WO2023244197A1

WO2023244197A1 - Extrusion-based and portable handheld three-dimensional bioprinter

Info

Publication number: WO2023244197A1
Application number: PCT/TR2023/050343
Authority: WO
Inventors: Ayça Bal ÖZTÜRK
Original assignee: Adbioink Bi̇yosi̇stem Teknoloji̇ Anoni̇m Şi̇rketi̇
Priority date: 2022-06-13
Filing date: 2023-04-12
Publication date: 2023-12-21
Also published as: TR2022009769A2

Abstract

The invention relates to a portable handheld three-dimensional bioprinter which is designed for use in tissue engineering applications, artificial organ and tissue production, musculoskeletal injuries, tissue damage, burn treatment, tissue regeneration, controlled release system and three-dimensional cell culture applications as well as cosmetics and drug discovery, which eliminates the need for PC connection, drawing file and professional user, which can be easily used with one hand, the printing speed and light source of which can be changed depending on the need, which can offer heating and cooling options during printing and can print directly to the desired area in the body.

Description

EXTRUSION-BASED AND PORTABLE HANDHELD THREE-DIMENSIONAL BIOPRINTER

TECHNICAL FIELD

The invention relates to an extrusion-based and portable handheld three- dimensional bioprinter designed for use in tispsue engineering applications, artificial organ and tissue production, musculoskeletal injuries, tissue damage, bum treatment, tissue regeneration, controlled release system and three- dimensional cell culture applications, as well as cosmetics and drug discovery.

PRIOR ART

Currently, three-dimensional bioprinters are the common solution for bioprinting. Such systems are high-tech biomedical devices that are used especially in the field of tissue engineering and enable the imitation of natural tissues by creating tissue-like structures via depositing biomaterials, also known as bio-ink, which can mimic natural tissues. In three-dimensional bioprinters, the material to be bioprinted (bioink) is mixed with the relevant cells depending on the area of use, transferred to the apparatus called barrel (cartridge) and placed in the three- dimensional bioprinter. The shape in the drawing file uploaded to the system is produced thanks to the motor connection moving in three axes. The bioprinted material can be cured and hardened thanks to the integrated light source. Bioprinting can be defined as the layer-by-layer creation of complex biological structures (tissues, organs) by precise positioning of living cells. When creating biological structures, the most important requirement is that cells can maintain their viability during the process. Three-dimensional bioprinters can also be used to reconstruct tissue in various parts of the body. In this sense, new applications are being realized every day in the medical field and the potential for the use of these systems is increasing. The application in the medical field is possible by first making a three-dimensional drawing of the body part to be filled with the biomaterial and uploading the drawing file to the three-dimensional bioprinter and realizing the production. System features do not allow for direct body application. The principle underlying bioprinting in three-dimensional bioprinters is divided into three main groups depending on the type of three-dimensional bioprinter: inkjet printer, laser-assisted printer and extrusion printers. In inkjet printers, the printhead is heated electrically and generates pulses of air pressure, causing the bio-ink to drip. Extrusion-based printers use either pneumatic systems or mechanical systems in which gas pressure is translated into mechanical motion. In laser-assisted printers, pulses are generated by using a focused laser on the absorbent surface, pushing the cells towards a collector surface at the bottom. In each case, the printing mechanism is based on different physical phenomena, but the goal is the same. Although such systems have significant advantages such as enabling the production of complex figures, they also have major disadvantages such as the need for a PC connection and drawing file, the need for professional users, high costs, the size and the inability to print directly on the human body due to the structure of the system.

In addition to the above-mentioned three-dimensional bioprinters, there are also patents for the production of hand-held three-dimensional bioprinters (biopencils/biopens) that have a similar purpose with the proposed invention. These patents and their technical problems are presented below.

US20160361867A1 “Blue-ray Photocuring 3D Printing Pen”

The proposed biopencil structure consists of a casing, an ink cartridge located in the casing, a blue-ray photocuring system located at the head of the ink cartridge and a pen tip located at the tip of the ink cartridge. In this system, there is a curing system consisting of a large number of LEDs with a wavelength of 465 nm to 485 nm, which is presented as green technology. It is an extrusion-based system, comprising a piston located inside the ink cartridge that can slide forward in the ink cartridge and a stepper motor configured to drive the piston to move forward. The lack of a heating/cooling system and the lack of information on whether different light sources can be integrated are considered as disadvantages thereof. In addition to design differences, our invention has a heating/cooling system, which is very important for bioprinting, and a modular structure where different light sources can be integrated.

US9731444B2 “Hand-held three-dimensional drawing device”

The proposed structure is a portable three-dimensional printer system. Bioprinting is not performed. Thanks to the heating system at the tip thereof, it melts and deposits the thermoplastic material and forms a three-dimensional structure by depositing layer by layer. Although similar in name, the printing mechanism is different. The melting/deposition concept is not used for printing biomaterials because of the negative impact on cell viability.

JP2020502994A “Organization printer”

Described here are bioprinters that enable in situ formation of structured biomaterials and tissues by rotating a printhead across a deposition surface (e.g. a skin wound, etc.). In a hand-held configuration of such a device, biopolymer solution containing the cell is dispensed by moving a micro-processed printhead and deposited onto a fixed flat surface or wound. This patent introduces a portable three-dimensional bioprinter system that allows cells to be printed directly at the injured site in the appropriate solution. This system refers to multiple extrusion channels and allows simultaneous printing of different solutions. When the biopolymer solution is dispensed from the printhead and deposited on the damaged area, the biopolymer solution is polymerized and solidified. Solidification can occur through different mechanisms, including coagulation, ion-induced, pH- induced and temperature-induced solidification, as well as enzymatic reactions and ultraviolet light-induced polymerization and combinations thereof. Although there is no light source integration for curing on the system, it is stated that UV light can be supplied from outside if needed. Furthermore, the outlet of the bioprinter is of a lamellar structure with a width. Finally, there is no heating and cooling system. Based on all these features, the disadvantages of the system are listed below.

- The lack of light integration leads to the need for a second device during use. In addition, the fact that UV light can be used limits its use, especially considering the interest in the visible light curing process in recent years. - The fact that it leads to a lamellar (strip) output rather than a fiber structure is a disadvantage, especially in the filling of wound areas in grift structure. However, the outputs in the fiber geometry will allow the entire region to be filled more easily and independently of the geometry.

- Lack of a heating/cooling system will cause cell death and negatively affect the printability of the solution due to the lack of a suitable temperature environment for printing solutions containing cells.

CA3050385A 1 “Devices and methods for wound-conformal guidance of bioprinter printhead”

Described herein is a device that enables in situ formation of structured planar biomaterials and tissues by rotating a printhead along a deposition surface in a burn-injured patient. In hand-held embodiments of the device, cell-loaded biopolymer solutions are perfused through a moving microfabricated printhead and deposited onto a fixed planar surface or a wound. The printhead can be turned by means of a drive mechanism. A soft deformable roller reduces further damage to the injured area of skin as it rotates over it, and a gimbal mechanism to which the printhead is attached is in contact with the injured tissue but does not exert excessive pressure on the area while the printhead is in contact with the tissue.

The system consists of the following components:

- Regardless of the contour of the surface, a series of extruded channels to be in physical contact with the first series of surface,

- A drive mechanism with a soft roller that, when activated by the operator, drives the biomaterial across the surface at a preselected V speed.

There is no light source integration for curing on the system, limiting its area of use. It can also be output with a roller rotation and applied to the injured area with gentle contact of the roller. Contact may cause pain. In addition, the roll outlet may not provide the desired closure rate, especially in the closure of wounds in the grift structure. The absence of a heating/cooling device will also result in the inability to obtain reproducible results for applications where the application quality may vary according to the ambient temperature, and will change the rheological behavior of the solution to be printed, thus adversely affecting the print quality and will damage cell viability if it is a cell-loaded application.

US US20200123485A1 “Bioprinter devices, systems and methods for printing soft gels for the treatment of musculoskeletal and skin disorders”

The present invention relates to medical devices for the treatment of musculoskeletal and skin disorders, and more particularly to devices, systems and methods that utilize bioprinters to create scaffolds in situ to facilitate the treatment of musculoskeletal and skin disorders in patients. The method includes extruding a hydrogel formulation and curing it in situ. It is an extrusion-based device capable of continuously extruding biomaterials and includes an integrated light source for crosslinking the extruded bioink. The platform can print photo-crosslinkable hydrogels such as gelatin methacryloyl (GelMA) for VML injuries instantly in situ. GelMA is a collagen-derived biomaterial that closely mimics the extracellular matrix (ECM) of natural skeletal muscles. It is noted here that the integrated light source is UV. It is also said to include a power supply placed into the casing. While the addition of the power supply to the bioprinter provides advantages such as being a wireless system, it creates disadvantages such as the fact that it causes additional weight, the batteries installed are not environmentally friendly, and similar quality curing cannot be realized with the decrease in battery power. This invention also does not mention heating/cooling apparatus and also does not mention the integration of a visible light source.

W0201 8166641 A1 “Handheld 3D bio printer”

The present invention relates to the additive manufacturing of biocompatible materials. In particular, it concerns hand-held three-dimensional printing of biocompatible materials for surgical biofabrication. This prototype system has two cartridge compartments that separately store two reagent containers (stem cells and biomaterial) as hydrogels, a mechanical extrusion system is used to extract the reagents from the three-dimensionally printed titanium extruder nozzle, and a UV light source is used. It is used to cross-link the hydrogels immediately after extrusion, thus forming a stable structure that encapsulates and supports the stem cells. A foot pedal is used to control the extrusion of the reagent and the extrusion speed is controlled using an electronic control interface. Each extruder has a circular cross-section and is coaxially arranged with a core material containing stem cells and a shell material encapsulating and supporting the core material. The disadvantages of this study are listed below.

- Prototype devices have reliability and consistency problems. The viscosity and hence the flow rate of the reagent is easily affected by temperature and the material properties are easily affected by the mixing ratio. This requires strict control of the extrusion speed.

- The prototype device has limited freedom of movement as it is connected to the foot pedal and the electronic control interface by a cable.

- The nozzle is a 3D printing titanium nozzle, which is expensive and not suitable for mass production.

The subject matter of the invention we have applied for is made of aluminum material. This selection of material is advantageous in terms of being cheaper, easier to process and having a high coefficient of thermal conductivity. In addition, it is stated in the relevant patent document that the problem in materials affected by temperature can be adjusted by speed control. This is an indirect control and the flow behaviors due to temperature differences cannot be fully controlled in this way. In our invention, there is a design that allows printing at the desired constant temperature thanks to its heating/cooling characteristics. On the other hand, ensuring the mixing within the prototype may lead to the appearance of inhomogeneous defects. In our invention, mixing is performed outside and a single-channel piston structure can be used to print the relevant region in a temperature-controlled manner. This makes the mechanism user-friendly and eliminates the problems of mixture homogenization.

Thus, the need to eliminate such shortcomings and disadvantages of the embodiments and practices employed in the prior art entails an improvement in the respective tehnical field. DESCRIPTION OF THE INVENTION

The present disclosure relates to an extrusion-based and portable handheld three-dimensional bioprinter developed for eliminating the aforementioned disadvantages and providing new advantages to the respective technical field.

With the system of our invention, a portable structure is obtained which eliminates the need for PC connection, drawing file and professional user, which can be easily used with one hand, the printing speed and light source of which can be changed depending on the need, which can offer heating and cooling options during printing and which can print directly to the desired area on the body.

The portable three-dimensional bioprinter system of the invention allows the biomaterial to be printed in the desired area by pushing the cartridge (barrel) containing the biomaterial consisting of a mixture of bioink and cells in a controlled manner with its sensitive piston structure that can move electrically. With the heating and cooling option that can be adjusted from the control panel, the biomaterial can be brought to the desired temperature between 20 and 40 °C, and the biomaterial temperature can be precisely monitored with the temperature meter integrated into the system. The light source has a modular plug-in mechanism and can be easily integrated by the user with light sources including, but not limited to, 405 nm or 445 nm light sources consisting of visible region LEDs and 360 nm UV light source. Thanks to the light on/off feature, it is possible to print by turning off the light source if desired.

The presence of the heating/cooling system in our invention can provide high cell viability in terms of the cells being able to mimic physiological temperature during printing and increase the efficacy of the treatment, especially if applied directly to the body. Since the printing properties (rheology) of bioinks are affected by very small temperature differences, temperature differences make it difficult to obtain reproducible data and lead to misinterpretations. Maintaining constant temperature by providing temperature control will offer significant advantages in the optimization process and will contribute to obtaining reproducible results.

Thanks to the fact that the invention does not require a PC connection and has a motor that can move in three axes, a portable size that can be controlled manually has been obtained. The system of the invention provides the advantage of being able to print directly on a desired area thanks to the control of the geometry of the material produced by hand movement.

The structural and characteristic features of the invention and all its advantages will be understood more clearly by the figures given below and the detailed description written with reference to these figures, and therefore and thus, the present disclosure should be evaluated by taking these figures and detailed description into consideration.

BRIEF DESCRIPTION OF THE DRAWINGS

The embodiments of the present disclosure which are summarized above and discussed in more detail below can be better understood by referring to exemplary embodiments of the present disclosure illustrated in the accompanying drawings. It should be noted, however, that the accompanying drawings only describe typical embodiments of the present invention and are not to be considered as limiting the scope of the invention.

Figure-1 : It is a representative view of the product of the invention.

Figure-2: It is representative view of the biopencil body and the bioink cartridge of the invention.

Figure-3: It is a representative view of the electrical cables, hot/cold liquid inlet/outlet lines and the tube casing enclosing them.

REFERENCE NUMBERS 16. Power switch

DETAILED DESCRIPTION OF THE INVENTION

The preferred alternatives of the present disclosure, which are mentioned in this detailed description, are only intended for providing a better understanding of the subject matter, and should not be construed in any restrictive sense.

The invention relates to an extrusion-based and portable handheld three- dimensional bioprinter designed for use in tissue engineering applications, artificial organ and tissue production, musculoskeletal injuries, tissue damage, bum treatment, tissue regeneration, controlled release system and three- dimensional cell culture applications, as well as cosmetics and drug discovery.

The bioprinter of the invention generally includes at least one biopencil body (1 ) with ergonomics to allow the bioprinter to be carried by hand, made of aluminum alloy to provide high thermal conductivity, anodized and ground on the outer surface to ensure a long service life and a modem appearance, at least one bioink cartridge (4) in which the bioink solution is placed and which has a hot/cold liquid inlet (4.1), a hot/cold liquid outlet (4.2) and an electrical cable inlet (4.3), at least one LCD display (2) to display the temperature information of the bioink contained in the bioink cartridge (4) and the pressure information applied by the bioink to the micro linear piston (3), at least one micro linear piston (3) to push the rear plug of the bioink cartridge (4) forward to allow the bioink solution contained in the bioink cartridge (4) to be ejected from the end of the bioink cartridge (4), at least one push button (5) to cause the micro linear piston (3) to move forward when bioprinting is to be performed so that the bioink solution contained in the bioink cartridge (4) is ejected from the end of the bioink cartridge (4), at least one pull button (6) for pulling back the micro linear piston (3) and removing the bioink cartridge (4) after the bioprinting process is completed, at least one infrared sensor (7) integrated in the light source body (8) for real-time measurement of the temperature of the bioink solution in the bioink cartridge (4), at least one light source body (8) containing light sources consisting of LEDs (9) and integrated into the biopencil body (1), which can be separated from the biopencil body (1 ) and replaced when light of different wavelengths is desired to be used, LEDs (9) to ensure simultaneous curing of the printed three-dimensional structure during bioprinting, at least one tube casing (10) containing the electrical cables (10.1) for the transmission of electrical energy required for the micro linear piston (3) and LEDs (9), as well as the hot/cold liquid inlet line (10.2) and the hot/cold liquid outlet line (10.3) for the transmission of closed circuit water in the heating/cooling unit used for heating and cooling the bioink cartridge (4), at least one control unit box (11) to provide the voltage adjustment required for the speed of the micro linear piston (3), to provide the adjustment of the voltage level suitable for the LEDs (9), to contain the heating and cooling unit, at least one micro linear piston driver (12) positioned on the control unit box (11 ) to adjust the movement speed of the micro linear piston (3) and thus optimize the printing of bioink solutions with different characteristics, at least one LED switch (13) positioned on the control unit box (11) to enable the LEDs (9) to be activated or deactivated, at least one heating control button (14) positioned on the control unit box (11 ) to increase the temperature to the desired level in case the temperature to be bioprinted is higher than the ambient temperature, at least one cooling control button (15) positioned on the control unit box (11 ) to reduce the temperature to the desired level in case the temperature to be bioprinted is lower than the ambient temperature, at least one power switch (16) positioned on the control unit box (11 ) to ensure that electrical power is supplied to or cut off from the entire system.

In our invention, the pressure exerted by the bio-ink solution in the bio-ink cartridge (4) on the micro linear piston (3) is calculated through the strain in the movement of the micro linear piston (3) and the change in the current value drawn by the micro linear piston (3) and with an algorithmic software.

In order to insert the bioink cartridge (4), the micro linear piston (3) must be pulled back by means of the pull button (6). If a continuous forward or backward movement is desired for the micro linear piston (3), the push or pull buttons (5, 6) are kept pressed. The LEDs (9) used in the invention have wavelengths of 360 nm, 405 nm and 445 nm and can be replaced with LEDs (9) having different wavelengths.

The control unit box (11 ) within the invention divides the electricity coming from the city network into two parts, one of which is used for the micro linear piston (3) and the other for the LEDs (9). The heating and cooling units are also located in the control unit box (11 ). The electrical energy required for the micro linear piston (3) is converted from 220V to 12V. When the movement speed of the micro linear piston (3) is desired to be changed, the voltage level is ranged between 0V and 12V. The present micro linear piston (3) used in the invention has a movement speed of 0-5 mm/s and can be made to operate in a wider range.

The biopencil structure proposed by the invention is suitable for bioprinting at room temperature even without features such as heating/cooling system, infrared sensor (7), LCD display (2) and pressure measurement.

Claims

CLAIMS portable handheld three-dimensional bioprinter which is designed for use in tissue engineering applications, artificial organ and tissue production, musculoskeletal injuries, tissue damage, bum treatment, tissue regeneration, controlled release system and three-dimensional cell culture applications as well as cosmetics and drug discovery, which eliminates the need for PC connection, drawing file and professional user, which can be easily used with one hand, the printing speed and light source of which can be changed depending on the need, which can offer heating and cooling options during printing and can print directly to the desired area in the body, characterized in that it includes:

- at least one biopencil body (1 ) with ergonomics to allow the bioprinter to be carried by hand, made of aluminum alloy to provide high thermal conductivity, anodized and ground on the outer surface to ensure a long service life and a modern appearance,

- at least one bioink cartridge (4) in which the bioink solution is placed and which has a hot/cold liquid inlet (4.1 ), a hot/cold liquid outlet (4.2) and an electrical cable inlet (4.3),

- at least one LCD display (2) to display the temperature information of the bioink contained in the bioink cartridge (4) and the pressure information applied by the bioink to the micro linear piston (3),

- at least one micro linear piston (3) to push the rear plug of the bioink cartridge (4) forward to allow the bioink solution contained in the bioink cartridge (4) to be ejected from the end of the bioink cartridge (4),

- at least one push button (5) to cause the micro linear piston (3) to move forward when bioprinting is to be performed so that the bioink solution contained in the bioink cartridge (4) is ejected from the end of the bioink cartridge (4), - at least one pull button (6) for pulling back the micro linear piston (3) and removing the bioink cartridge (4) after the bioprinting process is completed,

- at least one infrared sensor (7) integrated in the light source body (8) for real-time measurement of the temperature of the bioink solution in the bioink cartridge (4),

- at least one light source body (8) containing light sources consisting of LEDs (9) and integrated into the biopencil body (1 ), which can be separated from the biopencil body (1 ) and replaced when light of different wavelengths is desired to be used,

- LEDs (9) to ensure simultaneous curing of the printed three- dimensional structure during bioprinting,

- at least one tube casing (10) containing the electrical cables (10.1 ) for the transmission of electrical energy required for the micro linear piston (3) and LEDs (9), as well as the hot/cold liquid inlet line (10.2) and the hot/cold liquid outlet line (10.3) for the transmission of closed circuit water in the heating/cooling unit used for heating and cooling the bioink cartridge (4),

- at least one control unit box (11 ) to provide the voltage adjustment required for the speed of the micro linear piston (3), to provide the adjustment of the voltage level suitable for the LEDs (9), to contain the heating and cooling unit,

- at least one micro linear piston driver (12) positioned on the control unit box (11 ) to adjust the movement speed of the micro linear piston (3) and thus optimize the printing of bioink solutions with different characteristics,

- at least one LED switch (13) positioned on the control unit box (11 ) to enable the LEDs (9) to be activated or deactivated,

- at least one heating control button (14) positioned on the control unit box (11 ) to increase the temperature to the desired level in case the temperature to be bioprinted is higher than the ambient temperature, - at least one cooling control button (15) positioned on the control unit box (11 ) to reduce the temperature to the desired level in case the temperature to be bioprinted is lower than the ambient temperature,

- at least one power switch (16) positioned on the control unit box (11 ) to ensure that electrical power is supplied to or cut off from the entire system. The bioprinter according to claim 1 , characterized in that it comprises an algorithmic software for calculating the pressure exerted by the bioink solution in the bioink cartridge (4) on the micro linear piston (3) through the strain in the movement of the micro linear piston (3) and the change in the current value drawn by the micro linear piston (3). The bioprinter according to any one of the preceding claims, characterized in that it comprises push and pull buttons (5, 6) which, when depressed, provide a continuous movement for the micro linear piston (3). The bioprinter according to any one of the preceding claims, characterized in that it comprises LEDs (9) which currently have wavelengths of 360 nm, 405 nm and 445 nm, but which can be replaced by LEDs with different wavelengths if required. A bioprinter according to any one of the preceding claims, characterized in that it comprises a micro-linear piston (3) which currently has a movement speed range of 0-5 mm/s, but wherein this speed range can be changed if required.