Layered collagen and ha scaffold suitable for osteochondral repair

• US 10,052,407 B2
• Filed: 02/24/2017
• Issued: 08/21/2018
• Est. Priority Date: 01/23/2009
• Status: Active Grant

• Current Assignee: Royal College of Surgeons in Ireland
• Original Assignee: Royal College of Surgeons in Ireland
• Inventors: Gleeson, John P., Levingstone, Tanya J., O'Brien, Fergal J.
• Primary Examiner(s): Azpuru, Carlos A
• Application Number:
  US15/442,390
• Publication Number:
  US 20170157288A1
• Time in Patent Office: 543 Days
• CPC Class Codes: A61F 2/02 Prostheses implantable into the body closure means for urethra or rectum or for artificial body openings therefor A61F2/0004
• CPC Class Codes: A61F 2/30756 Cartilage endoprostheses A61F2/3603 takes precedence
• CPC Class Codes: A61F 2002/30762 Means for culturing cartilage
• CPC Class Codes: A61F 2002/30766 Scaffolds for cartilage ingrowth and regeneration
• CPC Class Codes: A61F 2210/0076 multilayered, e.g. laminated structures
• CPC Class Codes: A61L 15/28 Polysaccharides or their derivatives A61L15/225 takes precedence
• CPC Class Codes: A61L 15/325 Collagen
• CPC Class Codes: A61L 2430/02 for reconstruction of bones; weight-bearing implants
• CPC Class Codes: A61L 2430/06 for cartilage reconstruction, e.g. meniscus
• CPC Class Codes: A61L 27/12 Phosphorus-containing materials, e.g. apatite
• CPC Class Codes: A61L 27/20 Polysaccharides
• CPC Class Codes: A61L 27/24 Collagen
• CPC Class Codes: A61L 27/26 Mixtures of macromolecular compounds
• CPC Class Codes: A61L 27/3612 Cartilage, synovial fluid
• CPC Class Codes: A61L 27/3687 characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
• CPC Class Codes: A61L 27/3691 characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
• CPC Class Codes: A61L 27/3821 Bone-forming cells, e.g. osteoblasts, osteocytes, osteoprogenitor cells
• CPC Class Codes: A61L 27/3895 using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: A61L 27/425 of phosphorus containing material, e.g. apatite
• CPC Class Codes: A61L 27/46 with phosphorus-containing inorganic fillers
• CPC Class Codes: A61L 27/54 : Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: A61L 27/56 : Porous materials, e.g. foams or sponges
• CPC Class Codes: A61P 19/00 : Drugs for skeletal disorders
• CPC Class Codes: C12N 2533/54 : Collagen; Gelatin
• CPC Class Codes: C12N 2535/00 : Supports or coatings for cell culture characterised by topography
• CPC Class Codes: C12N 5/0654 : Osteocytes, Osteoblasts, Odontocytes; Bones, Teeth

Abstract

The invention relates to a method for producing a multi-layer collagen scaffold. The method generally comprises the steps of: preparing a first suspension of collagen and freezing or lyophilizing the suspension to provide a first layer; optionally preparing a further suspension of collagen and adding the further suspension onto the layer formed in the previous step to form a further layer, and freezing or lyophilizing the layers, wherein when the layer formed in the previous step is formed by lyophilization the lyophilized layer is re-hydrated prior to addition of the next layer; optionally, repeating the aforementioned step to form one or more further layers; and preparing a final suspension of collagen and pouring the final suspension onto the uppermost layer to form a final layer, and freeze-drying the layers to form the multi-layer collagen composite scaffold.

Claims

1. A cell seeded tissue engineering construct comprising cells from a host seeded onto a multi-layer collagen scaffold, wherein the cells are disposed within pores of the multi-layer scaffold and the multi-layer scaffold comprises a three freeze-dried layers in which an optional first layer comprises Type I collagen and HA;

a second layer comprises Type I collagen, a Type II collagen, a polymer and/or biologic, and HA; and

a third layer comprises Type I collagen, Type II collagen, and a polymer and/or biologic.

2. The cell seeded tissue engineering construct of claim 1, wherein the biologic is selected from the group consisting of a nucleic acid, a protein, a peptide, a cytokine, a hormone, a cell, and a growth factor.

3. The cell seeded tissue engineering construct of claim 1, wherein the polymer comprises a glycosaminoglycan (GAG).

9. A cell seeded tissue engineering construct of claim 1, wherein the multi-layer scaffold comprises:

(i) a plurality of porous freeze-dried layers in which the scaffold has a continuous pore architecture extending across the layers, and in which the pore architecture of at least two of the layers is typically different;

(ii) a plurality of porous freeze-dried layers in which the interface between each layer is seamless, and in which the pore architecture of at least two of the layers is typically different;

or (iii) continuous physical integration at the interface of each adjacent layer, wherein each layer comprises a pore architecture characteristic that is different to the other layers.

4. A cell seeded tissue engineering construct comprising cells from a host seeded onto a multi-layer collagen scaffold, wherein the cells are disposed within pores of the multi-layer scaffold and the multi-layer scaffold comprises a plurality of freeze-dried layers in which an optional first layer consists of collagen and HA;

a second layer consists essentially of a collagen, a polymer and/or biologic and HA; and

a third layer consisting essentially of collagen, wherein a ratio of HA in the first layer to HA in the second layer is at least 1;

1 (w/w), wherein the collagen component in the first layer comprises Type I collagen, and the collagen component of the second and third layers comprises Type I and II collagen.

5. The cell seeded tissue engineering construct of claim 4, wherein the ratio of HA in the first layer to HA in the second layer is at least 3:

1 (w/w).

6. The cell seeded tissue engineering construct of claim 4, wherein the scaffold has a continuous pore architecture extending across the layers.

7. The cell seeded tissue engineering construct of claim 6, wherein the porous scaffold layers are seamlessly integrated.

8. The cell seeded tissue engineering construct of claim 6, wherein scaffold comprises a pore architecture gradient extending across the layers.

10. A method for producing a cell seeded tissue engineering construct, the method comprising seeding cells from a host onto a multi-layer collagen scaffold and culturing the cells on the multi-layer scaffold prior to implantation into a defect, wherein the cells are disposed within pores of the multi-layer scaffold and the multi-layer scaffold comprises:

(i) a plurality of freeze-dried layers in which an optional first layer comprises Type I collagen and HA;

a second layer comprises Type I collagen, a Type II collagen, a polymer and/or biologic, and HA; and

a third layer comprises Type I collagen, Type II collagen, and a polymer and/or biologic;

(ii) a plurality of freeze-dried layers in an optional first layer consists of collagen and HA;

a second layer a collagen, a polymer and/or biologic and HA; and

a third layer consisting essentially of collagen, wherein a ratio of HA in the first layer to HA in the second layer is at least 1;

1 (w/w), wherein the collagen component in the first layer comprises Type I collagen, and the collagen component of the second and third layers comprises Type I and II collagen;

(iii) a plurality of porous freeze-dried layers in which the scaffold has a continuous pore architecture extending across the layers, and in which the pore architecture of at least two of the layers is typically different;

(iv) a plurality of porous freeze-dried layers in which the interface between each layer is seamless, and in which the pore architecture of at least two of the layers is typically different;

or (v) continuous physical integration at the interface of each adjacent layer, wherein each layer comprises a pore architecture characteristic that is different to the other layers.

11. The method of claim 10, further comprising preparing the multi-layer scaffold by an iterative layering technique in which each layer is independently formed by freezing, or lyophilisation followed by rehydration with an acidic solvent, prior to addition of a following layer, wherein the final layer is formed by lyophilisation.

12. The method of claim 11, wherein all layers in the scaffold are formed by lyophilisation.

13. The method of claim 10, further comprising cross-linking one or more of the layers of the scaffold.