Methods and compositions for tissue regeneration

• US 7,144,729 B2
• Filed: 12/19/2002
• Issued: 12/05/2006
• Est. Priority Date: 09/01/2000
• Status: Expired due to Fees

• Current Assignee: Smith & Nephew Incorporated (Smith & Nephew PLC)
• Original Assignee: DFB Pharmaceuticals, Inc.
• Inventors: Rinsch, Christopher, Hunziker, Thomas, Mis, Beatrice, Rolland, Eric
• Primary Examiner(s): Romeo, David S
• Assistant Examiner(s): Gamett, Daniel C
• Application Number:
  US10/324,257
• Publication Number:
  US 20030165482A1
• Time in Patent Office: 1,447 Days
• Field of Search: None
• US Class Current: 435/325
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 35/33 Fibroblasts
• CPC Class Codes: A61K 35/36 Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells islets of Langerhans A61K35/39
• CPC Class Codes: A61K 38/18 Growth factors; Growth regulators
• CPC Class Codes: A61K 38/19 Cytokines; Lymphokines; Interferons
• CPC Class Codes: A61K 38/363 Fibrinogen
• CPC Class Codes: A61K 38/4833 Thrombin (3.4.21.5)
• CPC Class Codes: A61K 41/0023 Agression treatment or altering
• CPC Class Codes: A61K 47/10 Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61K 9/0014 Skin, i.e. galenical aspects of topical compositions non-active ingredients are additionally classified in A61K47/00; A61K9/0009, A61K9/0021, A61K9/7015, A61K9/7023 take precedence; cosmetic preparations A61K8/00, A61Q; preparations for wound dressings or bandages A61L26/00
• CPC Class Codes: A61K 9/10 Dispersions; Emulsions A61K9/06 takes precedence; composition of dispersions, emulsions A61K47/00
• CPC Class Codes: A61K 9/19 lyophilised , i.e. freeze-dried, solutions or dispersions lyophilised products with subsequent particle size reduction A61K9/14; granules or pellets made by lyphilisation A61K9/1682; solid oral dosage forms made by lyophilisation A61K9/2095; lyophilisation additives A61K47/00
• CPC Class Codes: A61K 9/7015 Drug-containing film-forming compositions, e.g. spray-on
• CPC Class Codes: A61L 15/40 containing ingredients of undetermined constitution or reaction products thereof , e.g. plant or animal extracts
• CPC Class Codes: A61L 26/0057 Ingredients of undetermined constitution or reaction products thereof
• CPC Class Codes: A61L 27/3616 Blood, e.g. platelet-rich plasma
• CPC Class Codes: A61L 27/3633 Extracellular matrix [ECM]
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3813 Epithelial cells, e.g. keratinocytes, urothelial cells
• CPC Class Codes: A61L 27/3886 : comprising two or more cell types
• CPC Class Codes: A61P 17/02 : for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/04 : for non-specific disorders of the connective tissue
• CPC Class Codes: C12N 2502/094 : keratinocytes
• CPC Class Codes: C12N 2502/1323 : Adult fibroblasts
• CPC Class Codes: C12N 2510/00 : Genetically modified cells
• CPC Class Codes: C12N 2510/04 : Immortalised cells
• CPC Class Codes: C12N 5/0656 : Adult fibroblasts
• CPC Class Codes: C12N 5/0698 : Skin equivalents

Abstract

The present invention provides the use and composition of matter of angiogenic or other growth/cytokine factors expressed by mixtures of allogeneic human cell strains or lines of various types and stages of differentiation. Also provided are unencapsulated preparations (mixed with or applied to extracellular matrix material or synthetic biocompatible substances) for the purpose of temporary application to wounds or defects in the skin or other tissues for the restoration of blood supplying connective tissue to enable organ-specific cells to reestablish organ integrity as well as to inhibit excessive scar formation.

Claims

1. A tissue regeneration system comprising:

(a) a first component comprising fibrinogen, wherein the first component is in the form of a spray; and

(b) a second component comprising thrombin and mitotically inactivated allogeneic cells, wherein the second component is in the form of a spray and wherein the cells comprise one or more keratinocytes and fibroblasts that secrete one or more biologically active molecules selected from the group consisting of GM-CSF, VEGF, KGF, bFGF, TGFβ

, angiopoietin, EGF, IL-Iβ

, IL-6, IL-8, TGFα

, and TNFα

.

2. The tissue regeneration system of claim 1, wherein the one or more keratinocytes and fibroblasts are differentiated fibroblasts and keratinocytes.

3. The tissue regeneration system of claim 1, wherein the one or more keratinocytes and fibroblasts are mitotically inactivated by administration of mitomycin C or other chemically-based mitotic inhibitors, irradiation with γ

-Rays, irradiation with X-Rays, or irradiation with UV light.

4. The tissue regeneration system of claim 1, wherein the first component further comprises a collagen, alginate, alginate beads, agarose, fibrin, fibrin glue, blood plasma, fibrin beads, laminin, proteoglycan, fibronectin, chitosan, or heparin.

5. The tissue regeneration system of claim 1, wherein the second component further comprises a cryoprotectant.

6. The tissue regeneration system of claim 5, wherein the cryoprotectant is selected from the group consisting of 10% by volume of a glycerol solution, 15% by volume of a glycerol solution, and 15% glycerol and 5% human serum albumin by volume of a solution.

7. The tissue regeneration system of claim 5, wherein the cryoprotectant is glycerol.

8. A kit comprising, in one or more containers, the tissue regeneration system of claim 1.

9. The kit of claim 8, wherein the first component and the second component are cryopreserved prior to shipping and subsequently thawed prior to use.

10. The kit of claim 9, wherein the first and second components are each contained in a separate vial having a removable screw cap, wherein the vial is sterile and is made of a material resistant to low temperatures and wherein the removable lid can be replaced with a spray pump following thawing of the first and second components prior to use.

11. The kit of claim 10, wherein the spray pump delivers a volume of approximately 130 μ

l per spray.

12. The kit of claim 10, wherein the material resistant to low temperatures is selected from the group consisting of glass, polypropylene, polyethylene, and ethylene vinyl acetate (EVA).

13. The kit of claim 10, wherein the vials are sealed within a pouch or container prior to cryopreservation, wherein the pouch or container is fabricated of a material capable of withstanding temperatures ranging from −

80°

to −

196°

C. and wherein the pouch or container protects the first and second components from contamination during cryopreservation and subsequent thawing.

14. The kit of claim 13, wherein the pouch or container is waterproof.

15. The tissue regeneration system of claim 1, wherein the second component comprises from about 1 ×

10³ cells/μ

l to about 50 ×

10³ cells/μ

l of the mitotically inactive allogenic cells.

16. The tissue regeneration system of claim 3, wherein the one or more keratinocytes and fibroblasts are mitotically inactivated by mitomycin C.

17. The tissue regeneration system of claim 3, wherein the one or more keratinocytes and fibroblasts are mitotically inactivated by irradiation with γ

-Rays.

18. The tissue regeneration system of claim 3, wherein the one or more keratinocytes and fibroblasts are mitotically inactivated by irradiation with X-Rays.

19. The tissue regeneration system of claim 3, wherein the one or more keratinocytes and fibroblasts are mitotically inactivated by irradiation with UV light.

20. The tissue regeneration system of claim 5, wherein the cryoprotectant is human serum albumin.