Multilayer device for tissue engineering

• US 7,371,400 B2
• Filed: 01/02/2002
• Issued: 05/13/2008
• Est. Priority Date: 01/02/2001
• Status: Expired due to Term

• Current Assignee: Charles Stark Draper Laboratory Incorporated, The General Hospital Corporation (Mass General Brigham, Inc.)
• Original Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Inventors: Borenstein, Jeffrey T., King, Kevin R., Terai, Hidetomi, Vacanti, Joseph P.
• Primary Examiner(s): Naff; David M.
• Application Number:
  US10/038,891
• Publication Number:
  US 20020182241A1
• Time in Patent Office: 2,323 Days
• Field of Search: 424/423, 424/426, 424/93.7, 435/174, 435/177, 435/180, 435/182, 435/395
• US Class Current: 424/423
• CPC Class Codes: A61F 2/0077 Special surfaces of prostheses, e.g. for improving ingrowth A61F2/30767 takes precedence
• CPC Class Codes: A61F 2/062 Apparatus for the production of blood vessels made from natural tissue or with layers of living cells prostheses made from natural tissue or living cells A61L27/507
• CPC Class Codes: A61F 2/3094 Designing or manufacturing processes
• CPC Class Codes: A61F 2002/0086 for preferentially controlling or promoting the growth of specific types of cells or tissues of bone tissue onto implantable joints A61F2002/3093
• CPC Class Codes: A61F 2002/30004 the prosthesis being made from materials having different values of a given property at different locations within the same prosthesis
• CPC Class Codes: A61F 2002/30062 (bio)absorbable, biodegradable, bioerodable, (bio)resorbable, resorptive
• CPC Class Codes: A61F 2002/30064 Coating or prosthesis-covering structure made of biodegradable material
• CPC Class Codes: A61F 2002/30199 Three-dimensional shapes
• CPC Class Codes: A61F 2002/30925 etched
• CPC Class Codes: A61F 2002/30929 having at least two superposed coatings
• CPC Class Codes: A61F 2002/3093 for promoting ingrowth of bone tissue
• CPC Class Codes: A61F 2002/30957 using a positive or a negative model, e.g. moulds
• CPC Class Codes: A61F 2002/30971 Laminates, i.e. layered products
• CPC Class Codes: A61F 2210/0004 bioabsorbable
• CPC Class Codes: A61F 2230/0063 Three-dimensional shapes
• CPC Class Codes: A61F 2240/001 Designing or manufacturing processes
• CPC Class Codes: A61F 2240/004 Using a positive or negative model, e.g. moulds
• CPC Class Codes: A61F 2250/0014 having different values of a given property or geometrical feature, e.g. mechanical property or material property, at different locations within the same prosthesis
• CPC Class Codes: A61F 2310/00179 Ceramics or ceramic-like structures
• CPC Class Codes: A61F 2310/00329 Glasses, e.g. bioglass
• CPC Class Codes: A61K 35/12 : Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61L 27/3604 : characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
• 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 27/3808 : Endothelial cells
• CPC Class Codes: A61L 27/3891 : as distinct cell layers
• CPC Class Codes: A61L 27/56 : Porous materials, e.g. foams or sponges
• CPC Class Codes: B01L 2200/12 : Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0874 : Three dimensional network
• CPC Class Codes: B01L 2300/163 : Biocompatibility
• CPC Class Codes: B01L 3/5027 : by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502707 : characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B29C 64/129 : characterised by the energy source therefor, e.g. by global irradiation combined with a mask
• CPC Class Codes: B29C 64/40 : Structures for supporting 3D objects during manufacture and intended to be sacrificed after completion thereof
• CPC Class Codes: B29C 65/00 : Joining or sealing of preformed parts , e.g. welding of plastics materials; Apparatus therefor general aspects of processes or apparatus for joining preformed parts B29C66/00; using porous material formed by internal pressure generated therein for joining preformed parts B29C44/1228, B29C44/326
• CPC Class Codes: B29C 65/002 : Joining methods not otherwise provided for
• CPC Class Codes: B29C 65/02 : by heating, with or without pressure
• CPC Class Codes: B29C 65/48 : using adhesives , i.e. using supplementary joining material; solvent bonding
• CPC Class Codes: B29C 65/4895 : Solvent bonding, i.e. the surfaces of the parts to be joined being treated with solvents, swelling or softening agents, without adhesives
• CPC Class Codes: B29C 65/56 : using mechanical means or mechanical connections, e.g. form-fits
• CPC Class Codes: B29C 65/562 : using extra joining elements, i.e. which are not integral with the parts to be joined using plastic snap elements B29C65/58; using plastic rivets B29C65/601
• CPC Class Codes: B29C 66/5346 : said single elements being substantially flat
• CPC Class Codes: B29C 66/54 : Joining several hollow-preforms, e.g. half-shells, to form hollow articles, e.g. for making balls, containers; Joining several hollow-preforms, e.g. half-cylinders, to form tubular articles
• CPC Class Codes: B29C 66/71 : characterised by the composition of the plastics material of the parts to be joined welding bar compositions B29C65/125
• CPC Class Codes: B29C 66/7379 : degradable
• CPC Class Codes: B29C 66/73791 : biodegradable
• CPC Class Codes: B29K 2001/12 : Cellulose acetate
• CPC Class Codes: B29K 2023/00 : Use of polyalkenes or derivatives thereof as moulding material
• CPC Class Codes: B29K 2023/083 : EVA, i.e. ethylene vinyl acetate copolymer
• CPC Class Codes: B29K 2025/06 : PS, i.e. polystyrene
• CPC Class Codes: B29K 2027/06 : PVC, i.e. polyvinylchloride
• CPC Class Codes: B29K 2027/08 : PVDC, i.e. polyvinylidene chloride
• CPC Class Codes: B29K 2027/14 : PVF, i.e. polyvinyl fluoride
• CPC Class Codes: B29K 2029/04 : PVOH, i.e. polyvinyl alcohol B29K2031/04 takes precedence
• CPC Class Codes: B29K 2031/00 : Use of polyvinylesters or derivatives thereof as moulding material
• CPC Class Codes: B29K 2033/08 : Polymers of acrylic acid esters, e.g. PMA, i.e. polymethylacrylate
• CPC Class Codes: B29K 2033/12 : Polymers of methacrylic acid esters, e.g. PMMA, i.e. polymethylmethacrylate
• CPC Class Codes: B29K 2067/00 : Use of polyesters or derivatives thereof, as moulding material
• CPC Class Codes: B29K 2067/003 : PET, i.e. poylethylene terephthalate
• CPC Class Codes: B29K 2067/043 : PGA, i.e. polyglycolic acid or polyglycolide
• CPC Class Codes: B29K 2067/046 : PLA, i.e. polylactic acid or polylactide
• CPC Class Codes: B29K 2069/00 : Use of PC, i.e. polycarbonates or derivatives thereof, as moulding material
• CPC Class Codes: B29K 2071/02 : Polyalkylene oxides, e.g. PEO, i.e. polyethylene oxide, or derivatives thereof
• CPC Class Codes: B29K 2075/00 : Use of PU, i.e. polyureas or polyurethanes or derivatives thereof, as moulding material
• CPC Class Codes: B29K 2077/00 : Use of PA, i.e. polyamides, e.g. polyesteramides or derivatives thereof, as moulding material
• CPC Class Codes: B29K 2083/00 : Use of polymers having silicon, with or without sulfur, nitrogen, oxygen, or carbon only, in the main chain, as moulding material
• CPC Class Codes: B29L 2031/756 : Microarticles, nanoarticles
• CPC Class Codes: B33Y 80/00 : Products made by additive manufacturing
• CPC Class Codes: C12M 21/08 : for producing artificial tissue or for ex-vivo cultivation of tissue prostheses A61F2/00, grafts A61L27/00
• CPC Class Codes: C12M 23/16 : Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093
• CPC Class Codes: C12N 2502/14 : hepatocytes
• CPC Class Codes: C12N 2502/28 : Vascular endothelial cells
• CPC Class Codes: C12N 2533/40 : Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 2535/10 : Patterned coating
• CPC Class Codes: C12N 5/0068 : General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0671 : Three-dimensional culture, tissue culture or organ culture; Encapsulated cells
• CPC Class Codes: C12N 5/069 : Vascular Endothelial cells
• CPC Class Codes: C12N 5/0697 : Artificial constructs associating cells of different lineages, e.g. tissue equivalents blood vessels C12N5/0691
• CPC Class Codes: Y10T 428/24744 : Longitudinal or transverse tubular cavity or cell

Abstract

The invention provides for translating two-dimensional microfabrication technology into the third dimension. Two-dimensional templates are fabricated using high-resolution molding processes. These templates are then bonded to form three-dimensional scaffold structures with closed lumens. The scaffolds can serve as the template for cell adhesion and growth by cells that are added to the scaffolds through the vessels, holes or pores. These scaffolds can be formed by layering techniques, to interconnect flat template sheets to build up a fully vascularized organ.

Claims

1. A multilayer device for use in tissue engineering, comprising:

(a) at least a first layer comprised of a polymer scaffold having a pattern of microchannels therein and

(i) wherein the microchannels are suitable for the attachment and culturing of animal cells within the microchannels, and

(ii) wherein the microchannels are connected for the circulation of fluid through the first layer, and

(b) at least a second layer comprised of a polymer scaffold,

wherein the first and second layers are joined or fastened together and the first layer is formed by forming a mold from a substrate material using a photoresist processing technique that includes;

i) coating the substrate material with a photoresist; and

ii) forming a pattern in the photoresist, and casting the first layer on the respective mold.

2. The multilayer device of claim 1, wherein the polymer of the first layer is biodegradeable.

3. The multilayer device of claim 1, wherein the polymers of all the layers of the multilayer device are biodegradable.

4. The multilayer device of claim 1, wherein the second layer has a pattern of microchannels therein.

5. The multilayer device of claim 4, wherein the pattern in the first and second layers are similar.

6. The multilayer device of claim 4, wherein the pattern in the first and second layers are different.

7. The multilayer device of claim 4, wherein the pattern in the first layer is suitable for the culturing of endothelial cells and the layer in the second layer is suitable for the culturing of parenchymal cells.

8. The multilayer device of claim 4, wherein the patterns of the first and second layers are aligned to form a vasculature.

9. The multilayer device of claim 1, wherein the channels of the first layer are connected beginning from one or more inlets, expanding into more channels, and then converging back into one or more outlets.

10. The multilayer device of claim 1, wherein the channels of all the layers of the multilayer device are connected beginning from one or more inlets, expanding into more channels, and then converging back into one or more outlets.

11. The multilayer device of claim 1, further comprising a third layer comprised of a micromachined polymer scaffold suitable for attachment and culturing of animal cells, wherein the first, second and third layers are joined or fastened together.

12. The multilayer device of claim 11,

(a) wherein the second layer is unpatterned;

and

(b) wherein the third layer has a pattern of channels therein,

(i) wherein the channels are suitable for the attachment and culturing of animal cells within the channels, and

(ii) wherein the channels are connected for the circulation of fluid through the layer.

13. The multilayer device of claim 12, wherein the pattern in the first and third layers are similar.

14. The multilayer device of claim 12, wherein the pattern in the first and third layers are different.

15. The multilayer device of claim 14, wherein the pattern in the first layer is suitable for the culturing of endothelial cells and the layer in the third layer is suitable for the culturing of parenchymal cells.

16. The multilayer device of claim 1, wherein the animal cells are selected from the group consisting of endothelial cells, parenchymal cells, bone marrow cells, osteoblasts, mesenchymal stem cells, satellite cells, and fibroblasts.

17. The multilayer device of claim 1, wherein the cells cultured in the channels of the first layer are endothelial cells.

18. The multilayer device of claim 1, wherein one or more of the layers comprise through-holes.

19. The multilayer device of claim 1, wherein one or more of the layers comprise an alignment indentation on the surface of a layer and an alignment protrusion on an opposing surface of a layer, the alignment indentations shaped to mate with the alignment protrusion.

20. The multilayer device of claim 1, wherein the first layer is subdivided into zones of animal cell support.

21. The multilayer device of claim 20, wherein the zones of animal cell support comprise cell adhesion molecules.

22. The multilayer device of claim 7 or 15, wherein the pattern in the first layer comprises microchannels that are about 30-200 microns in width.

23. The multilayer device of claim 1, wherein the substrate is selected from the group consisting of silicon, ceramic, and glass.

24. The multilayer device of claim 1, wherein the second layer is formed by forming a mold from a substrate selected from the group consisting of silicon, ceramic, and glass using a photoresist processing technique, and casting the second layer on the respective mold, wherein the photoresist processing technique for the second layer includes coating the substrate with a photoresist, forming a pattern in the photoresist to form a second mold and casting the second layer on the second mold.

25. The multilayer device of claim 24, wherein the molds for the first and second layers are the same.

26. The multilayer device of claim 24, wherein the molds for the first and second layers are different.

27. The multilayer device of claim 1, wherein the polymer scaffold is selected from a material consisting of a biocompatible material, a biodegradable material, a porous material, a non-porous material and combinations thereof.

28. The multilayer device of claim 1, wherein the microchannels are a branched pattern.

29. The multilayer device of claim 1, wherein the microchannels have a height and width of about 30-200 microns.

30. The multilayer device of claim 1, wherein the photoresist in combination with the substrate material form the mold.

31. The multilayer device of claim 1, wherein forming the mold includes etching the substrate.

32. The multilayer device of claim 1, wherein forming the mold includes etching the substrate in a pattern exposed through the photoresist and stripping the photoresist therefrom.

33. The multilayer device of claim 1, wherein the photoresist processing technique further includes the step of etching the pattern into the substrate.

34. A multilayer device for use in tissue engineering, comprising:

(a) at least a first layer comprised of a polymer scaffold having a pattern of microchannels therein and

(i) wherein the microchannels are suitable for the attachment and culturing of animal cells within the microchannels,

(ii) wherein the microchannels are connected for the circulation of fluid through the first layer, and

(iii) wherein the at least a first layer is fabricated by forming a mold from a semiconductor substrate material using a photoresist processing technique that includes coating the semiconductor substrate material with a photoresist, and forming a pattern in the photoresist, and then casting the at least a first layer on the mold; and

(b) at least a second layer comprised of a polymer scaffold, wherein the first and second layers are joined or fastened together.

35. A method of making a multilayer device, comprising the steps of:

(a) fabricating at least a first layer comprised of a polymer scaffold suitable for attachment and culturing of animal cells and having a pattern of channels therein, wherein the at least a first layer is fabricated by forming a mold from a semiconductor substrate material using a photoresist processing technique that includes coating the semiconductor substrate material with a photoresist, and forming a pattern in the photoresist, and then casting the at least a first layer on the mold, and

(i) wherein the channels are suitable for the attachment and culturing of animal cells within the channels,

(ii) wherein the channels are connected for the circulation of fluid through the layer, and

(iii) wherein at least one of the channels is about 30-200 microns in width;

(b) obtaining at least a second layer for supporting animal cell growth wherein the second layer is comprised of a polymer scaffold suitable for attachment and culturing of animal cells; and

(c) fastening together the first and second layers to form lumens of the channels.

36. The method of claim 35, wherein the joining or fastening is by a method selected from the group consisting of the methods of solvent bonding;

reflow by heating;

treating the surface of the layer with oxygen plasma;

polymer flow at the surface of the layer, mechanically fastening the layers with fasteners selected from the group comprising barbs, pins, screws, clamps, staples, wires, string, and sutures; and

adhering the layers by the use of adhesives, adhesive films or adhesive layers.

37. The method of claim 35, wherein the lumens are substantially rectangular.

38. A method of making a multilayer device comprising the steps of:

(a) obtaining layers of a multilayer device, comprising;

(i) at least a first layer comprised of a polymer scaffold having a pattern of microchannels therein, and

(A) wherein the pattern of microchannels are suitable for the attachment and culturing of animal cells within the channels,

(B) wherein the pattern of microchannels are connected for the circulation of fluid through the layer; and

(C) wherein the first layer is formed by forming a first mold from a substrate material using a photoresist processing technique that includes coating the substrate material with a photoresist, and forming a pattern in the photoresist, and then casting an elastomer on the mold such that the pattern in the photoresist is transferred to the elastomer which acts as a second mold for the first layer, and

(ii) at least a second layer, wherein the second layer is comprised of a polymer scaffold for attachment and culturing of animal cells; and

(b) fastening the at least a first layer and the at least a second layer together.

39. The method of claim 38, wherein the animal cells are selected from the group consisting of endothelial cells, parenchymal cells, bone marrow cells, osteoblasts, mesenchymal stem cells, satellite cells, and fibroblasts.

40. The method of claim 38, further comprising the step of (c) seeding animal cells to into the channels.

41. The method of claim 38, wherein the animal cells are endothelial cells.

42. The method of claim 38, wherein the first layer is compression molded on the mold.

43. The method of claim 38, wherein the polymer scaffold is about 200 microns thick.

44. The method of claim 38, wherein the microchannels are about 2 microns in width.

45. The method of claim 38, further comprising the steps of implanting the multilayer device and seeding animal cells onto at least the first layer.

46. The method of claim 38, wherein the elastomer is a polydimethylsiloxane elastomer.

47. A method of implanting a bioartificial organ into a recipient, comprising:

(a) obtaining a multilayer device, comprising;

(i) at least a first layer comprised of a polymer scaffold having a pattern of microchannels therein, and

(A) wherein the microchannels are suitable for the attachment and culturing of animal cells within the channels,

(B) wherein the microchannels are connected for the circulation of fluid through the layer, and

(C) wherein the first layer is formed by forming a mold from a semiconductor substrate material using a photoresist processing technique that includes coating the semiconductor substrate material with a light sensitive photoresist, and forming a pattern in the photoresist by exposing the photoresist to short-wavelength light through a semi-transparent mask to create a microfluidic pattern in the photoresist, and then casting the first layer on the mold, and removing the first layer from the mold; and

(ii) at least a second layer wherein the second layer is comprised of a polymer scaffold, the layers of the multilayer device being fastened together; and

(b) implanting the multilayer device into the recipient, wherein the implanted multilayer device is a bioartificial organ.

48. The method of claim 47, wherein the multilayer device is seeded with animal cells.

49. The method of claim 48, wherein the multilayer device is seeded with animal cells after implanting.

50. A method as recited in claim 47, further comprising the step of etching the pattern of microchannels in the substrate.