Sterile cover

Abstract

The invention relates to a sterile mask (2) intended for printing a biological three-dimensional structure, comprising:-a rigid top plate (4), 5-a rigid base plate (6) comprising a main face (16) intended to receive a biological three-dimensional structure,-a flexible side wall (8), the present invention relates to a syringe comprising a top plate, a base plate, a flexible side wall fixed to the top plate and to the base plate to form a hermetically closed housing (10) defining an inner chamber (11), a pierceable membrane (22) or needle adapter arranged on the top plate, a stretching arm (36) hinged to the top plate and to the base plate, the flexible side walls and the extension arms conform to allow the top plate to move in three orthogonal directions (X, Y, Z) relative to the base plate; the at least one stretching arm is resilient and pushes the top plate away from the base plate.

Description

Technical Field

[0001] The present invention relates to a sterile enclosure intended for printing biological three-dimensional structures. Background Art

[0002] It is known to print biological three-dimensional structures on a printing platform of a three-dimensional printer. After the structure is formed, the printed component is immersed in a culture medium for a cell culture process. Therefore, once printed, the biological three-dimensional structure must be transferred to a separate bioreactor for a cell culture process. However, it is difficult to maintain sterile conditions during the transfer of the biological three-dimensional structure to the bioreactor.

[0003] In order to avoid this difficulty, it is also known to print out biological three-dimensional structures in a container suitable for containing and controlling the culture medium, and the container also includes a printer or a movable robotic arm equipped with a print head. After printing, it is no longer necessary to transfer the biological three-dimensional structure to a separate bioreactor for the cell culture process.

[0004] The disadvantage of this mode of operation is that the container is complex and bulky, because the container must contain the printer or the movable robotic arm. Therefore, when the biological three-dimensional structure has been formed, the printer or the movable robotic arm equipped with the print head cannot be used to print out another structure, because the container must remain hermetically closed during the cell culture process. Due to the size of the printer or the movable robotic arm and the fact that its mechanical parts are lubricated, it may be difficult to sterilize the printer or the movable robotic arm and keep it sterile. Because the container is large and complex, it is difficult to move the container during the cell culture process to place it in another room. Summary of the invention

[0005] A first object of the invention is to propose a sterile cover that allows the print head to be freely displaced in at least two orthogonal directions and in particular in three orthogonal directions without generating stress on the needle and without the risk of the needle bending or breaking. Thus, the shape of the biological three-dimensional structure can be better controlled. Another result is that the printing platform does not need to be movable in the vertical direction, because the sterile cover allows the print head to be freely moved in this direction. Thus, a less complex printer can be used. In the same way, the printer can use a smaller motor because the resistance applied to the robot arm is smaller.

[0006] A second object of the invention is to propose a sterile hood that is not too bulky, complex and cumbersome.Since the printer or part of it is not contained in the sterile hood, its size and weight are significantly reduced.

[0007] A third object of the present invention is to provide a sterile cover which can be freely moved and stored after the printing process.

[0008] A fourth object of the invention is to propose a sterile hood that allows the printer to be reused immediately after the biological three-dimensional structure has been formed.

[0009] A fifth object of the present invention is to propose a sterile hood that simplifies the printing of biological three-dimensional structures and their cultivation in a single container.

[0010] The sixth object of the present invention is to provide a method for printing and culturing three-dimensional biological structures.

[0011] These objects have been achieved by a sterile hood as defined in the independent claim. Secondary characteristics of the sterile hood are set out in the dependent claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 is a perspective view of a sterile cover according to a first embodiment of the present invention;

[0013] Figure 2 yes Figure 1 A top view of the top plate of the sterile cover shown in FIG.

[0014] Figure 3 yes Figure 1 A side view of the top plate of the sterile cover shown in FIG.

[0015] Figure 4 yes Figure 1 A perspective view of the diaphragm of the sterile cover shown in FIG.

[0016] Figure 5 yes Figure 1 A side view of a base plate of a sterile cover shown in FIG.

[0017] Figure 6 yes Figure 1 A side view of the extension arm of the sterile cover shown in FIG.

[0018] Figure 7 is a perspective view of a sterile cover according to a second embodiment of the present invention. DETAILED DESCRIPTION

[0019] The sterile cover 2 according to the invention is intended to be used together with a printing cartridge or a printing syringe for printing biological three-dimensional structures in the sterile cover.

[0020] The printing syringe or printing cartridge may include a bio-ink composition. A bio-ink composition is a biological material that can form hydrogels and cell groups in a controlled three-dimensional shape. An example of a bio-ink is described in WO 2017115056.

[0021] Preferably, for the printing operation, the printing syringe or ink cartridge is mounted on a displaceable printer, for example on a displaceable robot arm. Printing can also be done manually with a printing nozzle.

[0022] In the following description, the term "printing" means to perform manual deposition or injection of bio-ink or 3D printing with a displaceable robotic arm. In a similar manner, the term "printing needle" is used to refer to a needle or a deposition nozzle.

[0023] refer to Figure 1 The sterile cover 2 according to the present invention comprises a rigid top plate 4 , a rigid base plate 6 , and flexible side walls 8 fixed to the base plate and the top plate to form a hermetically closed housing 10 defining an inner chamber 11 .

[0024] In the embodiment shown, the top plate 4 presents a generally disc-like shape having two opposite main faces 12 14. The surfaces of the main faces are substantially equal to the surface of the upper main face 16 of the base plate.

[0025] Advantageously, this large surface of the top plate makes it possible to accommodate a large number of sterile connectors and / or ports.

[0026] Alternatively, the main faces 12 , 14 of the top plate have a surface that is 50% to 100% of the surface of the upper main face 16 of the base plate.

[0027] In the embodiment shown, the top plate 4 includes a port 18 that allows, for example, connection. In the embodiment shown, the port is a sterile exhaust port. The sterile exhaust port allows air to circulate during printing. Thus, the pressure in the inner chamber does not change significantly. Alternatively, the top plate 4 may include one or several sterile connectors, such as the commercialized 1000- ... Connector or one or more hose connectors.

[0028] Alternatively, the top plate 4 may comprise one or several sterile ports.

[0029] The top plate 4 is made of one of the following materials: biocompatible plastics; metals such as stainless steel; and inorganic materials such as glass and ceramics; polymer rigid materials, preferably polyethylene (PE), polyethylene terephthalate (PET), polyurethane (PU), polyvinyl chloride (PVC) or acid polylactide (acide polylactique).

[0030] The top plate 4 comprises a hole 20 covered by a pierceable membrane 22. The pierceable membrane 22 is intended to be pierced by a printing needle. The pierceable membrane is a septum.

[0031] exist Figure 4An example of a diaphragm is shown in FIG. The diaphragm takes the form of a cover 24. The cover 24 is fixed to the top plate in a sealed manner. The cover can be glued or otherwise fitted into the hole 20 with a certain strength.

[0032] Preferably, the hole 20 is located in the center of the top plate 4 .

[0033] Alternatively, the diaphragm 22 is replaced by a needle adapter 26 that holds a printing needle, such as Figure 7 as shown in .

[0034] The top plate 4 further comprises four fasteners 28. The fasteners are regularly angularly spaced around the top plate. Each fastener 28 comprises a pivot 29 on which an extension arm 36 is hinged about an upper hinge axis 38.

[0035] The base plate 6 comprises a main upper face 16 facing the top plate 4. In the example shown, the base plate 6 has the shape of a circular disk.

[0036] The main upper surface 16 is flat. The main upper surface 16 of the top plate is intended to receive a biological three-dimensional structure.

[0037] The base plate 6 comprises at least one fluid inlet port 32 and at least one fluid outlet port 34 for circulating the cell culture medium.

[0038] The base plate 6 further comprises four fasteners 28. The fasteners are regularly angularly spaced around the base plate in front of the fasteners 28 of the top plate. Each fastener 28 comprises a pivot 29 on which an extension arm 36 is articulated about a lower articulation axis 38.

[0039] The substrate 6 is made of one of the following materials: biocompatible plastic; metal, such as stainless steel; and inorganic materials, such as glass and ceramics; polymer rigid material, preferably polyethylene (PE), polyethylene terephthalate (ET), polyurethane (PU), polyvinyl chloride (PVC) or acid polylactide.

[0040] Flexible side walls 8 are secured to top panel 4 and base panel 6 to form container 10 .

[0041] In the embodiment shown, the flexible side wall 8 presents an annular shape. Advantageously, this annular shape limits the movement of the flexible side wall towards the inner chamber during printing.

[0042] Alternatively, the flexible side wall may have a cylindrical shape.

[0043] Preferably, the flexible side wall 8 is transparent.

[0044] The Shore hardness of the flexible side wall 8 is between A0 and A100 and preferably between A0 and A30. The Shore hardness is measured by a Shore hardness meter.

[0045] The elongation of the flexible side wall 8 is between 300% and 2000% and preferably between 800% and 1100%.In the present patent application, the term "elongation" is the ratio of the elongation of a material to the length of the material before stretching.

[0046] The flexible wall 8 is made of biocompatible plastic. Specifically, the flexible wall 8 can be made of one of the following materials:

[0047] - Unsaturated rubbers that can be cured by sulfur vulcanization:

[0048] Natural polyisoprene: cis-1,4-polyisoprene natural rubber (NR) and trans-1,4-polyisoprene eucommia gum

[0049] Synthetic polyisoprene (IR stands for isoprene rubber)

[0050] Polybutadiene (BR stands for butadiene rubber)

[0051] Chloroprene rubber (CR), polychloroprene, chloroprene (Neoprene), its Baypren, etc.

[0052] Butyl rubber (copolymer of isobutylene and isoprene, IIR)

[0053] o Halogenated butyl rubber (chlorobutyl rubber: CIIR; bromobutyl rubber: BIIR)

[0054] Styrene-butadiene rubber (copolymer of styrene and butadiene, SBR)

[0055] Nitrile rubber (copolymer of butadiene and acrylonitrile, NBR), also known as Buna N rubber

[0056] o Hydrogenated nitrile rubber (HNBR) Therban and Zetpol

[0057] - Saturated rubbers which cannot be cured by sulfur vulcanization:

[0058] EPM (ethylene propylene rubber, a copolymer of ethylene and propylene) and EPDM rubber (ethylene propylene diene rubber, a terpolymer of ethylene, propylene and a diene component)

[0059] ·Epichlorohydrin rubber (ECO)

[0060] Acrylic rubber (ACM, ABR)

[0061] Silicone rubber (SI, Q, VMQ)

[0062] Fluorosilicone rubber (FVMQ)

[0063] Fluoroelastomers (FKM and FEPM) Viton, Tecnoflon, Fluorel, Aflas and Dai-El

[0064] Perfluoroelastomer (FFKM) Tecnoflon PFR, Kalrez, Chemraz, Perlast

[0065] Polyether block amide (PEBA)

[0066] Chlorosulfonated polyethylene (CSM), (Hypalon)

[0067] Ethylene vinyl acetate (EVA)

[0068] - Various other types of 4S elastomers:

[0069] Thermoplastic elastomer (TPE)

[0070] Proteins: elastin and elastin

[0071] Polysulfide rubber

[0072] Elastomeric olefins, elastic fibers for fabric production

[0073] Poly(dichlorophosphazene), an "inorganic rubber" derived from the polymerization of hexachlorophosphazene

[0074] Alternatively, the flexible wall 8 can be made of a multilayer film. For example, a multilayer film as described in EP 20945799 can be used. For example, the multilayer film comprises a contact layer in contact with the medium of the filling container, a barrier layer and an outer layer in contact with the external environment. These three layers are connected to each other by a tie layer. The material can be selected from polyolefins, such as polyethylene. The barrier layer provides a barrier for the passage of gases (such as oxygen, carbon dioxide), and is typically made of ethylene vinyl alcohol (EVOH). The outer layer can contain a single polyolefin or a mixture of a copolymer of a polyolefin and ethylene and an alpha-olefin.

[0075] The sterile cover 2 further comprises four extension arms 36 which are articulated to the top plate 4 and the base plate 6 about articulation axes 38 .

[0076] Specifically, each extension arm 36 is hinged to the pivot 29 of the fastener 28 of the top plate 4 and the pivot 29 of the fastener 28 of the base plate 6. The flexible side wall 8 and the extension arm 36 are compliant to allow the top plate 4 to move relative to the base plate 6 in three orthogonal directions (X, Y, Z) while preventing the flexible side wall from contacting the biological three-dimensional structure deposited on the upper major surface 16 during printing.

[0077] The stretcher arm 36 is elastic. The stretcher arm 36 pushes the top plate 4 away from the substrate 6. Advantageously, the stretcher arm 36 makes it possible to keep the top plate 4 more or less parallel to the substrate 6 during printing.

[0078] Advantageously, the extension arms 36 allow the mask to return to its original shape after printing.

[0079] The extension arm 36 is curved with a radius of curvature R between 50% and 90% of the length of the vertical section S passing through the upper and lower articulation axes 38 of the extension arm. The greater the radius of curvature, the greater the possibility of movement of the top plate 4 relative to the base plate 6.

[0080] Preferably, the extension arm 36 is located outside the housing 10 .

[0081] Preferably, the extension arm 36 is made of silicone or a thermoplastic elastomer.

[0082] The Shore hardness of the extension arm 36 is between A0 and A100 and preferably between A40 and A100.

[0083] The extension rate of the extension arm 36 is between 100% and 2000% and preferably between 300% and 600%.

[0084] Preferably, the sterile cover 2 comprises a sensor for monitoring at least one parameter among Ph, temperature and oxygen level. This sensor is not shown in the drawings.

[0085] Alternatively, the top plate 4 comprises two or three fasteners or a number greater than four fasteners. In a similar manner, alternatively, the sterile cover comprises two or three extension arms or a number greater than four extension arms.

[0086] refer to Figure 7 The sterile cover 40 according to the second embodiment is similar to the sterile cover 2 according to the first embodiment of the invention, except that the pierceable membrane 22 is replaced by a needle adapter 26. The needle adapter 26 is sealed to the top plate 4.

[0087] The needle adapter 26 used herein is a tip that is typically connected to the open end of the barrel of a medical syringe. The needle adapter 26 includes a needle hub that holds the needle and a connector portion 42 that is configured to connect to a corresponding connector portion of the barrel of the syringe. The connector portion 42 and the corresponding connector portion are, for example, Luer lock connectors.

[0088] Other technical elements of the second embodiment that are the same as those of the first embodiment are denoted by the same reference numerals and will not be described again.

[0089] Alternatively, the extension arm 36 is located inside the housing 10 .

[0090] Alternatively, the top panel and the base panel are square or rectangular and the sterile cover includes four flexible side walls.

[0091] exist Figure 1 The method for printing a biological three-dimensional structure in a sterile cover shown in the figure begins with a step during which a printing needle of a 3D printing syringe drills a hole in a pierceable membrane 22. The printing needle of the 3D printing syringe is not shown in the drawings. The printing needle is introduced into the inner chamber 11 and contacts the main surface 16 of the substrate. During this step, the top plate 4 moves along the vertical direction Z. The top plate 4 is brought close to the substrate 6.

[0092] The deposition material, in particular bio-ink, is deposited on the main surface 16 by means of the inserted printing needle.

[0093] The printing syringe and top plate 4 are moved relative to the substrate in a horizontal plane (X, Y) while the bio-ink is deposited into the inner chamber 11. During this step, the biological three-dimensional structure is printed on the upper main surface 16 of the top plate. The flexible side walls 8 and the extension arm 36 allow the printing needle of the syringe and the top plate to be moved relative to the substrate while keeping the top plate 4 and the side walls 8 at a certain distance from the already printed biological three-dimensional structure during the printing operation.

[0094] exist Figure 7 The method for printing a biological three-dimensional structure in a sterile cover shown in begins with a step during which a printing syringe or a printing cartridge is fixed to a needle adapter via a connector portion 42. The subsequent steps are the same as the one of the above-described methods and will not be described again.

Claims

1. A sterile cover (2, 40) intended for printing biological three-dimensional structures, the sterile cover (2) comprising: - a rigid top plate (4), a rigid substrate (6) comprising at least one main face (16) intended to receive a biological three-dimensional structure, - at least one flexible side wall (8) fixed to the top plate and the base plate (6) to form a hermetically closed housing (10) delimiting an inner chamber (11), - a pierceable membrane (22) or a needle adapter (26) arranged on the top plate, the pierceable membrane (22) being intended to be pierced by a printing needle, - at least one extension arm (36) hinged to the top plate (4) and the base plate (6), the flexible side wall (8) and the at least one extension arm (36) being adapted to allow the top plate (4) to be The at least one extension arm (36) is movable in three orthogonal directions (X, Y, Z) relative to the base plate (6); the at least one extension arm (36) is elastic and pushes the top plate (4) away from the base plate (6).

2. The sterile cover (2, 40) according to claim 1, characterized in that The Shore hardness of the at least one flexible side wall (8) is between A0 and A100 and preferably between A0 and A30.

3. The sterile cover (2, 40) according to any one of claims 1 and 2, characterized in that The elongation of the at least one flexible side wall (8) is between 300% and 2000% and preferably between 800% and 1100%.

4. The sterile cover (240) according to any one of claims 1 to 3, characterized in that: The at least one extension arm (36) has a Shore hardness between A0 and A100 and preferably between A40 and A100.

5. The sterile cover (240) according to any one of claims 1 to 4, characterized in that: The at least one extension arm (36) has an elongation between 100% and 2000% and preferably between 300% and 600%.

6. The sterile cover (2, 40) according to any one of claims 1 to 5, characterized in that The at least one extension arm (36) is hinged to the top plate (4) and the base plate (6) about an articulation axis (38), the at least one extension arm (36) being curved and having a radius of curvature (R) between 50% and 90% of the length of a section (S) passing through the articulation axis (38) of the at least one extension arm and defined by the articulation axis.

7. The sterile cover (2, 40) according to any one of claims 1 to 6, characterized in that The at least one extension arm (36) is made of silicon or a thermoplastic elastomer.

8. The sterile cover (2, 40) according to any one of claims 1 to 7, characterized in that The flexible side wall (8) is made of a biocompatible plastic selected from the group consisting of multilayer films, polydimethylsiloxane, silicone, unsaturated rubbers that can be cured by sulfur vulcanization, saturated rubbers that cannot be cured by sulfur vulcanization, thermoplastic elastomers (TPE), the proteins elastin and elastin, polysulfide rubbers, elastic olefins, elastic fibers for textile production, poly(dichlorophosphazene) and inorganic rubbers from the polymerization of hexachlorophosphazene.

9. The sterile cover (2, 40) according to any one of claims 1 to 8, characterized in that The base plate (6) or the top plate (4) is made of one of the following materials: biocompatible plastic; metal; and inorganic material; polymer rigid material, preferably polyethylene (PE).

10. The sterile cover (2, 40) according to any one of claims 1 to 9, characterized in that The top plate (4) comprises at least one port (18) and / or at least one sterile connector.

11. The sterile cover (2, 40) according to any one of claims 1 to 10, characterized in that The top plate (4) and the base plate (6) are in the shape of disks, and the flexible side wall (8) is cylindrical or annular.

12. The sterile cover (2, 40) according to any one of claims 1 to 11, characterized in that The flexible side wall (8) is transparent.

13. The sterile cover (240) according to any one of claims 1 to 12, comprising at least one sensor for monitoring at least one parameter among Ph, temperature and oxygen level.

14. The sterile cover (2, 40) according to any one of claims 1 to 13, characterized in that The at least one extension arm (36) extends outside the housing (10).

15. The sterile cover (2, 40) according to any one of claims 1 to 14, characterized in that The surface of at least one main surface (12, 14) of the top plate (4) is 50% to 100% of the surface of at least one main surface (16) of the base plate (6).

16. The sterile cover (2, 40) according to any one of claims 1 to 15, characterized in that The base plate (6) comprises at least one fluid inlet port (32) and at least one fluid outlet port (34) for circulating a cell culture medium.

Images