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1. (WO2018006037) INNERVATED ARTIFICIAL SKIN
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CLAIMS

We Claim:

1. A multi-layer silk composition comprising

a first layer comprising silk fibroin and keratinocytes;

a second layer comprising silk fibroin and fibroblasts;

a third layer comprising silk fibroin and adipocytes; and

a plurality of nervous system cells,

wherein at least some of the plurality of nervous system cells span at least two layers.

2. The composition of claim 1, wherein all of the cells in each of the first, second and third layers are human cells.

3. The composition of claim 2, wherein the human cells are primary cells or cell lines.

4. The composition of any one of claims 1-3, wherein the nervous systems cells comprise at least one of neurons, glia, and neural stem cells.

5. The composition of any one of claims 1-4, wherein at least a plurality of the nervous system cells are functional.

6. The composition of any one of claims 1-5, further comprising a plurality of at least one type of immune cells.

7. The composition of claim 6, wherein the at least one type of immune cells includes macrophages.

8. The composition of any one of claims 1-7, further comprising a plurality of at least one type of endothelial cell.

9. The composition of claim 8, wherein the endothelial cells comprise a monolayer.

10. The composition of any one of the above claims, further comprising an electrical device that is functionally connected to at least some of the plurality nervous system cells.

11. The composition of claim 10, wherein activation of the electrical device results in the firing of one or more neurons.

12. The composition of claim 10 or claim 11, wherein the electrical device comprises at least one electrode.

13. The composition of any one of claims 10-12, wherein the electrical device comprises silk fibroin.

14. The composition of any one of the above claims, further comprising at least one of collagen, laminin, fibronectin, hyaluronic acid, fibrinogen, sulfated glycosaminoglycans, and/or one or more growth factors, in at least one layer.

15. The composition of claim 14, wherein the one or more growth factors comprise epidermal growth factor, fibroblast growth factor, nerve growth factor, platelet-derived growth actor, insulin-like growth factor, or tumor necrosis factor-β, and combinations thereof.

16. The composition of any one of the above claims, wherein at least one layer is exposed to an air-liquid interface.

17. The composition of any one of the above claims, wherein the third layer comprises lipoaspirate.

18. A method of making a multi-layer silk composition comprising

providing a first layer comprising silk fibroin and keratinocytes;

providing a second layer comprising silk fibroin and fibroblasts;

providing a third layer comprising silk fibroin and adipocytes;

providing a plurality of nervous system cells; and

associating the first layer, second layer, third layer, and plurality of nervous system cells to form a multi-layer silk composition, wherein at least some of the plurality of nervous system cells span at least two layers.

19. The method of claim 18, wherein all of the cells in each of the first, second and third layers are human cells.

20. The method of claim 19, wherein the human cells are primary cells or cell lines.

21. The method of any one of claims 18-20, wherein the nervous system cells comprise at least one of neurons, glia, and neural stem cells.

22. The method of any one of claims 18-21, wherein at least a plurality of the nervous system cells are functional.

23. The method of any one of claims 18-22, further comprising providing a plurality of at least one type of immune cell, and associating the plurality of at least one type of immune cell with the multi-layer silk composition.

24. The method of claim 23, wherein the at least one type of immune cell includes macrophages.

25. The method of any one of claims 18-24, further comprising providing a plurality of at least one type of endothelial cell, and associating the plurality of at least one type of endothelial cell with the multi-layer silk composition.

26. The method of claim 25, wherein the endothelial cells comprise a monolayer.

27. The method of any one of claims 18-26, further comprising providing an electrical device, and functionally associating the electrical device to at least some of the plurality nervous system cells.

28. The method of claim 27, wherein activation of the electrical device results in the firing of one or more neurons.

29. The method of claim 27 or claim 28, wherein the electrical device comprises at least one electrode.

30. The method of any one of claims 27-29, wherein the electrical device comprises silk fibroin.

31. The method of any one of claims 18-30, further comprising at least one of collagen, laminin, fibronectin, hyaluronic acid, fibrinogen, sulfated glycosaminoglycans, and/or one or more growth factors, in at least one layer.

32. The method of claim 31, wherein the one or more growth factors comprise epidermal growth factor, fibroblast growth factor, nerve growth factor, platelet-derived growth actor, insulin-like growth factor, or tumor necrosis factor-β, and combinations thereof.

33. The method of any one of claims 18-32, wherein at least one layer is exposed to an air-liquid interface.

34. The method of any one of claims 18-33, wherein the third layer comprises lipoaspirate.

35. A method compri sing

providing a composition according to any one of claims 1-16, wherein at least some of the cells in the composition are cancer cells;

exposing the composition to one or more therapeutic agents and/or inflammatory agents;

characterizing the response of the cells in the composition to the one or more therapeutic agents.

36. The method of claim 35, wherein the one or more therapeutics agents comprise at least one cancer therapeutic agent.

37. The method of claim 35, wherein the one or more inflammatory agents are selected from the group consisting of lipopolysaccharide, an allergen, and a toxin.

38. The method of claim 35, wherein the inflammatory agents comprise a pro-inflammatory agent.

39. The method of claim 35, wherein the inflammatory agents comprise an anti-inflammatory agent.