Systems and methods for screening pharmaceutical chemicals

• US 7,449,306 B2
• Filed: 08/14/2002
• Issued: 11/11/2008
• Est. Priority Date: 08/14/2001
• Status: Expired due to Term

• Current Assignee: Washington University In St Louis
• Original Assignee: Washington University In St Louis
• Inventors: McConnaughey, William B., Elson, Elliot, Wakatsuki, Tetsuro
• Primary Examiner(s): Marx; Irene
• Assistant Examiner(s): HANLEY, SUSAN MARIE
• Application Number:
  US10/219,097
• Publication Number:
  US 20030064358A1
• Time in Patent Office: 2,281 Days
• Field of Search: 435/40.5, 435/40.52, 435/371, 435/366, 435/325
• US Class Current: 435/40.52
• CPC Class Codes: G01N 2203/0005 Repeated or cyclic
• CPC Class Codes: G01N 2203/0016 Tensile or compressive
• CPC Class Codes: G01N 2203/0073 Fatigue
• CPC Class Codes: G01N 2203/0282 Two dimensional, e.g. tapes, webs, sheets, strips, disks or membranes
• CPC Class Codes: G01N 3/38 generated by electromagnetic means
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/502 for testing non-proliferative effects
• CPC Class Codes: G01N 33/5044 involving specific cell types
• CPC Class Codes: G01N 33/5061 Muscle cells
• CPC Class Codes: G01N 33/5064 Endothelial cells
• CPC Class Codes: G01N 33/5088 of vertebrates

Abstract

A method for obtaining a response of a tissue model system to an activator includes contacting a bio-artificial tissue model system with an activator and measuring cellular mechanical response thereto of at least one of contractile force and tissue stiffness. A method for obtaining a response of a tissue model system to an activator includes contacting a bio-artificial tissue model system with an activator and measuring cellular mechanical response thereto of at least one of contractile force and hysteresis.

Claims

1. A method of measuring a tissue response comprising:

a) forming and suspending a bio-artificial tissue on a scaffold to yield a bio-artificial tissue overlaying the scaffold without a fastener to facilitate tissue adhesion;

b) contacting a bio-artificial tissue with a test agent;

c) applying a mechanical force to the contacted bio-artificial tissue of step (b); and

d) measuring a mechanical response of the contacted bio-artificial tissue on the scaffold to the test agent, the mechanical response being at least one of tissue stiffness and contractile force of the bio-artificial issue.

2. The method of claim 1, wherein the mechanical response comprises tissue stiffness and contractile force.

3. The method of claim 1, wherein measuring the tissue stiffness includes measuring at least one of hysteresis, phase lag and dynamic stiffness.

4. The method of claim 1, wherein the bio-artificial tissue comprises cells and collagen.

5. The method of claim 1, wherein the bio-artificial tissue comprises muscle cells.

6. The method of claim 1, wherein the bio-artificial tissue comprises non-muscle cells.

7. The method of claim 1, wherein the bio-artificial tissue comprises endothelial cells.

8. The method of claim 1, wherein the bio-artificial tissue comprises cardiac cells.

9. The method of claim 1, wherein the bio-artificial tissue comprises extracellular matrix.

10. The method of claim 1, wherein the bio-artificial tissue comprises muscle cells and non-muscle cells.

11. The method of claim 1, wherein the bio-artificial tissue is suspended from a scaffold positioned within a multi-well plate to provide an array of locations in which responses are measured.

12. The method of claim 1, wherein the mechanical response is measured using a tissue indentation system.

13. The method of claim 12, wherein the tissue indentation system comprises an indentation probe coupled to a force transducer and a computer.

14. The method of claim 1, wherein a plurality of bio-artificial tissues are contacted with varying concentrations of the test agent to generate a dose response profile of the bio-artificial tissue to the test agent.

15. The method of claim 1, further comprising comparing the mechanical response of the bio-artificial tissue to contact with the test agent to the mechanical response of the bio-artificial tissue to contact with a second test agent.

16. The method of claim 15, wherein the mechanical responses are ranked based on a comparison of the mechanical responses of the bio-artificial tissue to a plurality of test agents.

17. The method of claim 1, wherein the bio-artificial tissue comprises cells known to be involved in a disease.

18. The method of claim 1, further comprising correlating the mechanical response with a cell present in the bio-artificial tissue.

19. The method of claim 1, further comprising:

e) applying a mechanical force to the bio-artificial tissue and measuring the mechanical response of the bio-artificial tissue prior to contact with the test agent in step (b); and

f) comparing the mechanical response of the bio-artificial tissue contacted with the test agent to the mechanical response of the bio-artificial tissue prior to contact with the test agent.

20. A method for screening the effects of pharmaceutical agents comprising:

a) forming and suspending a bio-artificial tissue on a scaffold to yield a bio-artificial tissue overlaying the scaffold without a fastener to facilitate tissue adhesion;

b) contacting the bio-artificial tissue with a pharmaceutical agent;

c) applying a mechanical force to the contacted bio-artificial tissue of step (b);

d) measuring a parameter of tissue stiffness or contractile force of the bio-artificial tissue on the scaffold;

e) repeating steps (a) through (d) with at least another pharmaceutical agent; and

f) identifying pharmaceutical agents that alter the tissue stiffness or contractile force of the bio-artificial tissue as compared to the tissue stiffness or contractile force of a bio-artificial tissue that is not contacted with the pharmaceutical agent.

21. The method of claim 20, wherein the parameter comprises tissue stiffness and contractile force.

22. The method of claim 20, wherein the tissue stiffness includes at least one of hysteresis, phase lag and dynamic stiffness.

23. The method of claim 20, wherein the bio-artificial tissue comprises cells and collagen.

24. The method of claim 20, wherein the bio-artificial tissue comprises muscle cells.

25. The method of claim 20, wherein the bio-artificial tissue comprises non-muscle cells.

26. The method of claim 20, wherein the bio-artificial tissue comprises endothelial cells.

27. The method of claim 20, wherein the bio-artificial tissue comprises cardiac cells.

28. The method of claim 20, wherein the bio-artificial tissue comprises muscle cells and non-muscle cells.

29. The method of claim 20, wherein the bio-artificial tissue comprises extracellular matrix.

30. The method of claim 20, wherein the bio-artificial tissue is suspended from a scaffold positioned within a well in a multi-well plate to provide an array of locations in which responses are measured.

31. The method of claim 20, wherein the tissue stiffness is measured using a tissue indentation system.

32. The method of claim 31, wherein the tissue indentation system comprises an indentation probe coupled to a force transducer and a computer.

33. The method of claim 20, wherein a plurality of bio-artificial tissues are contacted with varying concentrations of the pharmaceutical agent to generate a dose response profile of the bio-artificial tissue to the pharmaceutical agent.

34. The method of claim 20, further comprising comparing the response of the bio-artificial tissue to contact with the pharmaceutical agent to the response of the bio-artificial tissue to contact with a second pharmaceutical agent.

35. The method of claim 34, wherein the pharmaceutical agents are ranked based on a comparison of the tissue stiffness or contractile force of the bio-artificial tissue to a plurality of pharmaceutical agents.

36. The method of claim 20, wherein the bio-artificial tissue comprises cells known to be involved in a disease.

37. The method of claim 20, further comprising correlating the tissue stiffness or contractile force with a cell present in the bio-artificial tissue.

38. The method of claim 20, further comprising:

g) applying a mechanical force to the bio-artificial tissue and measuring the mechanical response of the bio-artificial tissue prior to contact with the pharmaceutical agent in step (b); and

h) comparing the mechanical response of the bio-artificial tissue contacted with the pharmaceutical agent to the mechanical response of the bio-artificial tissue prior to contact with the pharmaceutical agent.

39. A tissue response system, comprising:

a) a plurality of bio-artificial tissues suspended from and overlaying a plurality of scaffolds without a fastener to facilitate tissue adhesion to the scaffold;

each of the plurality of scaffolds positioned within a well in a multi-well plate to provide an array of locations in which responses of the bio-artificial tissue on the scaffold to a test agent can be measured; and

b) a tissue indentation system for applying a mechanical force and measuring a mechanical response of the bio-artificial tissue, the system comprising a probe coupled to a force transducer and a computer, the probe capable of contacting the bio-artificial tissue and measuring a response of the bio-artificial tissue to the test agent, the response being at least one of contractile force and tissue stiffness.

40. A tissue response system, comprising:

a) a bio-artificial tissue formed on and overlaying a scaffold, the bio-artificial tissue suspended without a fastener to facilitate tissue adhesion to the scaffold;

b) a well for holding the scaffold, wherein the suspended bio-artificial tissue can be contacted with a test agent; and

c) a tissue indentation system for applying a mechanical force and measuring a mechanical response of the bio-artificial tissue, the system comprising an indentation probe coupled to a force transducer and a computer, the probe capable of contacting the bio-artificial tissue on the scaffold and measuring a response of the bio-artificial tissue to the test agent, the response being at least one of contractile force and tissue stiffness.

41. The system of claim 40, wherein the response comprises tissue stiffhess and contractile force.

42. The system of claim 40, wherein tissue stififiess includes at least one of hysteresis, phase lag and dynamic stiffness.

43. The system of claim 40, wherein the bio-artificial tissue comprises cells and collagen.

44. The system of claim 40, wherein the bio-artificial tissue comprises muscle cells.

45. The system of claim 40, wherein the bio-artificial tissue comprises non-muscle cells.

46. The system of claim 40, wherein the bio-artificial tissue comprises endothelial cells.

47. The system of claim 40, wherein the bio-artificial tissue comprises cardiac cells.

48. The system of claim 40, wherein the bio-artificial tissue comprises muscle cells and non-muscle cells.

49. The system of claim 40, wherein the bio-artificial tissue comprises extracellular matrix.

50. The system of claim 40, wherein the scaffold is positioned within a well in a multi-well plate to provide an array of locations in which responses are measured.

51. The system of claim 40, wherein a plurality of bio-artificial tissues suspended from a plurality of scaffolds is contacted with varying concentrations of the test agent, and the measured response is used to generate a dose response profile of the bio-artificial tissue to the test agent.

52. The system of claim 40, wherein the response of the bio-artificial tissue to contact with the test agent is compared to the response of the bio-artificial tissue to contact with a second test agent.

53. The system of claim 52, wherein the test agents are ranked based on a comparison of the tissue stiffness or contractile force of the bio-artificial tissue to a plurality of agents.

54. The system of claim 40, wherein the bio-artificial tissue comprises cells known to be involved in a disease.

55. The system of claim 40, wherein the response is associated with a cell present in the bio-artificial tissue.

56. A method of measuring a tissue response comprising:

a) forming and suspending a bio-artificial tissue on a scaffold to yield a bio-artificial tissue overlaying the scaffold without a fastener to facilitate tissue adhesion;

b) contacting the bio-artificial tissue with an agent known to alter a mechanical response of a tissue;

c) contacting the bio-artificial tissue of step (b) with a test agent;

d) applying a mechanical force to the contacted bio-artificial tissue of step (c);

e) measuring a mechanical response of the contacted bio-artificial tissue on the scaffold, the mechanical response being at least one of contractile force and tissue stiffness of the bio-artificial tissue; and

f) comparing the mechanical response of the bio-artificial tissue contacted with both the agent known to alter a mechanical response of a tissue and the test agent to the mechanical response of at least one of (1) the bio-artificial tissue prior to contact with any agent;

(2) a bio-artificial tissue only contacted with an agent known to alter the mechanical response of a tissue;

or (3) a bio-artificial tissue only contacted with the test agent.

57. The method of claim 56, wherein the bio-artificial tissue is contacted with the test agent prior to being contacted with the agent known to alter a mechanical response.

58. The method of claim 56, wherein the bio-artificial tissue is contacted with the test agent at the same time as being contacted with the agent known to alter a mechanical response.

59. The method of claim 56, wherein the bio-artificial tissue is contacted with the test agent after being contacted with the agent known to alter a mechanical response.

60. The method of claim 56, wherein the mechanical response comprises tissue stiffness and contractile force.

61. The method of claim 56, wherein the tissue stiffness includes at least one of hysteresis, phase lag and dynamic stiffness.

62. The method of claim 56, wherein the bio-artificial tissue comprises cells and collagen.

63. The method of claim 56, wherein the bio-artificial tissue comprises muscle cells.

64. The method of claim 56, wherein the bio-artificial tissue comprises non-muscle cells.

65. The method of claim 56, wherein the bio-artificial tissue comprises endothelial cells.

66. The method of claim 56, wherein the bio-artificial tissue comprises cardiac cells.

67. The method of claim 56, wherein the bio-artificial tissue comprises muscle cells and non-muscle cells.

68. The method of claim 56, wherein the bio-artificial tissue comprises extracellular matrix.

69. The method of claim 56, wherein the bio-artificial tissue is suspended from a scaffold positioned within a well in a multi-well plate to provide an array of locations in which responses are measured.

70. The method of claim 56, wherein the tissue stiffness is measured using a tissue indentation system.

71. The method of claim 70, wherein the tissue indentation system comprises an indentation probe coupled to a force transducer and a computer.

72. A method of measuring a tissue response comprising:

a) positioning a scaffold within a well in a multi-well plate;

b) forming and suspending a bio-artificial tissue on the scaffold to yield a bio-artificial tissue overlaying the scaffold without a fastener to facilitate tissue adhesion;

c) contacting the bio-artificial tissue with a test agent;

d) applying a mechanical force to the contacted bio-artificial tissue of step (c); and

e) measuring a mechanical response of the contacted bio-artificial tissue on the scaffold to the test agent, the mechanical response being at least one of tissue stiffness and contractile force and comparing the mechanical response to a mechanical response of a bio-artificial tissue that is not contacted with the test agent.

73. A method of measuring a tissue response comprising:

a) forming and suspending a bio-artificial tissue on a scaffold to yield a bio-artificial tissue overlaying the scaffold without a fastener to facilitate tissue adhesion;

b) contacting the bio-artificial tissue with a test agent;

c) applying a mechanical force to the contacted bio-artificial tissue of step (b) using a tissue indentation system; and

d) measuring a mechanical response of the contacted bio-artificial tissue on the scaffold to the test agent using the tissue indentation system, the mechanical response being at least one of tissue stiffness and contractile force and comparing the mechanical response to a mechanical response of a bio-artificial tissue that is not contacted with the test agent.

74. The method of claim 73, wherein the tissue indentation system comprises an indentation probe coupled to a force transducer and a computer.

75. A method for screening the effects of pharmaceutical agents comprising:

a) forming and suspending a bio-artificial tissue on a scaffold to yield a bio-artificial tissue overlaying the scaffold without a fastener to facilitate tissue adhesion;

b) contacting the bio-artificial tissue with a pharmaceutical agent;

c) applying a mechanical force to the contacted bio-artificial tissue of step (b) using a tissue indentation system;

d) measuring a parameter of tissue stiffness or contractile force of the bio-artificial tissue on the scaffold to the pharmaceutical agent using the tissue indentation system;

e) repeating steps (a) through (d) with at least another pharmaceutical agent; and

f) identifying pharmaceutical agents that alter the tissue stiffness or contractile force of the contacted bio-artificial tissue as compared to the tissue stiffness or contractile force of a bio-artificial tissue that is not contacted with the pharmaceutical agent.

76. The method of claim 75, wherein the tissue indentation system comprises an indentation probe coupled to a force transducer and a computer.

77. A method of measuring a tissue response comprising:

a) positioning a scaffold within a well in a multi-well plate;

b) forming and suspending a bio-artificial tissue on the scaffold to yield a bio-artificial tissue overlaying the scaffold without a fastener to facilitate tissue adhesion;

c) contacting the bio-artificial tissue with an agent known to alter a mechanical response of a tissue;

d) applying a mechanical force to the contacted bio-artificial tissue of step (c);

e) measuring a mechanical response of the contacted bio-artificial tissue on the scaffold, the mechanical response being at least one of contractile force and tissue stiffness of the bio-artificial tissue;

f) contacting the bio-artificial tissue of step (e) with a test agent;

g) applying a mechanical force to the contacted bio-artificial tissue of step (f);

h) measuring a mechanical response of the contacted bio-artificial tissue of step (f) on the scaffold, the mechanical response being at least one of contractile force and tissue stiffness of the bio-artificial tissue; and

i) comparing the response of the bio-artificial tissue contacted with the agent known to alter a mechanical response from step (e) to the response of the bio-artificial tissue further contacted with the test agent from step (h).

78. A method of measuring a tissue response comprising:

a) forming and suspending a bio-artificial tissue on a scaffold to yield a bio-artificial tissue overlaying the scaffold without a fastener to facilitate tissue adhesion;

b) contacting the bio-artificial tissue with an agent known to alter a mechanical response;

c) contacting the bio-artificial tissue with a test agent;

d) applying a mechanical force to the contacted bio-artificial tissue of step (c);

e) measuring tissue stiffness of the bio-artificial tissue of step (d) using a tissue indentation system, wherein said bio-artificial tissue remains suspended without a fastener to facilitate tissue adhesion from said scaffold; and

f) comparing the tissue stiffness of a bio-artificial tissue contacted with the agent known to alter a mechanical response to the tissue stiffness of the bio-artificial tissue contacted with both the agent known to alter a mechanical response and the test agent.

79. The method of claim 78, wherein the tissue indentation system comprises an indentation probe coupled to a force transducer and a computer.