• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    system for producing additive of three-dimensional structures and method for the same. it is a system and method for producing additive from three-dimensional structures, which includes three-dimensional cell structures. the system comprises at least a printhead for receiving and dispensing materials, the materials comprising a coating fluid and a hydrogel, the printhead comprising an orifice for dispensing the materials, microfluidic channels for receiving and directing the materials, the fluidic switches that correspond to one of the microfluidic channels in the printhead and configured to allow or disallow fluid flow in the microfluidic channels; a receiving surface for receiving a first layer of materials dispensed from the orifice; a positioning unit for positioning the printhead hole in three-dimensional space; and a dispensing means for dispensing the materials from the orifice of the printhead.
    公开号:BR112015031320B1
    申请号:R112015031320-5
    申请日:2014-06-13
    公开日:2022-01-18
    发明作者:Simon Travis Beyer；Konrad WALUS；Tamer Mohamed；Anas Amjad Mohammed Bsoul
    申请人:Aspect Biosystems Ltd；
    IPC主号:
    专利说明:

    CROSS REFERENCE TO RELATED ORDERS
    [001] This application claims priority under the Paris Convention of Application No. US 61/834,420, filed on June 13, 2013, the full contents of which are hereby incorporated by reference. FIELD OF THE INVENTION:
    [002] The present invention generally relates to three-dimensional (3D) printing and generation of three-dimensional biological structures from digital files. Specifically, the invention relates to a system, apparatus and method for manufacturing 3D cell-loaded hydrogel structures. BACKGROUND OF THE INVENTION:
    [003] 3D printing, a form of additive manufacturing (AM), is a process for creating three-dimensional objects directly from digital files. Software is used to cut a computer-aided design (CAD) model or 3D scan of an object into a multitude of thin cross-sectional layers. This collection of layers is sent to the AM system where the system creates the three-dimensional object layer by layer. Each layer is deposited on top of the previous layer until the object has been completely constructed. Support material can be used to support prominent and complex features of the object. There are several AM processes that can create parts in plastic, metal, ceramic and/or biological materials.
    [004] Additive manufacturing would have applications in biological systems. For example, until recently, most cell culture studies were performed on two-dimensional (2D) surfaces, such as microwell plates and Petri dishes. However, 2D culture systems do not mimic the 3D environment in which cells exist in vivo. The researchers found 3D cell cultures behave more like natural biological tissue than 2D cell cultures, at least in part, due to the fact that the 3D arrangement of cells in natural tissue influences cell-cell interactions, which, in turn, influences cell physiology and growth.
    [005] Additive manufacturing devices and systems for manufacturing cellular constructs are known. For example, known fused fiber deposition techniques have been applied to biological materials. In fused fiber deposition, high viscosity liquids are dispensed from a relatively hollow orifice and then rapidly solidified by a variety of means. Biocompatible plastics, thermal gelling hydrogels, UV crosslinkable polymers and high concentration alginates have been used as scaffolds for 3D cell structures, with cells being added to the scaffold after they have solidified. A disadvantage of these techniques is that they require cells to be added to the frame after printing, making it difficult to control cell displacement. Furthermore, the composition of the scaffolding substrates may not be suitable for facilitating cell proliferation and growth.
    [006] Systems for 3D printing structures that comprise direct printing of cellular materials are known and desired, at least in part, due to the fact that they can allow cells to be deposited within a 3D scaffold. For example, inkjet printing technology has been used to print biological materials. However, the shear force involved with propelling droplets of fluid over a substrate can damage cells dispersed in the fluid. In addition, inkjet printing is a slow process, which makes it difficult to adapt to biological materials, which require specific environmental conditions for survival.
    [007] Other systems for directly printing cells within a structure in 3D include US Patent No: 8,639,484, which refers to the use of a CAD model and a 3D positioning unit to deposit cellular materials through a multitude of nozzles, layer by layer, to create a 3D object. Multiple nozzles allow multiple different materials to be included in the 3D object. Patent Application Publication No: 2012/0089238 discloses a multi-cartridge printing system for producing 3D organic composite structures whereby the structure is created using at least two syringes, one comprising a structural support polymer. and another comprising a living cell composition, which iteratively deposits the structural support polymer and the living cell composition on a surface. Patent Application Publication No: 2014/0012407 discloses a device comprising one or more printheads, each of which is configured to receive and retain one or more cartridges. Each cartridge comprises a fluid, such as a bio-ink comprising cells or support material, and an orifice from which the fluid can be dispensed from the cartridge.
    [008] Prior art methods generally require multiple nozzles and/or cartridge holes in order to facilitate printing of multiple different materials (i.e., one material is dispensed through a nozzle or cartridge hole). The use of multiple nozzles to dispense different materials requires a corresponding increase in movement of the printing system in order to position the appropriate nozzle or cartridge orifice in a controlled sequence to deliver a sequence of different materials. Such increased movement decreases printing speed and efficiency.
    [009] It is desirable to obviate or mitigate one or more of the above deficiencies. SUMMARY OF THE INVENTION
    [010] In a first aspect, a system for additive manufacturing of three-dimensional structures is provided. The system comprises at least one printhead for receiving and dispensing materials, the materials comprising a coating fluid and a hydrogel. In one embodiment, the printhead comprises an orifice for dispensing materials; microfluidic channels comprising one or more first channels for receiving and directing the coating fluid and one or more respective second channels for receiving and directing the hydrogel, wherein the second channels intersect at a first point of intersection with the first channels, in that the second and first channels join at the first point of intersection to form a distributor channel that extends to the orifice; and fluidic switches, each fluidic switch corresponding to one of the microfluidic channels in the printhead and configured to allow or disallow fluid flow in the microfluidic channels of the printhead when activated. In one embodiment, the system further comprises a receiving surface for receiving a first layer of materials dispensed from the orifice; a positioning unit for positioning the printhead hole in three-dimensional space, the positioning unit operably coupled to the printhead; and a dispensing means for dispensing the materials from the orifice of the printhead.
    [011] In an embodiment of the first aspect, the system comprises a programmable control processor for controlling the positioning unit and for controlling the distribution of printhead materials on the receiving surface.
    [012] In an embodiment of the first aspect, the one or more first channels comprise at least two channels, the one or more first channels being configured to flank respective second channels at the first point of intersection.
    [013] In an embodiment of the first aspect, the coating fluid comprises a crosslinking agent to solidify the hydrogel upon contact with it at the point of intersection and/or in the distributor channel.
    [014] In an embodiment of the first aspect, each second channel has a smaller diameter than that of the first channels and the distributor channel, whereby the flow from the first channels forms a coaxial coating around the hydrogel in the distributor channel.
    [015] In an embodiment of the first aspect, the hydrogel comprises living cells.
    [016] In an embodiment of the first aspect, the system additionally comprises a fluid removal feature to remove excess coating fluid from that delivered from the printhead.
    [017] In an embodiment of the first aspect, a receiving surface comprises a porous membrane comprising pores sized to allow excess coating fluid to pass through.
    [018] In an embodiment of the first aspect, the fluid removal feature comprises the absorbent material or a vacuum to extract excess coating fluid in the opposite direction to the receiving surface.
    [019] In an embodiment of the first aspect, the absorbent material or vacuum is applied below a porous membrane. In an embodiment of the first aspect, a vacuum is applied above the receiving surface.
    [020] In an embodiment of the first aspect, the vacuum is applied through one or more vacuum channels provided in the printhead, the one or more vacuum channels having an orifice located next to the orifice of the printhead.
    [021] In an embodiment of the first aspect, the system additionally comprises reservoirs to contain the materials, the reservoirs being fluidly coupled respectively to the microfluidic channels in the printhead.
    [022] In an embodiment of the first aspect, the printhead additionally comprises at least two inlets for receiving materials from the reservoirs, each of the inlets being in fluid communication with respective microfluidic channels and respective reservoirs.
    [023] In an embodiment of the first aspect, the distribution means comprises a pressure control unit.
    [024] In an embodiment of the first aspect, the fluidic switches comprise valves.
    [025] In an embodiment of the first aspect, the printhead additionally comprises a hollow projection configured to extend from the hole towards the receiving surface.
    [026] In an embodiment of the first aspect, the printhead comprises two second channels, each of the second channels being adapted to conduct respective hydrogels, with two second channels intersecting at a second intersection and joining at the second intersection. to form a third channel that extends to the first intersection point.
    [027] In a second aspect, a system for additive manufacturing of three-dimensional structures is provided, wherein the system comprises at least one printhead for receiving and distributing materials, wherein the materials comprise a coating fluid and a hydrogel. In one embodiment, the printhead comprises an orifice for dispensing materials; microfluidic channels to receive and direct materials to the orifice; and fluidic switches, each fluidic switch corresponding to one of the microfluidic channels in the printhead and configured to allow or disallow fluid flow in the microfluidic channels in the printhead when actuated. In one embodiment, the system further comprises a receiving surface for receiving materials dispensed from the orifice; a fluid removal feature to remove excess coating fluid dispensed from the orifice; a positioning unit for positioning the printhead hole in three-dimensional space, the positioning unit operably coupled to the printhead; and a dispensing means for dispensing the printhead orifice materials.
    [028] In an embodiment of the second aspect, the fluid removal facility comprises a vacuum to extract excess coating fluid away from or across the receiving surface and/or from the hydrogel distributed on the receiving surface.
    [029] In an embodiment of the second aspect, the receiving surface comprises a porous membrane comprising pores sized to allow excess coating fluid to pass therethrough.
    [030] In an embodiment of the second aspect, the vacuum is applied below the porous membrane. In an embodiment of the second aspect, a vacuum is applied above the receiving surface.
    [031] In an embodiment of the second aspect, the vacuum is applied through one or more vacuum channels provided in the printhead, one or more vacuum channels having an orifice situated close to the orifice of the printhead.
    [032] In an embodiment of the second aspect, the fluid removal feature comprises an absorbent material to extract, in the opposite direction to the receiving surface, the excess coating fluid.
    [033] In an embodiment of the second aspect, the system additionally comprises a programmable control processor for controlling the positioning unit and for controlling the distribution of materials from the printhead onto the receiving surface.
    [034] In an embodiment of the second aspect, the printhead additionally comprises a hollow projection configured to extend from the hole towards the receiving surface.
    [035] In an embodiment of the second aspect, the printhead comprises one or more first channels for receiving and directing the coating fluid and one or more respective second channels for receiving and directing the hydrogel, the second channels intersecting in a first point of intersection with the first channels, the second and first channels joining at the first point of intersection to form a distributor channel that extends into the orifice.
    [036] In an embodiment of the second aspect, the printhead comprises two second channels, each of the second channels being adapted to conduct respective hydrogels, with the two second channels crossing at a second intersection and joining at the second. intersection to form a third channel that extends to the first intersection point.
    [037] In a third aspect, a method of printing a three-dimensional (3D) structure is provided, the method comprising providing a 3D printer, the printer comprising: a printhead comprising an orifice for dispensing materials ; a receiving surface for receiving a first layer of materials dispensed from the orifice of the printhead; and a positioning unit operably coupled to the printhead, the positioning unit for positioning the printhead in three-dimensional space. In one embodiment, the method comprises providing the materials to be delivered, the materials to be delivered comprising a coating fluid and one or more hydrogels; that encode the printer with a 3D structure to be printed; distributing, from the printhead hole, the materials to be distributed; depositing a first layer of the distributed materials on the receiving surface; repeating the deposition step by deposition of the subsequent distributed material on the first and any subsequent layers of deposited material, thereby depositing layer after layer of distributed materials in a geometric arrangement in accordance with the 3D structure; and removing excess coating fluid delivered from the printhead orifice at one or more points in time during or between deposition steps.
    [038] In an embodiment of the third aspect, the coating fluid comprises a crosslinking agent suitable for crosslinking and solidifying the hydrogel upon contact with it, the contact creating a hydrogel fiber.
    [039] In an embodiment of the third aspect, the coating fluid and the hydrogel are distributed in a coaxial arrangement, with the coating fluid surrounding the hydrogel.
    [040] In an embodiment of the third aspect, the deposition step and the removal step are performed continuously, thereby continuously removing excess coating fluid as layers of distributed materials are deposited.
    [041] In an embodiment of the third aspect, the stripping step is performed intermittently between and/or at the same time as the deposition step, thereby intermittently removing excess coating fluid as layers of distributed materials are deposited.
    [042] In an embodiment of the third aspect, the one or more hydrogels are adapted to support the growth and/or proliferation of living cells dispersed therein. BRIEF DESCRIPTION OF THE DRAWINGS
    [043] The features of the invention will become more apparent in the following detailed description in which reference is made to the accompanying drawings in which:
    [044] Figure 1 is a perspective view of an embodiment of the printing system of the present invention.
    [045] Figure 2 is a perspective view of objects designed by software and corresponding objects printed using an embodiment of the printing system of the present invention.
    [046] Figure 3 is a perspective view of an embodiment of the printhead of the present invention.
    [047] Figure 4 is a cross-section of a valve in the printhead of Figure 3, including deflection of a valve membrane when the valve is activated.
    [048] Figure 5 is a cross-section of an alternative embodiment of the printhead of Figure 3.
    [049] Figure 6 is a top view of an alternative embodiment of the printhead of Figure 3.
    [050] Figure 7 is an exploded perspective view of an embodiment of the print bed assembly of the present invention.
    [051] Figure 8 is a cross-section of the assembled print bed of Figure 9.
    [052] Figure 9 is a cross-section of an alternative embodiment of the print bed of Figure 9.
    [053] Figure 10 is a perspective view of an embodiment of the printhead of the present invention. DETAILED DESCRIPTION OF THE INVENTION:
    [054] Definitions of certain terms as used in this descriptive report are provided below. Unless otherwise defined, all technical and scientific terms used herein generally have the same meaning as commonly understood by an individual of ordinary skill in the art to which the invention pertains.
    [055] As used herein, the term "about" will be understood by the person of ordinary skill in the art and will vary somewhat depending on the context in which it is used. If there are uses of the term that are not clear to the person of ordinary skill in the art, given the context in which it is used, "about" will mean up to more or less than 10% of the enumerated value.
    [056] As used herein, the term "hydrogel" refers to a composition comprising water and a network or lattices of polymer chains that are hydrophilic. Examples of natural hydrogels include, for example, alginate, agarose, collagen, fibrinogen, gelatin, chitosan, hyaluronic acid-based gels or any combination thereof. A variety of synthetic hydrogels are known and would be used in embodiments of the systems and methods provided herein. For example, in embodiments of the systems and method provided herein, one or more hydrogels form the structural basis for printed three-dimensional structures. In some embodiments, the hydrogel has the ability to support the growth and/or proliferation of one or more cell types, which may be dispersed within the hydrogel or added to the hydrogel after they have been printed in a three-dimensional configuration. In some embodiments, the hydrogel is crosslinkable by a chemical crosslinking agent. For example, a hydrogel comprising alginate may be crosslinkable in the presence of a divalent cation, where a hydrogel comprising fibrinogen may be crosslinkable in the presence of thrombin and a hydrogel comprising collagen or chitosan may be crosslinkable in the presence of heat or a basic solution. Crosslinking the hydrogel will increase the hardness of the hydrogel, in some embodiments, allowing the formation of a hydrogel that behaves like a solid.
    [057] As used herein, the term "coating fluid" refers to a liquid that is used, at least in part, to envelop or "coat" a material to be dispensed, such as, for example, a hydrogel. In some embodiments, the coating fluid comprises one or more of an aqueous solvent, e.g., water or glycerol, and a chemical cross-linking agent, e.g., materials comprising divalent cations (e.g., Ca<2+>, Ba< 2+>, Sr<2+>, etc.), thrombin, or pH modifying chemicals such as sodium bicarbonate.
    [058] As used herein, the term "excess coating fluid" refers to a portion of the coating fluid that is distributed from the printhead orifice and does not form part of a three-dimensional structure printed using one or more embodiments of the systems or methods provided herein. For example, excess coating fluid can be useful in passing hydrogel lubrication through a distributor channel in the printhead and through the printhead orifice. Once delivered from the printhead orifice, excess coating fluid can run off the surface of a distributed hydrogel layer and onto a receiving surface, where it can be collected or pooled.
    [059] As used herein, the term "receiving surface" refers to the surface upon which a first layer of material dispensed from a printhead hole is deposited. The receiving surface also receives excess coating fluid which is dispensed from the printhead orifice and which flows from one or more layers of material dispensed from the printhead orifice. In some embodiments, the receiving surface is produced from a solid material. In some embodiments, the receiving surface is produced from a porous material. For example, in some embodiments, the porosity of the porous material is sufficient to allow coating fluid to pass therethrough. In some embodiments, the receiving surface is substantially flat, thereby providing a smooth surface upon which a first layer of distributed material can be deposited. In some embodiments, the receiving surface has a topography that corresponds to the three-dimensional structure to be printed, thereby facilitating the printing of a three-dimensional structure that has a non-smooth first layer.
    [060] In one aspect, the present invention generally relates to an apparatus, system and method for additive manufacturing of three-dimensional (3D) biological structures. GENERAL DESCRIPTION OF THE PRINTING SYSTEM
    [061] In one aspect, the invention provides a system for additive manufacturing of three-dimensional structures (also referred to herein as a "printer", a "3D printer", or a "printing system" or "the system". ). The system comprises a microfluidic printhead, which is a microfluidic liquid handling device comprising one or more microfluidic channels for receiving and directing materials to be distributed, wherein the fluidic switches correspond to the microfluidic channels to regulate the flow of materials to be delivered. to be printed, and a single hole to distribute the materials to be distributed.
    [062] The materials to be distributed comprise a coating fluid and at least one hydrogel. In a preferred embodiment, the coating fluid comprises a chemical cross-linking agent suitable for solidifying the hydrogel upon contact therewith. In a preferred embodiment, the coating fluid also serves as a lubricant for the solidified hydrogel.
    [063] The microfluidic channels serve as conduits to direct and combine the materials to be distributed in a controlled manner. The microfluidic channels are arranged within the printhead so that one or more first channels for receiving and directing the coating fluid and a second channel for receiving and directing the hydrogel cross at a first point of intersection and join to form a distributor channel that extends into the printhead hole. In a preferred embodiment, the first channels are configured to flank the second channel at the first point of intersection. In this manner, the coating fluid is directed to flow along either side of the hydrogel in the distributor channel.
    [064] In a preferred embodiment, the materials in the distributor channel are directed coaxially, with the hydrogel being focused in the center of the distributor channel and with the coating fluid surrounding the hydrogel fluid, thereby forming a coating. around the hydrogel. In preferred embodiments where the coating fluid also comprises a chemical crosslinking agent suitable for crosslinking the hydrogel, a solidified hydrogel fiber is formed in the distributor channel and delivered from the orifice of the printhead.
    [065] In one aspect, the system further comprises a receiving surface for receiving a first layer of materials dispensed from the orifice and a positioning unit for positioning the printhead orifice in three-dimensional space, wherein the positioning unit is operably attached to the printhead. For example, the printhead can be coupled to a commercially available mechanized positioning system with three degrees of movement so that the printhead can be positioned above the receiving surface and oriented to direct the dispensed material downward toward the surface. receiver.
    [066] In one aspect, the system comprises means for dispensing materials from the printhead orifice and may additionally comprise and/or be in data communication with a programmable control processor to regulate the placement of the orifice. of printhead. The programmable control processor can also be used to regulate the dispensing of materials to be dispensed from the printhead hole.
    [067] Figure 1 shows a schematic perspective view of an embodiment of the 3D printing system provided in this document.
    [068] Referring to Figure 1, the system comprises a microfluidic printhead [100], which comprises a printhead hole [114] and at least one inlet to receive the material to be distributed from the printhead [100]. The material to be dispensed is stored in printed material reservoirs [110] and delivered to the printhead through respective first connection tubes [122], which provide fluid communication between the printhead and the printed material reservoirs. In the illustrated embodiment, the means for dispensing material to be dispensed from the printhead orifice is a pressure control unit [112] which is fluidly coupled to the printed material reservoirs [110] by respective second connecting tubes. [120]. The pressure control unit is a means to provide a force to distribute the materials to be distributed. The pressure control unit supplies pneumatic pressure to the printed material reservoirs [110] through respective second connecting tubes [120]. Pressure applied to the printed material reservoirs forces the fluid out of the reservoirs and into the printhead through respective first connecting tubes [122]. Alternative means for distributing the material to be distributed would be used in the illustrated embodiment. For example, a series of electronically controlled syringe pumps would be used to provide force to deliver the material to be dispensed from the printhead orifice.
    [069] Referring to Figure 1, the microfluidic printhead [100] is coupled to a 3D mechanized stage comprising three arms [102, 103 and 104] to position the printhead [100] and the head hole [114] in the three-dimensional space above a print bed [108], which comprises a surface [109] for receiving the printed material. In one embodiment, the 3D mechanized stage (i.e., the positioning unit) can be controlled to position a vertical arm [104], which extends along the z-axis of the 3D mechanized stage so that the printhead [114] is directed downwards. A first horizontal arm [102], which extends along the x-axis of the mechanized stage, is attached to an immovable base platform 116. A second horizontal arm [103], which extends along the y-axis of the mechanized stage is movably coupled to an upper surface of the first horizontal arm [102] so that the longitudinal directions of the first and second horizontal arms [102 and 103] are perpendicular to each other. It will be understood that the terms "vertical" and "horizontal" as used above in relation to the arms are intended to describe the manner in which the printhead is moved and do not necessarily limit the physical orientation of the arms themselves.
    [070] In the embodiment illustrated in Figure 1, the print bed [108] is positioned on top of a platform [118], and the platform is coupled to an upper surface of the second horizontal arm [103]. In one embodiment, the 3D mechanized stage arms [102, 103, and 104] are driven by three corresponding motors [105, 106, and 107], respectively, and controlled by a programmable control processor, such as a computer (not shown). In a preferred embodiment, the printhead [100] and the print bed [108] are collectively movable along all three primary geometric axes of a Cartesian coordinate system by the 3D mechanized stage and the stage's motion is defined. using computer software.
    [071] It will be understood that the invention is not limited only to the positioning system described and that other positioning systems are known in the art.
    [072] In the embodiment illustrated in Figure 1, as material is dispensed from the printhead hole [114], the positioning unit is moved in a software-controlled pattern, thereby creating a first layer. [109] Additional layers of distributed material are stacked on top of each other so that the final 3D geometry of the distributed material layers is generally a replica of the 3D geometry design provided by the software. The 3D design can be created using typical 3D CAD software (computer aided design) or generated from digital images as known in the art. Furthermore, if the software generated geometry contains information about the specific materials to be used, it is possible, according to an embodiment of the invention, to assign a specific material type to different geometric locations. For example, Figure 2 shows three 3D-printed structures using one embodiment of the system provided herein: a cube [128], a hollow cylinder [129] and a hollow coaxial cylinder [130]. The software was used to generate cube, hollow cylinder and hollow coaxial cylinder designs ([125], [126] and [127], respectively), with each design comprising two different types of materials (dyed alginate), the which were dyed different colors to provide visual clarity of the materials used to generate the printed cube and hollow cylinder.
    [073] Any software, application or module referred to in this document may be deployed using computer-readable/executable instructions that may be stored or otherwise owned by such computer-readable means. PRINTHEAD
    [074] Figure 3 shows a schematic perspective view of an embodiment of a microfluidic printhead [100] for use in the provided system.
    [075] Referring to Figure 3, the illustrated embodiment describes a microfluidic printhead [100] that comprises microfluidic channels to transport various fluids. In the illustrated embodiment, the microfluidic channels are cylindrical in shape. However, channel shapes other than cylindrical shapes would also be used in the print head provided in this document. Channel [200] is a conduit for a crosslinking agent, channel [202] is a conduit for water. In the illustrated embodiment, the crosslinking agent and water, separately or together, serve as the "coating fluid". Channel [204] is a conduit for a first hydrogel composition (referred to as "hydrogel A") and channel [206] is a conduit for a second hydrogel composition (referred to as "hydrogel B"). In a preferred embodiment, one or more live cell types are compatible with and optionally dispersed within hydrogels A and/or B. In the illustrated embodiment, each microfluidic channel comprises a fluid inlet [208a, 208b, 208c, 208d], which allows the fluid contained in the connection tubes [122] to pass through the respective channels of the printhead [100]. Downstream of the fluid inlets [208a, 208b, 208c, 208d] are valves [210, 212, 214, 216] that correspond to each channel. In the illustrated embodiment, the valves serve as "fluidic switches", which can be activated to allow and disallow fluid flow through a channel, with each valve having a corresponding inlet [218, 218a, 218b, 218c, 218d] , which facilitates the activation and deactivation of the valve. In one embodiment, valves [210, 212, 214, 216] may be electronically activated. In another embodiment, the valves [210, 212, 214, 216] can be activated by a change in applied pressure, for example, by means of solenoid pistons. The pressure or electronic activation of different valves facilitates rapid change of the dispensed material, thereby allowing the dispensed materials to be composed of a controlled sequence of different materials.
    [076] Referring further to Figure 3, in the illustrated embodiment, the crosslinking agent channels [200] and the water channels [202] intersect at points of intersection [203], such as in a "y-shaped" configuration. ", joining together to form channels referred to herein as "coating flow channels" [224] immediately downstream of the crosslinking agent and water channels [200, 202]. The channels of hydrogel A and hydrogel B [204, 206] intersect at an intersection point [207], such as in a "y-shaped" configuration, joining together to form a channel referred to herein as a "y-shaped channel". focusing" [226] immediately downstream of the two hydrogel channels. The coating flow channels [224] and the focusing channel [226] intersect at an intersection point [228] in a three-phase configuration, for the described embodiment, where the focusing channel [226] is flanked by the coating flow channels [224], which join to form a channel referred to herein as a distributor channel [220]. The distributor channel [220] ends at the distributor hole [222]. In a preferred embodiment illustrated in Figure 1, the distributor channel protrudes from the printhead [100] that ends in the distributor hole [1 14].
    [077] In addition to Figure 3, in the illustrated embodiment, the coating flow channels [224] and the distributor channel [220] have larger diameters than the focusing channel [226]. When hydraulic pressure is applied to the coating stream [224] and focusing channels [226], the liquid in the focusing channel [226] is laterally compressed and "focused" in a narrow stream along the central axis of the channel. of focusing [226]. Upon intersection with the focusing channel [226] at the intersection point [228], fluid from the larger diameter coating flow channels [224] surrounds and surrounds the narrower focused stream of hydrogel distributed from the focusing channel. - tion [226].
    [078] In a preferred embodiment, the liquid in the coating flow channels [224] comprises a chemical crosslinking agent and the liquid in the focusing channel [226] comprises one or more chemically crosslinkable hydrogels comprising one or more types of living cell. When the one or more chemically crosslinkable hydrogels are focused in a narrow stream in the focusing channel [226] and then enveloped by the crosslinking agent in the distributing channel [220], at least the outer surface of the one or more chemically crosslinkable hydrogels is solidified in the distributor channel [220], thereby creating a "solid" or cross-linked hydrogel fiber. The hydrogel fiber is then distributed from the delivery hole [222] onto the receiving surface in a controlled manner that creates a 3D structure, layer by layer.
    [079] In a particularly preferred embodiment, the coating fluid that surrounds the hydrogel fiber can also act to lubricate the passage of the hydrogel fiber through the distributor channel [220] until it is distributed from the cylinder head orifice. print [222].
    [080] In one embodiment, the coating fluid comprises a chemical crosslinking agent, water or a combination thereof. In embodiments where the coating fluid lacks a chemical crosslinking agent, the hydrogel will not solidify and would be delivered as a liquid. In order to adjust the coating fluid composition and start and/or stop solidification of the hydrogel, a crosslinking agent channel valve [210] and water channel valve [212] can be activated. It is contemplated that dispensing a liquid rather than a solid hydrogel, or dispensing the coating fluid alone, may be desirable in order to construct some aspects of various three-dimensional objects.
    [081] In one embodiment, the printhead [100] can be configured to receive and deliver only one hydrogel material. In one embodiment, the printhead may be configured to receive and dispense two or more hydrogel materials. For example, in an embodiment where the printhead [100] is configured to receive two hydrogel materials, each, for example, comprising a different cell type, the system provided herein may be programmed to deliver a heterogeneous cell structure, with the first and second cell types being placed in controlled patterns within and between layers, alone and/or in combination with each other. The contours between the two materials are controlled, for example, by software, and the programmable control processor is used to instruct the switched fluidic material (e.g., one or more valves [210], [212], [214], [ 216]) to alter the flow of material within one or more microfluidic channels, thereby altering the content of the material that is delivered from the printhead orifice. The number of hydrogel materials that can be received by and dispensed from the printhead provided herein is limited only by the size of the printhead that the user considers to be practical.
    [082] Referring to Figure 4, in one embodiment, the fluidic switch is a valve comprising a membrane [332] disposed over a bowl-shaped feature [318] shaped in a microfluidic channel [308]. By applying pneumatic pressure (represented by arrows in Figure 4) to the exposed surface of the valve diaphragm [332], the valve diaphragm [332] will be deflected in the bowl-shaped feature [318], thereby blocking the passage of fluid through the microfluidic channel [308]. In a preferred embodiment, the thickness of the valve membrane [332] is about 150 µm. In embodiments where the valve membrane thickness is increased, a skilled person would understand that the applied pneumatic valve activation pressure must be increased accordingly. Similarly, a valve membrane formed of less resilient material would require a higher activation pressure. A knowledgeable person would understand how to adjust the activation pressure to suit the specific valve membrane material.
    [083] In one embodiment, the printhead comprises alternate fluidic switches for regulating materials to be dispensed from the printhead orifice. For example, instead of using valves, a mechanism for engaging or disengaging the pressure applied to each channel would be used to regulate the flow of material in the microfluidic channels.
    [084] In one embodiment, the printhead further comprises an extension tip that comprises a hole for dispensing materials from the printhead. Such an extension tip facilitates accurate material dispensing and material deposition in confined areas such as, for example, a well in a multi-well plate (e.g., a standard microtiter plate, microwell plate, or microplate that has 6 , 24, 96 etc. wells) or a Petri dish. Referring to the embodiment illustrated in Figure 5, a portion [500] of the distribution channel [220] most adjacent to the distribution orifice [222] has a larger diameter than the upstream portion of the distribution channel [220]. The extension tip [502] comprises a tube (e.g. made of plastic, glass or metal) that has an outer part configured to fit the larger diameter portion [500] of the distributor channel and an inner surface (which defines a hollow space in the tube) configured to align with the distributor channel [220]. The extension tip [502] can be inserted into the larger diameter portion [500] of the distributor channel, thereby extending the length of the distributor channel [220], which facilitates the deposition of material delivered from an orifice [ 503] on the extension tip [502] in confined spaces, such as a cavity plate insert [504] or Petri dish (not shown).
    [085] In reference to the modality illustrated in Figure 1, the extension tip [130] is a projection that extends from the printhead [100], with the extension tip [130] ending in the printhead hole [085] print [114]. In this embodiment, the extension tip [130] is integral with the printhead.
    [086] In one embodiment, two or more hydrogel materials may be coaxially arranged in a hydrogel fiber distributed from the system provided herein. Referring to Figure 6, in the illustrated embodiment, the printhead [100] comprises microfluidic channels arranged to produce a coaxial hydrogel fiber comprising a hydrogel core material and hydrogel shell material. In the illustrated embodiment, the channel-carried shell material [508] is a rapidly gelling hydrogel such as alginate, and the channel-carried core material [506] is a different hydrogel chosen by the user ( e.g. collagen or fibrinogen). Channels [508] and channel [506] intersect at a hydrogel focus intersection point [510], e.g. in a "y-shaped" configuration (similar to intersection [528] shown in Figure 3 ) that join to form a focusing channel [226] downstream of the channels [506] and [508]. At the hydrogel focusing intersection [510], the shell material coaxially focuses the core material so that the shell material forms a coating around the core material. In preferred embodiments, channels [508] and [226] have a larger diameter than channel [506] to facilitate coaxial focusing of the core and shell materials. In a preferred embodiment, the purpose of the shell material is to provide the core material with physical structural support so that it can be formed into a 3D geometry. The core can be solidified after the material is deposited, and the precise solidification method is specific to different core materials. For example, the core may comprise a material that solidifies very slowly. In another embodiment, the core and shell materials comprise the same materials. In yet another embodiment, the shell material comprises a hydrogel that solidifies quickly and the core material comprises a material that will not clump together, thereby facilitating the generation of a hollow fiber.
    [087] In one embodiment, the printhead [100] depicted in Figure 6 would additionally comprise additional core material channels, each with a corresponding fluidic switch, e.g. a valve, to regulate the flow of material therein. The fluidic switch facilitates frequent and quick adjustments to the composition of the core material in the fiber that is distributed, for example, through commands provided by the programmable control processor.
    [088] In one embodiment, several printheads would be arranged, for example, in parallel, to allow simultaneous printing of multiple structures. This would increase yield production.
    [089] In some embodiments, the printhead is disposable. Using disposable printheads can reduce the likelihood of contamination from materials used in different print jobs.
    [090] The printhead can be manufactured, for example, using known microfluidic molding techniques (eg casting, printing or injection molding) and one or more moldable polymers, for example polydimethylsiloxane (PDMS). Alternatively, commercially available 3D printing technology would be used to manufacture the printhead. FLUID REMOVAL FEATURE
    [091] In one aspect, the invention provides a system for additive manufacturing of three-dimensional structures that comprises a facility for removing excess coating fluid from the receiving surface in which a first layer of material dispenses from the orifice of the cylinder head. print is deposited and optionally from a distributed hydrogel surface. During printing, it is possible for excess coating fluid to collect or "pool" on the receiving surface or on a distributed hydrogel surface. Such clumping can interfere with the deposition of distributed hydrogel from the printhead hole onto the receiving surface and/or onto one or more layers of distributed hydrogel. For example, pooled coating fluid can cause a distributed hydrogel fiber to slide from its intended position on the 3D printed structure. Therefore, in system embodiments, removal of excess coating fluid from the receiving surface and optionally from a surface of the distributed hydrogel via a fluidic removal feature can enhance additive manufacturing of three-dimensional structures. .
    [092] Excess coating fluid can be removed from the receiving surface or from a surface of one or more layers of distributed hydrogel by extracting the fluid out of those surfaces, allowing or facilitating evaporating coating fluid from those surfaces or, in embodiments, where the receiving surface is porous, excess coating fluid may be removed by extracting it through the porous surface.
    [093] In a preferred embodiment, the receiving surface comprises a porous material, the pores being sized to facilitate the passage of coating fluid through them and sized to support one or more layers of hydrogel deposited thereon.
    [094] Referring to Figures 7 and 8, in the illustrated embodiment, a print bed [108] comprises a porous membrane [400], which serves as the surface to receive a first layer of distributed material (i.e., the receiving surface). The porous membrane [400] is held in place on the print bed [108] between a housing part [408] and a lid part [402]. The box piece [408] is a container, which can be of any shape suitable for receiving and containing the liquid (eg square, round). The internal space of the box piece [408] is referred to as a chamber [404]. The box piece [408] has a top surface [409] that comprises a recessed cover [412] that extends from the perimeter of the top surface [409] of the box piece [408]. The upper surface [409] comprises an opening defined by one or more walls [410], wherein the opening is surrounded by the recessed lid [412] and which extends into the housing part [408].
    [095] In further reference to the embodiments illustrated in Figures 7 and 8, the cover piece [402] comprises an upper surface [403] having an opening [416] extending therethrough and side walls [418] configured to fit around the recessed lid [412] of the housing part [408] thereby facilitating placement of the lid part [402] on the top surface [409] of the housing part [408]. When the lid part [402] is placed on the housing part [408], the openings in the housing and the lid part [416] align. In operation, the porous membrane [400] is placed on the upper surface [409] of the box part [408] so that it extends over the opening in the upper surface [409] of the box part [408], the cap [402] is then placed on top of the box piece [408] and pressed down. The downward pressure of the lid piece [402] stretches the porous membrane [400] over the opening in the upper surface [409] of the box piece [408] thereby retaining the porous membrane [400] between the box piece [408] and the cover piece [402]. In preferred embodiments, the lid part [402] and the box part [408] fit together tightly, thereby providing a connection that will remain secure during operation of the system provided herein.
    [096] In further reference to the modalities illustrated in Figures 7 and 8, the box part [408] comprises a solid base [414] and at least one outlet duct [406] to direct the fluid in the opposite direction to the chamber [404] ] and a vacuum source (not shown) in fluid communication with the outlet duct [406] of the chamber [404]. The porous membrane [400] comprises pores sized to facilitate the passage of coating fluid. The vacuum source (not shown) coupled to the outlet duct [406] can be actuated to extract excess coating fluid collected in the porous membrane [400] through the porous membrane [400] in chamber [404] and chamber [ 404] through the outlet [406], leaving the hydrogel fiber in its distributed configuration on top of the porous membrane [400].
    [097] In a preferred embodiment, a feature to remove excess coating fluid from the receiving surface and, optionally, a distributed hydrogel surface can be included in a system configured to distribute materials in a multi-walled dish or Petri dish. For example, referring to Figure 9, in the illustrated embodiment, a commercially available cavity plate insert [504] is placed on top of the box part [408]. Some cavity plate inserts [504] have a gasket shape with a base produced from a porous membrane material [512]. In the illustrated embodiment, a gasket [514] is placed between the cavity plate [512] and box part [408] inserts to improve the seal between the two parts [504 and 408]. In such embodiments, the porous membrane [512] of the cavity plate insert [504] would serve as the "receiving surface" and any excess coating fluid could be removed therefrom using a vacuum coupled to the outlet duct [406 ], as described above, or using one of the other fluid removal features described below.
    [098] In one embodiment (not shown), the receiving surface on the print bed comprises or is placed adjacent to an absorption material, which facilitates the absorption of excess coating fluid from the receiving surface. For example, a cavity plate insert that has a base produced from a porous membrane material (e.g., as shown in Figure 9) or any other porous membrane substrate, could be placed on top of or adjacent to a absorbent material, such as a sponge. The absorbing material would act to draw excess coating fluid away from the receiving surface. In embodiments where the absorbent material is disposed below a porous receiving surface, excess coating fluid on the receiving surface would be drawn through the porous receiving surface and into the absorption material, thereby preventing pooling of excess coating fluid on the surface. receiving surface. In embodiments where the absorbent material is disposed immediately beside or on top of a portion of the receiving surface (e.g., at the periphery of the receiving surface so as not to interfere with the deposition of distributed material), the coating fluid in excess would be drawn off the receiving surface and onto the absorbent material.
    [099] In one embodiment (not shown), instead of using one of the print beds described above, one or more tubes may be provided in an area close to the receiving surface and close to the printhead hole. The one or more tubes can be fluidly coupled to a vacuum source (not shown), which can provide suction to remove excess coating fluid from the receiving surface and, optionally, a distributed hydrogel surface. In such embodiments, a solid or porous receiving surface may also be used.
    [0100] In one embodiment, illustrated in Figure 10, the printhead is configured to further comprise one or more vacuum channels [700a, 700b], wherein the one or more vacuum channels each have an orifice [0100] 702a, 702b] located next to the printhead hole [222]. The one or more vacuum channels [700a, 700b] each have an input [704a, 704b] configured to facilitate fluid communication with one or more vacuums (not shown). When the printhead [100] is in fluid communication with a vacuum, the one or more vacuum channels [702a, 702b] direct negative pressure to an area of the receiving surface where materials are being dispensed or have been dispensed from the orifice. printhead [222] and/or for a portion of the surface area of the dispensed hydrogel, thereby extracting excess coating fluid from the receiving surface and, optionally, a surface of the dispensed hydrogel, thereby eliminating pooling of coating fluid on the receiving surface and/or on the dispensed hydrogel.
    [0101] In one embodiment, the one or more vacuum tubes are provided, at least in part, in one or more extensions projecting from the printhead, the extensions projecting in the same general direction as the extension comprising the orifice printhead and distributor channel (see, for example, Figure 10). In such embodiments, the one or more extensions comprising vacuum tubes do not extend beyond the extension comprising the printhead orifice and dispensing channel so as not to interfere with the dispensed and deposited hydrogel.
    [0102] It is contemplated that, in some embodiments, the fluid removal feature may be a feature of the coating fluid composition itself. For example, the coating fluid composition can be designed to evaporate after it is dispensed from the printhead orifice, thereby eliminating excess coating fluid pooling on the receiving surface or dispensed hydrogel surfaces. For example, pot coating fluid will have a boiling point that results in evaporation after being dispensed, while remaining in a liquid state before being dispensed. PRINTING METHOD OF A THREE-DIMENSIONAL STRUCTURE
    [0103] In one aspect, a method of printing a three-dimensional (3D) structure is provided.
    [0104] The method first comprises providing a design for a 3D structure to be printed. The design can be created using commercially available CAD software. In one embodiment, the design comprises information regarding specific materials (eg, for heterogeneous structures comprising multiple materials) to be assigned to specific geometric locations in the design.
    [0105] The method comprises the use of a 3D printer, wherein the printer comprises: a printhead, a receiving surface for receiving material distributed by the printhead and a positioning unit operably coupled to the receiving surface, the positioning unit for positioning the printhead at a location in three-dimensional space above the receiving surface. For example, various embodiments of the printing system provided in the present document can be used in the method of printing a 3D structure.
    [0106] The method comprises providing at least two materials to be distributed by the printhead, such as a coating fluid and a hydrogel fluid. In preferred embodiments, one or more cell types are compatible with and, optionally, distributed within the hydrogel. In a preferred embodiment, the coating fluid serves as a lubricating agent to lubricate the hydrogel movement in and out of the printhead. In a preferred embodiment, the coating fluid comprises a crosslinking agent to solidify at least a portion of the hydrogel before or while it is delivered from the printhead.
    [0107] The method comprises communicating the design to the 3D printer. Communication can be achieved, for example, by a programmable control processor.
    [0108] The method comprises controlling the relative positioning of the printhead and the receiving surface in three-dimensional space and, simultaneously, distributing, from the printhead, the coating fluid and the hydrogel, alone or in combination. In preferred embodiments, the materials delivered from the preprint are delivered coaxially so that the coating fluid surrounds the hydrogel. Such a coaxial arrangement allows the crosslinking agent to solidify the hydrogel, thereby resulting in a solid hydrogel fiber, which is distributed from the print head.
    [0109] The method comprises depositing a first layer of the materials distributed on the receiving surface, the first layer comprising a specific material arrangement by the project and iteratively repeating the deposition step, depositing the subsequent material on the first and subsequent layers of material , thereby depositing layer after layer of materials distributed in a design-specific geometric arrangement to produce the cell-loaded 3D structure.
    [0110] In preferred embodiments, a plurality of materials, e.g. multiple hydrogels, at least some of which comprise one or more cell types, are deposited in a controlled sequence, thereby allowing a controlled array of hydrogels and types of cells to be deposited in a specific geometric arrangement by the project.
    [0111] In preferred embodiments, the method comprises removing excess coating fluid from the receiving surface and optionally from the surface of the delivered hydrogel. For example, the step of removing excess coating fluid can be done continuously throughout the printing process, thereby removing excess fluid that might otherwise interfere with the layering of materials distributed in the geometric arrangement. provided by the project. Alternatively, the step of removing excess coating fluid may be done intermittently throughout the printing process in sequence with or simultaneously with one or more deposition steps. In some embodiments, removal of excess coating fluid is achieved by drawing the fluid off the receiving surface and optionally off a surface of the delivered hydrogel. In another embodiment, removal of excess coating fluid is achieved by drawing excess fluid through the receiving surface, wherein the receiving surface comprises pores sized to allow passage of the coating fluid. In another embodiment, removal of excess coating fluid is achieved by providing a coating fluid which evaporates after being dispensed from the printhead orifice. EXEMPLARY USES OF SYSTEM MODALITIES AND PRINTING METHOD THREE-DIMENSIONAL CELL LOADED STRUCTURES
    [0112] In some embodiments, structures generated using the system and method provided herein may be useful in the field of drug discovery, where, for example, determining cellular responses to various compounds and chemical compositions is of interest. The use of 3D cell cultures fabricated using modalities of the systems and methods provided herein can provide experimental conditions that more closely resemble tissue and cell conditions in vivo relative to 2D cell cultures. The 3D arrangement of cells can more closely mimic in vivo cell-cell interactions and responses to external stimuli, and the heterogeneous nature of the 3D structures that can be generated using the device and methods provided allow for the study of tissues and organs. in potential. It is contemplated that 3D cell-loaded structures fabricated using modalities of the systems and methods provided herein may provide a similar benefit to the cosmetics industry by offering an alternative means of testing cosmetic products.
    [0113] In some embodiments, various system embodiments and the method provided herein are compatible with standard cavity plate technology. Cavity plates or cavity plate inserts can be used with or as part of the print bed in the methods and systems provided herein. Various embodiments of the system and method provided herein are thus compatible with instruments and practices using cavity plates, allowing them to be readily integrated into existing process flows.
    [0114] In some embodiments, the microfluidic channels within the printhead are compatible with other microfluidic modules. For example, known microfluidic modules can be included in the printhead of the systems provided herein upstream of the printhead orifice. Such modules may include, for example, cell counting, cell storage, cell analysis and/or concentration gradient generation modules.
    [0115] In some embodiments, 3D printing throughput can be increased by adding additional printheads in parallel to the system. Each printhead comprising all the elements required to print a multi-material structure thus allows multiple 3D structures to be printed simultaneously by adding additional printheads to the system.
    [0116] Although the invention has been described with reference to certain specific embodiments, various modifications thereof will be evident to those skilled in the art without departing from the purpose and scope of the invention as highlighted in the appended claims thereof. Any examples provided herein are included for the purpose of illustrating the invention only and are not intended to limit the invention in any way. Any drawings provided herein are for the purpose of illustrating various aspects of the invention only and are not intended to be drawn to scale or limit the invention in any way. The disclosures of all prior art cited herein are incorporated herein by reference as set forth in their entirety. REFERENCES The following references are provided as examples of the known art in connection with the present invention. The following listing is not intended to be a comprehensive list of all relevant technique. The complete contents of all references listed in this specification, which includes the following documents, are incorporated herein by reference as if presented in their entirety. 1. Su-Jung Shin, Ji-Young Park, Jin-Young Lee, Ho Park, Yong-Doo Park, Kyu-Back Lee, Chang-Mo Whang, and Sang-Hoon Lee, ""On the fly" continuous generation of alginate fibers using a microfluidic device", Langmuir, Volume 23, 2007, pages 9104 to 9108. 2. Saif Khalil, and Wei Sun, "Bioprinting endothelial cells with alginate for 3D tissue constructs", Journal of Biomechanical Engineering, Volume 131, 2009 , pages 11 1002-1 - 1 1 1002-8 3. Min Hu, Rensheng Deng, Karl M. Schumacher, Motoichi Kurisawa, Hongye Ye, Kristy Purnamawati, and Jackie Y. Ying, "Hydrodynamic spinning of hydrogel fibers", Biomaterials , Volume 31, 2010, pages 863 to 869. 4. Byung Kim, Intae Kim, Wooseok Choi, Sun Won Kim, JooSung Kim, and Geunbae Lim, "Fabrication of cell-encapsulated alginate microfiber scaffold using microfluidic channel", Journal of Manufacturing Science and Engineering, Volume 130, 2008, pages 021016-1 - 021016-6. 5. Edward Kang, Su-Jung Shin, Kwang Ho Lee, and Sang-Hoon Lee, "Novel PDMS cylindrical c hannels that generate coaxial flow, and application to fabrication of microfibers and particles", Lab on a Chip, Volume 10, 2010, page 1856 to 1861. 6. Hiroaki Onoe, Riho Gojo, Yukiko Tsuda, Daisuke Kiriyaand, and Shoji Takeuchi, " Core-shell gel wires for the construction of large area heterogeneous structures with biomaterials", IEEE MEMS Conference, 2010, pages 248 to 251. 7. Setareh Ghorbanian (2010), Microfluidic probe for direct write of soft cell scaffolds, M.Eng. Thesis. McGill University: Canada. 8. Edward Kang, Gi Seok Jeong, Yoon Young Choi, Kwang Ho Lee, AN Khademhosseini, and Sang-Hoon Lee, "Digitally tunable physicochemical coding of material composition and topography in continuous microfibers", Nature Materials, Volume 10, 201 1, pages 877 to 883. 9. EP 2489779 A1 10. US 2006/010501 1 A1 1 1. US 201 1/0136162 A1 12. US 2012/0089238 A1 2.
    权利要求:
    Claims (42)
    [0001]
    1. System for additive manufacturing of three-dimensional structures, the system CHARACTERIZED in that it comprises: - at least one print head (100) for receiving and distributing materials, the materials comprising at least one first material and at least one second material, the print head print (100) comprising: - an orifice (114) for distributing the materials; - microfluidic channels (200, 202, 204, 206) arranged inside the print head (100) comprising one or more first channels for receiving and directing the hair at least one first material and one or more second channels for receiving and directing the at least one second material, the one or more second channels intersecting at a first point of intersection (228) with the one or more first channels, the one or more second channels and the one or more first channels joining at the first point of intersection (228) to form a distributor channel (220) that extends into the orifice (114); e-fluidic switches (210, 212, 214, 216), each fluidic switch corresponding to one of the microfluidic channels (200, 202, 204, 206) in the printhead (100) and configured to allow or not allow flow of fluid in the microfluidic channels (200, 202, 204, 206) of the printhead (100) when actuated; - a receiving surface (109) for receiving a first layer of materials dispensed from the orifice (114); - a positioning to position the hole (114) of the printhead (100) in three-dimensional space, the positioning unit operably coupled to the printhead (100); and - a dispensing means for dispensing the materials from the orifice (114) of the printhead (100).
    [0002]
    2. System according to claim 1, CHARACTERIZED in that the at least one first material comprises a coating fluid and/or a crosslinking agent to solidify the at least one second material upon contact with it at the point of intersection (228) and/or the distributor channel (220).
    [0003]
    3. System, according to claim 1 or 2, CHARACTERIZED by the fact that each second channel has a smaller diameter than each of the first channels and the distributor channel (220), in which the flow of the first channels forms a coaxial coating in around the at least a second material in the distributor channel (220).
    [0004]
    4. System according to any one of claims 1 to 3, CHARACTERIZED in that the at least one second material comprises a hydrogel and/or living cells.
    [0005]
    A system as claimed in any one of claims 1 to 4, CHARACTERIZED in that it further comprises a fluid removal facility for first removing excess material delivered from the printhead (100).
    [0006]
    6. System, according to claim 5, CHARACTERIZED by the fact that the receiving surface (109) comprises a porous membrane (400) comprising pores sized to allow passage of the first excess material through them.
    [0007]
    7. System according to claim 6, CHARACTERIZED in that the fluid removal feature comprises absorbent material or a vacuum to extract the first excess material away from the receiving surface (109) and/or away from the fur least a second material distributed on the receiving surface (109).
    [0008]
    8. System, according to claim 7, CHARACTERIZED by the fact that the absorbent material or vacuum is applied below the porous membrane (400).
    [0009]
    9. System, according to claim 7, CHARACTERIZED by the fact that the vacuum is applied above the receiving surface (109).
    [0010]
    10. System, according to claim 9, CHARACTERIZED by the fact that the vacuum is applied through one or more vacuum channels (700a, 700b) provided in the printhead (100), the one or more vacuum channels ( 700a, 700b) having an orifice (702a, 702b) situated close to the orifice (114) of the printhead (100).
    [0011]
    11. System, according to any one of the preceding claims, CHARACTERIZED by the fact that the system comprises reservoirs (110) to contain the materials, the reservoirs (110) being fluidly coupled respectively to the microfluidic channels (200, 202, 204, 206 ) on the printhead (100).
    [0012]
    12. System, according to claim 11, CHARACTERIZED by the fact that the print head (100) comprises at least two inputs (208a, 208b, 208c, 208d) to receive the materials from the reservoirs (110), each of the inlets (208a, 208b, 208c, 208d) being in fluid communication with respective microfluidic channels (200, 202, 204, 206) and respective reservoirs (110).
    [0013]
    13. System, according to any one of the preceding claims, CHARACTERIZED by the fact that the distribution means comprises a pressure control unit (112).
    [0014]
    14. System, according to any one of the preceding claims, CHARACTERIZED by the fact that the fluidic switches (210, 212, 214, 216) comprise valves.
    [0015]
    15. System, according to any one of the preceding claims, CHARACTERIZED by the fact that the print head comprises a hollow projection configured to extend from the hole (114) towards the receiving surface (109).
    [0016]
    16. System, according to any one of the preceding claims, CHARACTERIZED in that the system comprises a programmable control processor for controlling the positioning unit and for controlling the distribution of materials from the printhead (100) on the surface receiver (109).
    [0017]
    17. System according to any one of the preceding claims, CHARACTERIZED in that each first channel is configured to flank the one or more second channels at the first intersection point (228).
    [0018]
    18. System according to any one of the preceding claims, CHARACTERIZED by the fact that the printhead (100) comprises two second channels, each of the second channels being adapted to transport respective second materials, the two second channels crossing each other in a second intersection point (207, 510) and joining at the second intersection point (207, 510) to form a third channel (226) that extends to the first intersection point (228).
    [0019]
    19. System according to claim 18, CHARACTERIZED in that the two second channels comprise a core channel (506) for receiving and directing a core material and a housing channel (508) for receiving and directing a material of carcass; wherein the housing channel (508) and the third channel (226) have larger diameters than the core channel (506) to facilitate coaxial focusing of the core and housing materials, wherein flow of housing material from the housing channel (508) forms a coaxial sheath around the core material in the third channel (226) and wherein the third channel (226) is flanked by the at least two first channels joining together to form the distributing channel (220) which extends to the hole (114).
    [0020]
    20. System for additive manufacturing of three-dimensional structures, the system CHARACTERIZED in that it comprises: - at least one printhead (100) for receiving and distributing materials, the materials comprising a first material and a second material, the printhead (100) comprising: - an orifice (114) for dispensing the materials; - microfluidic channels (200, 202, 204, 206) for receiving and directing the materials to the orifice (114); e-fluidic switches (210, 212, 214, 216), each fluidic switch corresponding to one of the microfluidic channels (200, 202, 204, 206) in the printhead (100) and configured to allow or not allow flow of fluid in the microfluidic channels (200, 202, 204, 206) in the printhead (100) when actuated; - a receiving surface (109) to receive the dispensed materials from the orifice; - a fluid removal feature to remove first excess material distributed from the orifice (114); - a positioning unit for positioning the orifice of the printhead in three-dimensional space, the positioning unit operably coupled to the printhead (100); and - a dispensing means for dispensing the materials from the orifice (114) of the printhead (100).
    [0021]
    21. System, according to claim 20, CHARACTERIZED in that the first material comprises a coating fluid and the second material comprises a hydrogel.
    [0022]
    22. System according to claim 20 or 21, CHARACTERIZED in that the fluid removal feature comprises: a vacuum to extract the first excess material away from or through the receiving surface (109) and/or the second material distributed on the receiving surface (109); or an absorbent material to draw away from the receiving surface (109) the first excess material.
    [0023]
    23. System, according to claim 22, CHARACTERIZED by the fact that the vacuum is applied above the receiving surface and through one or more vacuum channels (700a, 700b) provided in the printhead (100), the one or more vacuum channels (700a, 700b) having an orifice (702a, 702b) situated close to the orifice (114) of the printhead (100).
    [0024]
    24. System according to any one of claims 20 to 23, CHARACTERIZED in that the printhead (100) comprises at least two first channels for receiving and directing the first material and one or more second channels for receiving and directing the second material, the one or more second channels and each of the first channels joining at a first point of intersection (228) to form a distributor channel (220) that extends into the orifice (114).
    [0025]
    25. System, according to claim 24, CHARACTERIZED by the fact that each second channel has a smaller diameter than each of the first channels and the distributor channel (220), in which the flow of the first channels forms a coaxial coating around of the second material in the distributor channel.
    [0026]
    26. System, according to claim 24, CHARACTERIZED by the fact that the printhead (100) comprises two second channels, each of the second channels being adapted to transport respective second materials, the two second channels crossing each other in a second intersection point (207, 510) and joining at the second intersection point (207, 510) to form a third channel (226) that extends to the first intersection point (228).
    [0027]
    27. System, according to claim 26, CHARACTERIZED in that the two second channels comprise a core channel (506) for receiving and directing a core material and a housing channel (508) for receiving and directing a material of carcass; wherein the housing channel (508) and the third channel (226) have larger diameters than the core channel (506) to facilitate coaxial focusing of the core and housing materials, wherein flow of housing material from the housing channel (508) forms a coaxial sheath around the core material in the third channel (226) and wherein the third channel (226) is flanked by the at least two first channels joining together to form the distributing channel (220) which extends to the hole (114).
    [0028]
    28. System, according to any one of claims 20 to 27, CHARACTERIZED by the fact that the print head (100) comprises reservoirs (110) to contain the materials, the reservoirs (110) being fluidly coupled respectively to the microfluidic channels ( 200, 202, 204, 206) on the printhead (100).
    [0029]
    29. System, according to claim 28, CHARACTERIZED by the fact that the print head (100) comprises at least two inputs (208a, 208b, 208c, 208d) to receive the materials from the reservoirs (110), each of the inputs being in fluid communication with respective microfluidic channels (200, 202, 204, 206) and respective reservoirs (110).
    [0030]
    30. System according to any one of claims 20 to 29, CHARACTERIZED by the fact that the fluidic switches (210, 212, 214, 216) comprise valves.
    [0031]
    31. System according to any one of claims 20 to 30, CHARACTERIZED in that the print head (100) comprises a programmable control processor to control the positioning unit and to control the distribution of materials from the head print (100) on the receiving surface (109).
    [0032]
    32. System according to any one of claims 20 to 31, CHARACTERIZED in that the print head (100) comprises a hollow projection configured to extend from the hole (114) towards the receiving surface (109).
    [0033]
    33. Method of printing a three-dimensional structure (3D), the method CHARACTERIZED in that it comprises: - providing a 3D printer, the printer comprising: - at least one print head (100) comprising an orifice (114) for dispensing materials; - a receiving surface (109) for receiving a first layer of materials distributed from the orifice (114) of the printhead (100); and - a positioning unit operably coupled to the printhead (100), the positioning unit for positioning the printhead (100) in three-dimensional space; - providing the materials to be dispensed, the materials to be dispensed comprising a coating fluid and one or more hydrogels;- encode the printer with a 3D structure to be printed;- dispense from the printhead hole (114) the materials to be dispensed, in which the coating fluid and the hydrogel are distributed in a coaxial arrangement, and in which the coating fluid surrounds the hydrogel;- depositing a first layer of the distributed materials on the receiving surface (109);- repeating the deposition step by deposition of subsequent distributed material on the first and any subsequent layers of deposited material, thereby depositing layer after layer of materials distributed in a geometric arrangement in accordance with the 3D structure; and - removing excess coating fluid delivered by the printhead orifice (114) at one or more points in time during or between deposition steps.
    [0034]
    34. Method according to claim 33, CHARACTERIZED in that the coating fluid comprises a crosslinking agent suitable for crosslinking and solidifying the hydrogel upon contact therewith, the contact creating a hydrogel fiber.
    [0035]
    35. Method according to claim 33 or 34, CHARACTERIZED in that the deposition step and the removal step are performed continuously, thereby continuously removing excess coating fluid as the layers of distributed materials are deposited.
    [0036]
    36. Method according to claim 33 or 34, CHARACTERIZED in that the removal step is performed intermittently between and/or at the same time as the deposition step, thereby intermittently removing excess coating fluid as layers of distributed materials are deposited.
    [0037]
    37. Method, according to any one of claims 33 to 36, CHARACTERIZED by the fact that the one or more hydrogels are adapted to support growth and/or proliferation of live cells dispersed therein.
    [0038]
    38. Method according to claim 33, CHARACTERIZED in that the printhead (100) comprises microfluidic channels (200, 202, 204, 206) comprising at least two first channels for receiving and directing the coating fluid and one or more second channels to receive and direct the one or more hydrogels, the second channels intersecting at a first point of intersection (228) with the first channels, the second and first channels merging at the first point of intersection (228) to form a distributor channel (220) that extends into the hole (114).
    [0039]
    39. Method, according to claim 38, CHARACTERIZED in that the materials to be distributed comprise a core hydrogel, a housing hydrogel, and a coating fluid, and wherein the housing hydrogel and the core hydrogel are distributed in a coaxial arrangement, in which the housing hydrogel surrounds the core hydrogel, and in which the coating fluid surrounds the housing hydrogel.
    [0040]
    40. Method according to claim 39, CHARACTERIZED in that the coating fluid comprises a crosslinking agent suitable for crosslinking and solidifying the carcass hydrogel, optionally additionally suitable for crosslinking and solidifying the core hydrogel, upon contact with the same, the contact creating a hydrogel fiber.
    [0041]
    41. Method according to claim 40, CHARACTERIZED by the fact that the core hydrogel is incapable of gelling upon contact with the crosslinking agent, thus facilitating the creation of a hollow hydrogel fiber.
    [0042]
    42. Method according to claim 39, CHARACTERIZED by the fact that the core hydrogel solidifies after deposition.
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    同族专利:
    公开号 | 公开日
    RU2016100442A|2017-07-18|
    KR20210097816A|2021-08-09|
    EP3670155A1|2020-06-24|
    EP3007882A1|2016-04-20|
    CN105579219A|2016-05-11|
    JP2020182474A|2020-11-12|
    JP6733014B2|2020-07-29|
    SG11201510138RA|2016-01-28|
    AU2014280825A1|2016-01-28|
    CA2915737A1|2014-12-18|
    PT3007882T|2020-02-20|
    KR102284531B1|2021-08-03|
    DK3007882T3|2020-02-24|
    WO2014197999A1|2014-12-18|
    BR112015031320A2|2017-07-25|
    CN105579219B|2017-09-01|
    JP2019201644A|2019-11-28|
    IL243014A|2019-08-29|
    EP3007882B1|2019-11-20|
    US20210317404A1|2021-10-14|
    AU2014280825B2|2018-01-04|
    EP3007882A4|2017-03-22|
    US20160136895A1|2016-05-19|
    JP2016530874A|2016-10-06|
    JP6550379B2|2019-07-24|
    PL3007882T3|2020-06-01|
    ES2770418T3|2020-07-01|
    US11046930B2|2021-06-29|
    KR20160030939A|2016-03-21|
    CA2915737C|2016-09-13|
    RU2643138C2|2018-01-30|
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    法律状态:
    2018-02-27| B06F| Objections, documents and/or translations needed after an examination request according [chapter 6.6 patent gazette]|
    2019-12-10| B06U| Preliminary requirement: requests with searches performed by other patent offices: procedure suspended [chapter 6.21 patent gazette]|
    2021-07-13| B07A| Application suspended after technical examination (opinion) [chapter 7.1 patent gazette]|
    2021-07-20| B350| Update of information on the portal [chapter 15.35 patent gazette]|
    2021-11-09| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|
    2022-01-18| B16A| Patent or certificate of addition of invention granted [chapter 16.1 patent gazette]|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 13/06/2014, OBSERVADAS AS CONDICOES LEGAIS. |
    优先权:
    申请号 | 申请日 | 专利标题
    US201361834420P| true| 2013-06-13|2013-06-13|
    US61/834.420|2013-06-13|
    PCT/CA2014/050556|WO2014197999A1|2013-06-13|2014-06-13|System for additive manufacturing of three-dimensional structures and method for same|
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