# US 10,307,513 B2 · Biomimetic hydrogel scaffolds and related methods

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- Number: US 10,307,513 B2
- Document ID: US10307513B2
- Status: Granted
- Kind: B2
- Application number: 15/179,505
- Filed: 2016-06-10
- Published: 2016-12-15
- Granted: 2019-06-04
- Earliest claimed: 2015-06-12
- Assignees: University of Pittsburgh—Of the Commonwealth System of Higher Education
- Applicants: University of Pittsburgh—Of the Commonwealth System of Higher Education
- Inventors: Shilpa Sant
- Web page: https://patentgrep.com/patent/US10307513B2

## CPC

- A61L 27/54: Biologically active materials, e.g. therapeutic substances {(A61L27/227 takes precedence)}
- A61K 9/0024: {Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue (compositions for intravenous administration, normal injectable solutions or dispersions for, e.g. subcutaneous administration A61K9/0019; brain implants A61K9/0085; (coated) prostheses, catheters or stents A61L)}
- A61K 31/00: Medicinal preparations containing organic active ingredients
- A61K 33/42: Phosphorus; Compounds thereof
- A61K 38/18: Growth factors; Growth regulators
- A61K 47/02: Inorganic compounds
- A61K 47/36: Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
- A61L 27/26: Mixtures of macromolecular compounds
- A61L 27/443: {with carbon fillers}
- A61L 27/46: with phosphorus-containing inorganic fillers
- A61L 27/52: Hydrogels or hydrocolloids
- A61L 2300/112: Phosphorus-containing compounds, e.g. phosphates, phosphonates
- A61L 2300/252: Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
- A61L 2300/414: Growth factors
- A61L 2430/02: for reconstruction of bones; weight-bearing implants
- A61L 2430/30: for muscle reconstruction
- A61L 2430/32: for nerve reconstruction
- 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)

## Abstract

Provided herein are methods of making a biomimetic hydrogel scaffold comprising a polycation and a polyanion. Also provided are anisotropic biomimetic hydrogel scaffold compositions suitable for use in tissue growth, including bone, muscle, and nerve growth an optionally comprising a carbon allotrope such as graphene. Also provided are methods of producing tissue comprising growing tissue on the biomimetic hydrogel scaffold comprising a polycation and a polyanion.

## Claims

### Claim 1

A method of producing a biotherapeutic composition, comprising feeding a first component comprising a positively-charged polyelectrolyte, and a second component comprising a negatively-charged polyelectrolyte through a fluid flow passage to produce a product comprising an anisotropic structure,
wherein the method further comprises feeding a therapeutic agent into the passage with the first and second components, wherein the therapeutic agent is one or more of: basic fibroblast growth factor, acidic fibroblast growth factor, vascular endothelial growth factor, hepatocyte growth factor, insulin-like growth factors 1 and 2, platelet derived growth factor, stromal derived factor 1 alpha, nerve growth factor, ciliary neurotrophic factor, neurotrophin-3, neurotrophin-4, neurotrophin-5, pleiotrophin protein, midkine protein, brain-derived neurotrophic factor, tumor angiogenesis factor, corticotrophin releasing factor, transforming growth factors α and β, granulocyte-macrophage colony stimulating factor, osteopontin, osteocalcin, an interleukin, and/or an interferon, and
wherein the positively-charged polyelectrolyte has a net positive zeta potential, and the negatively-charged polyelectrolyte has a net negative zeta potential.

### Claim 2 (depends on 1)

The method of claim 1, wherein the passage is cylindrical.

### Claim 3 (depends on 1)

The method of claim 1, wherein the passage has a largest cross section of 10 mm or less.

### Claim 4 (depends on 1)

The method of claim 1, further comprising collecting the product onto a surface, and drying the collected material to produce a sheet or other form.

### Claim 5 (depends on 1)

The method of claim 1, wherein the negatively-charged polyelectrolyte is one or more of gellan gum and alginate, Kappa carrageenan, hyaluronic acid, chondroitin sulfate, poly(aspartic acid), and poly(glutamic acid); and wherein the positively-charged polyelectrolyte is chosen from one or more of: chitosan, glucosamine sulfate, chondroitin sulfate, or a positively-charged peptide.

### Claim 6 (depends on 1)

The method of claim 1, wherein the positively-charged electrolyte is chitosan and/or the negatively-charged electrolyte is alginate, gellan gum, and/or kappa-carageenan.

### Claim 7 (depends on 1)

The method of claim 1, wherein the positively-charged polyelectrolyte and/or the negatively-charged polyelectrolyte are a polyurethane, a polyester, or a polyether.

### Claim 8 (depends on 1)

The method of claim 1, further comprising feeding a cationic or anionic composition into the passage with the first and second components.

### Claim 9 (depends on 1)

The method claim 1, further comprising feeding a carbon allotrope into the passage with the first and second components.

### Claim 10

A composition comprising an anisotropic assembly of a positively-charged polyelectrolyte and a negatively-charged polyelectrolyte,
wherein the composition is complexed with a therapeutic agent, and wherein the therapeutic agent is one or more of: basic fibroblast growth factor, acidic fibroblast growth factor, vascular endothelial growth factor, hepatocyte growth factor, insulin-like growth factors 1 and 2, platelet derived growth factor, stromal derived factor 1 alpha, nerve growth factor, ciliary neurotrophic factor, neurotrophin-3, neurotrophin-4, neurotrophin-5, pleiotrophin protein, midkine protein, brain-derived neurotrophic factor, tumor angiogenesis factor, corticotrophin releasing factor, transforming growth factors α and β, granulocyte-macrophage colony stimulating factor, osteopontin, osteocalcin, an interleukin, and/or an interferon, and
wherein the positively-charged polyelectrolyte has a net positive zeta potential, and the negatively-charged polyelectrolyte has a net negative zeta potential.

### Claim 11 (depends on 10)

The composition of claim 10, wherein the negatively-charged polyelectrolyte is one or more of gellan gum and alginate, Kappa carrageenan, hyaluronic acid, chondroitin sulfate, poly(aspartic acid), and poly(glutamic acid); and wherein the positively-charged polyelectrolyte is chosen from one or more of: chitosan, glucosamine sulfate, chondroitin sulfate, or a positively-charged peptide.

### Claim 12 (depends on 10)

The composition of claim 10, wherein the positively-charged polyelectrolyte and/or the negatively-charged polyelectrolyte are a polyurethane, a polyester, or a polyether.

### Claim 13 (depends on 12)

The composition of claim 12, in which one or both of the positively-charged polyelectrolyte and the negatively-charged polyelectrolyte is conjugated to a peptide, and wherein the peptide is one or more of IKLLI (SEQ ID NO: 1), REDV (SEQ ID NO: 2), LDV, RGDSP (SEQ ID NO: 3), RGDV (SEQ ID NO: 4), LRGDN (SEQ ID NO: 5), RGDT (SEQ ID NO: 6), YIGSR (SEQ ID NO: 7), TTSWSQ (SEQ ID NO: 8), AEIDGIEL (SEQ ID NO: 9), WYRGRL (SEQ ID NO: 10), SIKVAVS (SEQ ID NO: 11), PDSGR (SEQ ID NO: 12), RNIAEIIKDI (SEQ ID NO: 13), DGEA (SEQ ID NO: 14), VTXG (SEQ ID NO: 15), PRRARV (SEQ ID NO: 16), YEKPGSPPREVVPRPRPGV (SEQ ID NO: 17), RPSLAKKQRFRHRNRKGYRSQRGHSRGR (SEQ ID NO: 18), RIQNLLKITNLRIKFVK (SEQ ID NO: 19), RGD, IKVAV (SEQ ID NO: 20) and IKVAVS (SEQ ID NO: 21).

### Claim 14 (depends on 10)

The composition of claim 10, complexed with a carbon allotrope.

### Claim 15 (depends on 10)

The composition of claim 10, complexed with a cationic composition.

### Claim 16 (depends on 10)

A method of delivering a therapeutic agent or cell to a patient, comprising administering to the patient the composition according to claim 10.

### Claim 17 (depends on 10)

A method of growing cells or tissue, comprising incubating, in an appropriate cell growth medium, cells placed in contact with the composition according to claim 10.

### Claim 18 (depends on 10)

A method of making tissue, comprising implanting a composition according to claim 10 into a patient.

### Claim 19 (depends on 18)

The method of claim 18, wherein the tissue is nerve or muscle, and the composition further comprises a carbon allotrope.

### Claim 20 (depends on 18)

The method of claim 18, wherein the composition further comprises a therapeutic agent.

### Claim 21 (depends on 5)

The method of claim 5, wherein the positively-charged peptide is one or more of poly-L-lysine, polycysteine and poly-L-arginine.

### Claim 22 (depends on 8)

The method of claim 8, wherein the cationic or anionic composition comprises Ca2+ and/or a calcium phosphate.

### Claim 23 (depends on 11)

The composition of claim 11, wherein the positively-charged peptide is one or more of poly-L-lysine, polycysteine and poly-L-arginine.

### Claim 24 (depends on 15)

The composition of claim 15, wherein the cationic or anionic composition comprises Ca2+ and/or a calcium phosphate.