Method for use of a double-structured tissue implant for treatment of tissue defects

1. A method for preparing an implant for tissue or cartilage repair, the method comprising the steps of:

providing an acellular, three-dimensional scaffold comprising collagen and defining a plurality of pores;

disposing a composition within the plurality of pores such that the composition fills the plurality of pores, wherein the composition has a basic pH and comprises a collagenous solution or gel and a surfactant; and

subjecting the scaffold and the composition disposed therein to a dehydrothermal treatment, thereby stabilizing the composition to form a fibrous and cross-linked collagen network within the plurality of pores, wherein, prior to the dehydrothermal treatment, the scaffold and composition are subjected to precipitation, freezing, and dehydration.

2. The method of claim 1 , wherein the acellular, three-dimensional collagen scaffold comprises a sponge, a honeycomb or honeycomb-like scaffold, or a thermo-reversible hydrogel.

3. The method of claim 1 , wherein the acellular, three-dimensional collagen scaffold is prepared from a compound selected from the group consisting of Type I collagen, Type II collagen, Type III collagen, Type IV collagen, Type V collagen.

4. The method of claim 1 , wherein the plurality of pores are randomly or non-randomly organized pores that are substantially uniform in size with a defined diameter.

5. The method of claim 1 , wherein the plurality of pores are vertically orientated and have a substantially homogenous defined diameter.

6. The method of claim 1 , wherein the collagenous solution or gel is selected from the group consisting of Type 1 collagen, Type II collagen, methylated collagen, gelatin, methylated gelatin, and any combination thereof.

7. The method of claim 1 , wherein the fibrous collagen network interdigitates within and among the pores.

8. The method of claim 1 , wherein the surfactant is ionic or non-ionic.

9. The method of claim 1 , wherein the surfactant is a derivatized polyethylene glycol or a block co-polymer of polyoxyethylene (PEO) and polyoxypropylene (PPO) having the generic organization of polymeric blocks PEG-PPO-PEG or PPO-PEG-PPO.

10. The method of claim 9 , wherein the derivatized polyethylene glycol is polyethylene glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether.

11. The method of claim 9 , wherein the block co-polymer is a polymer of polyoxyethylene (PEO) and polyoxypropylene (PPO) with two 96-unit hydrophilic PEO chains surrounding one 69-unit hydrophobic PPO chain.

12. The method of claim 1 , further comprising seeding the acellular, three-dimensional scaffold, the composition, or both with one or more cells.

13. The method of claim 12 , wherein the cells are differentiated cells or undifferentiated cells.

14. The method of claim 13 , wherein the differentiated cells comprise chondrocytes, osteoblasts, tenocytes, fibroblasts, fibrochondrocytes, and ligament cells.

15. The method of claim 13 , wherein the undifferentiated cells comprise mesenchymal cells derived from bone marrow aspirate, cells derived from an iliac crest needle biopsy, an immortalized cell line, hematopoietic stem cells, neural stem cells, embryonic stem cells or stem cells derived from a dissociable mesenchymal tissue.

16. The method of claim 15 , wherein the dissociable mesenchymal tissue is a somite, a muscle, or an interstitial connective tissue.

17. The method of claim 1 , further comprising

introducing a cell modulator into the acellular, three-dimensional collagen scaffold, the composition, or both.

18. The method of claim 17 , wherein the cell modulator is selected from a group consisting of a pharmaceutical agent, a growth factor, a growth hormone, a mediator, an enzyme promoting cell incorporation, an enzyme promoting cell proliferation, an enzyme promoting cell division, a pharmaceutically acceptable excipient, an additive, and a buffer.

19. The method of claim 1 , wherein the dehydrothermal treatment comprises subjecting the scaffold and composition disposed therein to a temperature between about 70° C. to about 200° C., for about 30 minutes to about 7 days, under a vacuum.