof.

6. The article of claim 1 wherein the polyelectrolyte film further comprises agents selected from the group consisting of paclitaxel, sirolimus, and a combination thereof.

7. The article of claim 1 wherein the polymer repeat unit having at least two fluorine atoms has a structure:

wherein

A₁, A₂, and A₃ are each independently —

(CH₂)_nH or —

(CH_xF_2-x)_nF;

m and n are independently 0 to 12, x is 0, 1, or 2; and

V is independently selected from the group consisting of;

(i) fluorinated hydrocarbons having the structure;

—

(CH₂)_p(CF₂)_qF;

—

(CH₂)_p(CF₂)_qCOOH;

—

(CH₂)_p(CF₂)_qOPO₃⁻

;

—

(CH₂)_p(CF₂)_qSO₃⁻

;

—

(CH₂)_p(CF₂)_qOSO₃⁻

;

—

O(CH₂)_p—

(CF₂)_q—

F;

or —

O(CH₂)_p(CF₂)_q—

SO₃⁻

;

(ii) fluorinated amides having the structure;

wherein B₁ is —

(CH₂)_p(CF₂)_qF;

—

(CH₂)_p(CF₂)_qCOOH;

—

(CH₂)_p(CF₂)_qOPO₃⁻

;

—

(CH₂)_p(CF₂)_qSO₃⁻

;

or —

(CH₂)_p(CF₂)_qOSO₃⁻

;

(iii) fluorinated esters having the structure;

wherein C₁ is —

(CH₂)_p(CF₂)_qF;

—

(CH₂)_p(CF₂)_qCOOH;

—

(CH₂)_p(CF₂)_qOPO₃⁻

;

—

(CH₂)_p(CF₂)_qSO₃—

;

or —

(CH₂)_p(CF₂)_qOSO₃⁻

;

(iv) fluorinated phenyl groups having the structure;

wherein D1 is 2 to 5;

or

wherein D₂ is —

(CH₂)_p(CF₂)_qF or —

O(CH₂)_p(CF₂)_qF;

(v) fluorinated pyridiniums having the structure;

wherein E₁ is —

(CH₂)_p(CF₂)_qF;

(vi) fluorinated imidazoliums having the structure;

wherein F₁ is —

(CH₂)_p(CF₂)_qF;

(vii) fluorinated quaternary nitrogens having the structure;

wherein G₁, G₂, and G₃ are each independently —

(CH₂)_p(CF₂)_qF or -arylF_z wherein z is 2 to 8;

(viii) fluorinated sulfoniums having the structure;

wherein H₁ and H₂ are each independently —

(CH₂)_p(CF₂)_qF or -arylF_z wherein z is 2 to 8; and

(ix) fluorinated phosphoniums having the structure;

wherein J₁, J₂, and J₃ are each independently;

—

(CH₂)_p(CF₂)_qF or -arylF_z wherein z is 2 to 8; and

(x) combinations of (i) through (ix); and

in each of (i) through (ix);

p is 0 to 6 and q is 1 to 21.

8. The article of claim 1 wherein the polymer repeat unit having at least two fluorine atoms comprises an allyl group having a structure:

wherein L₁ and L₂ are —

(CH₂)_p(CF₂)_qF, p and q are independently selected for L₁ and L₂, and p is 0 to 6 and q is 1 to 21.

9. The article of claim 1 wherein the net positively charged composition comprises a polymer repeat unit having at least two fluorine atoms having the structure:

wherein M is a mole fraction from about 0.1 to about 1.0.

10. The article of claim 1 wherein the net negatively charged composition comprises a polymer repeat unit having at least two fluorine atoms having the structure:

wherein X is from about 6 parts to about 10 parts, Y is about 1 part, and Z is from about 1 parts to about 3 parts.

11. The article of claim 1 wherein the net negatively charged composition comprises a polynucleic acid selected from the group consisting of ribonucleic acid and deoxyribonucleic acid.

12. The article of claim 1 wherein the polymer repeat unit having a zwitterion group is selected from the group consisting of:

N,N-dimethyl-N-acryloyloxyethyl-N-(3-sulfopropyl)-ammonium betaine, N,N-dimethyl-N-acrylamidopropyl-N-(2-carboxymethyl)-ammonium betaine, N,N-dimethyl-N-acrylamidopropyl-N-(3-sulfopropyl)-ammonium betaine, N,N-dimethyl-N-acrylamidopropyl-N-(2-carboxymethyl)-ammonium betaine, 2-(methylthio)ethyl methacryloyl-S-(sulfopropyl)-sulfonium betaine, 2-[(2-acryloylethyl)dimethylammonio]ethyl 2-methyl phosphate, 2-(acryloyloxyethyl)-2′

-(trimethylammonium)ethyl phosphate, [(2-acryloylethyl)dimethylammonio]methyl phosphonic acid, 2-methacryloyloxyethyl phosphorylcholine (MPC), 2-[(3-acrylamidopropyl)dimethylammonio]ethyl 2′

-isopropyl phosphate, 1-vinyl-3-(3-sulfopropyl)imidazolium hydroxide, (2-acryloxyethyl) carboxymethyl methylsulfonium chloride, 1-(3-sulfopropyl)-2-vinylpyridinium betaine, N-(4-sulfobutyl)-N-methyl-N,N-diallylamine ammonium betaine, N,N-diallyl-N-methyl-N-(2-sulfoethyl) ammonium betaine, N,N-dimethyl-N-(3-methacrylamidopropyl)-N-(3-sulfopropyl) ammonium betaine, N,N-dimethyl-N-acryloyloxyethyl-N-(3-sulfopropyl)-ammonium betaine, N,N-dimethyl-N-acrylamidopropyl-N-(2-carboxymethyl)-ammonium betaine, N,N-dimethyl-N-methacryloyloxyethyl-N-(3-sulfopropyl)-ammonium betaine, N,N-dimethyl-N-(3-methacrylamidopropyl)-N-(3-sulfopropyl) ammonium betaine, and copolymers thereof.

13. The article of claim 1 wherein the net positively charged composition comprises a polyallylamine repeat unit and the net negatively charged composition a copolymer of acrylic acid and at least about 15 mole % 3-[2-(acrylamido)-ethyldimethyl ammonio] propane sulfonate.

14. An article adapted for use in combination with living tissue or organisms, the article comprising a polyelectrolyte film, the film comprising an interpenetrating network of a net positively charged polyelectrolyte and a net negatively charged polyelectrolyte, the film having first and second surface regions with a net positively charged or net negatively charged polyelectrolyte exposed at each of said first and second surface regions wherein (1) the net positively charged or negatively charged polyelectrolyte exposed in said first surface region contains a polymer repeat unit having at least two fluorine atoms and (2) the net positively charged or negatively charged polyelectrolyte exposed in said second surface region contains a polymer repeat unit having a zwitterion group.

15. The article of claim 14 wherein the article comprises the polyelectrolyte film and a metal substratum with the first surface region contacting the metal substratum.

16. The article of claim 14 wherein the article is a membrane having opposing sides, the first surface region being one of the opposing sides and the second surface region being the other opposing side.

17. The article of claim 16 further comprising a synthetic elastomeric polymer selected from the group consisting of polyvinyl chloride, polyurethane, polysiloxanes, and combinations thereof.

18. The article of claim 14 wherein the first surface region and the second surface region have substantially planar exposed surfaces wherein the first and second surface regions are substantially contiguous so as to define a pattern of regions having water contact angles that differ by at least 30 degrees.

19. The article of claim 18 wherein the first surface region has a dimensional aspect between 1 micrometer and 1000 micrometers.

20. The article of claim 18 wherein the first surface region is stamped onto the second surface region.

21. A method for controlling the attachment and growth of cells on a surface of an article, the method comprising contacting the article with living tissue, living organisms, or with water in an aqueous system comprising living organisms wherein the article comprises a substratum having a surface and a film on the surface, the film comprising a network of a net positively charged composition and a net negatively charged composition, wherein the net positively charged composition comprises a net positively charged polyelectrolyte or the net negatively charged composition comprises a net negatively charged polyelectrolyte, and the net positively charged polyelectrolyte or the net negatively charged polyelectrolyte contain (i) a polymer repeat unit having at least two fluorine atoms, or (ii) a polymer repeat unit having a zwitterion group.

22. The method of claim 21 wherein the substratum is selected from the group consisting of stents, catheters, vascular grafts, vascular prostheses, contact lenses, intraocular implants, artificial valves for in vivo use, artificial organs, dental implants, metal implants into bone, corneal implants, Petri dishes, roller bottles, microcarriers, porous structural supports for three dimensional cell growth, synthetic elastomeric polymers, and metal objects adapted for use in aqueous systems containing living organisms.

23. The method of claim 21 wherein the film is deposited by alternately layering the net positively charged composition and the net negatively charged composition and wherein layers 0 through n of the film comprise polyelectrolyte comprising non-fluorinated polymer repeat units and layers n+1 through n+2 of the film comprise polyelectrolyte comprising the polymer repeat unit having at least two fluorine atoms.

24. The method of claim 23 wherein the film promotes the attachment and growth of cells on the surface of the substratum.

25. The method of claim 21 wherein the film comprises a first surface region and a second surface region with a net positively charged or net negatively charged polyelectrolyte exposed at each of said first surface region and second surface region, wherein (1) the net positively charged or net negatively charged polyelectrolyte exposed in said first surface region contains polyelectrolyte comprising non-fluorinated polymer repeat units and (2) the net positively charged or net negatively charged polyelectrolyte exposed in said second surface region contains polyelectrolyte comprising the polymer repeat unit having at least two fluorine atoms.

26. The method of claim 25 wherein the film promotes the attachment and growth of cells on the surface of the substratum.

27. The method of claim 21 wherein the film is deposited by alternately layering the net positively charged composition and the net negatively charged composition and wherein layers 0 through n of the film comprise polyelectrolyte comprising non-zwitterion polymer repeat units and layers n+1 through n+2 of the film comprise polyelectrolyte comprising polymer repeat unit having a zwitterion group.

28. The method of claim 27 wherein the film inhibits the attachment and growth of cells on the surface of the substratum.

29. The method of claim 21 wherein the film comprises a first surface region and a second surface region with a net positively charged or net negatively charged polyelectrolyte exposed at each of said first surface region and second surface region, wherein (1) the net positively charged or net negatively charged polyelectrolyte exposed in said first surface region contains polyelectrolyte comprising non-zwitterion polymer repeat units and (2) the net positively charged or net negatively charged polyelectrolyte exposed in said second surface region contains polyelectrolyte comprising the polymer repeat unit having a zwitterion group.

30. The method of claims 29 wherein the film inhibits the attachment and growth of cells on the surface of the substratum.

31. The method of claim 21 wherein the film comprises a first surface region and a second surface region with a net positively charged or net negatively charged polyelectrolyte exposed at each of said first surface region and second surface region, wherein (1) the net positively charged or net negatively charged polyelectrolyte exposed in said first surface region contains polyelectrolyte comprising polymer repeat units having at least two fluorine atoms and (2) the net positively charged or net negatively charged polyelectrolyte exposed in said second surface region contains polyelectrolyte comprising the polymer repeat unit having a zwitterion group.

32. The method of claim 31 wherein the first stratum of the film promotes the attachment and growth of cells and the second stratum inhibits the attachment and growth of cells.