TISSUE ENGINEERING SCAFFOLDS

Abstract

A scaffold (12) for tissue engineering comprises an inner portion (14), an outer portion (16), and a base portion (22) connecting the inner portion and the outer portion. The inner portion (14) comprises a channel (18) surrounded by a first set of one or more walls. The outer portion (16) comprises a second set of one or more walls. The portions are arranged such that the second set of one or more walls substantially surrounds the first set of one or more walls with a spacing between the first and second sets of walls defining a cavity (20) between the inner portion (14) and the outer portion (16). The inner portion (14) and the outer portion (16) may have different shapes; and/or the scaffold (12) may further comprise a filler material in the cavity (20) defined between the inner and outer portions.

Claims

1.-40. (canceled)

41. A device for tissue engineering comprising a scaffold, the scaffold comprising: an inner portion comprising a channel surrounded by a first set of one or more ways an outer portion comprising a second set of one or more walls and arranged such that the second set of one or more walls substantially surrounds the first set of one or more walls with a spacing between the first and second sets of one or more walls defining a cavity between the inner portion and the outer portion; and a base portion connecting the inner portion and the outer portion; wherein the device comprises at least one of the following features (i) or (ii): (i) the inner portion and the outer portion have different shapes; or (ii) a filler material is arranged in the cavity defined between the inner and outer portions.

42. The device of claim 41, wherein the inner portion consists of a substantially solid material comprising a plurality of apertures in the first set of one or more walls.

43. The device of claim 41, wherein the outer portion consists of a substantially solid material comprising a plurality of apertures in the second set of one or more walls.

44. The device of claim 41, wherein: the first set of one or more walls of the inner portion comprises a plurality of apertures arranged in a. first set of layers spaced along the length of the first set of one or more walls by a first interval; the second set of one or more walls of the outer portion comprises a plurality of apertures arranged in a second set of layers spaced along the length of the second set of one or more walls by a second interval; the first set of layers is arranged to coincide with the second intervals between the second set of layers; and the second set of layers is arranged to coincide with the first intervals between the first set of layers.

45. The device of claim 41, wherein the first set of one or more walls of the inner portion comprises a plurality of first apertures, and the second set of one or more walls of the outer portion comprises a plurality of second apertures formed in alternate layers to the first apertures in the outer portion.

46. The device of claim 41, wherein at least one of the outerportion or the inner portion comprises a continuous annular wall.

47. The device of claim 41, wherein the inner portion comprises a triangular channel surrounded by the first set of walls, and the outer portion comprises a cylindrical wall surrounding the triangular channel.

48. The device of claim 41, wherein the second set of one or more walls is thicker than the first set of one or more walls,

49. The device of claim 41, wherein the base portion extends substantially continuously between the inner portion and the outer portion.

50. The device of claim 41, wherein the base portion comprises a window at least partially aligned with the channel, and the window comprises an opening in the base portion.

51. The device of claim 41, wherein the base portion consists of a substantially porous material, and wherein the base portion comprises a micro-filament mesh.

52. The device of claim 41, wherein the cavity is at least partially filled with a porous filler material; wherein the filler material comprises one or more hydrogel materials.

53. The device of claim 52, wherein the filler material comprises a cellulose nanofibril (CNF) hydrogel or nanocellulose hydrogel.

54. The device of claim 41, wherein the scaffold is made using a computer-controlled fabrication technique

55. The device of claim 41, wherein the scaffold is made using a 3D fiber deposition (3DF) method.

56. The device of claim 41, wherein the scaffold is customized to match the dimensions of a particular osseous environment in a human or animal target.

57. A method of tissue engineering using the device of claim 41.

58. The method of claim 57, comprising: filling the cavity defined between the inner and outer portions of the scaffold with a filler material; and seeding the scaffold with tissue cells.

59. The method of claim 58, further comprising: growing tissue on the scaffold in vitro or implanting the scaffold in a living target.

60. A method of osseous generation or regeneration comprising: making a device according to claim 41; filling the cavity defined between the inner and outer portions of the scaffold with a filler material; seeding the scaffold with tissue cells; and implanting the scaffold in an osseous environment in a living human or animal patient.

Description

[0062] Some embodiments of the present invention will now be described, by way of example only, and with reference to the accompanying drawings, in which:

[0063] FIGS. 1a-1c schematically show how a scaffold shape can be modelled on a long bone such as a femur;

[0064] FIG. 2 is perspective view of a scaffold according to an embodiment of the present invention;

[0065] FIG. 3 is an exploded view of the components making up the scaffold of FIG. 2;

[0066] FIGS. 4a-4c are micro-CT images of the scaffold, showing the architecture of different layers at different heights in the scaffold; and

[0067] FIG. 5 is a top view image of a scaffold made and filled according to the Example given below.

[0068] FIGS. 1a-1c give an overview of how a long bone such as a femur can be modelled when fabricating a customised scaffold. The basic anatomy of the bone, as seen in the perspective view of FIG. 1a and cross-sectional view of FIG. 1b, comprises the outer compact bone 2, the inner spongy bone 4 and the medullary cavity 6 in the centre where the bone marrow is located. This basic structure can be modelled, as shown schematically by FIG. 1c, as two generally concentric sets of walls. A central channel 8 can be dimensioned to match the medullary cavity 6. The average human bone medullary cavity is 20 mm in diameter. A cavity 10 between the two sets of walls can be filled with a porous material to match the spongy bone 4. One example of a scaffold modelled in such a way is seen in more details in FIGS. 2 and 3.

[0069] FIG. 2 shows a scaffold 12 comprising an inner portion in the form of a centric tube 14 and an outer portion in the form of a contour tube 16. In this example the centric tube 14 is shown as a triangular tube but the inner portion may be fabricated to have any suitable shape, for example cylindrical. Also, in this example the contour tube 16 is shown as a cylindrical tube but it may have any suitable cross-sectional shape. In particular, where the scaffold 12 has been customised to substantially match a target bone then its shape and/or dimensions may be tailored accordingly. Furthermore, the scaffold may include more than one contour tube 16, for example a number of concentric cylinders arranged inside one another. The walls of the centric tube 14 surround a central channel 18 extending through the scaffold 12. An annular cavity 20 is defined between the inner and outer portions 14, 16. The main function of the contour tube 16 is to support material that may be contained and/or grown in the cavity 20.

[0070] The concentric tube portions 14, 16 are connected by a base plate 22. As is seen more clearly from FIG. 3, the base plate 22 is a generally circular disc comprising a central window 24 that is shaped to match the central channel 18. In this example the window 24 comprises an opening in the material of the base plate 22. Fluid can therefore flow longitudinally through the scaffold 12, i.e. through the window 24 and along the central channel 18. The base plate 22 can be made to different diameters, for example to help incorporate the scaffold 12 into a particular device or fit an intended container such as a cell culture plate or bioreactor chamber.

[0071] It can be seen in both FIGS. 2 and 3 that circumferential apertures 26 are provided in both the inner and outer tubes 14, 16. The apertures 26 are opened on alternate layers. For example, there are apertures 26 open in layers 1 and 2 of the inner tube 14 while layers 1 and 2 of the outer tube 16 are continuous, then there are apertures 26 open in layers 3 and 4 of the outer tube 16 while layers 3 and 4 of the inner tube 14 are continuous. This may be understood more clearly with reference to FIGS. 4a-4c. This alternating arrangement of the apertures 26 ensures that there is sufficient radial diffusion available through the scaffold 12 at any given height. It can also be seen from FIGS. 4a-4c that the outer tube 16 may be fabricated with a double wall thickness as compared to the inner tube 14. This helps to provide the outer tube 16 with increased mechanical strength. The apertures 26 may be formed by an interruption in the layering process used to deposit the polymer material of the inner and outer tubes 14, 16. Accordingly each aperture 26 may have a height corresponding to an integer number of the fabrication layers, for example a height of two layers as deposited by a 3D printer.

[0072] The inner and outer tubes 14, 16 are both made from biocompatibale polymer materials. The same material may be used for both tubes 14, 16, or different materials may be used so as to provide different mechanical strengths. The base plate 22 is formed from a pattern of polymeric microfilaments, for example a layering pattern of 60 degrees incrementally (0/60/120 degrees alternatively). Of course the angle and/or distance between the filaments can be controlled to give a desired degree of porosity. The base plate 22 therefore provides a mesh that can contain filler material in the cavity 20 while also connecting the inner and outer tubes 14, 16.

[0073] In such a scaffold 12 the inner channel 18 can provide for blood vessels and nutritional transfer/diffusion, allowing a culture medium to penetrate through the scaffold 12. The cavity 20 between the inner and outer tubes 14, 16 can be filled with one or more porous materials e.g. to form a sponge structure that mimics the spongy bone.

[0074] Note that the top of the scaffold 12 may be left open, as shown, or optionally be closed by a cover plate similar to the base plate 22.

EXAMPLE

[0075] A scaffold was made from a customised reproducible design recorded in a CAD model using standard CAD software such as Solidworks, AutoCAD, ProE, Magics, etc.. The design was recorded in STereoLithography (STL) file format and the dimensions confirmed with reference to images of a real bone. Alternatively the design could have been recorded according to the Additive Manufacturing File Format (AMF) standard or OBJ geometry format instead of the STereoLithography (STL) file format. The STL file was then read and transferred to a 3D slicer software (such as Slic3r, Bioplotter RP, Cura, etc.) to be divided to a layer-by-layer structure. The sliced designs were then imported into RP software for fabrication by a rapid prototyping (RP) machine such as a 3D printer.

[0076] Two polyesters were used to manufacture the scaffold 12. The inner portion 14 was made from a copolymer of poly(lactide-co-caprolactone) ordered from Purac. The outer portion 16 and the base portion 22 were made from polylactide. Polylactide was chosen for the main load-bearing parts of the scaffold 12 as it has a modulus much higher than that of the copolymer used to make the inner portion 14 defining the central channel 18. The cavity 20 between the inner and outer portions 14, 16 was filled with nanofibrous (TEMPO-oxidized nanocellulose) sponge fabricated by freeze-drying an injected hydrogel solution of gelatin/CNF. Post-modification and cleaning steps prepared the scaffold for cell seeding. For cell culture, L929 (fibroblasts) was mixed with the freeze-dried nanocellulous sponge.

[0077] Scanning electron microscope images taken 1 and 3 days after cell seeding showed positive attachment of the cells and proliferation behaviour. FIG. 5 shows the cell culture growth on the scaffold 12.

[0078] In this example the scaffold 12 was made to fit a 48-well plate in a bioreactor. The outer contour tube 16 had an outer diameter of 11.4 mm and a height of 5.0 mm. The inner triangular tube 14 had a side length of 3.5 mm and the same height of 5.0 mm. The apertures 26 in both the inner and outer tubes 14, 16 had a length of 1.5 mm and a height of 0.68 mm, corresponding to two layers of thickness 0.34 mm (as set by the RP machine). The base plate 22 included a central triangular window 24 of side length 3.5 mm to match the inner triangular tube 14. The outer diameter of the base plate 22 was 11.4 mm. The base plate 22 was made 0.68 mm thick i.e. two layers thick. It will be appreciated that the scaffold could be scaled up or down to fit any particular environment. For example, the outer diameter could be set up to 15 mm to fit a 24-well plate in a bioreactor.