ORGAN-ON-A-CHIP (OOC) DEVICE

Description

DESCRIPTION of the industrial invention entitled: "Organ-on-a-Chip (OoC) Device" by: University of Eastern Piedmont "Amedeo Avogadro", Italian nationality, via Duomo, 6 - 13100 Vercelli (40%); National Research Council, Italian nationality, Piazzale Aldo Moro, 7 - 00185 Rome (60%) Designated inventors: Alessandro POLINI, Francesca GERVASO, Annalisa CHIOCCHETTI, Francesco BISCONTI, Alessandro DE GIORGI, Giuseppe GIGLI Filed on: August 2, 2024 *** DESCRIPTION TEXT Technical field of the invention The present invention relates to a chip device for an organ-on-a-chip (OoC). As already known in the state of the art, an OoC is a system of cell culture that can recreate a microenvironment complex and simulate human physiology for cultivation mobile phone. A favorite application of the chip device described here is for 3D cell cultivation. In particular, the present invention relates to a chip device of the type comprising: - a chip that includes a culture chamber for host a cellular structure; - a pressure generating system to exert a compressive action on the hosted cellular structure in the culture chamber; wherein the pressure generating system comprises: - a chamber body having a pressure chamber; - a membrane having a first side facing the culture chamber and a second side facing the chamber of pressure, and - a pressure generator to supply a fluid working on the pressure chamber and create a pressure in the pressure chamber which moves the membrane in the direction of the culture chamber, thus performing an action of compression on the cellular structure contained within it. A chip device of the above type is already known, for example, from WO2016174607A1. The present invention aims to realize a chip device improved over chip devices of the prior art, in particular from the point of view of the effectiveness and controllability of the action of compression exerted on the cellular structure. Summary of the invention The present invention relates in general to a chip device according to Claim 1. The present invention also relates to a method according to Claim 16. The claims form part integral to the teachings provided here. Brief description of the drawings and detailed description of one or more ways of realizing the invention Further features and advantages of the invention will be evident from the following description, which refers to the The attached drawings are provided for illustrative purposes only. and not limited to, where: - Figure 1 shows one embodiment preferred version of the device described here, according to a view perspective; - Figure 2 shows the device of Figure 1 in disassembled condition; - Figure 3 shows a cross-sectional view of the device in Figure 1, along the section plane III-III illustrated in Figure 1; - Figure 4 is a plan view of a component of the device in Figure 1. The following description illustrates various details specific ones intended to provide a deeper understanding of the implementation methods. The methods of realization can be done without one or more of the specific details, or with other methods, components or materials, etc. In other cases, structures, materials or known operations are not shown or described in detail, to avoid obscuring various aspects of the modality of realization. The references used here are purely for convenience and therefore do not define the scope of protection or the extension of the implementation methods. As mentioned above, the device described here is a chip device for an organ-on-a-chip (OoC). Referring to Figures 1-3, the chip device – indicated as a whole with the numerical reference 10 – comprehends: - a chip 20, which includes a culture chamber 22 to accommodate a 3D cellular structure; - a chamber body 40 having a pressure chamber 42; And - a membrane 80 having a first side 82 facing the culture chamber 22 and a second side 84 facing pressure chamber 42. In the chip device 10 described here, the membrane 80 is supported by a support element 60 arranged between the chip 20 and the camera body 40. A cover 50 is arranged against the chamber body 40, at a side opposite the support element 60, to close the pressure chamber 42 (the lid 50 (not shown in figure 2). The above components are assembled together according to a layered formation, in the following sequence (starting from the bottom with reference to figure 3): chip 20, support element 60 with membrane 80, chamber body 40 and lid 50. The chip device 10 includes clamping members 30 which engage the above-mentioned members to exercise on them clamping forces that press together the components. The clamping forces also have the function of create a watertight connection between the contacting regions mutual interaction of the components, so as to avoid losses from the culture chamber 22 and from the pressure chamber 42 towards the outside. The chip device 10 also includes a generator of pressure 90 to supply a working fluid to the chamber of pressure 42 and create a pressure in it that acts on the second side 84 of the membrane 80 to move the latter in the direction of the culture chamber 22. The expert in the field will understand that the movement of the membrane 80 is actually the result of the flexion that the membrane undergoes due to the pressure created in the pressure chamber 42. In operation, this movement of the membrane 80 it is able to subject the structure to compression cell contained in the culture chamber 22 of the chip 20. The pressure generator 90 may comprise, for example example, a microfluidic flow controller that can control the flow of fluid fed to the chamber pressure 42 based on pressure-indicating signals present inside the same pressure chamber 42. In one or more preferred embodiments, such as the one illustrated, the chip 20 is in the form of a plate presenting a cavity 21 in a central region, which is opens on a top surface 26 of the chip (with reference (Figure 3). In one or more preferred embodiments, such as as illustrated, cavity 21 includes a chamber 22 defining the culture chamber, which is delimited on three sides from corresponding internal edges of the chip 20, and a channel 23 which extends next to room 22 and along the open side of the same and which is in fluid communication with the same room 22. Channel 23 has an end 23A which communicates with a entrance 24 and an opposite end 23B which communicates with an output 25, both input 24 and output 25 being located on a surface less than 28 (with reference to the figure 4) of chip 20. Entrance 24 and exit 25 are intended to be connected to a power system to power a culture fluid. In operation, the culture fluid fed by the power supply enters chip 20 via input 24, flows along channel 23 passing at least partially to the culture chamber 22, and its remaining part comes out of the chip 20 via output 25; possibly, a part of the culture fluid already present in the culture chamber 22 returns to channel 23 to be replaced by the new fluid of powered culture. In one or more preferred embodiments, such as the one illustrated, in the culture chamber 22 are arranged 29 boundary elements that divide the room culture 22 in at least one first compartment 22A and a second compartment 22B. Preferably, the two compartments 22A and 22B are divided along a T-line running parallel to the canal 23, so that a compartment (i.e. compartment 22A in the example shown) is located next to channel 23, and the other compartment (i.e. compartment 22B in the example illustrated) is located next to the compartment previous to and, with respect to it, opposite to channel 23. The delimiting elements 29 are such as to delimit the two compartments 22A, 22B without determining a complete separation between the two, which means that the culture fluid can flow from one compartment to another without any particular restrictions. In one or more preferred embodiments, such as the one illustrated, the 29 delimitation elements are cylindrical columns erected from the bottom of the culture chamber 22 and arranged in a row along the T line and equidistant from each other on the other. In one or more preferred embodiments, such as that illustrated, delimitation elements are also provided 29 to separate channel 23 from culture chamber 22. Preferably, the chip 20 is made by printing 3D, for example by stereolithography or Digital Light Processing (DLP). A resin can be used photopolymerizable with UV light. Please note that the manufacturing through Digital Light Processing (DLP) allows for rapid and relatively inexpensive customization and prototyping of the chip design. Preferably, the chip 20 is transparent. In one or more preferred embodiments, such as the one illustrated, the camera body 40 is also in shape of a plate, and has a central opening through 41, which runs through the entire thickness of the chamber body 40 and defines pressure chamber 42. The opening 41 is positioned in the chamber body 40 so that to be superimposed on the cavity 21 of the chip 20. Preferably, the opening 41 covers, in planimetric view, an area enclosing the area of ​​cavity 21. In one or more preferred embodiments, such as the one illustrated, the chamber body 40 includes an input 43 and an exit 44 which both communicate with the chamber pressure 42 and which are connected to the pressure generator 90. The cover 50 is arranged against a surface upper 45 of the chamber body 40 to close the opening 41 and define an upper wall of the pressure chamber 42. Preferably, the lid 50 is transparent. Preferably, the camera body 40 is also made by 3D printing. In one or more preferred embodiments, such as the one illustrated, the support element 60 is also in shape of a plate and has a central region 62 on where the membrane 80 lies in an extended configuration and to which it is fixed. Preferably, the membrane 80 is attached to a lower surface 61 of the support element 60 (with (refer to figure 3). In its central region 62, the support element 60 has a through opening 63 which is completely covered from membrane 80. The opening 63 places the second side 84 of the membrane 80 in fluid communication with the pressure chamber 42 of the chamber body 40. In operation, the pressure created in the chamber pressure 42 flexes the membrane 80 to move it in the direction towards the culture chamber 22. Since aperture 63 is smaller in area occupied in a planimetric view with respect to the membrane 80, only a selected portion 80' of the membrane 80 - the one that covers the opening 63 - is subject to movement above indicated. The selected portion 80' is determined by the configuration of the aperture 63, in particular from its position, shape and size. In one or more preferred embodiments, such as the one illustrated, the selected portion 80' is such as extend exclusively above one of the two compartments 22A, 22B of culture chamber 22. Preferably, the selected portion 80' extends above compartment 22A, i.e. on compartment located next to channel 23. In one or more preferred embodiments, such as as illustrated, the support element 60 comprises a outer region 64 which at least partially surrounds the central region 62 and has a contrasting surface upper 65 and a lower contrast surface 66. Incidentally, by contrast surface we mean here a surface intended to bear loads and forces which act on the component of which the surface is part. In the assembled condition of the device, the upper contrast surface 65 is in contact with a lower surface 46 of the chamber body 40, while the lower contrast surface 66 is in contact with the upper surface 26 of chip 20. Let the surface lower surface 46 of the chamber body 40 that the upper surface 26 of chip 20 define contrast surfaces themselves which cooperate with the contrast surfaces 65 and 66 of the support element 60. In one or more preferred embodiments, such as the one illustrated, in the external region 64, the element of support 60 has a plurality of through openings 67 which extend from the contact surface 65 to the surface of contact 66. Preferably, the plurality of holes 67 comprises two parallel rows of holes 67 equidistant from each other which are arranged on opposite sides to the opening 63. Furthermore, the other members of the training also layers of the chip device 10, i.e. chip 20, the body chamber 40 and the lid 50, have respective pluralities of through holes, 27, 47 and 57, all in corresponding positions such that they align with the holes 67 of the element of support 60. The above holes are configured to receive the organs tightening 30. The support element 60 can be made, for example for example, with polystyrene sheets. The opening 63 and the holes 67 can be made by micro-milling. The membrane 80 can have a thickness between 150 and 350 microns and can be manufactured by spin coating directly on the support element 60; the latter can be subjected to oxygen plasma activation for facilitate membrane adhesion 80. It can now be noted that the support element 60 is such whereby the clamping forces exerted by the organs of tightening 30 act exclusively on the external region 64 of the support element 60; on the contrary, the region central 62 and membrane 80 are subjected to forces of pressures much lower than the clamping forces. This way, there is no risk of membrane 80 flexes from its flat configuration due to forces excessive pressure acting on it. In this regard, the Applicant has verified that, without the support element 60 (see for example the chip device described in the prior art document WO2016174607A1 cited at the beginning), if the membrane is crushed with pressing clamping forces, the membrane assumes a deformed state even when at rest, that is, when no pressure is generated in the pressure chamber pressure from the pressure generator, which can affect the operability of the membrane. It is therefore clear that the chip device described here can operate the 80 membrane more precisely and effectively. Referring to figure 3, the internal region 62 of the support member 60 is illustrated as having a recess 68 configured to receive the membrane 80 in so that the first side 82 of the latter is positioned at the same level as the contrast surface 66 of the support element. It follows that the surface of contrast 66 is able to come into contact with the surface top 26 of chip 20. However, it should be noted that, to achieve this, the the aforementioned recess is not essential; in fact, the The applicant has verified that, even in the absence of the recess 68 and with the membrane 80 lying underneath the contrast surface 66 before assembly, the forces of tightening generated by the tightening organs 30 are such to bend the support element 60 to finally bring the contrast surface 66 in contact with the surface top 26 of chip 20. In one or more preferred embodiments, such as the one illustrated, the clamping organs 30 are organs of releasable clamps, preferably of the type quick connection. Preferably, the clamping members 30 include bolts and nuts. Thanks to the features described above, the 10-chip device can be easily disassembled, for example, to allow the extraction of the structure cell phone or part of it, and can subsequently be reassembled for further operations. Since the membrane 80 is supported by the element of support 60 and through the latter it is assembled to the others device components, there is no risk that the membrane 80 may be damaged during assembly operations and disassembly. In one or more preferred embodiments, such as the one illustrated, the 10-chip device also includes a control unit 70 for controlling the generator pressure 90. In particular, the control unit 70 is configured to selectively activate and deactivate the generator pressure 90 according to a mechanical stimulation routine to be performed on the cellular structure. The expert in the field will understand that the intensity of the mechanical stimulation depends on the length of the movement performed by the selected portion 80' of the membrane 80. In fact, the deeper this portion enters the culture chamber 22, the greater the compression exerted on the cellular structure. In one or more preferred embodiments, the 10 chip device can also adjust the intensity of the mechanical stimulation performed on the cellular structure. For this purpose, the control unit is configured to check the final position of the portion movement selected 80' in the direction of the culture chamber 22 regulating the pressure generated in the pressure chamber 42 from pressure generator 90. The stimulation interval through which the device can operate may vary according to needs of specific applications. Furthermore, as already indicated above, the opening 63 of the support element 60 allows to determine a selected portion 80' of membrane 80 to act on a specific region or compartment of the culture chamber 22 based on the needs of specific applications. Now, referring again to WO2016174607A1, note that the solution described in this document provides for contrary to the final position of the movement of the membrane is defined by the same delimiting elements present in the culture chamber, which, for this purpose, they act as arresting organs to stop the movement of the membrane. In this case, the stimulation performed on the cell structure cannot be regulated but is actually fixed, since it is dictated by the geometry of the boundary organs. Furthermore, the action of the membrane necessarily involves the entire culture chamber, while cannot be limited to a specific region of the room of cultivation. Furthermore, this prior art solution suffers also of the inconvenience that the membrane can be damaged by the delimitation bodies. In view of the above, it is now evident how the chip device described here surpasses many of the drawbacks of the prior art. Of course, as long as the principle is maintained of the invention, the construction details and the forms of realization may vary, even in size significant, compared to what is illustrated here purely exemplary title, without therefore going beyond the scope of the invention, as defined in the claims attached.

Claims

CLAIMS 1. Chip device (10) for an organ-on-a-chip (OOC), comprising: - a chip (20) comprising a culture chamber (22) for hosting a cellular structure; - a pressure generation system for exerting a compressive action on the cellular structure hosted in the culture chamber (22); wherein the pressure generation system comprises: - a chamber body (40) having a pressure chamber (42); - a membrane (80) having a first side (82) facing the culture chamber (22) and a second side (84) facing the pressure chamber (42); - a pressure generator (90) for supplying a working fluid to the pressure chamber (22) and generating a pressure in the pressure chamber (22) that moves said membrane in the direction of the culture chamber (22) performing a compressive action on the cellular structure contained therein;said chip being characterised in that it comprises a support element (60) supporting the membrane (80), which is arranged between the camera body (40) and the chip (20).; 2. The chip device of claim 1, wherein the support member (60) has an internal region (62) to which the membrane (80) is attached and which has a through opening (63) configured to allow only a selected portion (80') of the membrane (80) to move in the direction of the culture chamber (22), and wherein the support member (60) has an external region (64) that at least partially surrounds the internal region (62); said chip device (10) comprising clamping members (30) configured to exert clamping forces that press the chamber body (40) and the chip (20) against the external region (62) of the support member (60).

3. Chip device according to claim 2, wherein the outer region (64) of the support member (60) comprises a first contrast surface (65) in contact with the camera body (40), and a second contrast surface (66) in contact with the chip (20), and wherein the pressing clamping forces exerted by the clamping members (30) press the camera body (40) against the first contrast surface (65), and the chip (20) against the second contrast surface (66).

4. The chip device of claim 3, wherein, in and along the outer region (64), the support member (60) has a plurality of holes (67) extending from the first abutment surface (65) to the second abutment surface (66), and wherein the chip (20) and the camera body (40) have respective pluralities of holes (47, 67) in corresponding positions such as to align with the plurality of holes (67) of the support member (60); and wherein clamping members (30) are received in the holes of each of the support member (60), the chip (20) and the camera body (40).

5. A chip device according to claim 4, wherein the chip (20) has a third contrast surface (26) in contact with the first contrast surface (66) of the support member, and the camera body has a fourth contrast surface (46) in contact with the second contrast surface (65) of the support member, and wherein the plurality of holes (27) of the chip (20) are arranged along and on the contrast surface (26) of the chip (20), and the plurality of holes (47) of the camera body (40) are arranged along and on the contrast surface (46) of the camera body (40).

6. A chip device according to any of the preceding claims, wherein the clamping members (30) are releasable clamping members that allow assembly and disassembly of the chip device.

7. A chip device according to any of the preceding claims, wherein the opening (63) of the support member (60) is such that the selected portion (80') of the membrane (80) extends exclusively over a partial region of the culture chamber (22).

8. Chip device according to any of the preceding claims, wherein, in said culture chamber (22), delimiting elements (29) divide the culture chamber (22) into at least a first compartment (22A) and a second compartment (22B), and wherein the opening (63) of the support member (60) is such that the selected portion (80') of the membrane (80) extends exclusively over the first compartment (22A) or the second compartment (22B).

9. Chip device according to any of the preceding claims, wherein the chamber body (22) comprises an inlet (43) and an outlet (44) in fluid communication with the pressure chamber (42) and wherein the pressure generator (90) is connected to the inlet (43) and the outlet (44) to supply the working fluid to the pressure chamber and generate a predetermined pressure therein.

10. A chip device according to any preceding claim, wherein the chip (20) comprises an inlet (24) and an outlet (25) that are in fluid communication with the culture chamber (22) and that are configured to be connected to a supply system for supplying a culture fluid to the culture chamber (22).

11. Chip device according to any of the preceding claims, wherein at least the camera body (40) and the chip (20) are made by 3D printing.

12. Chip device according to any of the preceding claims, comprising a control unit (70) for controlling the pressure generator (90).

13. A chip device according to claim 12, wherein the control unit (70) is configured to selectively activate and deactivate the pressure generator according to a mechanical stimulation routine to be performed by the membrane (80) on the cellular structure.

14. Chip device according to claim 12 or 13, wherein the control unit (70) is configured to control the final position of the movement of the selected portion (80') of the membrane (80) in the direction of the culture chamber (22) by regulating the pressure generated in the pressure chamber (42) by the pressure generator (90).

15. A chip device according to any preceding claim, wherein at least said chip is transparent.

16. A method of operating a chip device (10) according to any preceding claim, comprising the following steps: - providing the chip (20) and depositing a cellular structure in the culture chamber (22) of the chip (20); - assembling the pressure generating system to the chip (20) having the cellular structure deposited in its culture chamber (22), wherein the assembly includes arranging the support member (60) with the membrane supported thereby (80) between the chip (20) and the chamber body (40) and by means of clamping members (30) exerting clamping forces that press the chip (20) and the chamber body (40) against the outer region (64) of the support member (60); said method further comprising: - actuating the movement of the selected portion (80') of the membrane (80) by the pressure generating system (90) to subject the cellular structure deposited in the culture chamber (22) to a mechanical stimulation routine.

17. The method of claim 16, wherein actuating the movement of the selected portion (80') of the membrane (80) includes controlling the end position of the movement of the selected portion (80') in the direction of the culture chamber (22) by adjusting the pressure generated in the pressure chamber (42) by the pressure generator (90).

18. A method according to claim 16 or 17, comprising: - disassembling the pressure generation system from the chip (20); and - extracting the cellular structure from the culture chamber (22) of the chip (20).

19. A method according to any of claims 16 to 18, comprising regulating the intensity of the mechanical stimulation exerted on the cellular structure.