# US 2013/0156862 A1 · Extracellular Matrix-Derived Gels and Related Methods

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- Number: US 2013/0156862 A1
- Document ID: US20130156862A1
- Status: Published application
- Kind: A1
- Application number: 13/684,830
- Filed: 2012-11-26
- Published: 2013-06-20
- Earliest claimed: 2007-03-02
- Assignees: University of Pittsburgh—Of the Commonwealth System of Higher Education
- Inventors: Stephen F. Badylak; Donald Freytes
- Web page: https://patentgrep.com/patent/US20130156862A1

## CPC

- A61L 27/3633: {Extracellular matrix [ECM]}
- 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)
- A61L 27/34: Macromolecular materials
- A61L 27/3687: {characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents}
- A61L 27/38: {containing added animal cells (organs or tissue containing native cells A61L27/36)}
- A61L 27/52: Hydrogels or hydrocolloids
- A61L 27/54: Biologically active materials, e.g. therapeutic substances {(A61L27/227 takes precedence)}
- A61P 19/00: Drugs for skeletal disorders
- C12P 21/06: produced by the hydrolysis of a peptide bond, e.g. hydrolysate products (preparing foodstuffs by protein hydrolysis A23J3/00)
- 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)
- A61L 2300/64: Animal cells
- A61L 2400/06: Flowable or injectable implant compositions
- A61L 2420/04: Coatings containing a composite material such as inorganic/organic, i.e. material comprising different phases
- A61L: METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES (preservation of bodies or disinfecting characterised by the agents employed A01N; preserving, e.g. sterilising, food or foodstuffs A23; preparations for medical, dental or toiletry purposes A61K)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}

## Abstract

Provided are methods for preparing gelled, solubilized extracellular matrix (ECM) compositions useful as cell growth scaffolds. Also provided are compositions prepared according to the methods as well as uses for the compositions. In one embodiment a device, such as a prosthesis, is provided which comprises an inorganic matrix into which the gelled, solubilized ECM is dispersed to facilitate in-growth of cells into the ECM and thus adaptation and/or attachment of the device to a patient.

## Claims

### Claim at position 1

1-96. (canceled)

### Claim 97

A method of preparing an extracellular matrix-derived gel comprising: (a) solubilizing decellularized extracellular matrix (ECM) derived from cardiac tissue by digestion with an acid protease in an acidic solution to produce digested cardiac ECM; and (b) raising the pH of the digested cardiac ECM to a pH between 7.2 and 7.8 to produce a neutralized digest solution, and (c) gelling the neutralized digest solution at a temperature greater than 25° C.

### Claim 98 (depends on 97)

The method of claim 97, wherein the ECM is not dialyzed or subjected to a cross-linking process prior to the gelling step (c).

### Claim 99 (depends on 97)

The method of claim 97, wherein the ECM is intact ECM.

### Claim 100 (depends on 97)

The method of claim 97, wherein the neutralized digest solution is maintained at or below 25° C. before the gelling step (c).

### Claim 101 (depends on 97)

The method of claim 97, wherein the protease is pepsin, trypsin or a combination thereof.

### Claim 102 (depends on 97)

The method of claim 97, wherein in the step of raising the pH of the digested cardiac ECM (b), a base or an isotonic buffer is added to raise the pH of the digested cardiac ECM.

### Claim 103 (depends on 102)

The method of claim 102, wherein the base or isotonic buffer is NaOH or phosphate buffered saline.

### Claim 104 (depends on 97)

The method of claim 97, wherein the pH is raised to 7.4 in the step of raising the pH of the digested cardiac ECM (b).

### Claim 105 (depends on 97)

The method of claim 97, wherein the digest solution is gelled at 30° C. or higher.

### Claim 106 (depends on 97)

The method of claim 97, wherein the digest solution is gelled at 37° C.

### Claim 107 (depends on 97)

The method of claim 97, wherein the gel is molded during the gelling step (c).

### Claim 108 (depends on 97)

The method of claim 97, further comprising integrating one or more of a cell, a drug, a cytokine and a growth factor into the gel prior to or after gelling step (c).

### Claim 109 (depends on 108)

The method of claim 108, in which the one or more of a cell, a drug, a cytokine and a growth factor is selected from the group consisting of an interferon, an interleukin, a chemokine, a monokine, a hormone, an angiogenic factor, and an antibiotic.

### Claim 110 (depends on 108)

The method of claim 108, comprising seeding the gel with cells prior to or after gelling step (c).

### Claim 111 (depends on 97)

The method of claim 97, further comprising coating a matrix of a biocompatible scaffold with the neutralized digest solution and gelling the matrix.

### Claim 112 (depends on 97)

The method of claim 97, further comprising dispersing the neutralized digest solution in an inorganic matrix.

### Claim 113 (depends on 112)

The method of claim 112, wherein the inorganic matrix comprises a metal.

### Claim 114 (depends on 112)

The method of claim 112, wherein the inorganic matrix comprises titanium.

### Claim 115

A composition comprising extracellular matrix (ECM) derived from cardiac tissue that is a liquid at 4° C. and is a gel at a temperature greater than 25° C. and an acid protease, the composition having a pH of from 7.2 to 7.8.

### Claim 116 (depends on 115)

The composition of claim 115, in which the ECM derived gel is prepared according to a method comprising: (a) solubilizing decellularized ECM derived from cardiac tissue by digestion with an acid protease in an acidic solution to produce digested cardiac ECM; and (b) raising the pH of the digested cardiac ECM to a pH between 7.2 and 7.8 to produce a neutralized digest solution, and (c) gelling the neutralized digest solution at a temperature greater than 25° C.

### Claim 117 (depends on 116)

The composition of claim 116, wherein the decellularized ECM is not dialyzed or subjected to a cross-linking process prior to the gelling step (c).

### Claim 118 (depends on 116)

The composition of claim 116, dispersed in an inorganic matrix.

### Claim 119 (depends on 118)

The composition of claim 118, wherein the inorganic matrix comprises a metal.

### Claim 120 (depends on 118)

The composition of claim 118, wherein the inorganic matrix comprises titanium.

### Claim 121 (depends on 1)

A method of producing tissue in a patient, comprising implanting into a patient a composition prepared according to the method of claim 1.

### Claim 122 (depends on 121)

The method of claim 121, wherein the neutralized digest solution is injected into the patient.

### Claim 123

A kit comprising either:
a) a vessel containing composition comprising extracellular matrix (ECM) derived from cardiac tissue that is a liquid at 4° C. and is a gel at a temperature greater than 25° C. and an acid protease, the composition having a pH of from 7.2 to 7.8; or
b) a plurality of vessels in which a first vessel of the plurality of vessels contains digested cardiac ECM prepared by solubilizing decellularized ECM derived from cardiac tissue by digestion with an acid protease in an acidic solution and a second vessel of the plurality of vessels contains a base or isotonic buffer for neutralizing the digested cardiac ECM.