Bioartificial extracellular matrix containing hydrogel matrix derivatized with cell adhesive peptide fragment

• US 6,156,572 A
• Filed: 09/25/1998
• Issued: 12/05/2000
• Est. Priority Date: 07/20/1994
• Status: Expired due to Term

• Current Assignee: Neurotech USA, Inc. (Woodbrook Group Holdings, Inc.)
• Original Assignee: Neurotech S.A. (LivaNova Plc)
• Inventors: Ranieri, John P., Bellamkonda, Ravi, Aebischer, Patrick
• Primary Examiner(s): Naff, David M.
• Application Number:
  US09/160,654
• Time in Patent Office: 802 Days
• Field of Search: 435/177, 435/178, 435/325, 435/368, 435/395, 435/397, 435/402, 424/570, 424/423, 424/93.7, 424/488, 530/326, 530/328, 530/329, 530/812, 530/402, 530/813, 606/152
• US Class Current: 435/395
• CPC Class Codes: A61B 17/1128 of nerves
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61L 2430/32 for nerve reconstruction
• CPC Class Codes: A61L 27/20 Polysaccharides
• CPC Class Codes: A61L 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• 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 27/52 Hydrogels or hydrocolloids
• CPC Class Codes: A61L 27/54 Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: A61L 31/005 Ingredients of undetermined constitution or reaction products thereof
• CPC Class Codes: A61L 31/042 Polysaccharides
• CPC Class Codes: A61L 31/047 Other specific proteins or polypeptides not covered by A61L31/044 - A61L31/046
• CPC Class Codes: A61L 31/145 Hydrogels or hydrocolloids
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: C08L 5/12 Agar or agar-agar , i.e. mixture of agarose and agaropectin; Derivatives thereof
• CPC Class Codes: Y10S 530/812 Peptides or proteins is immobilized on, or in, an organic carrier
• CPC Class Codes: Y10S 530/813 Carrier is a saccharide

Abstract

A bioartificial extracellular matrix for use in tissue regeneration or replacement is provided by derivatizing a three-dimensional hydrogel matrix with a cell adhesive extracellular matrix protein or cell adhesive peptide fragment of the protein. Preferably, derivatizing is by covalent immobilization of a cell adhesive peptide fragment having the amino acid sequence, ArgGlyAsp, TyrIleGlySerArg or IleLysValAlaVal. Cartilage or tendon can be regenerated by implanting a matrix containing an adhesive peptide fragment that favors chondrocyte invasion. The matrix can be pre-seeded with cells, and tissue can be reconstituted in vitro and then implanted. A cell-seeded matrix can be encapsulated in a semi-permeable membrane to form a bioartificial organ. An agarose hydrogel matrix having an agarose concentration of 0.5-1.25% (w/v) and an average pore radius between 120 nm and 290 nm is preferred. A nerve guidance channel for use in regenerating severed nerve is prepared containing a tubular semi-permeable membrane having openings adapted to receive ends of a severed nerve, and an inner lumen containing the hydrogel matrix having a bound cell adhesive peptide fragment through which the nerve can regenerate.

Claims

1. A bioartificial extracellular matrix comprising a three-dimensional high water content derivatized hydrogel matrix having a hydrogel matrix core

(a) wherein the hydrogel matrix is derivatized through the matrix by covalent-immobilization of at least one cell adhesive peptide fragment, homogeneously dispersed throughout the hydrogel matrix, and

(b) wherein the hydrogel matrix has an average pore radius greater than 120 nm.

2. The matrix according to claim 1, wherein the amino acid sequence is SEQ ID NO:

1.

3. The matrix according to claim 1, wherein the amino acid sequence is SEQ ID NO:

2.

4. The matrix according to claim 1, wherein the amino acid sequence is SEQ ID NO:

3.

5. The matrix according to any one of claims 1 and 2-4, wherein the hydrogel matrix is a polysaccharide hydrogel matrix.

6. The matrix according to claim 1, wherein the hydrogel matrix is an agarose hydrogel matrix.

7. The matrix according to claim 6, wherein the agarose concentration in the hydrogel matrix ranges between 0.5-1.25% (w/v) and the hydrogel matrix has an average pore radius ranging between 120-290 nm.

8. The matrix according to claim 6, wherein the agarose concentration in the hydrogel matrix is 1.0% (w/v) and the hydrogel matrix has an average pore radius of approximately 150 nm.