Liquid collagen bioinks and methods to make and use collagen structures

Patent metadata

Field	Value
Publication number	US-12167965-B2
Application number	US-202117643046-A
Country	US
Kind code	B2
Filing date	Dec 7, 2021
Priority date	Dec 7, 2020
Publication date	Dec 17, 2024
Grant date	Dec 17, 2024

Abstract

The present disclosure provides collagen bioink compositions and chemically uncrosslinked and crosslinked collagen structures including collagen microparticles and scaffolds. Also provided are methods of their fabrication and use. Applications for using these collagen structures include treatments of damaged tissue, particularly those caused by osteoarthritis.

First claim

The invention claimed is:

1. A method to fabricate chemically uncrosslinked collagen microparticles, wherein the method comprises:
a) combining an oil and either sorbitan monooleate or sorbitan monostearate and stirring to create a printing solution;
b) adding a collagen bioink composition dropwise to the printing solution to create a combined solution, wherein the collagen bioink composition comprises collagen, a polar solvent, and a stabilizing divalent ion chelating agent, wherein the collagen bioink fibrilizes when introduced into the printing solution;
c) stirring the combined solution;
d) increasing the temperature of the combined solution to 37° C. while continuing stirring; and
e) centrifuging the combined solution to collect the microparticles, wherein the combined solution is free of crosslinker or up to 0.05 mM of crosslinker is present.

2. A method to fabricate a chemically crosslinked collagen structure, wherein the method comprises the method of claim 1 and further comprises incubating the chemically uncrosslinked collagen microparticles or structure with a crosslinker and optionally an antibody, a peptide, an aptamer, a functionalized nanoparticle, or a combination thereof.

3. The method of claim 2 , wherein the method comprises adding an antibody, a peptide, an aptamer, a functionalized nanoparticle, or a combination thereof to the incubation step.

4. The method of claim 1 , wherein the oil is olive oil.

5. The method of claim 1 , wherein the sorbitan monooleate concentration is from about 0.02 to about 0.35% by volume.

6. The method of claim 1 , wherein the sorbitan monostearate concentration is from about 0.01 to about 2.50% by volume.

7. The method of claim 1 , wherein step c) comprises stirring for 1 hour.

8. The method of claim 1 , wherein step d) comprises stirring for up to 16 hours.

9. The method of claim 1 , wherein the method further comprises washing the collected microparticles.

10. The method of claim 1 , wherein the collected microparticles are uniform in size.

11. The method of claim 1 , wherein the collected microparticles have a standard deviation in size of less than 15 μm.

12. The method of claim 1 , wherein printing solution further comprises nanoparticles, an anti-inflammatory agent, or a combination thereof.

13. A method to fabricate a chemically crosslinked collagen structure, wherein the method comprises:
a) combining an oil and either sorbitan monooleate or sorbitan monostearate and stirring to create a printing solution;
b) adding a collagen bioink composition dropwise to the printing solution to create a combined solution, wherein the collagen bioink composition comprises collagen, a polar solvent, and a stabilizing divalent ion chelating agent, wherein the collagen bioink fibrilizes when introduced into the printing solution;
c) stirring the combined solution;
d) increasing the temperature of the combined solution to 37° C. while continuing stirring; and
e) centrifuging the combined solution to collect the microparticles, wherein the combined solution is substantially free of crosslinker;
and further comprising incubating the chemically uncrosslinked collagen microparticles with a crosslinker and a functionalized nanoparticle.

14. The method of claim 13 , wherein the functionalized nanoparticle comprises gold, silver, silicon carbide, silicon, silica, or a combination thereof.

15. The method of claim 14 , wherein the crosslinker is EDC/NHS (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide) or genipin.

16. The method of claim 13 , wherein the functionalized nanoparticle comprises gold.

17. The method of claim 13 , wherein the crosslinker has a concentration of from about 2 mM to about 10 mM.

Assignees

• Univ Missouri

Inventors

• Grant Sheila
• Snider Colten
• Grant David

Classifications

• B33Y10/00

  Processes of additive manufacturing · CPC title

• A61F2002/30062

  (bio)absorbable, biodegradable, bioerodable, (bio)resorbable, resorptive · CPC title

• B82Y40/00

  Manufacture or treatment of nanostructures · CPC title

• B82Y5/00

  Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery · CPC title

• A61F2310/00371

  Collagen · CPC title