Interpenetrating polymer network structure for use with sewable flexible materials

Abstract

An advanced manufacturing interpenetrating polymer network structure (AM-IPN) comprising a primary polymer network and a secondary polymer network coupled to the primary polymer network via one or more crosslinks, wherein one or more of the primary polymer network, the secondary polymer network, and the one or more crosslinks are printed using a synthetic bioink, is disclosed. Manufacturing methods and methods of use are also disclosed.

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 392,250, filed on July 26, 2022, the entire content of which is incorporated herein by reference.

[0002] This application relates to 3D - printed structures, namely, 3D - printed interpenetrating polymer network (IPN) structures for use in suture - able flexible hydrogel materials.

Background Art

[0003] Interpenetrating polymer network (IPN) enables the development of hydrogel materials that can be sutured and manipulated (e.g., bent, twisted, flexed) without breaking and can be used for perfusion and ventilation.

[0004] There is a need for IPNs that can be 3D - printed as suture - able flexible hydrogel materials for organ additive manufacturing.

Summary of the Invention

[0005] The systems and methods of the present disclosure include 3D - printed interpenetrating polymer network (IPN) structures for use in suture - able flexible hydrogel materials using synthetic bio - ink.

[0006] In embodiments, an advanced manufacturing interpenetrating polymer network (AM - IPN) comprises a primary polymer network and a secondary polymer network coupled to the primary polymer network via one or more cross - links. In embodiments, one or more of the primary polymer network, the secondary polymer network, and the one or more cross - links are printed using synthetic bio - ink.

[0007] In an embodiment, the synthetic bioink contains one or more of HPA in an amount of about 5% to about 20%, and any range or value therebetween; PEGDA 6000 in an amount of about 5% to about 20%, and any range or value therebetween; TMPTA 912 in an amount of about 0.5% to about 3%, and any range or value therebetween; NAP in an amount of about 0.5% to about 3%, and any range or value therebetween; UV386A in an amount of about 0.1% to about 0.5%, and any range or value therebetween; polymer in an amount of about 0.001% to about 2.0%, and any range or value therebetween; and water in an amount as the balance.

[0008] In an embodiment, the polymer contains one or more of polyethylene oxide (PEO), polyethyleneimine (PEI), polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), and combinations thereof.

[0009] In an embodiment, the polymer is PEO having a molecular weight of about 100,000 to about 4,000,000, and any range or value therebetween.

[0010] In an embodiment, the polymer is PEO in an amount of about 0.005% to about 1%, and any range or value therebetween.

[0011] In an embodiment, the polymer is PEI having a molecular weight of about 25,000 to about 75,000, and any range or value therebetween.

[0012] In an embodiment, the polymer is PEI in an amount of about 0.005% to about 1%, and any range or value therebetween.

[0013] In an embodiment, the polymer is PVP having a molecular weight of about 1,000,000 to about 1,300,000, and any range or value therebetween.

[0014] In an embodiment, the polymer is PVP in an amount of about 0.005% to about 2%, and any range or value therebetween.

[0015] In an embodiment, the polymer is PVA having a molecular weight of about 89,000 to about 98,000, and any range or value therebetween.

[0016] In an embodiment, the polymer is PVA in an amount of about 0.005% to about 2%, and any range or value therebetween.

[0017] In an embodiment, the synthetic bioink comprises one or more of: HPA in an amount of about 5% to about 20%, and any range or value therebetween; PEGDA 3400 in an amount of about 5% to about 20%, and any range or value therebetween; NAP in an amount of about 0.5% to about 3%; UV386A in an amount of about 0.1% to about 0.5%, and any range or value therebetween; a polymer in an amount of about 0.001% to about 2.0%, and any range or value therebetween; and water in the balance.

[0018] In an embodiment, the polymer comprises one or more of polyethylene oxide (PEO), polyethyleneimine (PEI), polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), and combinations thereof.

[0019] In an embodiment, the polymer is PEO having a molecular weight of about 100,000 to about 4,000,000, and any range or value therebetween.

[0020] In an embodiment, the polymer is PEO in an amount of about 0.005% to about 1%, and any range or value therebetween.

[0021] In an embodiment, the polymer is PEI having a molecular weight of about 25,000 to about 75,000, and any range or value therebetween.

[0022] In an embodiment, the polymer is PEI in an amount of about 0.005% to about 1%, and any range or value therebetween.

[0023] In an embodiment, the polymer is PVP having a molecular weight of about 1,000,000 to about 1,300,000, and any range or value therebetween.

[0024] In an embodiment, the polymer is PVP in an amount of about 0.005% to about 2%, and any range or value therebetween.

[0025] In an embodiment, the polymer is PVA having a molecular weight of about 89,000 to about 98,000, and any range or value therebetween.

[0026] In an embodiment, the polymer is PVA in an amount of about 0.005% to about 2%, and any range or value therebetween.

[0027] In an embodiment, a method for manufacturing an advanced manufacturing interpenetrating polymer network structure (AM-IPN) includes printing one or more of a primary polymer network, a secondary polymer network, and one or more of one or more crosslinks using a synthetic bioink and 3D printing technology; and assembling and / or printing the AM-IPN to form an assembled AM-IPN.

[0028] In an embodiment, the synthetic bioink includes one or more of polyethylene oxide (PEO), polyethyleneimine (PEI), polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), and combinations thereof.

[0029] In an embodiment, the AM-IPN is printed at about 50 mW / cm 2 ~ about 400 mW / cm 2 and any range or value therebetween.

[0030] In an embodiment, the synthetic bioink includes polyethylene oxide (PEO) in an amount of about 0.005% to about 1%, and any range or value therebetween, and the AM-IPN is printed at 50 mW / cm 2 ~ about 300 mW / cm 2 and any range or value therebetween.

[0031] In an embodiment, the synthetic bioink contains polyethyleneimine (PEI) in an amount of about 0.005% to about 1%, and any range or value therebetween, and the AM-IPN is printed at about 150 mW / cm 2 to about 300 mW / cm 2 , and any range or value therebetween.

[0032] In an embodiment, the synthetic bioink contains polyvinylpyrrolidone (PVP) in an amount of about 0.005% to about 2%, and any range or value therebetween, and the AM-IPN is printed at about 50 mW / cm 2 to about 300 mW / cm 2 , and any range or value therebetween.

[0033] In an embodiment, the synthetic bioink contains polyvinyl alcohol (PVA) in an amount of about 0.005% to about 2%, and any range or value therebetween, and the AM-IPN is printed at about 100 mW / cm 2 to about 300 mW / cm 2 , and any range or value therebetween.

[0034] In an embodiment, the 3D printing technology is one or more of digital light projection printing (DLP), stereolithography (SLA) printing technology, extrusion 3D printing technology, or selective laser sintering 3D printing technology, or a combination thereof.

[0035] In an embodiment, the 3D printing technology is digital light printing (DLP) printing technology.

[0036] In an embodiment, the method of using the advanced manufacturing interpenetrating polymer network (AM-IPN) includes modifying the surface of the AM-IPN to adhere small airway epithelial cells (SAECs) to form a modified AM-IPN. In an embodiment, the method further includes using the modified AM-IPM for perfusion and ventilation.

[0037] Those skilled in the art will understand that this summary is merely illustrative and is not intended to be limiting in any way. Other aspects, inventive features, and advantages of the devices and / or processes described herein, which are defined only by the claims, will become apparent in the detailed description set forth herein and considered in conjunction with the accompanying drawings.

[0038] This patent or patent application file contains drawings created in at least one color. Copies of this patent or patent application publication with color drawings can be obtained from the United States Patent and Trademark Office by requesting and paying the necessary fees.

[0039] Details of one or more implementations of the subject matter described herein are set forth in the accompanying drawings and the following description. Other features, aspects, and advantages of the subject matter will become apparent from the specification, drawings, and claims.

Brief Description of the Drawings

[0040]

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Embodiments for Carrying Out the Invention

[0041] The following detailed description of various embodiments of the present invention refers to the accompanying drawings that illustrate specific embodiments in which the present invention can be practiced. While the preferred embodiments of the present invention have been specifically described, it will be understood that various other modifications will be apparent to those skilled in the art and can be readily made by those skilled in the art without departing from the spirit and scope of the present invention. Accordingly, the scope of the claims appended hereto is not intended to be limited to the examples and descriptions set forth herein, but rather, the claims are intended to include all features of patentable novelty existing in the present invention, including all features that would be treated as equivalents by those skilled in the art to which the present invention pertains. Accordingly, the scope of the present invention is defined only by the appended claims, together with the full scope of equivalents to which such claims are entitled.

[0042] Advanced Manufacturing Interpenetrating Polymer Network Structure (AM-IPN) An interpenetrating polymer network structure is a primary network bonded to a secondary network via non-covalent bonds.

[0043] Figure 1A is a diagram showing the interpenetrating polymer network structure in a synthetic AA42 bioink using 8% PEGDA 3400, indicating a high crosslink density of the interpenetrating polymer network structure in the bioink.

[0044] Figure 1B is a diagram showing the interpenetrating polymer network structure in a synthetic AI28 bioink using 5% PEGDA 6000 and 3% triacetate, indicating a medium crosslink density of the interpenetrating polymer network structure in the bioink.

[0045] Figure 1C is a diagram showing the interpenetrating polymer network structure in a synthetic 602 bioink using 5% PEGDA 6000, indicating a low crosslink density of the interpenetrating polymer network structure in the bioink.

[0046] Any suitable synthetic bioink may be used. For example, suitable synthetic bioinks include, but are not limited to, the AA42, AI15, AI28, and AJ55 bioinks described herein.

[0047] Preferred Bioinks

Table 1

Table 2

Table 3

Table 4

[0048] Any suitable polymer may be incorporated into the synthetic bioink. For example, suitable polymers include, but are not limited to, polyethylene oxide (PEO), polyethyleneimine (PEI), polyvinylpyrrolidone (PVP), and polyvinyl alcohol (PVA).

Table 5

[0049] Preferred Bioink Formulations and Printing Settings for the FS20 DLP 3D System Printer Polyethylene Oxide (PEO)

[0050] In these examples, the polyethylene oxide (PEO) polymer was incorporated into the AA42 bioink (stock) as follows.

[0051] Polyethylene Oxide 4000000 (0.5% PEO Stock Solution)

[0052] The polyethylene oxide 4000000 polymer was added to water, heated at 100 °C, mixed at 600 RPM, and then incorporated into the bioink.

[0053] Polyethylene oxide 1000000 (0.5% PEO stock solution)

[0054] The polyethylene oxide 1000000 polymer was added to water, heated at 100 °C, mixed at 600 RPM, and then incorporated into the bioink. [Table 6]

[0055] In the AA42 bioink (stock) solution, the concentration of water was slightly decreased (compared to the AA42 bioink) so that the polyethylene oxide polymer stock solution could be added to prepare the AL35, AL36, AL37, AL38, and AL39 bioinks. [Table 7] [Table 8] [Table 9] [Table 10] [Table 11] [Table 12] [Table 13] [Table 14] [Table 15]

Table 16

Table 17

[0056] Rheology test

[0057] Figure 2A is a diagram showing a photograph of the air supply for a TA device equipped with an OmniCure light source and a TRIOS software package. The air supply to the TA device should be air at about 20 psi to about 40 psi.

[0058] Figure 2B is a diagram showing a photograph of the OmniCure light source for the TA device of Figure 2A.

[0059] Figure 2C-1 is a diagram showing a first screenshot of the TRIOS software package for the TA device of Figures 2A to 2B, showing the geometry icon under the experiment tab.

[0060] Figure 2C-2 is a diagram showing a second screenshot of the TRIOS software package for the TA device of Figures 2A to 2B, showing the device tab, and Figure 2C-3 is a diagram showing a third screenshot of the TRIOS software package for the TA device of Figure 2A, showing the inertia (calibration) and vibration (new mapping) settings under the device tab.

[0061] Under the inertia setting, a new calibration reading was approved because the reading was within the acceptable range (e.g., close to the previous calibration reading).

[0062] Under the vibration setting, the new mapping was calibrated.

[0063] The light source was calibrated with the black cover of a UV radiometer placed on the surface of the light source where the sample can be placed.

[0064] Figure 2C-4 is a diagram showing the fourth screenshot of the TRIOS software package for the TA device in Figures 2A - 2B, showing the accessory settings under the calibration tab. The UV radiation should be set to 50%. To calibrate the light source, the value measured with a UV radiometer was entered into the software (e.g., 56.2) and approved.

[0065] Figure 2C-5 is a diagram showing the fifth screenshot of the TRIOS software package for the TA device in Figures 2A - 2B, showing the geometry tab. The geometry of the 20 mm parallel plate was selected. The appropriate plate was placed on the rheometer.

[0066] Under the inertia settings, a new calibration reading was approved because the reading was within the acceptable range (e.g., close to the previous reading).

[0067] Under the friction settings, a new calibration reading was approved because the reading was within the acceptable range (e.g., close to the previous calibration reading).

[0068] Lower the rheometer slightly above the sample placement area and select the zero gap icon.

[0069] Pull the rheometer up to the loading gap.

[0070] Under the geometry tab, set the gap to 200 μm and the minimum sample volume to 0.062 mL.

[0071] Figure 2C-6 is a diagram showing the sixth screenshot of the TRIOS software package for the TA device in Figures 2A - 2B, showing the condition options and settings.

[0072] Figure 2C-7 is a diagram showing the seventh screenshot of the TRIOS software package for the TA device in Figures 2A - 2B, showing the UV settings for other events.

[0073] Figure 2C-8 shows the eighth screenshot of the TRIOS software package of the TA device in FIGS. 2A to 2B, showing the vibration high-speed sampling setting.

[0074] 75 μL of the sample was loaded at the center of the rheometry plate. All bubbles were removed by gently piercing the bubbles with a needle. The gap was set at 200 μm. Before each experiment, the black UV cover was lowered to shield the UV light.

[0075] Figure 2D is a graph showing the step time ts (seconds) versus the storage modulus G' (kPa) of the advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethylene oxide (PEO) in the synthetic AL35, AL36, AL37, and AL38 bioinks. [Table 18] [Table 19]

[0076] Compression test

[0077] Dynamic mechanical analysis (DMA) is a technique often used to evaluate the properties of materials as a function of atmosphere, frequency, stress, temperature, time, and combinations thereof.

[0078] The compression test is performed using a DMA 850 device designed to measure the viscoelastic properties (e.g., modulus, attenuation) of hard and soft solid materials. The compression test can be used to measure the properties of low to medium modulus materials including gels and weak elastomers. During the compression test, the sample needs to support a static (preload) force. The sample should have the highest possible thickness-to-diameter ratio, depending on sample preparation and the limitations of the device.

[0079] A typical sample is a printed disk (e.g., about 8 mm in diameter and about 3 mm in thickness).

[0080] The printed disk is attached to the clamp, one side of the clamp is fixed, and the other side is connected to the drive motor and is movable. This motor directly applies deformation to the printed disk.

[0081] The compressive modulus of elasticity is calculated from the slope in the linear region of stress (kPa) versus strain (%), typically in the region of initial strain from 0% to 10%. The R 2 value of the modulus of elasticity should be from about 0.95 to about 0.99.

[0082] The compressive failure strength and strain are the values of the maximum stress (kPa) and maximum strain (%) at which the printed disk breaks.

[0083] Figure 3A is a diagram showing the air supply for the DMA 850 device equipped with the TRIOS software package. The air supply to the DMA device should be about 60 psi.

[0084] Figure 3B is a diagram showing a photograph of calibrating the bottom clamp for the DMA 850 device of Figure 3A. The bottom clamp for the DMA device should be calibrated before the compression test. Attach the bottom clamp to the DMA device and tighten it with a hex wrench until the bottom clamp is aligned and stable.

[0085] Figure 3C is a diagram showing the first screenshot of the TRIOS software package for the DMA 850 device of Figures 3A - 3B, showing the clamp calibration tab. The furnace of the DMA device was in a closed state. Calibrate and approve the mass of the bottom clamp.

[0086] Figure 3D is a diagram showing a photograph of assembling the bottom part of the compression equipment for the DMA 850 device of Figures 3A - 3B. After calibrating the mass of the bottom clamp, assemble the bottom part of the compression equipment and tighten it to a predetermined position with a hex wrench.

[0087] Figure 3E is a diagram showing a photograph of aligning the top clamp with respect to the bottom clamp for the DMA apparatus of FIGS. 3A to 3B. After attaching the bottom plate, the top clamp was aligned with respect to the bottom clamp and tightened to a predetermined position using a hex wrench.

[0088] Figure 3F is a diagram showing a second screen shot of the TRIOS software package for the DMA 850 apparatus of FIGS. 3A to 3B and FIGS. 3D to 3E, showing the clamp calibration tab. When the plates are fully aligned, the clamps are calibrated for compliance and approved. For approval, the compliance value should be less than 1 μN / m.

[0089] Figure 3G-1 is a diagram showing a screen shot of the DMA 850 apparatus, showing a floating icon. After setting up the clamp, the clamp should be calibrated. Pressing the floating icon causes the top clamp to float. This ensures that the top clamp is in a loose state with respect to the printed disk.

[0090] The printed disk should be stored in 1×DPBS overnight before the compression test. Immediately before placing the printed disk in the clamp, the excess 1×DPBS should be gently wiped off using a Kimwipe.

[0091] Figure 3G-2 is a diagram showing a third screen shot of the TRIOS software package for the DMA 850 apparatus of FIGS. 3A to 3B and FIGS. 3D to 3E, showing the sample dimensions. The diameter and thickness of the printed disk are measured using calipers. The measured values are entered into the TRIOS software package.

[0092] Place the printed disk between the bottom clamp and the top clamp, and ensure that the printed disk is placed exactly in the center between the clamps. Lower the movable top clamp so that it just touches the printed disk.

[0093] Under the Procedure tab, the desired compression parameters (e.g., final strain percent, strain rate) should be entered. A typical compression test is performed at about 20% strain / min.

[0094] Figure 3G-3 is a diagram showing a fourth screenshot of the TRIOS software package for the DMA 850 apparatus of FIGS. 3A-3B and 3D-3E, showing the DMA Control tab. For a printed disk (e.g., hydrogel), a preload force of 0.05 N was entered. The compression test may be performed using an open furnace. The compression test can be stopped when the printed disk begins to break (e.g., fracture).

[0095] After stopping the compression test, the Young's modulus can be calculated from the slope of the stress-strain curve in the linear region (e.g., about 0% to about 10% strain). The ultimate compression strain (%) and ultimate compression strength (kPa) at the break point can be calculated. For the Young's modulus and break strain, the average of about 6 printed disks should be calculated.

[0096] Figure 3H is a diagram showing a graph of strain ε (%) versus stress σε (kPa) of advanced manufacturing interpenetrating polymer network structures (AM-IPN) and lung G1 in synthetic AA42, AL35 (0.5% PEO), AL36 (0.1% PEO), AA42, AL43 (0.5% PEO), AL44 (0.1% PEO), AL45 (0.05% PEO), and AI28 bioinks, showing a comparison of PEO concentrations in the bioinks.

[0097] Figure 3I is a diagram showing a graph of strain ε (%) versus stress σε (kPa) of advanced manufacturing interpenetrating polymer network structures (AM-IPN) in synthetic AL35, AL36, and AA42 bioinks with polyethylene oxide (PEO) 4000000, showing a comparison of PEO concentrations in the bioinks.

[0098] Figure 3J is a graph showing the advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethylene oxide (PEO) in synthetic AL42, AL45, and AI28 bioinks, and the strain ε (%) vs. stress σε (kPa) of lung G1, showing a comparison of the PEO concentrations in the bioinks.

[0099] Figure 3K is a graph showing the advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethylene oxide (PEO) 4000000 in synthetic AL45 (3% triacrylate, 0.05% PEO), AL48 (4% triacrylate, 0.05% PEO), and AL49 (3% triacrylate, 0.01% PEO) bioinks, and the strain ε (%) vs. stress σε (kPa) of lung G1, showing a comparison of the PEO concentrations in the bioinks.

[0100] Figure 4A is a diagram showing a photograph of the advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethylene oxide (PEO) in synthetic AL35 bioink.

[0101] Figure 4B is a photograph of the advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethylene oxide (PEO) in synthetic AL36 bioink.

[0102] Figure 4C is a photograph of the advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethylene oxide (PEO) in synthetic AL37 bioink.

Table 20

Table 21

[0103] Figure 5 is a graph showing the strain ε (%) versus stress σε (kPa) of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethylene oxide (PEO) in synthetic AL45 (energy 210 mJ), AL49 (energy 90 mJ), and AI28 (parent ink) (energy 48 mJ) bioinks.

[0104] Figure 6A is a diagram showing a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethylene oxide (PEO) in synthetic AL45 bioink (3% triacrylate, 0.05% PEO4000000), showing 100.100 dense rings with an average vascular structure thickness of 95.4 ± 8.7 μm.

[0105] Figure 6B is a diagram showing a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethylene oxide (PEO) in synthetic AL45 bioink, showing 50.80 dense rings with an average vascular structure thickness of 48.8 ± 10.1 μm.

[0106] Figure 7A is a diagram showing a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in an advanced manufacturing hydrogel material of synthetic AL49 bioink (3% triacrylate, 0.1% PEO4000000), showing the state where the hydrogel material is placed in a bioreactor and preparations for perfusion and ventilation are being made.

Table 22

[0107] Place the hydrogel material in a bioreactor and insert tubes for the vascular structure and airways through the blue cap of the bioreactor. This is to test the performance of the hydrogel material in expanding around the tubes.

[0108] Figure 7B is a diagram showing a photograph of the AM-IPN in the advanced manufacturing hydrogel material of Figure 7A, showing the hydrogel material during perfusion and ventilation.

[0109] Biocompatibility test

[0110] The printed disk (e.g., hydrogel) of AL36 bioink was printed (e.g., with a diameter of about 8 cm and a thickness of about 3 mm).

[0111] Small airway epithelial cells were seeded onto the printed disk (e.g., hydrogel). This was performed in a transparent-bottom cell culture plate.

[0112] At each time point, the printed disk (e.g., hydrogel) was placed under a microscope to determine whether the adhesion of small airway epithelial cells was still occurring and whether the small airway epithelial cells remained unstained.

[0113] Figure 8A is a diagram showing a photograph of a biocompatibility test (glass control) in the case of polyethylene oxide (PEO) in synthetic AL36 bioink.

[0114] Figure 8B is a diagram showing a photograph of a biocompatibility test (day 1) of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethylene oxide (PEO) in synthetic AL36 bioink, showing the adhesion of small airway epithelial cells (SAEC).

[0115] Figure 8C is a diagram showing a photograph of a biocompatibility test (day 4) of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethylene oxide (PEO) in synthetic AL36 bioink, showing a decrease in the adhesion of small airway epithelial cells (SAEC).

[0116] Suturing test

[0117] Using a 45 mm gauge needle, the needle was passed through the solid part (capsulent) of the advanced manufacturing hydrogel material. The hydrogel material (capsulent) was inspected for cracks. The purpose of this test was to confirm whether the hydrogel material could be sutured without cracking or falling apart.

[0118] Figure 9A is a diagram showing a photograph of a stitching test of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethylene oxide (PEO) in a synthetic AL48 (4% triacrylate, 0.05% PEO) bioink, indicating a high crosslinking density of the interpenetrating polymer network structure in the bioink.

[0119] Figure 9B is a diagram showing a photograph of a stitching test of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethylene oxide (PEO) in a synthetic AL49 (3% triacrylate, 0.1% PEO) bioink, indicating a medium crosslinking density of the interpenetrating polymer network structure in the bioink.

[0120] AL35 bioink swelled and disintegrated during the surface modification process. However, although AL36 bioink was maintained intact during the surface modification, it showed a decrease in the adhesion of small airway epithelial cells (SAEC) by the fourth day.

[0121] AA42 bioink forms an interpenetrating polymer network structure with a high crosslinking density in the bioink. Therefore, the polyethylene oxide (PEO) polymer may be strongly bound in the interpenetrating polymer network structure.

[0122] Polyethyleneimine (PEI)

[0123] In these examples, the polyethyleneimine (PEI) polymer was incorporated into the AI28 bioink (stock) as follows.

[0124] Polyethyleneimine (PEI) 750000 (50% PEI stock solution)

[0125] Polyethyleneimine (PEI) 750000 was dissolved in water and mixed at 600 RPM before adding the other components.

[0126] Polyethyleneimine (PEI) 25000 (50% PEI stock solution)

[0127] Polyethyleneimine (PEI) 25000 was dissolved in water and mixed at 600 RPM before adding the other components.

Table 23

Table 24

Table 25

Table 26

Table 27

Table 28

Table 29

Table 30

Table 31

[0128] Rheology test

[0129] The rheology tests discussed above and shown in FIGS. 2A to 2C-8.

[0130] FIG. 10 is a graph showing the step time ts (seconds) versus the storage modulus G’ (kPa) of the advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethyleneimine (PEI) in synthetic AL60, AL62, AL364, AL65, and AL66 bioinks.

Table 32

Table 33

Table 34

Table 35

Table 36

Table 37

[0131] Swelling test

[0132] A printed disk (e.g., hydrogel) of AL36 bioink was printed (e.g., with a diameter of about 8 cm and a thickness of about 3 mm).

[0133] The printed disk was weighed before immersion (i.e., the initial weight). Just before weighing, any excess resin needs to be gently wiped from the printed disk using a Kimwipe.

[0134] The printed disk was placed in a 24-well plate. 1×DPBS++ was added to each well to approximately five times the volume of the printed disk (i.e., since the volume of an 8 mm × 3 mm printed disk is approximately 150 μL, approximately 1 mL of 1×DPBS++ needs to be added to each well).

[0135] The printed disk needs to be weighed after being immersed in 1×DPBS++ at 37 °C for 24 hours (i.e., the final weight after 24 hours). Just before weighing, any excess 1×DPBS++ needs to be gently wiped from the printed disk using a Kimwipe.

[0136] The swelling rate (%) can be calculated as follows.

Equation

[0137] Fresh 1×DPBS++ was added to each well. To continue the swelling test, the printed disks need to be weighed after immersion in 1×DPBS++ at 37 °C for up to 4 days or up to 7 days (i.e., the final weight after 4 or 7 days).

Table 38

[0138] Compression test

[0139] The compression test was discussed above and is shown in FIGS. 3A to 3G-3.

[0140] FIG. 11 is a graph showing the advanced manufacturing interpenetrating polymer network structure (AM-IPN) and the strain ε (%) vs. stress σε (kPa) of lung G1 in the case of polyethyleneimine (PEI) 750,000 in synthetic AL60 (output 50 mW / cm 2 , time 1900 ms), AL62 (output 50 mW / cm 2 , time 750 ms), AL62 (output 150 mW / cm 2 , time 500 ms), AL62 (output 200 mW / cm 2 , time 500 ms) bioinks.

[0141] FIG. 12A is a diagram showing a photograph of the advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethyleneimine (PEI) in synthetic AL60 bioink.

[0142] FIG. 12B is a photograph of the advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethyleneimine (PEI) in synthetic AL61 bioink.

[0143] FIG. 12C is a photograph of the advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethyleneimine (PEI) in synthetic AL62 (output 50 mW / cm 2 ) bioink, showing a low crosslink density of the interpenetrating polymer network structure of PEI in the bioink.

[0144] Figure 12D is a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethyleneimine (PEI) in the synthetic AL62 (output 150 mW / cm 2 ) bioink, showing a medium crosslink density of the interpenetrating polymer network structure of PEI in the bioink.

[0145] Figure 12E is a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyethyleneimine (PEI) in the synthetic AL62 (output 200 mW / cm 2 ) bioink, showing a low crosslink density of the interpenetrating polymer network structure of PEI in the bioink.

Table 39

Table 40

Table 41

Table 42

[0146] When printed at 50 mW / cm 2 , the AL62 bioink results in a softer hydrogel than the AL60 bioink (stock), and when printed at 150 mW / cm 2 , the AL62 bioink results in a tougher hydrogel than the AL60 bioink (stock).

[0147] When printed at 150 mW / cm 2 and 200 mW / cm 2 , the polyethyleneimine (PEI) polymer was incorporated into the AL60 bioink (stock).

[0148] Suture test

[0149] Using a 45 mm gauge needle, the needle was passed through the solid part (encapsulating agent) of the advanced manufacturing hydrogel material. The hydrogel material (encapsulating agent) was inspected for cracks. The purpose of this test was to confirm whether the hydrogel material could be sutured without cracking or falling apart.

[0150] Figure 13A is a diagram showing a photograph of a stitching test of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in polyethyleneimine (PEI) in a synthetic AL62 (output 50 mW / cm 2 , time 750 ms) bioink, showing a low crosslink density of the interpenetrating polymer network structure of PEI in the bioink.

[0151] Figure 13B is a diagram showing a photograph of a stitching test of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in polyethyleneimine (PEI) in a synthetic AL62 (output 100 mW / cm 2 , time 500 ms) bioink, showing a low crosslink density of the interpenetrating polymer network structure of PEI in the bioink.

[0152] Figure 13C is a diagram showing a photograph of a stitching test of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in polyethyleneimine (PEI) in a synthetic AL62 (output 150 mW / cm 2 , time 450 ms) bioink, showing a medium crosslink density of the interpenetrating polymer network structure of PEI in the bioink.

[0153] The AL62 bioink failed the stitching test when printed at 50 mW / cm 2 and 100 mW / cm 2 . However, the AL62 bioink passed the stitching test when printed at 150 mW / cm 2 and 200 mW / cm 2 .

[0154] Polyvinylpyrrolidone (PVP)

[0155] In these examples, the polyvinylpyrrolidone (PVP) polymer was incorporated into the AI28 and AJ55 bioinks as follows.

[0156] Since the PVP polymer has bulky side chains, this polymer may impart rigidity to the AI28 bioink. [Table 43] [Table 44] [Table 45] [Table 46] [Table 47] [Table 48] [Table 49] [Table 50] [Table 51] [Table 52] [Table 53] [Table 54] [Table 55] [Table 56] [Table 57]

Table 58

[0157] Rheology test

[0158] The rheology test was considered above and is shown in FIGS. 2A to 2C-8.

[0159] FIG. 14 is a graph showing the step time ts (seconds) versus the storage modulus G' (kPa) of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyvinylpyrrolidone (PVP) in synthetic AL68, AL69, AL70, AL72, AL73, AL74, and AL75 bioinks.

Table 59

Table 60

[0160] Compression test

[0161] The compression test was considered above and is shown in FIGS. 3A to 3G-3.

[0162] FIG. 15 is a graph showing the advanced manufacturing interpenetrating polymer network structure (AM-IPN) and the strain ε (%) versus stress σε (kPa) of lung G1 in the case of polyvinylpyrrolidone (PVP) in synthetic AL68 (3% triacrylate, 0.5% PVP), AL70 (3% triacrylate, 0.05% PVP), AL72 (2% triacrylate, 1% PVP), AL73 (2% triacrylate, 0.5% PVP), and AL74 (2% triacrylate, 0.1% PVP) bioinks.

[0163] FIG. 16A is a diagram showing a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyvinylpyrrolidone (PVP) in synthetic AL68 bioink.

[0164] Figure 16B is a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyvinylpyrrolidone (PVP) in the synthetic AL70 bioink.

[0165] Figure 16C is a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyvinylpyrrolidone (PVP) in the synthetic AL72 bioink.

[0166] Figure 16D is a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyvinylpyrrolidone (PVP) in the synthetic AL73 bioink.

[0167] Figure 16E is a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyvinylpyrrolidone (PVP) in the synthetic AL74 bioink. [Table 61] [Table 62] [Table 63] [Table 64] [Table 65]

[0168] The polyvinylpyrrolidone (PVP) polymer was incorporated into the AI28 bioink (stock) and the AJ55 bioink (stock).

[0169] The AL73 bioink showed the best toughness and the highest maximum strain while maintaining flexibility.

[0170] Suture test

[0171] Using a 45 mm gauge needle, the needle was passed through the solid part (encapsulating agent) of the advanced manufacturing hydrogel material. The hydrogel material (encapsulating agent) was inspected for cracks. The purpose of this test was to confirm whether the hydrogel material could be sutured without cracking or falling apart.

[0172] Figure 17A is a diagram showing a photograph of the suturing test of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyvinylpyrrolidone (PVP) in synthetic AL68 bioink.

[0173] Figure 17B is a diagram showing a photograph of the suturing test of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyvinylpyrrolidone (PVP) in synthetic AL70 bioink.

[0174] Figure 17C is a diagram showing a photograph of the suturing test of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyvinylpyrrolidone (PVP) in synthetic AL73 bioink.

[0175] AL70 bioink failed the suturing test. However, AL68 and AL73 bioinks passed the suturing test.

[0176] Polyvinyl alcohol (PVA)

[0177] In these examples, the polyvinyl alcohol (PVA) polymer was incorporated into AI28 bioink (stock) as follows.

[0178] Polyvinyl alcohol (PVA) (2% PVP stock solution)

[0179] Polyvinyl alcohol (PVA) was added to water, heated at 200 °C, mixed at 200 RPM, and then incorporated into the bioink. [Table 66] [Table 67]

Table 68

[0180] Rheology test

[0181] The rheology test was considered above and is shown in FIGS. 2A to 2C-8.

[0182] FIG. 18 is a graph showing the step time ts (seconds) versus the storage modulus G' (kPa) of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyvinyl alcohol (PVA) in synthetic AL76 and AL77 bioinks.

Table 69

Table 70

[0183] Compression test

[0184] The compression test was considered above and is shown in FIGS. 3A to 3G-3.

[0185] FIG. 19 is a graph showing the advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyvinyl alcohol (PVA) in synthetic AL76 and AL77 bioinks, and the strain ε (%) versus the stress σε (kPa) of lung G1.

[0186] FIG. 20A is a diagram showing a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyvinyl alcohol (PVA) in synthetic AL76 (output 50 mW / cm 2 , time 1000 ms) bioink, showing a low crosslink density of the interpenetrating polymer network structure of PVA in the bioink. As shown in FIG. 20A, 50 mW / cm 2When printed, the polyvinyl alcohol (PVA) polymer was not incorporated into the AI28 bioink (stock).

[0187] Figure 20B is a diagram showing a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in polyvinyl alcohol (PVA) in a synthetic AL76 (output 100 mW / cm 2 , time 700 ms) bioink, showing a low to medium crosslink density of the interpenetrating polymer network structure of PVA in the bioink. As shown in Figure 20B, when printed at 100 W / cm 2 , the polyvinyl alcohol (PVA) is incorporated into the AI28 bioink (stock).

[0188] Figure 20C is a diagram showing a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in polyvinyl alcohol (PVA) in a synthetic AL77 (output 100 mW / cm 2 , time 700 ms) bioink, showing a medium crosslink density of the interpenetrating polymer network structure of PVA in the bioink. As shown in Figure 19C, when printed at 100 W / cm 2 , the polyvinyl alcohol (PVA) is incorporated into the AI28 bioink (stock).

[0189] Figure 21A is a diagram showing a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in polyvinyl alcohol (PVA) in a synthetic AL76 (output 50 mW / cm 2 , time 1000 ms) bioink, showing a low crosslink density of the interpenetrating polymer network structure of PVA in the bioink. As shown in Figure 21A, when printed at 50 mW / cm 2 , the polyvinyl alcohol (PVA) polymer was not incorporated into the AI28 bioink (stock).

[0190] Figure 21B is a diagram showing a synthetic AL76 (output 50 mW / cm 2, it is a diagram showing a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyvinyl alcohol (PVA) in the bioink, indicating a low to medium crosslink density of the interpenetrating polymer network structure of PVA in the bioink. As shown in Figure 21B, when printed at 100 W / cm 2 , polyvinyl alcohol (PVA) is incorporated into the AI28 bioink (stock).

[0191] Figure 21C is a diagram showing a photograph of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in the case of polyvinyl alcohol (PVA) in the bioink at a synthesis of AL76 (output 100 mW / cm 2 , time 700 ms), indicating a medium crosslink density of the interpenetrating polymer network structure of PVA in the bioink. As shown in Figure 21C, when printed at 100 W / cm 2 , polyvinyl alcohol (PVA) is incorporated into the AI28 bioink (stock).

Table 71

Table 72

[0192] When printed at 50 mW / cm 2 , the polyvinyl alcohol (PVA) polymer was not incorporated into the AI28 bioink (stock). However, when printed at 100 mW / cm 2 , the polyvinyl alcohol (PVA) polymer was incorporated into the AI28 bioink (stock).

[0193] Suture test

[0194] Using a 45 mm gauge needle, the needle was passed through the solid portion (encapsulant) of the advanced manufacturing hydrogel material. The hydrogel material (encapsulant) was inspected for cracks. The purpose of this test was to confirm whether the hydrogel material could be sutured without cracking or falling apart.

[0195] Figure 22A is a diagram showing a photograph of a suturing test of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in polyvinyl alcohol (PVA) in a synthetic AL76 (output 100 mW / cm 2 , time 700 ms) bioink, showing a medium crosslink density of the interpenetrating polymer network structure of PVA in the bioink.

[0196] Figure 22B is a diagram showing a photograph of a suturing test of an advanced manufacturing interpenetrating polymer network structure (AM-IPN) in polyvinyl alcohol (PVA) in a synthetic AL77 (output 100 mW / cm 2 , time 700 ms) bioink, showing a medium crosslink density of the interpenetrating polymer network structure of PVA in the bioink.

[0197] The AL76 and AL77 bioinks passed the suturing test.

[0198] Interpenetrating polymer network structure mixture

[0199] In these examples, one or more of the polyethylene oxide (PEO), polyvinylpyrrolidone (PVP), and polyvinyl alcohol (PVA) polymers were incorporated into the synthetic bioink as follows.

[0200] Polymer

[0201] Polyethylene oxide (PEO)

[0202] In these examples, the polyethylene oxide (PEO) polymer was incorporated into the AW55 bioink (stock) as follows.

[0203] Polyethylene oxide 4000000 (0.5% PEO stock solution)

[0204] The polyethylene oxide 4000000 polymer was added to water, heated at 100 °C, mixed at 600 RPM, and then incorporated into the bioink.

[0205] Polyethylene oxide 400000 (0.5% PEO stock solution)

[0206] The polyethylene oxide 400000 polymer was added to water, heated at 100 °C, mixed at 600 RPM, and then incorporated into the bioink.

[0207] Polyethylene 100000 (0.5% PEO stock solution)

[0208] The polyethylene 100000 polymer was added to water, heated at 100 °C, and mixed at 600 RPM.

[0209] Polyvinyl alcohol (PVA)

[0210] Polyvinyl alcohol 100000 (4% PVA stock solution)

[0211] Polyvinyl alcohol (PVA) was added to water, heated at 200 °C, mixed at 200 RPM, and then incorporated into the bioink.

[0212] Polyethylene oxide 100000 (2% PEO stock solution)

[0213] The polyethylene oxide 100000 polymer was added to water, heated at 100 °C, mixed at 600 RPM, and then incorporated into the bioink.

[0214] Polyethylene oxide 200000 (2% PEO stock solution)

[0215] The polyethylene oxide 200,000 polymer was added to water, heated at 100 °C, mixed at 600 RPM, and then incorporated into the bioink.

[0216] Polyethylene oxide 4,000,000 (0.4% PEO stock solution)

[0217] The polyethylene oxide 1,000,000 polymer was added to water, heated at 100 °C, mixed at 600 RPM, and then incorporated into the bioink.

Table 73

Table 74

Table 75

Table 76

Table 77

Table 78

Table 79

Table 80

Table 81

Table 82

Table 83

Table 84

Table 85

Table 86

Table 87

Table 88

Table 89

[0218] Swelling test

[0219] The swelling of the bioink hydrogel (dog bone type) was observed for 24 hours.

[0220] The AW93 bioink hydrogel (dog bone type) swelled by about 30%, so the HPA was decreased from 10% to 7%.

[0221] Compression test

[0222] The compression test was considered above and is shown in FIGS. 3A to 3G-3.

[0223] The compression failure strength and strain are the values of the maximum stress (kPa) and maximum strain (%) at which the printed disk breaks.

Table 90

Table 91

[0224] FIG. 23A is a diagram showing a chart of stress (kPa) in the case of polyvinylpyrrolidone (PVP) in synthetic AW55, AW99, and AI28 bioinks.

[0225] FIG. 23B is a diagram showing a chart of strain (%) in the case of polyvinylpyrrolidone (PVP) in synthetic AW55, AW99, and AI28 bioinks.

[0226] Figure 23C is a chart showing the Young's modulus (kPa) in the case of polyvinylpyrrolidone (PVP) in synthetic AW55, AW99, and AI28 bioinks.

[0227] Figure 24 is a figure showing a photograph of a hydrogel (dog bone type) material.

[0228] Method for manufacturing an advanced manufacturing interpenetrating polymer network structure (AM-IPN) The method for manufacturing an advanced manufacturing interpenetrating polymer network structure (AM-IPN) includes: a) using a synthetic bioink and 3D printing technology to print one or more of a primary polymer network, a secondary polymer network, and one or more crosslinks; b) assembling and / or printing the AM-IPN as described herein to form an assembled AM-IPN.

[0229] In an embodiment, the 3D printing technology is one or more of digital light projection printing (DLP), stereolithography (SLA) printing technology, extrusion 3D printing technology, or selective laser sintering 3D printing technology, or a combination thereof. In an embodiment, the 3D printing technology is digital light printing (DLP) printing technology.

[0230] Method for using an advanced manufacturing interpenetrating polymer network structure (AM-IPN) The method for using an advanced manufacturing interpenetrating network structure (AM-IPN) includes: a) modifying the surface of the AM-IPN to adhere small airway epithelial cells (SAECs) to form a modified AM-IPN.

[0231] In an embodiment, this method further includes: b) using the modified AM-IPM for perfusion and ventilation as described herein.

[0232] The embodiments and examples described in this specification are presented in order to best explain the invention and its practical applications, and thereby enable those skilled in the art to manufacture and use the invention. However, those skilled in the art will recognize that the foregoing descriptions and examples are presented for illustrative and exemplary purposes only. The described content is not intended to be exhaustive or to limit the invention to the form disclosed. Many modifications and variations are possible in light of the above teachings without departing from the spirit and scope of the following claims. The invention is particularly intended to be as broad as the following claims and their equivalents.

[0233] Any reference in this specification to an implementation or an element or an execution of a system and method in the singular may include implementations that include a plurality of these elements, and any reference in this specification to an implementation or an element or an execution in the plural may include implementations that include only a single element. References in the singular or plural are not intended to limit the systems or methods, their components, executions, or elements disclosed in the invention to a single or plural configuration. Any reference to an execution or an element being based on any information, execution, or element may include implementations where the execution or element is at least partially based on any information, execution, or element.

[0234] As used herein, the terms "about," "approximately," "substantially," and similar terms are intended to have a broad meaning consistent with the ordinary and accepted usage by those skilled in the art to which the subject matter of this disclosure pertains. Those skilled in the art considering this disclosure will understand that these terms are intended to allow a description of a particular feature described and claimed without limiting the scope of such feature to the numerical ranges given. Accordingly, these terms are to be interpreted as indicating that minor or insignificant modifications or variations of the described and claimed subject matter are considered to be within the scope of the disclosure as set forth in the appended claims.

[0235] As used herein to describe various embodiments, the term "exemplary" and variations thereof are intended to indicate that such embodiments are intended to be possible examples, representatives, or illustrations of possible embodiments (and such terms are not intended to imply that such embodiments are necessarily special examples or best examples).

[0236] As used herein, the term "coupled" and variations thereof mean that two members are joined to each other either directly or indirectly. Such joining may be stationary (e.g., permanent or fixed) or movable (e.g., removable or dissociable). Such joining can be achieved in a state where two members are directly coupled to each other, a state where two members are coupled to each other using separate intervening members and any additional intermediate members coupled to each other, or a state where two members are coupled to each other using an intervening member integrally formed as a single entity with one of the two members. When "coupled" or variations thereof are modified by additional terms (e.g., directly coupled), the general definition of "coupled" given above is modified by the literal meaning of that additional term (e.g., "directly coupled" means the joining of two members without separate intervening members), resulting in a definition narrower than the general definition of "coupled" given above. Such coupling may be mechanical, electrical, or fluidic.

[0237] Any of the embodiments disclosed herein may be combined with any other embodiment, and references to "one embodiment", "some embodiments", "an alternative embodiment", "various embodiments", "one implementation", or the like are not necessarily mutually exclusive, and are intended to indicate that the particular features, structures, or characteristics described in connection with that embodiment may be included in at least one embodiment. Such terms, as used herein, do not necessarily all refer to the same embodiment. Any embodiment may be combined with any other embodiment, inclusively or exclusively, in any manner consistent with the aspects and embodiments disclosed herein.

[0238] References to "or" may be construed inclusively, so that any terms listed using "or" may indicate any one, more than one, or all of the recited terms. References to at least one of a list of conjunctive terms may be construed as an inclusive "or" indicating any one, more than one, or all of the recited terms. For example, a reference to "at least one of 'A' and 'B'" may include only 'A', only 'B', or both 'A' and 'B'. Elements other than 'A' and 'B' may also be included.

[0239] References to the position of elements in this specification (e.g., "top", "bottom", "above", "below") are used merely to describe the orientation of various elements in the drawings. It should be noted that the orientation of the various elements may vary according to other preferred embodiments, and such variations are intended to be encompassed by this disclosure.

[0240] Although a particular order of method steps may be shown in the drawings and description, such order of steps may be different from that shown and described, unless otherwise specified above. Also, two or more steps may be performed simultaneously or partially simultaneously, unless otherwise specified above. Such variations may depend, for example, on the selected software and hardware systems and on the designer's choices. All such variations are within the scope of this disclosure. Similarly, the execution of the software of the described method may be accomplished by standard programming techniques using rule-based logic and other logic to achieve various connection steps, processing steps, comparison steps, and decision steps.

[0241] If a reference sign is attached following a technical feature in the drawings, the detailed description, or any of the claims, the reference sign is included to enhance the understandability of the drawings, the detailed description, and the claims. Therefore, the presence or absence of a reference sign has no limiting effect on the scope of the elements of any of the claims.

[0242] The systems and methods described herein may be embodied in other specific forms without departing from their characteristics. The foregoing embodiments are illustrative rather than limiting of the described systems and methods. Accordingly, the scope of the systems and methods described herein is indicated not by the foregoing description but by the appended claims, and changes within the meaning and scope of the equivalence of the claims are embraced by the claims.

Claims

**Claim 1** An advanced manufacturing interpenetrating polymer network structure (AM-IPN), comprising: A primary polymer network; A secondary polymer network coupled to the primary polymer network via one or more crosslinks, wherein one or more of the primary polymer network, the secondary polymer network, and the one or more crosslinks are printed using a synthetic bioink. An advanced manufacturing interpenetrating polymer network structure (AM-IPN). **Claim 2** The synthetic bioink comprises one or more of: HPA in an amount of about 0.5% to about 20%; PEGDA 6000 in an amount of about 0.5% to about 20%; TMPTA 912 in an amount of about 0.05% to about 3%; NAP in an amount of about 0.05% to about 3%; UV386A in an amount of about 0.0001% to about 0.5%; A polymer in an amount of about 0.001% to about 2.0%; and Water in the balance, The AM-IPN according to claim 1. **Claim 3** The polymer comprises one or more of polyethylene oxide (PEO), polyethyleneimine (PEI), polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), and combinations thereof, the AM-IPN according to claim 2. **Claim 4** The polymer is PEO having a molecular weight of about 100,000 to about 4,000,000, the AM-IPN according to claim 3. **Claim 5** The polymer is PEO in an amount of about 0.005% to about 1%, the AM-IPN according to claim 3. **Claim 6** The polymer is PEI having a molecular weight of about 25,000 to about 75,000, the AM-IPN according to claim 3. **Claim 7** The polymer is PEI in an amount of about 0.005% to about 1%, the AM-IPN according to claim 3. **Claim 8** The polymer is PVP having a molecular weight of about 1,000,000 to about 1,300,000, the AM-IPN according to claim 3. **Claim 9** The polymer is PVP in an amount of about 0.005% to about 2%, the AM-IPN according to claim 3. **Claim 10** The polymer is PVA having a molecular weight of about 89,000 to about 98,000, the AM-IPN according to claim 3. **Claim 11** The polymer is PVA in an amount of about 0.005% to about 2%, the AM-IPN according to claim 3. **Claim 12** The synthetic bioink comprises one or more of: HPA in an amount of about 0.5% to about 20%; PEGDA 6000 in an amount of about 0.5% to about 20%; PEGDA 3400 in an amount of from about 0.2% to about 15%; NAP in an amount of from about 0.05% to about 3%; UV386A in an amount of from about 0.0001% to about 0.5%; a polymer in an amount of from about 0.001% to about 2.0%; and water in an amount as the balance, The interpenetrating network structure according to claim 1, comprising one or more of the foregoing.

13. The AM-IPN according to claim 12, wherein the polymer comprises one or more of polyethylene oxide (PEO), polyethyleneimine (PEI), polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), and combinations thereof.

14. The AM-IPN according to claim 13, wherein the polymer is PEO having a molecular weight of from about 100,000 to about 4,000,000.

15. The AM-IPN according to claim 13, wherein the polymer is PEO in an amount of from about 0.005% to about 1%.

16. The AM-IPN according to claim 13, wherein the polymer is PEI having a molecular weight of from about 25,000 to about 75,000.

17. The AM-IPN according to claim 13, wherein the polymer is PEI in an amount of from about 0.005% to about 1%.

18. The AM-IPN according to claim 13, wherein the polymer is PVP having a molecular weight of from about 1,000,000 to about 1,300,000.

19. The AM-IPN according to claim 13, wherein the polymer is PVP in an amount of from about 0.005% to about 2%.

20. The AM-IPN according to claim 13, wherein the polymer is PVA having a molecular weight of from about 89,000 to about 98,000.

21. The AM-IPN according to claim 13, wherein the polymer is PVA in an amount of from about 0.005% to about 2%.

22. A method for manufacturing an advanced manufacturing interpenetrating polymer network structure (AM-IPN), comprising: printing one or more of a primary polymer network, a secondary polymer network, and one or more crosslinks using a synthetic bioink and 3D printing technology; and assembling and / or printing the AM-IPN to form an assembled AM-IPN. The manufacturing method comprising the foregoing.

23. The method according to claim 22, wherein the synthetic bioink comprises one or more of polyethylene oxide (PEO), polyethyleneimine (PEI), polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), and combinations thereof.

24. wherein the AM-IPN is printed at about 50 mW / cm ２ to about 400 mW / cm ２ The method according to claim 23, wherein the printing is carried out. Claim 25 The synthetic bioink contains polyethylene oxide (PEO) in an amount of from about 0.005% to about 1%, and the AM-IPN is printed at 50 mW / cm ２ to about 300 mW / cm ２ The method according to claim 24, wherein the printing is carried out at ２ to ２ . Claim 26 The synthetic bioink contains polyethyleneimine (PEI) in an amount of from about 0.005% to about 1%, and the AM-IPN is printed at about 150 mW / cm ２ to about 300 mW / cm ２ The method according to claim 24, wherein the printing is carried out. Claim 27 The synthetic bioink contains polyvinylpyrrolidone (PVP) in an amount of from about 0.005% to about 2%, and the AM-IPN is from about 50 mW / cm ２ to about 300 mW / cm ２ The method according to claim 24, wherein the method is printed at. Claim 28 The synthetic bioink contains polyvinyl alcohol (PVA) in an amount of from about 0.005% to about 2%, and the AM-IPN is from about 100 mW / cm ２ to about 300 mW / cm ２ The method according to claim 24, wherein the method is printed at. Claim 29 The method according to claim 13, wherein the 3D printing technology is one or more of digital light projection printing (DLP), stereolithography (SLA) printing technology, extrusion 3D printing technology, or selective laser sintering 3D printing technology, or a combination thereof. Claim 30 The method according to claim 13, wherein the 3D printing technology is digital light printing (DLP) printing technology. Claim 31 A method of using an advanced manufacturing interpenetrating polymer network structure (AM-IPN), comprising: modifying the surface of the AM-IPN to adhere small airway epithelial cells (SAECs) to form a modified AM-IPN. The method of use comprising the above. Claim 32 The method according to claim 31, further comprising using the modified AM-IPN for perfusion and ventilation.

Images