Compositions for regenerating tissue that has deteriorated, and methods for using such compositions

• US 20070207131A1
• Filed: 04/30/2007
• Published: 09/06/2007
• Est. Priority Date: 05/22/1998
• Status: Abandoned Application

• Current Assignee: Isolagen Technologies Incorporated
• Original Assignee: Isolagen Technologies Incorporated
• Inventors: Marko, Olga, Boss, William
• Application Number:
  US11/799,133
• Publication Number:
  US 20070207131A1
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: A61F 2013/00374 against wrinkles, e.g. face lift
• CPC Class Codes: A61F 2310/00293 containing a phosphorus-containing compound, e.g. apatite
• CPC Class Codes: A61F 2310/00365 Proteins; Polypeptides; Degradation products thereof
• CPC Class Codes: A61F 2310/00383 Gelatin
• 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 35/34 Muscles; Smooth muscle cells; Heart; Cardiac stem cells; Myoblasts; Myocytes; Cardiomyocytes vascular smooth muscle A61K35/44
• CPC Class Codes: A61L 2400/06 Flowable or injectable implant compositions
• CPC Class Codes: A61L 27/12 Phosphorus-containing materials, e.g. apatite
• CPC Class Codes: A61L 27/22 Polypeptides or derivatives thereof , e.g. degradation products
• CPC Class Codes: A61L 27/222 Gelatin
• CPC Class Codes: A61L 27/24 Collagen
• CPC Class Codes: A61L 27/3608 Bone, e.g. demineralised bone matrix [DBM], bone powder
• CPC Class Codes: A61L 27/3629 Intestinal tissue, e.g. small intestinal submucosa
• CPC Class Codes: A61L 27/3641 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: A61L 27/365 Bones
• CPC Class Codes: A61L 27/3683 subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
• CPC Class Codes: A61L 27/3687 characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
• 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 27/3839 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: A61L 27/3847 Bones
• CPC Class Codes: A61L 27/3895 : using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: A61L 27/58 : Materials at least partially resorbable by the body
• CPC Class Codes: A61L 31/005 : Ingredients of undetermined constitution or reaction products thereof
• CPC Class Codes: A61L 31/026 : Ceramic or ceramic-like structures, e.g. glasses
• CPC Class Codes: A61L 31/043 : Proteins; Polypeptides; Degradation products thereof
• CPC Class Codes: A61L 31/044 : Collagen
• CPC Class Codes: A61L 31/045 : Gelatin
• CPC Class Codes: A61L 31/148 : Materials at least partially resorbable by the body
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 2533/18 : Calcium salts, e.g. apatite, Mineral components from bones, teeth, shells
• CPC Class Codes: C12N 2533/54 : Collagen; Gelatin
• CPC Class Codes: C12N 2533/90 : Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 5/0068 : General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0656 : Adult fibroblasts

Abstract

A composition for promoting regeneration of tissue which has degenerated in a subject as a result of a disease or disorder and a method of using the composition is provided. The composition comprises a biodegradable acellular matrix, and passaged autologous fibroblasts substantially free of immunogenic proteins, e.g., culture medium serum-derived proteins, integrated within the matrix. The method of using the composition to promote regeneration of tissue involves placing the composition on a site of degenerated tissue in a subject so that the composition promotes tissue regeneration at the site. The composition and the method of its use have applications promoting regeneration of tissue (i) that has degenerated as a result of numerous diseases or disorders or (ii) that has a defect, including, but not limited to, defects of the oral mucosa, trauma to the oral mucosa (e.g., extraction of a tooth), periodontal disease, diabetes, cutaneous ulcers, venous stasis, scars of the skin, or wrinkles of the skin. Also provided is an injectable composition comprising any type of collagen and passaged autologous fibroblasts substantially free of immunogenic proteins, e.g., culture medium serum-derived proteins, for correcting defects in skin, such as wrinkles or scars, and for augmenting tissue in the subject, particularly facial tissue.

Claims

1. A method for repairing tissue that has been damaged in a subject, wherein the method comprises:

a) providing a pharmaceutical composition comprising;

(i) autologous passaged fibroblasts substantially free of proteins derived from xenogeneic serum of xenogeneic serum-containing culture medium; and

(ii) a pharmaceutically acceptable carrier;

b) identifying a site in the subject of;

(i) tissue degeneration due to a disease or disorder;

or (ii) a tissue defect, wherein the tissue degeneration or the tissue defect in the subject comprises a defect of an oral mucosa, trauma to an oral mucosa, trauma to an oral bone, periodontal disease, diabetes, cutaneous ulcers, or venous stasis; and

c) injecting a therapeutically effective amount of the pharmaceutical composition subadjacent to the tissue degeneration or defect,

wherein the injecting results in repair of the tissue degeneration or the defect

2. The method of claim 1, wherein the tissue degeneration or the tissue defect comprises periodontal disease.

3. The method of claim 2, wherein the periodontal disease comprises periodontal degeneration.

4. The method of claim 2, wherein the periodontal disease comprises gingivitis.

5. The method of claim 2, wherein the periodontal disease comprises a non-healing wound of a palatal mucosa.

6. The method of claim 2, wherein the periodontal disease comprises a non-healing wound of a gingival mucosa.

7. The method of claim 1, wherein the tissue degeneration or the tissue defect comprises a defect of an oral mucosa.

8. The method of claim 7, wherein the defect of an oral mucosa is caused by trauma.

9. The method of claim 7, wherein the defect of an oral mucosa is caused by a dermatosis.

10. The method of claim 7, wherein the defect of an oral mucosa is caused by aphthous stomatitis.

11. The method of claim 7, wherein the defect of an oral mucosa is caused by an infection.

12. The method of claim 1, wherein the tissue degeneration or the tissue defect comprises trauma to an oral mucosa.

13. The method of claim 12, wherein the trauma to an oral mucosa is caused by extraction of a tooth.

14. The method of claim 1, wherein the tissue degeneration or the tissue defect comprises trauma to an oral bone.

15. The method of claim 14, wherein the trauma to an oral bone is caused by extraction of a tooth.

16. The method of claim 14, wherein the oral bone is a maxillary bone.

17. The method of claim 14, wherein the oral bone is a mandibular bone

18. The method of claim 1, wherein the step of providing a pharmaceutical composition comprises:

a) taking a biopsy of dermis comprising fibroblasts from a subject;

b) separating fibroblasts from said biopsy so as to provide fibroblasts substantially free of extracellular matrix and non-fibroblast cells;

c) placing the fibroblasts in a culture medium comprising between 0.0% and about 20% serum in order to grow fibroblasts;

d) incubating the fibroblasts in a serum-free medium for at least 2 hours at between about 30°

C. and about 37.5°

C. to form passaged fibroblasts;

e) exposing the passaged fibroblasts to a proteolytic enzyme so as to suspend the passaged fibroblasts; and

f) adding a pharmaceutically acceptable carrier to the suspended passaged fibroblasts to form the pharmaceutical composition.

19. The method of claim 18, wherein the biopsy is taken from gums or palate of the subject.

20. The method of claim 18, wherein the biopsy is taken from skin of the subject.

21. The method of claim 18, wherein the proteolytic enzyme is trypsin.