# US 12,005,158 B2 · Methods for preparation of a terminally sterilized hydrogel derived from extracellular matrix

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- Number: US 12,005,158 B2
- Document ID: US12005158B2
- Status: Granted
- Kind: B2
- Application number: 17/728,399
- Filed: 2022-04-25
- Published: 2022-08-11
- Granted: 2024-06-11
- Earliest claimed: 2014-03-21
- Assignees: University of Pittsburgh—Of the Commonwealth System of Higher Education
- Applicants: University of Pittsburgh—Of the Commonwealth System of Higher Education
- Inventors: Stephen F. Badylak; Timothy Joseph Keane, Jr.; Neill Jordon Turner; Christopher Lee Dearth
- Web page: https://patentgrep.com/patent/US12005158B2

## CPC

- A61L 27/52: Hydrogels or hydrocolloids
- A61L 27/36: containing ingredients of undetermined constitution or reaction products thereof {, e.g. transplant tissue, natural bone, extracellular matrix (isolated natural compounds, e.g. collagen A61L27/24)}
- A61L 27/3633: {Extracellular matrix [ECM]}
- A61L 27/38: {containing added animal cells (organs or tissue containing native cells A61L27/36)}
- A61L 27/50: Materials characterised by their function or physical properties {, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials}
- A61L 27/54: Biologically active materials, e.g. therapeutic substances {(A61L27/227 takes precedence)}
- A61L 2430/02: for reconstruction of bones; weight-bearing implants
- 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)

## Abstract

Provided are methods for preparing sterilized, gelled, solubilized extracellular matrix (ECM) compositions useful as cell growth substrates. 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 1

A gelable extracellular matrix (ECM) composition comprising a decellularized, enzymatically digested, dried, electron-beam sterilized, intact extracellular matrix, wherein said composition is capable of forming a gel upon hydration, neutralization to pH 7.2-7.8, and warming to a temperature greater than 25° C.

### Claim 2 (depends on 1)

The composition of claim 1, wherein the composition comprises a protease.

### Claim 3 (depends on 2)

The composition of claim 2, wherein the protease is pepsin or trypsin.

### Claim 4 (depends on 1)

The composition of claim 1, wherein the extracellular matrix is bioactive.

### Claim 5 (depends on 1)

The composition of claim 1, wherein the extracellular matrix is derived from urinary bladder, spleen, liver, heart, pancreas, ovary, small intestine, large intestine, colon, esophagus, central nervous system tissue, adipose tissue, dermis, or bone.

### Claim 6 (depends on 1)

The composition of claim 1, wherein the extracellular matrix is from a human, monkey, pig, cow, or sheep.

### Claim 7 (depends on 1)

The composition of claim 1, wherein said composition is capable of forming a gel when warmed to 37° C.

### Claim 8 (depends on 1)

A biocompatible scaffold coated with the composition according to claim 1.

### Claim 9 (depends on 8)

The biocompatible scaffold of claim 8, wherein the scaffold comprises: (a) one or more of a cobalt-chrome alloy, stainless steel, titanium, tantalum, and a titanium alloy; (b) an inorganic mineral; (c) a ceramic; and/or (d) a polymer.

### Claim 10

A method of preparing an extracellular matrix-derived gelable composition comprising, in order: (i) solubilizing intact extracellular matrix (ECM) by digestion with an acid protease in an acidic solution to produce a digest solution; (ii) drying the digest solution; and (iii) terminally sterilizing the dried digest with electron beam radiation.

### Claim 11 (depends on 10)

The method of claim 10, further comprising: (iv) hydrating and neutralizing the sterilized, dried digest to a pH between 7.2 and 7.8 to produce a neutralized digest solution; and (v) gelling the solution at a temperature greater than 25° C.

### Claim 12 (depends on 11)

The method of claim 11, wherein the neutralized digest solution is maintained at or below 25° C. before gelation.

### Claim 13 (depends on 11)

The method of claim 11, further comprising coating a matrix of a biocompatible scaffold with the solubilized ECM and gelling the matrix.

### Claim 14 (depends on 13)

The method of claim 13, wherein the biocompatible scaffold comprises: (a) one or more of a cobalt-chrome alloy, stainless steel, titanium, tantalum, and a titanium alloy; (b) an inorganic mineral; (c) a ceramic; and/or (d) a polymer.

### Claim 15 (depends on 10)

The method of claim 10, wherein the ECM is not terminally sterilized, dialyzed, or subjected to a cross-linking process prior to the solubilizing step.

### Claim 16 (depends on 10)

The method of claim 10, wherein the digest solution is lyophilized.

### Claim 17 (depends on 10)

The method of claim 10, wherein the ECM is derived from mammalian tissue.

### Claim 18 (depends on 17)

The method of claim 17, wherein the mammalian tissue is derived from one of urinary bladder, spleen, liver, heart, central nervous system tissue, adipose tissue, bone, pancreas, ovary, small intestine, large intestine, or colon.

### Claim 19 (depends on 10)

The method of claim 10, wherein the ECM is comminuted.

### Claim 20 (depends on 10)

The method of claim 10, wherein the acid protease is pepsin.