Methods and compositions for culturing a biological tooth

• US 6,899,915 B2
• Filed: 11/29/2001
• Issued: 05/31/2005
• Est. Priority Date: 11/29/2000
• Status: Expired due to Fees

• Current Assignee: Forsyth Dental Infirmary For Children, President and Fellows of Harvard College (Harvard Corporation), The General Hospital Corporation (Mass General Brigham, Inc.)
• Original Assignee: Forsyth Dental Infirmary For Children, President and Fellows of Harvard College (Harvard Corporation), The General Hospital Corporation (Mass General Brigham, Inc.)
• Inventors: Stashenko, Phillip, Bartlett, John D., Vacanti, Joseph P., Yelick, Pamela C., Olsen, Bjorn R.
• Primary Examiner(s): Seidleck, James J.
• Assistant Examiner(s): BISSETT, MELANIE D
• Application Number:
  US09/997,734
• Publication Number:
  US 20020119180A1
• Time in Patent Office: 1,279 Days
• Field of Search: 427/2.26, 427/2.27, 433/202.1, 433/204, 433/215, 433/223, 264/19, 523/115, 521/50, 521/51, 521/55, 514/21, 424/435, 623/23.58, 623/23.72, 623/23.75
• US Class Current: 427/2.26
• CPC Class Codes: A61K 6/891 Compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: A61K 6/90 Compositions for taking dental impressions
• CPC Class Codes: A61L 27/18 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3865 Dental/periodontal tissues
• CPC Class Codes: C08L 51/085 on to polysiloxanes
• CPC Class Codes: C08L 67/04 Polyesters derived from hydroxycarboxylic acids, e.g. lactones C08L67/06 takes precedence

Abstract

Tooth tissues include the pulp mesenchyme that forms the dentin and an epithelium that is responsible for enamel formation. Cells from these tissues were obtained from porcine third molars and were seeded onto a biodegradable scaffold composed of a polyglycolic acid—polylactic acid copolymer. Cell polymer constructs were then surgically implanted into the omentum of athymic nude rats so that the constructs would have a blood supply and these tissues were allowed to develop inside the rats. Infrequently, columnar epithelial cells were observed as a single layer on the outside of the dentin-like matrix similar to the actual arrangement of ameloblasts over dentin during early tooth development. Developing tooth tissues derived from such cell polymer constructs could eventually be surgically implanted into the gum of an edentulous recipient where the construct would receive a blood supply and develop to maturity, providing the recipient with a biological tooth replacement.

Claims

1. A method to make new tooth tissue comprising, applying tooth germ cells onto a biodegradable polymer scaffold and allowing the tooth germ cells to develop into a tooth for implantation, wherein the tooth germ cells comprise cells from an enamel organ and a pulp organ.

2. The method of generating tooth tissue of claim 1, wherein the tooth germ cells are mammalian.

3. The method of generating tooth tissue of claim 1, wherein the tooth germ cells are porcine.

23. The method of generating tooth tissue of claim 1, wherein the tooth germ cells comprise cells dissociated from an enamel organ, a pulp organ, and from tissue cultured cells derived from tooth tissues.

24. The method of generating tooth tissue of claim 1, wherein the biodegradable polymer scaffold is in the shape of a human tooth.

25. The method of generating tooth tissue of claim 1, wherein the biodegradable polymer scaffold is in the shape of a tooth.

26. The method of generating tooth tissue of claim 1, wherein the scaffold is implanted into an omentum of a host animal.

27. The method of generating tooth tissue of claim 1, wherein the scaffold is implanted into the omentum of a rat.

28. The method of generating tooth tissue of claim 1, wherein the tooth germ cells are applied to the biodegradable polymer scaffold with between about 20 to 50 million cells per square inch of scaffold.

29. The method of generating tooth tissue of claim 1, wherein the biodegradable polymer scaffold is selected from the group consisting of poly(lactide), poly(glycolide), and poly(L-lactide-co-glycolide).

30. The method of generating tooth tissue of claim 1, wherein the tooth germ cells are allowed to attach to the scaffold for at least one hour prior to implanting.

31. The method of generating tooth tissue of claim 1, wherein the biodegradable polymer scaffold is coated in collagen prior to applying.

4. A method of generating tooth tissue comprising, applying tooth germ cells onto a biodegradable polymer scaffold and allowing the tooth germ cells to develop into a tooth for implantation, wherein the tooth germ cells comprise cells dissociated from an enamel organ, a pulp organ, and from tissue cultured cells derived from tooth tissues.

32. The method of generating tooth tissue of claim 4, further comprising forming a tooth mold, wherein the biodegradable polymer scaffold is formed in the tooth mold.

33. The method of generating tooth tissue of claim 4, wherein the biodegradable polymer scaffold is in the shape of a tooth.

34. The method of generating tooth tissue of claim 4, wherein the biodegradable polymer scaffold is in the shape of a human tooth.

35. The method of generating tooth tissue of claim 4, wherein the tooth germ cells are mammalian or porcine.

36. The method of generating tooth tissue of claim 4, wherein the scaffold is implanted into an omentum of a host animal.

37. The method of generating tooth tissue of claim 4, wherein the scaffold is implanted into the omentum of a rat.

38. The method of generating tooth tissue of claim 4, wherein the tooth germ cells are applied to the biodegradable polymer scaffold with between about 20 to 50 million cells per square inch of scaffold.

39. The method of generating tooth tissue of claim 4, wherein the biodegradable polymer scaffold is selected from the group consisting of poly(lactide), poly(glycolide), and poly(L-lactide-co-glycolide).

40. The method of generating tooth tissue of claim 4, wherein the tooth germ cells are allowed to attach to the scaffold for at least one hour prior to implanting.

41. The method of generating tooth tissue of claim 4, wherein the biodegradable polymer scaffold is coated in collagen prior to applying.

5. A method of generating tooth tissue comprising, applying tooth germ cells onto a biodegradable polymer scaffold and allowing the tooth germ cells to develop into a tooth for implantation, wherein the scaffold is implanted into an omentum of a host animal.

6. The method of generating tooth tissue of claim 5, wherein the scaffold is implanted into the omentum of a rat.

42. The method of generating tooth tissue of claim 5, further comprising forming a tooth mold, wherein the biodegradable polymer scaffold is formed in the tooth mold.

43. The method of generating tooth tissue of claim 5, wherein the biodegradable polymer scaffold is in the shape of a tooth.

44. The method of generating tooth tissue of claim 5, wherein the biodegradable polymer scaffold is in the shape of a human tooth.

45. The method of generating tooth tissue of claim 5, wherein the tooth germ cells are mammalian.

46. The method of generating tooth tissue of claim 5, wherein the tooth germ cells are porcine.

47. The method of generating tooth tissue of claim 5, wherein the scaffold is implanted into the omentum of a rat.

48. The method of generating tooth tissue of claim 5, wherein the tooth germ cells are applied to the biodegradable polymer scaffold with between about 20 to 50 million cells per square inch of scaffold.

49. The method of generating tooth tissue of claim 5, wherein the biodegradable polymer scaffold is selected from the group consisting of poly(lactide), poly(glycolide), and poly(L-lactide-co-glycolide).

50. The method of generating tooth tissue of claim 5, wherein the tooth germ cells are allowed to attach to the scaffold for at least one hour prior to implanting.

51. The method of generating tooth tissue of claim 5, wherein the biodegradable polymer scaffold is coated in collagen prior to applying.

7. A The method of generating tooth tissue comprising, forming a biodegradable polymer scaffold, applying tooth germ cells onto the biodegradable polymer scaffold, and implanting the scaffold into a host animal, wherein the tooth germ cells comprise cells dissociated from an enamel organ and a pulp organ.

8. The method of generating tooth tissue of claim 7, wherein the tooth germ cells are mammalian.

9. The method of generating tooth tissue of claim 7, wherein the tooth germ cells are porcine.

10. The method of generating tooth tissue of claim 7, wherein the tooth germ cells comprise cells dissociated from an enamel organ, a pulp organ, and from tissue cultured cells derived from tooth tissues.

11. The method of generating tooth tissue of claim 7, wherein the scaffold is implanted into an omentum of a host animal.

12. The method of generating tooth tissue of claim 11, wherein the scaffold is implanted into the omentum of a rat.

52. The method of generating tooth tissue of claim 7, further comprising preparing a tooth mold wherein the biodegradable polymer scaffold is formed in the tooth mold.

53. The method of generating tooth tissue of claim 7, wherein the biodegradable polymer scaffold is in the shape of a tooth.

54. The method of generating tooth tissue of claim 7, wherein the biodegradable polymer scaffold is in the shape of a human tooth.

13. A method of generating tooth tissue comprising preparing a tooth mold in the shape of a human tooth, forming a biodegradable polymer scaffold in the tooth mold, applying tooth germ cells onto the biodegradable polymer scaffold, and implanting the scaffold into an omentum of a host animal.

14. The method of generating tooth tissue of claim 13, wherein the tooth germ cells comprise cells from an enamel organ and a pulp organ.

15. The method of generating tooth tissue of claim 13, wherein the tooth germ cells are mammalian.

16. The method of generating tooth tissue of claim 13, wherein the tooth germ cells are porcine.

17. The method of generating tooth tissue of claim 13, wherein the tooth germ cells comprise cells dissociated from an enamel organ, a pulp organ, and from tissue cultured cells derived from tooth tissues.

18. The method of generating tooth tissue of claim 13, wherein the scaffold is implanted into the omentum of a rat.

19. The method of generating tooth tissue of claim 13, wherein the tooth germ cells are applied to the biodegradable polymer scaffold with between about 20 to 50 million cells per square inch of scaffold.

20. The method of generating tooth tissue of claim 13, wherein the biodegradable polymer scaffold is selected from the group consisting of poly(lactide), poly(glycolide), and poly(L-lactide-co-glycolide).

21. The method of generating tooth tissue of claim 13, wherein the tooth germ cells are allowed to attach to the scaffold for at least one hour prior to implanting.

22. The method of generating tooth tissue of claim 13, wherein the biodegradable polymer scaffold is coated in collagen prior to applying.