Co-delivery of stimulatory and inhibitory factors to create temporally stable and spatially restricted zones

1. A method for promoting cartilage regeneration, comprising:

contacting a target site of a subject with a device comprising a polymeric scaffold comprising a first and a second spatially distinct restricted zones, wherein the first zone comprises a purified morphogen that promotes cartilage regeneration, and wherein the second zone comprises a purified inhibitor of the morphogen;

wherein the morphogen and the inhibitor are released from the device into the target site;

wherein the morphogen promotes cartilage regeneration over a sustained period of time at a spatially restricted first region of the target site contacted by the first zone, thereby creating a field of action; and

wherein the inhibitor inhibits the activity of the morphogen at a spatially restricted second region of the target site contacted by the second zone, thereby creating a boundary between the first region and the second region of the target site defining where cartilage regeneration is promoted.

2. The method of claim 1 , wherein said morphogen is released at a first rate and said inhibitor is released at a second rate.

3. The method of claim 1 , wherein the morphogen that promotes cartilage regeneration is a transforming growth factor-β (TGF-β).

4. The method of claim 3 , wherein the TGF-β is selected from the group consisting of TGF-β1, TGF-β2, TGF-β3, and Latent TGF-β1 (LTGF-β1).

5. The method of claim 3 , wherein the inhibitor is an anti-TGF-β antibody.

6. The method of claim 5 , wherein the anti-TGF-β antibody is an anti-TGF-β1 antibody.

7. The method of claim 3 , wherein the inhibitor is the small molecule inhibitor SB431542.

8. The method of claim 1 , wherein the purified morphogen that promotes cartilage regeneration is Latent TGF-β1 and the purified inhibitor is the small molecule inhibitor SB431542.

9. The method of claim 1 , wherein the morphogen is encapsulated within a microsphere.

10. The method of claim 1 , wherein the inhibitor is encapsulated within a microsphere.

11. The method of claim 1 , wherein the morphogen and the inhibitor are released from the device by diffusion.

12. The method of claim 1 , wherein the polymeric scaffold comprises poly(lactic-co-glycolic) acid (PLGA).

13. The method of claim 1 , wherein said device further comprises cells.

14. The method of claim 13 , wherein said cells comprise mesenchymal stem cells, embryonic stem cells, or induced pluripotent stem cells.

15. The method of claim 1 , wherein said device further comprises:

a) a second purified morphogen, wherein the second purified morphogen promotes bone regeneration, and wherein the second purified morphogen is present in a spatially distinct restricted zone from the purified morphogen that promotes cartilage regeneration; and

b) a second purified inhibitor, wherein the second purified inhibitor inhibits the activity of the second purified morphogen, and wherein the second purified inhibitor is present in a spatially distinct restricted zone from the second purified morphogen.

16. The method of claim 15 , wherein the second purified morphogen is a bone morphogenetic protein (BMP).

17. The method of claim 16 , wherein the bone morphogenetic protein is BMP4.

18. The method of claim 16 , wherein the second purified inhibitor is an anti-BMP antibody.

19. The method of claim 18 , wherein the anti-BMP antibody is an anti-BMP4 antibody.

20. The method of claim 1 , wherein said method is performed in vivo.

21. A method of inducing cartilage regeneration in a spatially restricted region of a target site, comprising:

contacting the target site with a device comprising a polymeric scaffold comprising a first and a second spatially distinct restricted zones, wherein the first zone comprises a purified morphogen that promotes cartilage regeneration, and wherein the second zone comprises a purified inhibitor of the morphogen;

wherein the morphogen and the inhibitor are released from the device into the target site;

wherein the morphogen promotes cartilage regeneration at a spatially restricted first region of the target site contacted by the first zone, thereby creating a field of action; and

wherein the inhibitor inhibits the activity of the morphogen at a spatially restricted second region of the target site contacted by the second zone, thereby creating a boundary between the first region and the second region of the target site defining where cartilage regeneration is promoted, and inducing cartilage regeneration in a spatially restricted region of the target site.

22. The method of claim 21 , wherein the morphogen that promotes cartilage regeneration is a transforming growth factor-β (TGF-β).

23. The method of claim 22 , wherein the transforming growth factor-β is selected from the group consisting of TGF-β1, TGF-β2, TGF-β3, and Latent TGF-β1 (LTGF-β1).

24. The method of claim 22 , wherein the inhibitor is an anti-TGF-β antibody.

25. The method of claim 24 , wherein the anti-TGF-β antibody is an anti-TGF-β1 antibody.

26. The method of claim 22 , wherein the inhibitor is the small molecule inhibitor SB431542.

27. The method of claim 21 , wherein the morphogen is encapsulated within a microsphere.

28. The method of claim 21 , wherein the inhibitor is encapsulated within a microsphere.

29. The method of claim 21 , wherein the morphogen and the inhibitor are released from the device by diffusion.

30. The method of claim 21 , wherein the polymeric scaffold comprises PLGA.

31. The method of claim 21 , wherein said device further comprises cells.

32. The method of claim 31 , wherein said cells comprise mesenchymal stem cells, embryonic stem cells, or induced pluripotent stem cells.

33. The method of claim 21 , wherein the device further comprises:

a) a second purified morphogen, wherein said second purified morphogen promotes bone regeneration, and wherein the second purified morphogen is present in a spatially distinct restricted zone from the purified morphogen that promotes cartilage regeneration; and

b) a second purified inhibitor, wherein said second purified inhibitor inhibits the activity of the second purified morphogen, and wherein the second purified inhibitor is present in a spatially distinct restricted zone from the second purified morphogen.

34. The method of claim 33 , wherein the second purified morphogen is a bone morphogenetic protein (BMP).

35. The method of claim 34 , wherein the bone morphogenetic protein is BMP4.

36. The method of claim 34 , wherein the second purified inhibitor is an anti-BMP antibody.

37. The method of claim 36 , wherein the anti-BMP antibody is an anti-BMP4 antibody.

38. The method of claim 21 , wherein said method is performed in vivo.