Biomaterial based on a hydrophilic polymeric carrier

• US 10,196,602 B2
• Filed: 01/11/2010
• Issued: 02/05/2019
• Est. Priority Date: 07/13/2007
• Status: Active Grant

• Current Assignee: NMI Naturwissenschaftliches und Medizinisches Institut an an der Universität Tübingen. (NMI Natural and Medical Sciences Institute at the University TÜBINGEN)
• Original Assignee: NMI Naturwissenschaftliches und Medizinisches Institut an an der Universität Tübingen. (NMI Natural and Medical Sciences Institute at the University TÜBINGEN)
• Inventors: Wurst, Helmut
• Primary Examiner(s): Long, Scott
• Assistant Examiner(s): Martin, Paul C
• Application Number:
  US12/655,983
• Publication Number:
  US 20100184223A1
• Time in Patent Office: 3,312 Days
• Field of Search: None
• 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: C12N 2533/30 Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 2533/50 Proteins
• CPC Class Codes: C12N 2537/10 Cross-linking
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06

Abstract

The present invention relates to a biomaterial for cell or tissue culture, based on a polymeric carrier, which contains at least one crosslinkable hydrophilic polymer. The polymer is functionalized with groups that are selected from maleimide, vinylsulfonic, acrylate, alkyl halide, azirine, pyridyl, thionitrobenzene acid groups, or arylating groups. The invention relates further to a method of production of said biomaterial, and the use of particular functionalizing groups for the production of a biomaterial for the cultivation of tissue and/or cells. The biomaterial can have biofactors that exert a particular action on cells.

Claims

1. A hydrogel for the cultivation of a cell or tissue, comprising:

a hydrophilic polymeric carrier, which comprises at least one crosslinkable hydrophilic polymer, wherein the polymer is functionalized with at least three maleimide groups which the non-functionalized polymer does not normally possess, and wherein the polymer is maleimide-serum albumin, and

a crosslinking agent comprising at least two thiol groups, wherein the crosslinking agent is dithio-PEG having a molecular weight of 3 kDa, and wherein the dithio-PEG crosslinks the polymer via covalent bonding between the thiol groups of the dithio-PEG and the maleimide groups of the maleimide-serum albumin polymer.

2. The hydrogel of claim 1, wherein the polymer is crosslinkable in the presence of cells.

3. The hydrogel of claim 1, further comprising at least one biofactor.

4. The hydrogel of claim 1, wherein the polymer is aminated.

5. A method comprising:

a) introducing a cell or tissue into the hydrogel of claim 1, and

b) culturing the cell or tissue in the hydrogel.

6. The method of claim 5, wherein the polymer is aminated.

7. The method of claim 5, wherein the hydrogel further comprises a biofactor.

8. The method of claim 7, wherein the biofactor is selected from the group consisting of peptides, proteins, nucleic acids, organic active substances, apoptosis-inducing active substances, adhesion-promoting active substances, growth-inhibiting active substances, anti-inflammatory active substances, receptor agonists, receptor antagonists, proteins of the extracellular matrix, cell surface proteins, antibodies, growth factors, sugars, lectins, carbohydrates, cytokines, DNA, RNA, siRNA, aptamers, and fragments thereof that are relevant to binding or action, and mixtures thereof.

9. The method of claim 7, wherein the biofactor has at least one thiol group, via which the biofactor is bound to a maleimide group of the polymer.

10. The method of claim 5, wherein the polymer is crosslinked using thiol-functionalized PEG.

11. A hydrogel for the cultivation of a cell or tissue, comprising:

a hydrophilic polymeric carrier, which comprises at least one crosslinkable hydrophilic polymer, wherein the polymer is functionalized with at least three maleimide groups which the non-functionalized polymer does not normally possess, and wherein the polymer is maleimide-serum albumin,

a crosslinking agent comprising at least two thiol groups, wherein the crosslinking agent is dithio-PEG having a molecular weight of 3 kDa which crosslinks the polymer, and

at least one biofactor, wherein the biofactor has at least one thiol group, via which the biofactor is bound to a maleimide group of the polymer.

12. The hydrogel of claim 11, wherein the biofactor is selected from the group consisting of peptides, proteins, nucleic acids, organic active substances, apoptosis-inducing active substances, adhesion-promoting active substances, growth-inhibiting active substances, anti-inflammatory active substances, receptor agonists, receptor antagonists, proteins of the extracellular matrix, cell surface proteins, antibodies, growth factors, sugars, lectins, carbohydrates, cytokines, DNA, RNA, siRNA, aptamers, and fragments thereof that are relevant to binding or action, and mixtures thereof.

13. The hydrogel of claim 11, wherein the polymer is aminated.

14. The hydrogel of claim 11, wherein the polymer is crosslinkable in the presence of cells.

15. A hydrogel for the cultivation of a cell or tissue, comprising:

a hydrophilic polymeric carrier, which comprises at least one crosslinkable hydrophilic polymer, wherein the polymer is functionalized with at least three maleimide groups which the non-functionalized polymer does not normally possess, and wherein the polymer is maleimide-serum albumin or maleimide-PVA (polyvinyl alcohol), and

a crosslinking agent comprising at least two thiol groups, wherein the crosslinking agent has a molecular weight of 3 kDa, and wherein the crosslinking agent crosslinks the polymer via covalent bonding between the thiol groups of the crosslinking agent and the maleimide groups of the maleimide-serum albumin polymer or maleimide-PVA polymer.

16. A hydrogel for the cultivation of a cell or tissue, comprising:

a hydrophilic polymeric carrier, which comprises at least one crosslinkable hydrophilic polymer, wherein the polymer is functionalized with at least three maleimide groups which the non-functionalized polymer does not normally possess, and wherein the polymer is maleimide-serum albumin or maleimide-PVA, and

a crosslinking agent comprising at least two thiol groups, wherein the crosslinking agent is dithio-PEG and has a molecular weight of 3 kDa, and wherein the crosslinking agent crosslinks the polymer via covalent bonding between the thiol groups of the crosslinking agent and the maleimide groups of the maleimide-serum albumin polymer or maleimide-PVA polymer.

17. The hydrogel of claim 16, further comprising at least one biofactor, wherein the biofactor has at least one thiol group, via which the biofactor is bound to a maleimide group of the polymer, wherein the biofactor comprises a polypeptide sequence selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

2, and SEQ ID NO;

3.

18. The hydrogel of claim 16, further comprising at least one biofactor, wherein the biofactor has at least one thiol group, via which the biofactor is bound to a maleimide group of the polymer, wherein the biofactor is peptide HW1 (acetyl-KGLQGCGLQGK-OH).

19. The hydrogel of claim 16, further comprising at least one biofactor, wherein the biofactor has at least one thiol group, via which the biofactor is bound to a maleimide group of the polymer, wherein the biofactor is peptide HW2 (acetyl-KGGLQGGK-OH).

20. The hydrogel of claim 16, further comprising at least one biofactor, wherein the biofactor has at least one thiol group, via which the biofactor is bound to a maleimide group of the polymer, wherein the biofactor is peptide HW9 (FITC-β

-Ala-GCGYGRGDSPGSGC).

21. The hydrogel of claim 16, further comprising at least one cell to be cultivated in the hydrogel, wherein the cell is selected from the group consisting of a human chondrocyte from osteoarthritic cartilage, a human mesenchymal stem cell, a human epithelial cell, and a human fibroblast.

22. A hydrogel for the cultivation of a cell or tissue, comprising:

a hydrophilic polymeric carrier, which comprises at least one crosslinkable hydrophilic polymer, wherein the polymer is functionalized with at least three maleimide groups which the non-functionalized polymer does not normally possess, and wherein the polymer is maleimide-serum albumin,

a crosslinking agent comprising two thiol groups, wherein the crosslinking agent is dithio-PEG and has a molecular weight of 3 kDa, and wherein the crosslinking agent crosslinks the polymer via covalent bonding between the two thiol groups of the crosslinking agent and the maleimide groups of the maleimide-serum albumin polymer.