METHODS AND COMPOSITIONS FOR REPAIR OF CARTILAGE USING AN IN VIVO BIOREACTOR

• US 20070184033A1
• Filed: 02/05/2007
• Published: 08/09/2007
• Est. Priority Date: 02/07/2006
• Status: Active Grant

• Current Assignee: Spinalcyte LLC
• Original Assignee: Spinalcyte LLC
• Inventors: Sevrain, Lionel C., Verdier-Sevrain, Sylvie Y.
• Granted Patent: US 7,850,983 B2
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: A61F 2/30756 Cartilage endoprostheses A61F2/3603 takes precedence
• CPC Class Codes: A61F 2/3099 for temporo-mandibular [TM, TMJ] joints endoprostheses for mandibular reconstruction A61F2/2803
• CPC Class Codes: A61F 2/32 for the hip
• CPC Class Codes: A61F 2/38 for elbows or knees
• CPC Class Codes: A61F 2/3804 for elbows
• CPC Class Codes: A61F 2/40 for shoulders
• CPC Class Codes: A61F 2/441 made of inflatable pockets or chambers filled with fluid, e.g. with hydrogel
• CPC Class Codes: A61F 2002/2817 Bone stimulation by chemical reactions or by osteogenic or biological products for enhancing ossification, e.g. by bone morphogenetic or morphogenic proteins [BMP] or by transforming growth factors [TGF]
• CPC Class Codes: A61F 2002/286 Bone stimulation by mechanical vibrations for enhancing ossification
• CPC Class Codes: A61F 2002/30062 (bio)absorbable, biodegradable, bioerodable, (bio)resorbable, resorptive
• CPC Class Codes: A61F 2002/30586 having two or more inflatable pockets or chambers
• CPC Class Codes: A61F 2002/30677 Means for introducing or releasing pharmaceutical products, e.g. antibiotics, into the body
• CPC Class Codes: A61F 2002/30762 Means for culturing cartilage
• CPC Class Codes: A61F 2002/30764 Cartilage harvest sites
• CPC Class Codes: A61F 2002/30766 Scaffolds for cartilage ingrowth and regeneration
• CPC Class Codes: A61F 2002/444 for replacing the nucleus pulposus
• CPC Class Codes: A61F 2002/4445 Means for culturing intervertebral disc tissue
• CPC Class Codes: A61F 2002/445 Intervertebral disc tissue harvest sites
• CPC Class Codes: A61F 2002/4635 using minimally invasive surgery
• CPC Class Codes: A61F 2210/0004 bioabsorbable
• CPC Class Codes: A61K 2300/00 : Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 35/32 : Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
• CPC Class Codes: A61K 35/33 : Fibroblasts
• CPC Class Codes: A61K 38/18 : Growth factors; Growth regulators
• CPC Class Codes: A61K 45/06 : Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61L 2430/06 : for cartilage reconstruction, e.g. meniscus
• CPC Class Codes: A61L 27/3817 : Cartilage-forming cells, e.g. pre-chondrocytes
• CPC Class Codes: A61L 27/3852 : Cartilage, e.g. meniscus
• CPC Class Codes: A61L 27/3895 : using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: A61P 19/00 : Drugs for skeletal disorders
• CPC Class Codes: A61P 19/04 : for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 31/04 : Antibacterial agents
• CPC Class Codes: A61P 31/10 : Antimycotics
• CPC Class Codes: A61P 31/12 : Antivirals
• CPC Class Codes: A61P 41/00 : Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 2500/02 : Atmosphere, e.g. low oxygen conditions
• CPC Class Codes: C12N 2500/38 : Vitamins
• CPC Class Codes: C12N 2501/155 : Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2506/1307 : from adult fibroblasts
• CPC Class Codes: C12N 2521/00 : Culture process characterised by the use of hydrostatic pressure, flow or shear forces
• CPC Class Codes: C12N 2533/74 : Alginate
• CPC Class Codes: C12N 5/0655 : Chondrocytes; Cartilage

Abstract

Methods and compositions for the biological repair of cartilage using a hybrid construct combining both an inert structure and living core are described. The inert structure is intended to act not only as a delivery system to feed and grow a living, core component, but also as an inducer of cell differentiation. The inert structure comprises concentric internal and external and inflatable/expandable balloon-like bio-polymers. The living core comprises the cell-matrix construct comprised of HDFs, for example, seeded in a scaffold. The method comprises surgically removing a damaged cartilage from a patient and inserting the hybrid construct into the cavity generated after the foregoing surgical intervention. The balloons of the inert structure are successively inflated within the target area, such as a joint, for example. Also disclosed herein are methods for growing and differentiating human fibroblasts into chondrocyte-like cells via mechanical strain.

Claims

1. An implantable device, comprising:

a cells/scaffold composition; and

an encapsulating device that comprises;

a first membrane having an inside and an outside;

a second membrane having an inside and an outside, wherein the first membrane is encapsulated inside the second membrane;

a first volume disposed inside the first membrane;

a second volume that is disposed outside the first membrane and that is disposed inside the second membrane; and

a structure for adding fluid to the second volume, removing fluid from the second volume, or both,

wherein the cells/scaffold composition is disposed inside the first membrane and the first membrane has one or more of the following characteristics;

semi-permeable;

biocompatible;

biodegradable; and

resorbable,

wherein the second membrane has one or more of the following characteristics;

biocompatible;

hermetic to fluid;

permeable to oxygen;

resorbable;

biodegradable; and

expandable.

2. The implantable device of claim 1, wherein the scaffold is comprised of a synthetic polymer, a natural hydrogel, or a synthetic hydrogel.

3. The implantable device of claim 2, wherein the synthetic polymer is polyglycolic acid, polylactic acid, polylactic-co-glycolic acid, poly-ε

-caprolactone, or poly(glycerol-Sebacate) (PGS).

4. The implantable device of claim 2, wherein the synthetic polymer is a polyphosphazene, a polyanhydride, or a poly(orthoester).

5. The implantable device of claim 2, wherein the natural hydrogel comprises collagen, hyaluronic acid, alginate, agarose, chitosan, fibrin, gelatin, or a copolymer thereof.

6. The implantable device of claim 2, wherein the synthetic hydrogel comprises poly(ethylene oxide), poly(vinyl alcohol), poly(acrylic acid), poly(propylene fumarate-co-ethylene glycol), or a copolymer thereof.

7. The implantable device of claim 1, wherein the cells are chondrocyte cells or chondrocyte-like cells.

8. The implantable device of claim 1, wherein the chondrocyte cells or chondrocyte-like cells secrete a molecule selected from the group consisting of aggrecan, type II collagen, Sox-9 protein, cartilage link protein, and perlecan.

9. The implantable device of claim 1, wherein the cells were differentiated from fibroblast cells and/or stem cells.

10. The implantable device of claim 9, wherein the fibroblast cells are dermal fibroblasts, tendon fibroblasts, ligament fibroblasts, synovial fibroblasts, foreskin fibroblasts, or a mixture thereof.

11. The implantable device of claim 1, wherein the first membrane is comprised of a biodegradable, biocompatible, and resorbable polymer.

12. The implantable device of claim 1, wherein the first membrane is comprised of a polyacrylate, a polyvinylidene, a polyvinyl chloride copolymer, a polyurethane, a polystyrene, a polyamide, a cellulose acetate, a cellulose nitrate, a polysulfone, a polyphosphazene, a polyacrylonitrile, a poly(acrylonitrile/covinyl chloride) or a derivative, copolymer or mixture thereof.

13. The implantable device of claim 1, wherein the first membrane is generated by polyelectrolyte complexation.

14. The implantable device of claim 1, wherein the second membrane is comprised of polyglycolic acid (PGA), polylactic acid (PLA), polylactic-co-glycolic acid (PLGA), poly-ε

-caprolactone (PCL), polyurethane (PU), polydioxanone (PDO), a polyethylene, poly(glycerol sebacate) (PGS), or a derivative, copolymer, or mixture thereof.

15. The implantable device of claim 1, wherein the rate of resorbability of the second membrane is slower than the rate of resorbability of the first membrane.

16. The implantable device of claim 1, wherein the implantable device comprises one or more nutrients, growth factors, and/or medicaments.

17. The implantable device of claim 16, further defined as comprising a basal cell culture medium comprising the one or more nutrients, growth factors, and/or medicaments.

18. The implantable device of claim 17, wherein the medium is supplemented with Fetal Bovine Serum (FBS), ascorbic acid, and/or dexamethasone.

19. The implantable device of claim 16, wherein the nutrients, growth factors, and/or medicaments are present in the scaffold, the first volume, the second volume, or a combination thereof.

20. The implantable device of claim 16, wherein the growth factor is selected from the group consisting of bone morphogenetic protein 2 (BMP-2), BMP-4, BMP-6, BMP-7, cartilage-derived morphogenetic protein (CDMP), transforming growth factor beta (TGF-β

), insulin growth factor one (IGF-I), fibroblast growth factors (FGFs), basic fibroblast growth factor (bFGF), FGF-2, platelet-derived growth factor (PDGF), and a mixture thereof.

21. The implantable device of claim 16, wherein the medicament is further defined as one or more of an antibiotic, antifungal agent, or antiviral agent.

22. The implantable device of claim 1, wherein the structure comprises one or more tubes.

23. The implantable device of claim 1, wherein the structure comprises one or more catheters and/or one or more reservoirs.

24. The implantable device of claim 1, wherein the structure is further defined as comprising one or more of:

a first tube;

a second tube;

optionally, a first reservoir; and

optionally, a second reservoir.

25. The implantable device of claim 24, wherein the first and second tubes respectively comprise first ends positioned within the second volume, wherein the first and second tubes respectively comprise second ends connected to first and second reservoirs, or both.

26. The implantable device of claim 24, wherein the first and/or second tubes are comprised of the same material as the second membrane.

27. The implantable device of claim 24, wherein the first and/or second tubes are comprised of silicone rubber.

28. A method of repairing damaged cartilage in a joint of an individual, comprising delivering a device in accordance with claim 1 to the respective joint of the individual.

29. The method of claim 28, wherein the method further comprises preparing the cells/scaffold composition under suitable ex vivo conditions.

30. The method of claim 29, wherein preparing the cells/scaffold composition is defined as subjecting one or more cells to a scaffold under suitable conditions.

31. The method of claim 29, wherein the method further comprises preparing the cells/scaffold composition for no less than about two to three days.

32. The method of claim 29, wherein the suitable conditions allow proliferation of the cells.

33. The method of claim 29, wherein the suitable conditions allow the stimulation of chondrogenic differentiation.

34. The method of claim 28, wherein the cells are chondrocytes or chondrocyte-like cells derived from fibroblasts or stem cells.

35. The method of claim 34, wherein the fibroblasts cells are dermal fibroblasts, tendon fibroblasts, ligament fibroblasts, synovial fibroblasts, or foreskin fibroblasts.

36. The method of claim 34, wherein the fibroblasts are dermal fibroblasts.

37. The method of claim 29, wherein the suitable conditions are further defined as being under high density micromass culture, being under low oxygen tension (between about 1.0%-7.5%), being under mechanical stress, and/or being fed by a medium supplemented with growth factors, ascorbic acid, and/or dexamethasone.

38. The method of claim 28, wherein the cells/scaffold composition is subjected to mechanical stress.

39. The method of claim 38, wherein the mechanical stress is defined as hydrostatic pressure, fluid shear stress, or a combination thereof.

40. The method of claim 38, wherein the mechanical stress is intermittent.

41. The method of claim 38, wherein the mechanical stress is fluid shear stress and the scaffold is microfluidic scaffold.

42. The method of claim 28, wherein the delivering step is defined as implanting the device using minimally invasive surgery.

43. The method of claim 28, wherein following implantation of the device into the individual, the second membrane is inflated to fill a void in the joint.

44. The method of claim 28, wherein the joint is an intervertebral disc and wherein prior to delivery of the device to the intervertebral disc of the individual, at least part of an endogenous intervertebral disc was removed from the individual.

45. The method of claim 28, wherein the joint is an intervertebral disc, a knee, a shoulder, an elbow, a hip, or a temporo-mandibular joint.

46. The method of claim 28, wherein at least part of the second volume is exchanged.

47. The method of claim 28, wherein the structure of the device comprises:

a first tube having first and second ends, said first end of the first tube disposed within the second volume;

a second tube having first and second ends, said first end of the second tube disposed within the second volume;

a first reservoir; and

a second reservoir,

wherein following delivery of the device to an intervertebral disc in the individual and following inflation of the second membrane, the second ends of the first and second tubes are respectively connected to the first and second reservoirs.

48. The method of claim 47, wherein the first and second reservoirs are subcutaneously positioned in the individual.

49. The method of claim 28, further comprising sealing the first membrane, sealing the second membrane, or both.

50. The method of claim 47, further comprising removing at least part of the second volume through the first reservoir.

51. The method of claim 47, further comprising removing fluid from the first or second reservoir, delivering a fluid to the respective second or first reservoir, or concomitantly removing fluid from the first or second reservoir and delivering a fluid to the respective second or first reservoir.

52. The method of claim 47, wherein the cells/scaffold composition is inserted into the first membrane prior to delivery of the device into the individual or wherein the cells/scaffold composition is inserted into the first membrane subsequent to delivery of the device into the individual.

53. The method of claim 47, wherein the first membrane is inserted into the second membrane prior to delivery of the device into the individual or wherein the first membrane is inserted into the second membrane subsequent to delivery of the device into the individual.

56. A kit comprising the device of claim 1, wherein the device is housed in one or more suitable containers.

57. The kit of claim 56, wherein the kit further comprises cells that are chondrocyte cells, chondrocyte-like cells, or cells that are capable of differentiating to chondrocyte cells or chondrocyte-like cells.

54. A method of preparing a cells/scaffold composition, wherein the cells are chondrocytes or chondrocyte-like cells, comprising:

subjecting cells capable of differentiating into a chondrocyte-like cell to the scaffold;

subjecting the cells to mechanical stress; and

optionally subjecting the cells to one or more growth factors suitable for differentiation to a chondrocyte or chondrocyte-like cell.

55. The method of claim 54, wherein the mechanical stress is intermittent.

58. An implantable device, comprising:

a cells/scaffold composition encapsulated inside a membrane, said membrane having an inside and an outside; and

a structure for exchanging at least part of fluid that is inside the membrane,

wherein the membrane has one or more of the following characteristics;

semi-permeable;

biocompatible;

biodegradable; and

resorbable.

59. A hybrid structure for cartilage repair, comprising:

an encapsulating device comprising inert material; and

a living core comprising chondrocyte-like cells,

wherein said encapsulating device encapsulates the living core.

60. An in vivo bioreactor for cartilage engineering, comprising a device that encapsulates cells, wherein said cells are capable of differentiating to chondrocytes or chondrocyte-like cells, wherein the encapsulation of said cells provides suitable conditions for in vivo growth and differentiation of said cells, wherein said conditions comprise providing a physiologic loading regimen on said cells.

61. The bioreactor of claim 60, wherein the physiologic loading regimen comprises force from a spine of an individual.