Novel tissue engineered scaffolds derived from copper capillary alginate gels

• US 20050196423A1
• Filed: 03/07/2005
• Published: 09/08/2005
• Est. Priority Date: 03/05/2004
• Status: Active Grant

• Current Assignee: University of Florida Research Foundation, Incorporated (State University System of Florida)
• Original Assignee: University of Florida Research Foundation, Incorporated (State University System of Florida)
• Inventors: Batich, Christopher D., Terada, Naohiro, Willenberg, Bradley Jay, Hamazaki, Takashi
• Granted Patent: US 7,601,525 B2
• Time in Patent Office: Days
• US Class Current: 424/423
• CPC Class Codes: A61L 27/20 Polysaccharides
• CPC Class Codes: C08L 5/04 Alginic acid; Derivatives thereof

Abstract

The present invention provides copper capillary alginate gels stabilized with barium, chitosan, its derivates, or a combination thereof. These stabilized gels are useful as scaffolds for containing, growing, or regenerating biological agents and cells for in vivo or in vitro use.

Claims

1. A biomaterial scaffold comprising an alginate gel and at least one stabilizing agent;

wherein the alginate gel comprises a plurality of continuous parallel microtubular copper capillaries having a first end and a second end; and

wherein the at least one stabilizing agent is cross-linked to the alginate gel.

2. The biomaterial scaffold according to claim 1, wherein the at least one stabilizing agent is selected from the group consisting of divalent cations, an electrostatically charged molecule, and a carbodiimide.

3. The biomaterial scaffold according to claim 1, wherein the at least one stabilizing agent comprises a barium cation.

4. The biomaterial scaffold according to claim 2, wherein the electrostatically charged molecule is selected from the group consisting of chitosan, chitosan derivatives, oligochitosan, poly-lysine, and combinations of any of the foregoing.

5. The biomaterial scaffold according to claim 1, wherein the diameter of each of the continuous parallel microtubular capillaries is within the range of about 3 μ

m to about 300 μ

m.

6. The biomaterial scaffold according to claim 1, wherein the diameters of each of the continuous parallel microtubular copper capillaries may be different;

and wherein any one diameter may vary between the first end and the second end.

7. The biomaterial scaffold according to claim 1, wherein the cross-section of each of the continuous parallel microtubular copper capillaries is non-circular.

8. The biomaterial scaffold according to claim 1, wherein the biomaterial scaffold possesses a positive charge.

9. The biomaterial scaffold according to claim 1, further comprising at least one biologically active agent.

10. The biomaterial scaffold according to claim 1, further comprising at least one biologically active agent, and wherein the at least one biologically active agent comprises a plurality of cells seeded within the continuous parallel microtubular copper capillaries.

11. The biomaterial scaffold according to claim 10, wherein the plurality of cells are selected from the group consisting of embryonic stem cells, adult stem cells, blast cells, cloned cells, fertilized ova, placental cells, keratinocytes, basal epidermal cells, hair shaft cells, hair-root sheath cells, surface epithelial cells, basal epithelial cells, urinary epithelial cells, salivary gland cells, mucous cells, serous cells, von Ebner'"'"'s gland cells, mammary gland cells, lacrimal gland cells, ceruminous gland cells, eccrine sweat gland cells, apocrine sweat gland cells, Moll gland cells, sebaceous gland cells, Bowman'"'"'s gland cells, Brunner'"'"'s gland cells, seminal vesicle cells, prostate gland cells, bulbourethral gland cells, Bartholin'"'"'s gland cells, Littre gland cells, uterine endometrial cells, goblet cells of the respiratory or digestive tracts, mucous cells of the stomach, zymogenic cells of the gastric gland, oxyntic cells of the gastric gland, insulin-producing β

cells, glucagon-producing α

cells, somatostatin-producing δ

cells, pancreatic polypeptide-producing cells, pancreatic ductal cells, Paneth cells of the small intestine, type II pneumocytes of the lung, Clara cells of the lung, anterior pituitary cells, intermediate pituitary cells, posterior pituitary cells, hormone secreting cells of the gut or respiratory tract, thyroid gland cells, parathyroid gland cells, adrenal gland cells, gonad cells, juxtaglomerular cells of the kidney, macula densa cells of the kidney, peripolar cells of the kidney, mesangial cells of the kidney, brush border cells of the intestine, striated duct cells of exocrine glands, gall bladder epithelial cells, brush border cells of the proximal tubule of the kidney, distal tubule cells of the kidney, nonciliated cells of ductulus efferens, epididymal principal cells, epididymal basal cells, hepatacytes, fat cells, type I pneumocytes, pancreatic duct cells, nonstriated duct cells of the sweat gland, nonstriated duct cells of the salivary gland, nonstriated duct cells of the mammary gland, parietal cells of the kidney glomerulus, podocytes of the kidney glomerulus, cells of the thin segment of the loop of Henle, collecting duct cells, duct cells of the seminal vesicle, duct cells of the prostate gland, vascular endothelial cells, synovial cells, serosal cells, squamous cells lining the perilymphatic space of the ear, cells lining the endolymphatic space of the ear, choroids plexus cells, squamous cells of the pia-arachnoid, ciliary epithelial cells of the eye, corneal endothelial cells, ciliated cells having propulsive function, ameloblasts, planum semilunatum cells of the vestibular apparatus of the ear, interdental cells of the organ of Corti, fibroblasts, pericytes of blood capillaries, nucleus pulposus cells of the intervertebral disc, cementoblasts, cementocytes, odontoblasts, odontocytes, chondrocytes, osteoblasts, osteocytes, osteoprogenitor cells, hyalocytes of the vitreous body of the eye, stellate cells of the perilymphatic space of the ear, skeletal muscle cells, heart muscle cells, smooth muscle cells, myoepithelial cells, red blood cells, megakaryocytes, monocytes, connective tissue macrophages, Langerhan'"'"'s cells, osteoclasts, dendritic cells, microglial cells, neutrophils, eosinophils, basophils, mast cells, plasma cells, helper T cells, suppressor T cells, killer T cells, immunoglobulin M, immunoglobulin G, immunoglobulin A, immunoglobulin E, killer cells, rod cells, cone cells, inner hair cells of the organ of Corti, outer hair cells of the organ of Corti, type I hair cells of the vestibular apparatus of the ear, type II cells of the vestibular apparatus of the ear, type II taste bud cells, olfactory neurons, basal cells of olfactory epithelium, type I carotid body cells, type II carotid body cells, Merkel cells, primary sensory neurons specialized for touch, primary sensory neurons specialized for temperature, primary neurons specialized for pain, proprioceptive primary sensory neurons, cholinergic neurons of the autonomic nervous system, adrenergic neurons of the autonomic nervous system, peptidergic neurons of the autonomic nervous system, inner pillar cells of the organ of Corti, outer pillar cells of the organ of Corti, inner phalangeal cells of the organ of Corti, outer phalangeal cells of the organ of Corti, border cells, Hensen cells, supporting cells of the vestibular apparatus, supporting cells of the taste bud, supporting cells of olfactory epithelium, Schwann cells, satellite cells, enteric glial cells, neurons of the central nervous system, astrocytes of the central nervous system, oligodendrocytes of the central nervous system, anterior lens epithelial cells, lens fiber cells, melanocytes, retinal pigmented epithelial cells, iris pigment epithelial cells, oogonium, oocytes, spermatocytes, spermatogonium, ovarian follicle cells, Sertoli cells, and thymus epithelial cells, or combinations thereof.

12. A method for synthesizing a biomaterial scaffold, comprising contacting a copper capillary alginate gel having a light blue color with a solution comprising at least one stabilizing agent;

washing any excess Cu²⁺ ions from the gel; and

producing the biomaterial scaffold of claim 1;

wherein the washing step occurs before the contacting step, after the contacting step, or before and after the contacting step.

13. The method according to claim 12, wherein the stabilizing solution comprises barium hydroxide and the contacting step comprises submerging the copper capillary alginate gel in the barium hydroxide until the gel color changes to deep royal blue.

14. The method according to claim 12, wherein the at least one stabilizing agent comprises chitosan and the contacting step comprises pumping the stabilizing solution through the copper capillary alginate gel.

15. The method according to claim 14, wherein the contacting step is repeated to form a multilayered gel.

16. The method according to claim 12, wherein the at least one stabilizing agent comprises oligochitosan or a carbodiimide.

17. A method of engineering tissue, comprising providing a biomaterial scaffold of claim 1;

and implanting the biomaterial scaffold within a patient.

18. The method according to claim 17, wherein the at least one stabilizing agent is selected from the group consisting of a divalent cation, an electrostatically charged molecule, and a carbodiimide.

19. The method according to claim 17, wherein the at least one stabilizing agent comprises a barium cation.

20. The method according to claim 18, wherein the electrostatically charged molecule is selected from the group consisting of chitosan, chitosan derivatives, oligochitosan, poly-lysine, and a combination of any of the foregoing.

21. The method according to claim 17, wherein the diameter of each of the continuous parallel microtubular copper capillaries is within the range of about 3 μ

m to about 300 μ

m.

22. The method according to claim 17, wherein the diameter of each of the continuous parallel microtubular copper capillaries may be different;

and wherein any one diameter may vary between the first end and the second end.

23. The method according to claim 17, wherein the cross-section of each of the continuous parallel microtubular copper capillaries is non-circular.

24. The method according to claim 17, wherein the biomaterial scaffold possesses a positive or negative charge.

25. The method according to claim 17, wherein the biomaterial scaffold of claim 1 further comprises at least one biologically active agent, wherein the at least one biologically active agent is released into the patient after implanting.

26. The method according to claim 17, wherein the biomaterial scaffold of claim 1 further comprises at least one biologically active agent, and wherein the at least one biologically active agent comprises a plurality of cells.

27. The method according to claim 26, wherein the plurality of cells are selected from the group consisting of embryonic stem cells, adult stem cells, blast cells, cloned cells, fertilized ova, placental cells, keratinocytes, basal epidermal cells, hair shaft cells, hair-root sheath cells, surface epithelial cells, basal epithelial cells, urinary epithelial cells, salivary gland cells, mucous cells, serous cells, von Ebner'"'"'s gland cells, mammary gland cells, lacrimal gland cells, ceruminous gland cells, eccrine sweat gland cells, apocrine sweat gland cells, Moll gland cells, sebaceous gland cells, Bowman'"'"'s gland cells, Brunner'"'"'s gland cells, seminal vesicle cells, prostate gland cells, bulbourethral gland cells, Bartholin'"'"'s gland cells, Littre gland cells, uterine endometrial cells, goblet cells of the respiratory or digestive tracts, mucous cells of the stomach, zymogenic cells of the gastric gland, oxyntic cells of the gastric gland, insulin-producing β

cells, glucagon-producing α

cells, somatostatin-producing δ

cells, pancreatic polypeptide-producing cells, pancreatic ductal cells, Paneth cells of the small intestine, type II pneumocytes of the lung, Clara cells of the lung, anterior pituitary cells, intermediate pituitary cells, posterior pituitary cells, hormone secreting cells of the gut or respiratory tract, thyroid gland cells, parathyroid gland cells, adrenal gland cells, gonad cells, juxtaglomerular cells of the kidney, macula densa cells of the kidney, peripolar cells of the kidney, mesangial cells of the kidney, brush border cells of the intestine, striated duct cells of exocrine glands, gall bladder epithelial cells, brush border cells of the proximal tubule of the kidney, distal tubule cells of the kidney, nonciliated cells of ductulus efferens, epididymal principal cells, epididymal basal cells, hepatacytes, fat cells, type I pneumocytes, pancreatic duct cells, nonstriated duct cells of the sweat gland, nonstriated duct cells of the salivary gland, nonstriated duct cells of the mammary gland, parietal cells of the kidney glomerulus, podocytes of the kidney glomerulus, cells of the thin segment of the loop of Henle, collecting duct cells, duct cells of the seminal vesicle, duct cells of the prostate gland, vascular endothelial cells, synovial cells, serosal cells, squamous cells lining the perilymphatic space of the ear, cells lining the endolymphatic space of the ear, choroids plexus cells, squamous cells of the pia-arachnoid, ciliary epithelial cells of the eye, corneal endothelial cells, ciliated cells having propulsive function, ameloblasts, planum semilunatum cells of the vestibular apparatus of the ear, interdental cells of the organ of Corti, fibroblasts, pericytes of blood capillaries, nucleus pulposus cells of the intervertebral disc, cementoblasts, cementocytes, odontoblasts, odontocytes, chondrocytes, osteoblasts, osteocytes, osteoprogenitor cells, hyalocytes of the vitreous body of the eye, stellate cells of the perilymphatic space of the ear, skeletal muscle cells, heart muscle cells, smooth muscle cells, myoepithelial cells, red blood cells, megakaryocytes, monocytes, connective tissue macrophages, Langerhan'"'"'s cells, osteoclasts, dendritic cells, microglial cells, neutrophils, eosinophils, basophils, mast cells, plasma cells, helper T cells, suppressor T cells, killer T cells, immunoglobulin M, immunoglobulin G, immunoglobulin A, immunoglobulin E, killer cells, rod cells, cone cells, inner hair cells of the organ of Corti, outer hair cells of the organ of Corti, type I hair cells of the vestibular apparatus of the ear, type II cells of the vestibular apparatus of the ear, type II taste bud cells, olfactory neurons, basal cells of olfactory epithelium, type I carotid body cells, type II carotid body cells, Merkel cells, primary sensory neurons specialized for touch, primary sensory neurons specialized for temperature, primary neurons specialized for pain, proprioceptive primary sensory neurons, cholinergic neurons of the autonomic nervous system, adrenergic neurons of the autonomic nervous system, peptidergic neurons of the autonomic nervous system, inner pillar cells of the organ of Corti, outer pillar cells of the organ of Corti, inner phalangeal cells of the organ of Corti, outer phalangeal cells of the organ of Corti, border cells, Hensen cells, supporting cells of the vestibular apparatus, supporting cells of the taste bud, supporting cells of olfactory epithelium, Schwann cells, satellite cells, enteric glial cells, neurons of the central nervous system, astrocytes of the central nervous system, oligodendrocytes of the central nervous system, anterior lens epithelial cells, lens fiber cells, melanocytes, retinal pigmented epithelial cells, iris pigment epithelial cells, oogonium, oocytes, spermatocytes, spermatogonium, ovarian follicle cells, Sertoli cells, and thymus epithelial cells, or combinations thereof.

28. A method for proliferating stem cells, comprising introducing a plurality of stem cells to a biomaterial scaffold of claim 1;

and cultivating the plurality of stem cells, wherein the plurality of stem cells seed the continuous parallel microtubular copper capillaries, and wherein each of the stem cells line up within the capillaries.

29. The method according to claim 28, wherein the plurality of stem cells are cultivated in a solution comprising leukemia inhibitory factor.

30. The method according to claim 28, wherein the stem cells are embryonic stem cells;

further comprising differentiating the embryonic stem cells.

31. The method according to claim 30, wherein the differentiating step comprises adjusting the diameter of the capillaries, adjusting the cross-section of the capillaries, diffusing differentiating agents across the alginate gel, or a combination of any of the foregoing.