Polymerizable emulsions for tissue engineering

• US 7,651,683 B2
• Filed: 05/01/2007
• Issued: 01/26/2010
• Est. Priority Date: 10/29/2003
• Status: Expired due to Fees

• Current Assignee: Gentis, Inc.
• Original Assignee: Gentis, Inc.
• Inventors: Cohen, Charles, Devore, David, Ducheyne, Paul
• Primary Examiner(s): Foley; Shanon A
• Application Number:
  US11/799,347
• Publication Number:
  US 20070218124A1
• Time in Patent Office: 1,001 Days
• Field of Search: None
• US Class Current: 424/78.08
• CPC Class Codes: A61K 31/00 Medicinal preparations containing organic active ingredients
• CPC Class Codes: A61K 31/74 Synthetic polymeric materials
• CPC Class Codes: A61L 2300/256 Antibodies, e.g. immunoglobulins, vaccines
• CPC Class Codes: A61L 2300/414 Growth factors
• CPC Class Codes: A61L 2300/43 Hormones, e.g. dexamethasone
• CPC Class Codes: A61L 2300/64 Animal cells
• CPC Class Codes: A61L 27/26 Mixtures of macromolecular compounds
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61L 27/50 Materials characterised by their function or physical properties , e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
• CPC Class Codes: A61L 27/52 Hydrogels or hydrocolloids
• CPC Class Codes: A61L 27/54 Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: C08F 290/048 Polymers of monomers having two or more carbon-to-carbon double bonds as defined in group C08F36/00

Abstract

Provided are biocompatible viscoelastic solid materials derived from polymerization of fluid water-in-oil emulsions, along with methods of their preparation and methods for their use for tissue engineering applications, including for reforming diseased, damaged or degenerated intervertebral discs by acceptably non-invasive means.

Claims

1. A method of preparing a nontoxic, viscoelastic material for tissue engineering, wherein the viscoelastic material comprises a polymerized emulsion having two discrete organic and aqueous phases, the method comprising:

a) preparing a nontoxic, inverse (“

water-in-oil”

) fluid emulsion comprising two discrete phases;

i) a continuous hydrophobic organic (“

oil”

) phase, comprising one or more reactive, ethylenically unsaturated, polymerizable acrylate monomers, and at least one activator- controlled polymerization initiator; and

ii) a discontinuous aqueous (“

water”

) phase comprising water,

wherein the aqueous phase is present in the emulsion in an amount of about 0.5% to about 90% weight/weight of the organic phase; and

b) polymerizing the fluid emulsion in situ to form the viscoelastic material, wherein the viscoelastic material comprises;

i) a continuous hydrophobic organic phase comprised of polymerized ethylenically unsaturated acrylate monomers;

ii) a discontinuous aqueous (“

water”

) phase comprising water.

2. The method of claim 1, wherein at least one of the reactive, ethylenically unsaturated, polymerizable acrylate monomers, is selected from the group consisting of methyl acrylate;

methyl methacrylate;

ethyl acrylate;

ethyl methacrylate;

stearyl acrylate and methacrylate;

lauryl acrylate and methacrylate;

ethoxylated and/or propoxylated acrylates and methacrylates;

alkoxylated acrylates and methacrylates;

allyl acrylates and methacrylates;

tetrahydrofurfuryl methacrylate;

isodecyl acrylate or methacrylate;

octyldecyl acrylate and methacrylate;

phenoxyethyl acrylate and methacrylate;

glycidyl acrylate and methacrylate;

isobornyl acrylate;

caprolactone acrylate;

ethoxylated nonyl phenol acrylates and methacrylates;

propylene glycol acrylate and methacrylate;

alkoxylated nonyl phenol acrylates and methacrylates;

alkoxylated diol diacrylates;

alkoxylated bisphenol diacrylates;

ethylene glycol diacrylate;

butylene glycoldiacrylate and dimethacrylate;

butanediol diacrylate and dimethacrylate;

neopentylglycol diacrylate and dimethacrylate;

polyethyleneglycol diacrylate and dimethacrylate;

ethoxylated bisphenol diacrylates;

saturated and unsaturated (C36) dimer diol diacrylates;

polyethyleneglycol diacrylate and dimethacrylate;

polybutadiene diacrylate and dimethacrylate;

polybutadiene urethane diacrylate;

triethylene glycol dimethacrylate;

acrylamide and methacrylamide.

3. The method of claim 1, further comprising pre-polymerizing at least one of the reactive, ethylenically unsaturated, polymerizable acrylate monomers.

4. The method of claim 3, wherein the pre-polymerized monomer is lauryl acrylate, and it replaces a fraction of the monomer in the organic phase of the emulsion.

5. The method of claim 1, wherein the aqueous phase of the fluid emulsion further comprises at least one additive selected from the group consisting of stabilizers, stabilizing salts, diluents, solubilizers, lubricants, suspending agents, encapsulating materials, solvents, thickeners, dispersants, buffers, anti-oxidants, preservatives, low molecular weight peptides, proteins immunoglobulins, hydrophilic polymers, amino acids, monosaccharides, disaccharides, carbohydrates, cellulose or its derivatives, glucose, mannose, dextrines, chelating agents, sugar alcohols, counter-ions, and non-ionic surfactants.

6. The method of claim 1, wherein the polymerization of the fluid emulsion is initiated by thermal, chemical or photoinitiation means.

7. The method of claim 1, wherein the polymerization initiator is a photoinitiator.

8. The method of claim 6, wherein the photoinitiation means comprises exposing the fluid emulsion in situ to electromagnetic radiation.

9. The method of claim 1, wherein the polymerization initiator is a chemical initiator.

10. The method of claim 1, wherein the fluid emulsion further comprises particles, solutions or dispersions of at least one inert, bioactive or biological material.

11. The method of claim 7, wherein the photoinitiator is 2-hydroxy-1-[4-(2-hydroxyethoxyphenyl]-2-methyl-1-propane or bisacyl phosphine oxide.

12. The method of claim 8, wherein the electromagnetic radiation is ultraviolet light, visible light, infrared light, or microwaves.

13. The method of claim 9, wherein the chemical initiator is a peroxide or a redox catalyst.

14. The method of claim 9, wherein the chemical initiator is a combination of benzoyl peroxide and a tertiary amine.

15. The method of claim 10, wherein the inert, bioactive or biological material is selected from the group consisting of biological cells or cell fragments, tissue fragments derived from in vitro or in vivo cell culture, biological cell or tissue growth factors, agents to facilitate cell delivery or cell growth and their delivery, antibodies, antibody fragments, hormones, cell targeting moieties, inert ceramics, inert glasses, bioactive ceramics and bioactive glasses.

16. The method of claim 1, wherein the aqueous phase of the fluid emulsion comprises droplets having a size of ≧

1 micron.

17. The method of claim 1, wherein the fluid emulsion further comprises an emulsifier.

18. The method of claim 17, wherein the emulsifier is present in an amount of about 0.1% to about 5.0% by weight of the emulsion.

19. The method of claim 1, wherein the aqueous phase of the fluid emulsion further comprises at least one polymer selected from the group consisting of polyclectrolytes, polysaccharides, proteins, proteoglycans, nucleic acids, hydroxyethylcellulose, and carboxymethylcellulose.

20. The method of claim 1, wherein the fluid emulsion further comprises at least one filler selected from the group consisting of radio-opacifiers, polymeric microspheres, glass, sugar crystals, and proteins.

21. The method of claim 1, wherein the fluid emulsion further comprises at least one medicament.

22. The method of claim 21, wherein the medicament is an anesthetic.

23. The method of claim 22, wherein the anesthetic is selected from the group consisting of lidocaine, xylocaine, novocaine, benzocaine, prilocaine, ripvacaine, and propofol.

24. The method of claim 21, wherein the medicament is selected from the group consisting of antibiotics, steroids, fibronectins, cytokines, growth factors, analgesics, antiseptics, glucagons, polynucleotides, proteins, peptides, cell attachment mediators, and osteoinductive substances.

25. The method of claim 24, wherein the polynucleotide is cDNA or DNA.

26. The method of claim 21, wherein the medicament is selected from the group consisting of alpha-, beta, or gamma-interferon, erythropoietin, calcitonin, heparin, interleukin-1, interleukin-2, filgrastim, HGH, luteinizing hormone, atrial natriuretic factor, Factor VIII, Factor IX, follicle-stimulating hormone, acyclovir, cephradine, malfalen, procaine, ephedrine, adriomycin, daunomycin, plumbagin, atropine, quanine, digoxin, quinidine, cephalothin, proline, cis-hydroxy-L-proline, penicillin V, aspirin, ibuprofen, nicotinic acid, chemodeoxycholic acid, chlorambucil, bone morphogenic proteins, epidermal growth factor, fibroblast growth factor, platelet-derived growth factor, insulin-like growth factor, and transforming growth factor-beta.

27. The emulsion of claim 21, further comprising at least one carrier selected from the group consisting of saline, starch, hydrogel, polyvinylpyrrolidone, polysaccharide, hyaluronic acid ester, and plasma.

32. The method of claim 1, wherein the fluid emulsion further comprises inert glass.

28. A method for preparing a nontoxic, viscoelastic material for tissue engineering, wherein the viscoelastic material comprises a polymerized emulsion having two discrete organic and aqueous phases, the method comprising:

a) preparing a nontoxic, inverse (“

water-in-oil”

) fluid emulsion comprising two discrete phases;

i) a continuous hydrophobic organic (“

oil”

) phase, comprising one or more reactive, ethylenically unsaturated, polymerizable acrylate monomers, and a peroxide free radical generator; and

ii) a discontinuous aqueous (“

water”

) phase, wherein the aqueous phase is present in an amount of about 0.5% to about 90% weight/weight of the organic phase;

b) preparing a nontoxic, inverse (“

water-in-oil”

) fluid emulsion comprising two discrete phases;

i) a continuous hydrophobic organic (“

oil”

) phase, comprising one or more reactive, ethylenically unsaturated, polymerizable acrylate monomers, and an amine catalyst; and

ii) a discontinuous aqueous (“

water”

) phase, wherein the aqueous phase is present in an amount of about 0.5% to about 90% weight/weight of the organic phase; and

c) combining the emulsions of steps a) and b) to initiate polymerization to form the viscoelastic material comprising of;

i) a continuous hydrophobic organic phase comprised of polymerized ethylenically unsaturated acrylate monomers; and

ii) a discontinuous aqueous (“

water”

) phase comprising water.

29. The method of claim 28, wherein the peroxide free radical generator is benzoyl peroxide.

30. The method of claim 28, wherein the amine catalyst is a tertiary amine catalyst.

31. The method of claim 30, wherein the tertiary amine catalyst is N,N-diethyl-p-toluidine.