Devices and methods to determine and / or terminate cardiac arrhythmias

Abstract

The present disclosure generally relates to devices and methods to determine and / or terminate cardiac arrhythmias. In some embodiments, the devices comprise meshes comprising nanoscale wires and / or nanoelectronics. In some case, meshes may be embedded in or placed upon a tissue at a target location within or on a heart. In some embodiments, the meshes may be connectable to an external device (e.g., to determine an electrical property at the target location and / or to apply an electrical stimulus to the target location). Other aspects relate to using the devices and methods disclosed herein for the treatment of cardiac arrythmia, such as atrial fibrillation. Additional non-limiting embodiments are generally directed to Al-driven systems to guide treatment of the arrythmia. For example, some embodiments are generally directed to predictive models that can be used to predict and / or control a pulse sequence of electroactive cells at the target location in the heart (e.g., to detect and correct an aberrant electrogram at the target location by delivering an appropriate pulse sequence).

Description

[0001] DEVICES AND METHODS TO DETERMINE AND / OR TERMINATE CARDIAC ARRHYTHMIAS

[0002] RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Patent Application Serial No. 63 / 681,576, filed August 9, 2024, entitled “Devices and Methods to Determine and / or Terminate Cardiac Arrhythmias,” by Liu, et al., incorporated herein by reference in its entirety.

[0004] FIELD

[0005] The present disclosure generally relates to devices and methods to determine and / or terminate cardiac arrhythmias.

[0006] BACKGROUND

[0007] Atrial fibrillation (“AF”) is the most frequent cardiac arrhythmia and is linked with remarkable mortality and morbidity, which are caused by thromboembolism, heart failure, and impaired cognitive function. The current standard of care includes lifestyle changes, medicines, and procedures to help prevent blood clots, slow the heart beat, and / or restore the hearts normal rhythm. Refractory AF is typically treated with cardiac ablation therapy, which aims to surround the arrhythmic tissue with scar tissue, thus electrically isolating it from the rest of the heart. However, despite medical intervention, AF often returns and may even worsen over time, thus improvements in the treatment of AF are needed.

[0008] SUMMARY

[0009] The subject matter of the present disclosure involves, in some cases, interrelated products, alternative solutions to a particular problem, and / or a plurality of different uses of one or more systems and / or articles.

[0010] Aspects of the disclosure generally relate to methods for treating an arrythmia. In some embodiments, the methods comprise placing a flexible scaffold comprising a nanoelectronic mesh on a tissue at a target location in a heart. In some embodiments, the methods further comprise applying electrical stimuli to portions of the tissue at the target location using one or more of electrodes within the nanoelectronic mesh of the flexible scaffold. In some cases ,the electrical stimuli are generated using a predictive model programmed by electrically stimulating the tissue at the target location using one or more of the electrodes and determining a tissue response based on the electrical stimulation.

[0011] In other embodiments, the methods comprise administering, to the heart of a subject having, or at risk of arrythmia, a device comprising a mesh of nanoelectrodes. In other

[0012] 1

[0013] 12713828.1 embodiments, the methods comprise applying an electrical stimulation to the heart of the subject having or at risk of having arrythmia using the device compromising a mesh of nanoelectrodes.

[0014] In other embodiments, still, the methods comprise determining an electrical signal of the heart of a subject having or at risk of arrythmia using a device comprising a mesh of nanoelectrodes and applying an electrical stimulation to the heart using the mesh of nanoelectrodes. In some cases, the electrical stimulation alters the heartbeat to treat the arrythmia.

[0015] In other embodiments, the methods relate to treating atrial fibrillation, the most common form of cardiac arrythmia. The methods comprise, according to some embodiments, placing a flexible scaffold defining at least a portion of an electrical circuit and comprising a plurality of electrodes on a tissue at one or more pulmonary vein-left atrial junctions in a heart. In some embodiments, the methods further comprise applying electrical stimuli to portions of the tissue at one or more pulmonary vein-left atrial junctions using one or more of the electrodes within the flexible scaffold. In some cases, the electrical stimuli are generated using a predictive model programmed by electrically stimulating the tissue at the target location using one or more of the electrodes and determining a tissue response based on the electrical stimulation.

[0016] Other aspects of the disclosure relate to devices. In some embodiments, the devices comprise a flexible scaffold comprising a nanoelectronic mesh and a catheter for delivering said flexible scaffold. In certain embodiments, the flexible scaffold is disposed within at least a portion of the catheter prior to delivery, and is configured to self-assemble into a mesh structure after release from the catheter. In some embodiments, the catheter is a thoracoscope.

[0017] In other embodiments, the devices comprise a thoracoscope and a device comprising a mesh of nanoelectrodes positionable using the thoracoscope.

[0018] Several methods are disclosed herein of administering a subject with a compound for prevention or treatment of a particular condition. It is to be understood that in each such aspect of the disclosure, the disclosure specifically includes, also, the compound for use in the treatment or prevention of that particular condition, as well as use of the compound for the manufacture of a medicament for the treatment or prevention of that particular condition.

[0019] In another aspect, the present disclosure encompasses methods of making one or more of the embodiments described herein. In still another aspect, the present disclosure encompasses methods of using one or more of the embodiments described herein.

[0020] 2

[0021] 12713828.1 Other advantages and novel features of the present disclosure will become apparent from the following detailed description of various non-limiting embodiments of the disclosure when considered in conjunction with the accompanying figures.

[0022] BRIEF DESCRIPTION OF DRAWINGS

[0023] Non-limiting embodiments of the present disclosure will be described by way of example with reference to the accompanying figures, which are schematic and are not intended to be drawn to scale. In the figures, each identical or nearly identical component illustrated is typically represented by a single numeral. For purposes of clarity, not every component is labeled in every figure, nor is every component of each embodiment of the disclosure shown where illustration is not necessary to allow those of ordinary skill in the art to understand the disclosure. In the figures:

[0024] Fig. 1A-1B illustrates a method of producing a flexible scaffold and characterization of stretchable mesh nanoelectronics in accordance with one embodiment;

[0025] Figs. 2A-2I illustrate electrophysiological mapping of an organoid, in another embodiment;

[0026] Figs. 3A-3B illustrate electrophysiology data for cardiac organoids containing flexible scaffold, in yet another embodiment

[0027] Fig. 4 is a block diagram of an example special purpose computer system improved by the functions and / or processes disclosed herein, in certain embodiments.

[0028] DETAILED DESCRIPTION

[0029] The present disclosure generally relates to devices and methods to determine and / or terminate cardiac arrhythmias. In some embodiments, the devices comprise meshes comprising nanoscale wires and / or nanoelectronics. In some case, meshes may be embedded in or placed upon a tissue at a target location within or on a heart. In some embodiments, the meshes may be connectable to an external device (e.g., to determine an electrical property at the target location and / or to apply an electrical stimulus to the target location). Other aspects relate to using the devices and methods disclosed herein for the treatment of cardiac arrythmia, such as atrial fibrillation. Additional non-limiting embodiments are generally directed to Al-driven systems to guide treatment of the arrythmia. For example, some embodiments are generally directed to predictive models that can be used to predict and / or control a pulse sequence of electroactive cells at the target location in the heart (e.g., to detect and correct an aberrant electrogram at the target location by delivering an appropriate pulse sequence).

[0030] 3

[0031] 12713828.1 Aspects of the present disclosure are directed toward methods of determining and / or treating cardiac arrhythmias in a subject. In some embodiments, the methods comprise placing a device comprising a flexible scaffold comprising a nanoelectronic mesh on a tissue at a target location in or on a heart. Once placed, the device may be used to monitor the electrical activity of the heart (e.g., much like an electrocardiogram). Thus, in some embodiments, the systems and methods comprise monitoring the electrical activity of the heart using the device (e.g., flexible scaffold). Additionally, the systems and methods may further comprise, according to some embodiments, applying electrical stimuli to portions of the tissue at the target location using one or more electrodes within the nanoelectronic mesh of the flexible scaffold of the device. In some cases, the electrical stimuli are generated using Al-driven predictive models programmed, for example, by electrically stimulating the tissue at the target location using one or more of the electrodes and determining a tissue response (e.g., cardiac conduction) based on the electrical stimulation.

[0032] As described in more detail elsewhere herein, a device according to certain embodiments may comprise a flexible scaffold comprising a nanoelectronic mesh comprising at least a portion of an electrical circuit. In some embodiments, at least a portion of the electrical circuit is electrically connected to an electrical device external of the target location. In some embodiments, nanoelectronic mesh is a rectangular mesh or an irregular mesh. In some cases, the nanoelectronic mesh comprises a plurality of nodes, at least some of which are connected by interconnects to from the mesh. The interconnects, in some embodiments, are serpentine, stretchable, and / or comprise at least one inflection point.

[0033] In some embodiments, a flexible scaffold may be placed on a tissue at a target location in a heart. In some instances, the target location is on an epicardial surface (e.g., on the outside of the pericardium). Any suitable technique known to the skilled artisan may be used to place the flexible scaffold on the tissue at the target location. For example, in some embodiments, a throrascopic procedure may be used to place the flexible scaffold around a pulmonary vein-left atrial junction. In some embodiments, multiple flexible scaffolds may be placed at multiple target regions (e.g., multiple flexible scaffolds placed at 4 pulmonary veinleft atrial junctions). In some embodiments, the one or more flexible scaffold is coupled to a device, positioned internally (e.g., within the thoracic cavity) or externally (e.g., subcutaneously) the thoracic cavity. In some embodiments, the target location is selected from the pulmonary veins, coronary sinus, the posterior wall of the heart, or the left atrial appendage.

[0034] 4

[0035] 12713828.1 In other cases, the target location is on an endocardial surface (e.g., on the inside surface of the heart). Any suitable technique known to the skilled artisan may be used to place the flexible scaffold on an endocardial surface of the heart. For example, in some embodiments, a cardiac catheterization procedure may be used to place the flexible scaffold on an endocardial surface of the heart (e.g., left atrium). In some embodiments, multiple flexible scaffolds may be placed at on the endocardial surface of the heart at different locations (e.g., left atrium, right atrium, ventricles, etc.). In some embodiments, the one or more flexible scaffold is coupled to a device, positioned internally (e.g., within the thoracic cavity) or externally (e.g., subcutaneously) the thoracic cavity. In some embodiments, the target location is selected from the pulmonary veins, coronary sinus, the posterior wall of the heart, or the left atrial appendage.

[0036] Without wishing to be bound by any particular theory, it is generally believed that once implanted at a target location a system comprising a flexible scaffold coupled to a device is capable of sensing normal and abnormal cardiac conduction. When the system detects an abnormal event, the system identifies the location of the abnormal signal and applies an appropriate pulse sequence to terminate said signal. For example, in some embodiments, the system is triggered when a microrotor electrogram is detected (e.g., small circuits that initiate atrial fibrillation that propagate to the rest of the atrial tissue). In some embodiments, upon sensing the microrotor, the device determines the location of the abnormal electrogram and computes and delivers an optimal pulse sequence to terminate the microrotor signal. It is believed that application of the appropriate electrical stimuli (e.g., pulse sequence) will place the heart back into normal sinus rhythm.

[0037] In some embodiments, the device comprises one or more computers configured to compute the appropriate pulse sequence to terminate a detected microrotor signal. In some embodiments, the device comprises one or more on-board algorithms configured to determine the optimal pulse sequence (e.g., optimal spatial sequence, temporal sequence, and / or energy) for each flexible scaffold at each target location. The on-board algorithms, in some embodiments, are determined using artificial intelligence and subsequently loaded into the one or more computers. Alternatively, or additionally, in some embodiments, the device is configured to perform the algorithm optimization (e.g., in real time). In some embodiments, the system is configured to learn (e.g., adaptive learning) over time, such that a previously calculated pulse sequence can be used to treat a recurring microrotor electrogram (e.g., the device will not have to re-compute the pulse sequence for previously encountered microrotors).

[0038] 5

[0039] 12713828.1 Additionally, it is herein contemplated that the optimization of the pulse sequence could be done externally and then uploaded to the device (e.g., similar to pacemakers receive firmware updates from external sources), in accordance with some embodiments.

[0040] In some embodiments, the systems and methods comprise administering to the heart of a subject having or at risk of arrythmia a device comprising a mesh of nanoelectrodes. In some embodiments, certain systems and methods comprise applying an electrical stimulation to the heart of a subject having or at risk of arrythmia, e.g., using a device comprising a mesh of nanoelectrodes. Additionally, in some embodiments, the systems and methods comprise determining an electrical signal of the heart of a subject having or at risk of arrythmia using a device comprising a mesh of nanoelectrodes and applying an electrical stimulation to the heart using the mesh of nanoelectrodes, such that upon applying the electrical stimulation alters the heartbeat to treat the arrythmia.

[0041] The systems and methods disclosed herein may be used to treat any type of arrhythmia including but not limited to, atrial fibrillation, atrial flutter, bradycardia, tachycardia, supraventricular tachycardia, ventricular arrhythmias, Wolff-Parkinson- White Syndrome, Ventricular fibrillation, ventricular premature complexes, atrial premature complexes, and sick sinus syndrome.

[0042] In some embodiments, the systems and methods disclosed herein may be used to treat atrial fibrillation. The atrial fibrillation may originate from any one of the four pulmonary veins, coronary sinus, left atrial appendage, or from a posterior wall of the heart. In some cases, the atrial fibrillation is paroxysmal atrial fibrillation (e.g., paroxysms of arial fibrillation). In other cases, however, the atrial fibrillation is persistent atrial fibrillation (e.g., episodes of atrial fibrillation lasting longer than a week). The systems and methods disclosed herein may be used to treat other types of atrial fibrillation as well, including for example, long-term persistent atrial fibrillation (e.g., episodes of abnormal heart rhythms lasting for more than a year) and permanent atrial fibrillation (e.g., atrial fibrillation that does not improve despite intervention).

[0043] Without wishing to be bound by any particular theory, it is also believed that the systems and methods disclosed herein may be useful for preventing strokes in patients on anticoagulation therapy for atrial fibrillation (AF), and may even reduce the need for anticoagulation therapy in patients with AF in some cases.

[0044] The devices and methods disclosed herein may also be useful of other arrhythmic conditions. For example, in some embodiments, the devices and methods disclosed herein are used to treat total electrical standstill (e.g., asystole). In such cases, the devices disclosed

[0045] 6

[0046] 12713828.1 herein may be configured to enable atrial pacing through the flexible scaffold. Additionally, in some embodiments, the devices and methods disclosed here in are used to treat intractable flutter or SVT (e.g., arrhythmias not arising from the pulmonary veins). In such cases, the devices disclosed herein would be configured to permit external control for enable burst pacing. It is contemplated that such configurations that having external control to give signals on command could be helpful to terminate the arrhythmia in certain embodiments.

[0047] Some embodiments are directed to systems and methods relating to treating atrial fibrillation by placing a flexible scaffold, defining at least a portion of an electrical circuit and comprising a plurality of electrodes, on a tissue at one or more pulmonary vein-left atrial junctions in or on a heart. The systems and methods may further comprise applying electrical stimuli to portions of the tissue at the one or more pulmonary vein-left atrial junctions using one or more of the electrodes within the flexible scaffold. In some embodiments, the electrical stimuli are generated, e.g., using a predictive model programmed by electrically stimulating the tissue at the target location using one or more of the electrodes and determining a tissue response based on the electrical stimulation. Additionally, or alternatively, in some embodiments, the systems and methods comprise placing the plurality of electrodes at a coronary sinus, a posterior wall of the heart or a left atrial appendage of the heart, and / or other suitable locations.

[0048] Other aspects of the disclosure relate to devices and methods useful for delivering flexible scaffold comprising a nanoelectronic mesh to a target location in a heart. Accordingly, some embodiments are directed to a flexible scaffold comprising a nanoelectronic mesh and a catheter for delivering said flexible scaffold. In some embodiments, the flexible scaffold is disposed within at least a portion of the catheter prior to delivery and configured to self-assemble into a mesh structure after release from the catheter. In some embodiments, the catheter is a thoracoscope. In certain embodiments, the devices comprise a thoracoscope and a device comprising a mesh of nanoelectrodes positionable using the thoracoscope.

[0049] Various embodiments described herein include a device able to interact with (e.g., electrically stimulate) one or more biological entities, such as cells, tissues, organoids, and / or organs. The device may be or comprise nano wires and / or nanoelectrodes, along with nanosized interconnects that electrically connect various components of the device. However, it should be understood that not every component of the device is of the nano scale and some components may be larger or smaller, as desired, for the fabrication or design of the device. In some embodiments, the device is or comprises a mesh, the mesh having an array of

[0050] 7

[0051] 12713828.1 nanowires. In some such embodiments, the mesh is integrally connected with portions of the biological system (e.g., tissues, organoids) such that the mesh can receive and / or send electrical signals to portions of the biological system. Details regarding electrical signals sent and / or received from the biological system are described elsewhere herein.

[0052] Accordingly, a component within the device may comprise an electrode (e.g., a nanoelectrode). The electrode may comprise any suitable material, for example, carbon, or metals such as gold, platinum, silver, or the like. In some cases, the electrode may be used to determine a property of a biological system (e.g., an electrical property, a chemical property, a mechanical property, etc.), and / or to apply a stimulus (e.g., an electrical stimulus) to the biological system. In some cases, a conductive polymer may also be used with the electrode. Non-limiting examples of conductive polymers include poly (3, 4-ethylenedioxy thiophene) (PEDOT), poly acetylene, polyphenylene vinylene, polypyrrole, poly thiophene (for example poly (3, 4-ethylenedioxy thiophene)), polyphenylene sulfide, or other conductive polymers such as those described herein.

[0053] In some embodiments, a device comprises one or more components such as nanowires, nanoelectrodes and / or interconnects of the nanoscale. In some embodiments, a component has a dimension (e.g., a length, a width, a diameter) of less than or equal to 1,000 nm, less than or equal to 500 nm, less than or equal to 250 nm, less than or equal to 100 nm, less than or equal to 75 nm, less than or equal to 50 nm, less than or equal to 25 nm, or less than or equal to 10 nm. In some embodiments, a component has a dimension of greater than or equal to 10 nm, greater than or equal to 25 nm, greater than or equal to 50 nm, greater than or equal to 75 nm, greater than or equal to 100 nm, greater than or equal to 250 nm, greater than or equal to 500 nm, or greater than or equal to 1,000 nm. Combinations of the abovereferenced ranges are also possible (e.g., greater than or equal to 10 nm and less than or equal to 1,000 nm). Other ranges are possible.

[0054] In some embodiments, a nanowire or nanoelectrode may be used to assist in determining a property of a biological system, e.g., when it is embedded within and / or placed on a biological structure (for example, the heart). For example, one or more locations within the biological system (for example, an electrode or a nanoscale wire, etc.) may be determined to determine a property, such as a chemical property, an electrical property, a mechanical property, or the like. Other examples include sensing Ca2+spikes, voltage changes, cell signaling pathways, ion concentrations, pH changes, sensing of biomolecules or reaction entities, etc. In some cases, the locations are defined as one or more nodes within the biological system, some or all of which may be individually addressable. For example, a

[0055] 8

[0056] 12713828.1 node within a flexible scaffold may comprise a nanoscale wire, such as those described in more detail elsewhere herein.

[0057] Devices (e.g., meshes, nanowire arrays) described herein may be integrally a part of biological systems. Accordingly, devices described herein may have a degree of flexibility and / or stretchability to move with the biological system during motion of biological system while not damaging components of the device (e.g., nanowires, interconnects).

[0058] Accordingly, one or more materials within device are stretchable and / or flexible. For example, in some cases, the device may comprise a mesh or portions thereof (e.g., interconnects) that can be stretchable and / or flexible, or can be manipulated or distorted in some fashion. It should be understood that the flexibility of a material is not purely an intrinsic material propriety; a thinner piece of material may offer more flexibility than a comparably thicker piece of the same material. In addition, in some cases, the flexibility of the material may also be a function of its shape, e.g., as described elsewhere herein.

[0059] Some embodiments are generally directed to flexible or stretchable devices, such as meshes or networks, that can be used as flexible scaffolds to grow structures, such as organoids, or tissues. In some cases, these may be included in an implant, e.g., for the heart. In general, flexible scaffolds are structures that cells can attach to and grow on, e.g., to form biological tissues, organoids, and other biological structures. For example, the biological structure may be the heart of an organism. The flexible scaffold may comprise biocompatible and / or biodegradable materials, and may in some embodiments also contain growth factors such as growth hormones, extracellular matrix proteins, specific metabolites or nutrients, or the like. The flexible scaffold typically is porous, e.g., to facilitate cell seeding therein, and / or diffusion into and out of the flexible scaffold, for example, of nutrients, waste products, etc.

[0060] In some embodiments, a device, or a component of the device (e.g., a nanowire, a mesh), is flexible and / or stretchable such that the device, or the component of the device, has an effective bending stiffness of greater than or equal to 0.01 n-Nm, greater than or equal to 0.02 n-Nm, greater than or equal to 0.03 n-Nm, greater than or equal to 0.05 n-Nm, greater than or equal to 0.1 n-Nm, greater than or equal to 0.3 n-Nm, greater than or equal to 0.5 n-Nm, greater than or equal to 1 n-Nm, greater than or equal to 2 n-Nm, greater than or equal to 3 n-Nm, or greater than or equal to 5 n-Nm. In some embodiments, the device, or a component of the device, has an effective bending stiffness of less than or equal to 5 n-Nm, less than or equal to 3 n-Nm, less than or equal to 2 n-Nm, less than or equal to 1 n-Nm, less than or equal to 0.5 n-Nm, less than or equal to 0.3 n-Nm, less than or equal to 0.1 n-Nm, less than or equal to 0.05, less than or equal to 0.03 n-Nm, less than or equal to 0.02 n-Nm, or less

[0061] 9

[0062] 12713828.1 than or equal to 0.01 n-Nm. Combinations of foregoing ranges are also possible (e.g., greater than or equal to 0.01 n-Nm and less than or equal to n-Nm). Other ranges are possible as this disclosure is not so limited.

[0063] In one set of embodiments, the device (e.g., nanowires, a mesh) may have a shape and / or may be formed from one or more materials that allow the device to be flexible and / or stretchable. For example, the scaffold may be formed of shapes, such as serpentine shapes, that can be extended. In some cases, the device can be formed of components that are not straight, and can be extended, e.g., when pulled on. For instance, the flexible scaffold may comprise one or more nodes that are connected by various interconnects, e.g., forming a mesh or a network. The nodes may be evenly or nonevenly distributed within the flexible scaffold, and the interconnects may connect them in a regular pattern (for example, in rectangular or triangular arrays of nodes), or in an irregular pattern. The nodes may represent points of connectivity, or there may be one or more electronic components at some or all of the nodes, such as conductive pathways, nanoscale wires, sensors, or the like. The same or different electronic components may independently be present at different nodes within a mesh or network.

[0064] Interconnects connecting two (or more) nodes together may have the same or different shapes or structure within a device (e.g., a mesh or network), and different interconnects within the device may independently have the same or different shapes. In some cases, an interconnect may have a shape that is extendible. For example, an interconnect may have a straight-line or linear shape, or have shapes that are non-linear, such as S shapes, serpentine shapes (e.g., having two, three, four, or more bends or inflection points), zigzag shapes (e.g., having two, three, four, or more vertices), coiled shapes, or the like. Such interconnect shapes may allow various manipulations to occur without disrupting the connection of the interconnect to the nodes, e.g., during stretching, compression, folding, etc.

[0065] In one set of embodiments, an interconnect (or some other component of the device) may comprise one or metal leads and one or more polymers, such as those discussed below. The polymers can include photoresist polymers (such as SU-8), and / or biocompatible polymers (such as Matrigel™). Other examples of photoresist polymers include, but are not limited to, those described.

[0066] In addition, in some embodiments, the device may have components, such as interconnects, that are sufficiently flexible or stretchable such that the device (or a component thereof, such as an interconnect) is foldable by at least 30°, at least 45°, at least 90°, at least 135°, at least 150°, at least 180°, etc. from an initial planar structure.

[0067] 10

[0068] 12713828.1 In some instances, a device may have components, such as interconnects, that are sufficiently flexible or stretchable such that the device (or a component thereof, such as an interconnect) may be stretchable in a linear direction by at least 10%, at least 20%, at least 30%, at least 50%, at least 75%, at least 100%, at least 150%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 450%, at least 500%, etc., for example, before catastrophic failure of the flexible scaffold, breakage, disruption of the connection of the interconnect to the nodes, loss of electrical connections, or the like.

[0069] In addition, in certain cases, the flexible scaffold may also exhibit some degree of elasticity, e.g., such that the flexible scaffold may return (at least partially) to its original structure prior to stretching. For instance, the device (or a component thereof, such as an interconnect) may return at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100% (perfectly elastic) back to its original structure, measured from when stretching of the material is stopped. For example, a 1 cm material stretched to 2 cm experiences a 100% stretch in a linear direction, and if it afterwards contracts to 1.5 cm, it exhibits a 50% recovery to its original structure (returning 0.5 cm from its stretch of 1 cm). However, it should be understood that in some embodiments, the device is not elastic.

[0070] In some cases, the device may have an overall filling ratio or area of less than or equal to 50%, less than or equal to 40%, less than or equal to 30%, less than or equal to 25%, less than or equal to 20%, less than or equal to 15%, less than or equal to 13%, less than or equal to 12%, less than or equal to 11%, less than or equal to 10%, less than or equal to 9%, less than or equal to 8%, less than or equal to 7%, less than or equal to 6%, less than or equal to 5%, etc. The filling ratio or area is the area of the physical components of the device, compared to the overall area of the device or biological system (including void spaces). Thus, this is a measure of the “porosity” in two dimensions. For example, in some cases, the device or the biological system may have a mesh structure or layout as described above, where the mesh is relatively open. Devices with smaller filling ratios thus would have greater “open space,” for example, to allow cells to penetrate.

[0071] As mentioned, in some cases, the device or biological system can be defined by one or more pores. Pores that are too small can hinder or restrict cell access. Thus, in some embodiments, the device or the biological system may have an average pore size of at least 100 micrometers, at least 200 micrometers, at least 300 micrometers, at least 400 micrometers, at least 500 micrometers, at least 600 micrometers, at least 700 micrometers, at least 800 micrometers, at least 900 micrometers, or at least 1 mm. However, in other

[0072] 11

[0073] 12713828.1 embodiments, pores that are too big may prevent cells from being able to satisfactorily use or even access the pore volume. Thus, in some cases, the device may have an average pore size of no more than 1.5 mm, no more than 1.4 mm, no more than 1.3 mm, no more than 1.2 mm, no more than 1.1 mm, no more than 1 mm, no more than 900 micrometers, no more than 800 micrometers, no more than 700 micrometers, no more than 600 micrometers, or no more than 500 micrometers. Combinations of these are also possible, e.g., in one embodiment, the average pore size is at least 100 micrometers and no more than 1.5 mm. In addition, larger or smaller pores than these can also be used in a flexible scaffold in certain cases. Pore sizes may be determined using any suitable technique, e.g., through BET measurements.

[0074] Devices (e.g., nanowires, meshes) described herein may comprise a variety of materials in different embodiments. For example, the device may comprise one or more polymers, such as photoresists, that define interconnects or other components within the device. In some cases, one or more portions of the device may comprise components, such as nanoelectric components, that may form electrical circuits within the device. For example, the device may contain metal or other conductive pathways, e.g., which define an electrical circuit, and / or can be connected to an external electrical device.

[0075] In one set of embodiments, the device may contain metal or other conductive pathways, e.g., within interconnects or nodes within the device (e.g., a flexible scaffold). Examples of metals for metal leads or pathways that can be used include, but are not limited to platinum, aluminum, gold, silver, copper, molybdenum, tantalum, titanium, nickel, tungsten, chromium, palladium, or the like, as well as any combinations of these and / or other metals. Other examples include conductive polymers such as poly (3,4- ethylenedioxythiophene) (PEDOT), polyacetylene, polyphenylene vinylene, polypyrrole, poly thiophene (for example poly (3, 4-ethylenedioxy thiophene)), polyphenylene sulfide, etc.

[0076] In certain embodiments, the device includes one or more polymers, e.g., photoresists, biocompatible polymers, biodegradable polymers, etc., as is described herein. For example, in various embodiments, one or more of the polymers may be a photoresist. Photoresists are typically used in lithographic techniques, which can be used as described herein. For example, the photoresist may be chosen for its ability to react to light to become substantially insoluble (or substantially soluble, in some cases) to a photoresist developer.

[0077] Photoresists that can be used include, but are not limited to, SU-8, SI 805, FOR 3A, poly(methyl methacrylate), poly(methyl glutarimide), phenol formaldehyde resin (diazonaphthoquinone / novolac), diazonaphthoquinone (DNQ), Hoechst AZ 4620, Hoechst

[0078] 12

[0079] 12713828.1 AZ 4562, Shipley 1400-17, Shipley 1400-27, Shipley 1400-37, or the like. These and many other photoresists are available commercially. Other examples of photoresist polymers include, but are not limited to, those described below, and those described in Int. Pat. Apl. Pub. No. WO 2019 / 084498, incorporated herein by reference. In some cases, the photoresist may be a soft material, for example, a hydrogel. In some embodiments, the photoresist comprises a polymer formed by photo-curing a fluorinated monomer including cross -linkable function groups using a photoinitiator. One example of such a polymer is perfluoropolyether dimethacrylate (PFPE-DMA). Other examples are discussed in more detail below.

[0080] In some cases, one or more of the polymers may be biocompatible and / or biodegradable. Examples of such biocompatible and / or biodegradable polymers include, but are not limited to, poly(lactic-co-glycolic acid), polylactic acid, polyglycolic acid, poly(methyl methacrylate), poly(trimethylene carbonate), collagen, fibrin, polysaccharidic materials such as chitosan or glycosaminoglycans, hyaluronic acid, polycaprolactone, and the like. Certain photoresists are also biocompatible and / or biodegradable in some cases.

[0081] Typically, a biocompatible material is one that does not illicit an immune response, or elicits a relatively low immune response, e.g., one that does not impair the flexible scaffold or the cells therein from continuing to function for its intended use. In some embodiments, the biocompatible material is able to perform its desired function without eliciting any undesirable local or systemic effects in a subject, e.g., when present within a subject. In some cases, the material is present without eliciting any undesirable local or systemic effects, or such that any biological response by the subject does not substantially affect the ability of the material from continuing to function for its intended use. For example, in a device that is a flexible scaffold, the flexible scaffold may be able to support appropriate cellular or tissue activity when implanted within a subject, e.g., including the facilitation of molecular and / or mechanical signaling systems, without substantially eliciting undesirable effects in those cells, or undesirable local or systemic responses, or without eliciting a response that causes the flexible scaffold to cease functioning for its intended use.

[0082] A biodegradable material typically degrades over time when exposed to a biological system, e.g., through oxidation, hydrolysis, enzymatic attack, phagocytosis, or the like. For example, a biodegradable material can degrade over time when exposed to water (e.g., hydrolysis) or enzymes. In some cases, a biodegradable material is one that exhibits degradation (e.g., loss of mass and / or structure) when exposed to physiological conditions for at least about a month, at least about 6 months, or at least about a year. For example, the biodegradable material may exhibit a loss of mass of at least 10%, at least 20%, at least 30%,

[0083] 13

[0084] 12713828.1 at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%. In certain cases, some or all of the degradation products may be resorbed or metabolized, e.g., into cells or tissues. For example, certain biodegradable materials, during degradation, release substances that can be metabolized by cells or tissues.

[0085] For many embodiments describes herein, the device comprises a stimulator. A stimulator provides an electrical signal to a portion of a biological system (e.g., a cell, a portion of tissue, an organoid). In some embodiments, the stimulator is a nanowire or nanoelectrode of the device configured to provide electrical current. Details describing nanowires and nanoelectrodes are provided elsewhere herein.

[0086] In some embodiments, a device includes a recording sensor. The recording sensor may comprise one or more nanowires or nanoelectrodes and is configured to receive an electrical signal (e.g., an electrical impedance). In some embodiments, the recording sensor is operatively associated with a computer-readable medium, and the compute readable-medium is configured to use the received electrical signal as data for a machine-learning and / or an Al process. As a non-limiting example, electrical signals received by the recording sensor can be used to weight how a subsequent signal should be provided to the biological system, e.g., to the heart. Details regarding machine learning and / or Al processes (e.g., predictive models) are provided elsewhere herein.

[0087] As mentioned above and elsewhere herein, a device (e.g., a stimulator of the device, a recording sensor of the device) may be configured to receive and / or transmit electrical signals within the biological system (e.g., the heart), in accordance with some embodiments. For example, in some embodiments, a first current is applied to the heart and / or a second signal is applied to the heart. In some embodiments, a first signal and / or a second signal is received by the recording signal, for example, as an impedance signal. Of course, additional signals (e.g., a third signal, a fourth signal, a fifth signal, and so forth) may be transmitted and / or received.

[0088] In some embodiments, a signal received and or transmitted may have a particular peak shape. That is to say, when a signal is transmitted and / or received, an intensity of the signal may vary over time such that a distribution of values is received and / or transmitted. In some such embodiments, a peak shape has a relatively Gaussian distribution. Of course, other distributions are possible, as this disclosure is not so limited.

[0089] Devices (e.g., a mesh comprises nanowires and / or interconnects) described herein may be a part of a biological system. The biological system may comprise a cell, cells, tissue, and / or an organ (e.g., the heart). As described elsewhere herein, the device may be integrated into the biological system and may also move or flex with the biological system. In some

[0090] 14

[0091] 12713828.1 embodiments, the device is a part of an in vivo biological system. However, in other embodiments, the device is a part of in vitro biological system.

[0092] In addition, certain embodiments are generally related to nanoscale wires and nanoelectronics, which, in some aspects, may be embedded in biological systems, such as cells, tissues, organoids, organs, organisms, and the like. In one embodiment, the biological structure is a heart of a subject.

[0093] Certain aspects described herein are directed to systems and methods for preparing flexible scaffolds. The flexible scaffolds can be fabricated, for example, using well-known lithographic techniques such as those discussed below.

[0094] In various embodiments, a flexible scaffold is constructed by assembling various polymers, metals, and other components (for example, nanoscale wires) together on a substrate. For example, lithographic techniques such as e-beam lithography, photolithography, X-ray lithography, extreme ultraviolet lithography, ion projection lithography, etc. may be used to pattern polymers, metals, etc. on the substrate. After assembly, at least a portion of the substrate (e.g., a sacrificial material) may be removed, allowing the scaffold to be partially or completely removed from the substrate. Other materials may also be added to the scaffold, e.g., to help stabilize the structure, to add additional agents to enhance its biocompatibility, etc. The scaffold can be used in vivo, e.g., by implanting it in a subject, and / or in vitro, e.g., by seeding cells, etc. on the scaffold. In addition, in some cases, cells may initially be grown or cultured on the scaffold, e.g., to form a biological structure, such as tissues, organoids, organs, organisms, and the like. In some cases, as discussed, the flexible scaffold may be sufficiently flexible such that the flexible scaffold becomes embedded within the biological structures as it forms.

[0095] For example, in one set of embodiments, a flexible scaffold may be constructed by providing a substrate, depositing a sacrificial layer on the substrate, then patterning a first photoresist on the sacrificial layer, a conductive pathway on the first photoresist, and a second photoresist on the conductive pathway, and removing the sacrificial layer to produce the flexible scaffold. See, e.g., Fig. 1. The first and second photoresists may comprise the same or different materials. Optionally, other components can also be added to the flexible scaffold, before or during formation, such as electrode components, nanoscale wires, connectors such as cables, or the like.

[0096] The substrate may be chosen to be one that can be used for lithographic techniques such as e-beam lithography or photolithography, or other lithographic techniques including those discussed herein. For example, the substrate may comprise or consist essentially of a

[0097] 15

[0098] 12713828.1 semiconductor material such as silicon, although other substrate materials (e.g., a metal) can also be used. Typically, the substrate is one that is substantially planar, e.g., so that polymers, metals, and the like can be patterned on the substrate. In some cases, a portion of the substrate can be oxidized, e.g., forming SiCh and / or ShN4 on a portion of the substrate, which may facilitate subsequent addition of materials (metals, polymers, etc.) to the substrate.

[0099] In certain embodiments, one or more polymers can also be deposited or otherwise formed prior to depositing the sacrificial material. In some cases, the polymers may be deposited or otherwise formed as a layer of material on the substrate. Deposition may be performed using any suitable technique, e.g., using lithographic techniques such as e-beam lithography, photolithography, X-ray lithography, extreme ultraviolet lithography, ion projection lithography, etc. In some cases, some or all of the polymers may be biocompatible and / or biodegradable. The polymers that are deposited may also comprise methyl methacrylate and / or poly (methyl methacrylate), in some embodiments.

[0100] Next, a sacrificial material may be deposited. The sacrificial material can be chosen to be one that can be removed without substantially altering other materials (e.g., polymers, other metals, nanoscale wires, etc.) deposited thereon. For example, in one embodiment, the sacrificial material may be a metal, e.g., one that is easily etchable. For instance, the sacrificial material can comprise germanium or nickel, which can be etched or otherwise removed, for example, using a peroxide (e.g., H2O2) or a nickel etchant (many of which are readily available commercially). In some cases, the sacrificial material may be deposited on oxidized portions or polymers previously deposited on the substrate. In some cases, the sacrificial material is deposited as a layer. The layer can have a thickness of less than about 5 micrometers, less than about 4 micrometers, less than about 3 micrometers, less than about 2 micrometers, less than about 1 micrometer, less than about 900 nm, less than about 800 nm, less than about 700 nm, less than about 600 nm, less than about 500 nm, less than about 400 nm, less than about 300 nm, less than about 200 nm, less than about 100 nm, etc.

[0101] In some embodiments, a first photoresist can be deposited, e.g., on the sacrificial material. The photoresist may include one or more polymers, which may be deposited as one or more layers. Examples of photoresist include, but are not limited to, SU-8, SI 805, LOR 3A, poly(methyl methacrylate), poly(methyl glutarimide), phenol formaldehyde resin (diazonaphthoquinone / novolac), diazonaphthoquinone (DNQ), Hoechst AZ 4620, Hoechst AZ 4562, Shipley 1400-17, Shipley 1400-27, Shipley 1400-37, etc., as well as any others discussed herein.

[0102] 16

[0103] 12713828.1 The photoresist can be used to at least partially define a flexible scaffold. In one set of embodiments, the photoresist may be deposited as a layer of material, such that portions of the photoresist may be subsequently removed. For example, the photoresist can be deposited using lithographic techniques such as e-beam lithography, photolithography, X-ray lithography, extreme ultraviolet lithography, ion projection lithography, etc., or using other techniques for removing polymer that are known to those of ordinary skill in the art. In some cases, more than one photoresist is used, e.g., deposited as more than one layer (e.g., sequentially), and each layer may independently have a thickness of less than about 5 micrometers, less than about 4 micrometers, less than about 3 micrometers, less than about 2 micrometers, less than about 1 micrometer, less than about 900 nm, less than about 800 nm, less than about 700 nm, less than about 600 nm, less than about 500 nm, less than about 400 nm, less than about 300 nm, less than about 200 nm, less than about 100 nm, etc. For example, in some embodiments, portions of the photoresist may be exposed to light (visible, UV, etc.), electrons, ions, X-rays, etc. (e.g., projected onto the photoresist), and the exposed portions can be etched away (e.g., using suitable etchants, plasma, etc.) to produce the pattern.

[0104] Accordingly, the photoresist may be formed into a particular pattern, e.g., in a grid or a mesh, e.g., as discussed herein. For instance, the pattern may include a mesh and interconnects that have a shape that allow the interconnects to be manipulated or distorted without disrupting their connections, e.g., during stretching, compression, folding, or the like. The pattern can be regular or irregular.

[0105] Next, a metal or other conductive material can be deposited e.g., on one of the previous materials, to form conductive pathways within the flexible scaffold. More than one metal can be used, which may be deposited as one or more layers. For example, a first metal may be deposited, and a second metal may be deposited on at least a portion of the first metal. Optionally, more metals can be used, e.g., a third metal may be deposited on at least a portion of the second metal, and the third metal may be the same or different from the first metal. In some cases, each metal may independently have a thickness of less than about 5 micrometers, less than about 4 micrometers, less than about 3 micrometers, less than about 2 micrometers, less than about 1 micrometer, less than about 900 nm, less than about 800 nm, less than about 700 nm, less than about 600 nm, less than about 500 nm, less than about 400 nm, less than about 300 nm, less than about 200 nm, less than about 100 nm, less than about 80 nm, less than about 60 nm, less than about 40 nm, less than about 30 nm, less than about

[0106] 17

[0107] 12713828.1 20 nm, less than about 10 nm, less than about 8 nm, less than about 6 nm, less than about 4 nm, or less than about 2 nm, etc., and the layers may be of the same or different thicknesses.

[0108] Any suitable technique can be used for depositing metals, and if more than one metal is used, the techniques for depositing each of the metals may independently be the same or different. For example, in one set of embodiments, deposition techniques such as sputtering can be used. Other examples include, but are not limited to, physical vapor deposition, vacuum deposition, chemical vapor deposition, cathodic arc deposition, evaporative deposition, e-beam PVD, pulsed laser deposition, ion-beam sputtering, reactive sputtering, ion-assisted deposition, high-target-utilization sputtering, high-power impulse magnetron sputtering, gas flow sputtering, or the like.

[0109] The metals can be chosen in some cases such that the deposition process yields a prestressed arrangement, e.g., due to atomic lattice mismatch, which causes the subsequent metal leads to warp or bend, for example, once released from the substrate. Although such processes were typically undesired in the prior art, in certain embodiments of the present disclosure, such pre-stressed arrangements may be used to cause the resulting flexible scaffold to form a 3-dimensional structure, in some cases spontaneously, upon release from the substrate. See, e.g., U.S. Pat. Apl. Pub. Nos. 2014 / 0073063, 2014 / 0074253, 2017 / 0069858, 2017 / 0072109, each of which is incorporated herein by reference in its entirety. However, it should be understood that in other embodiments, the metals may not necessary be deposited in a pre-stressed arrangement.

[0110] Examples of metals that can be deposited (stressed or unstressed) include, but are not limited to, aluminum, gold, silver, copper, molybdenum, tantalum, titanium, nickel, tungsten, chromium, palladium, as well as any combinations of these and / or other metals. For example, a chromium / gold / chromium deposition process can be used, as is shown in Fig. 1.

[0111] In certain embodiments, a second photoresist can be deposited on the previous materials. The second photoresist may be the same or different from the first photoresist, and may include any of the photoresist materials discussed herein, including any of those described with reference to the first photoresist. The second photoresist may include one or more polymers, which may be deposited as one or more layers. In some embodiments, the second photoresist may be deposited on one or more portions of a substrate, e.g., as a layer of material such that portions of the second photoresist can be subsequently removed, e.g., using lithographic techniques such as e-beam lithography, photolithography, X-ray lithography, extreme ultraviolet lithography, ion projection lithography, etc., or using other techniques for removing photoresist that are known to those of ordinary skill in the art. In some cases, more

[0112] 18

[0113] 12713828.1 than one photoresist may be used, e.g., deposited as more than one layer (e.g., sequentially), and each layer may independently have a thickness of less than about 5 micrometers, less than about 4 micrometers, less than about 3 micrometers, less than about 2 micrometers, less than about 1 micrometer, less than about 900 nm, less than about 800 nm, less than about 700 nm, less than about 600 nm, less than about 500 nm, less than about 400 nm, less than about 300 nm, less than about 200 nm, less than about 100 nm, etc.

[0114] After formation of the flexible scaffold, some or all of the sacrificial material may then be removed in some cases. In one set of embodiments, for example, at least a portion of the sacrificial material is exposed to an etchant able to remove the sacrificial material. For example, if the sacrificial material is a metal such as nickel, a suitable etchant (for example, a metal etchant such as a nickel etchant, acetone, etc.) can be used to remove the sacrificial metal. Many such etchants may be readily obtained commercially. In addition, in some embodiments, the flexible scaffold can also be dried, e.g., in air (e.g., passively), by using a heat source, by using a critical point dryer, etc.

[0115] Other materials may be also added to the flexible scaffold, e.g., before or after it forms a 3-dimensional structure, for example, to help stabilize the structure, to add additional agents to enhance its biocompatibility (e.g., growth hormones, extracellular matrix protein, Matrigel™, etc.), to cause it to form a suitable 3-dimension structure, to control pore sizes, etc. Non-limiting examples of such materials have been previously discussed above, and include other polymers, growth hormones, extracellular matrix protein, specific metabolites or nutrients, additional scaffold materials, or the like.

[0116] In addition, in some cases, the flexible scaffold is exposed to cells, which can be cultured or allowed to grow, e.g., to form a biological structure. In some cases, the cells are plated or seeded as individual cells, although in certain cases, larger cell assemblies (tissues, embryos, etc.) may be used. In one set embodiments, the flexible scaffold may be exposed to cells in vitro, and / or the flexible scaffold may be exposed or even submerged within a suitable cell growth medium. Such media are widely available commercially. In some embodiments, the flexible scaffold can be subsequently implanted in vivo into a subject, e.g., upon the growth of tissue, an organ, an organoid, etc. However, it should be understood that implantation is not required in all embodiments, for example, in cases where an entire organism develops from the cells.

[0117] In addition, it should be understood that exposure to cells is not necessarily required in all embodiments. For instance, in one set of embodiments, the flexible scaffold may be

[0118] 19

[0119] 12713828.1 prepared without the presence of cells. For example, the flexible scaffold may be sold as part of a kit, and the user may expose the flexible scaffold to cells (or use it for other purposes).

[0120] In addition, the flexible scaffold can be interfaced in some embodiments with one or more electronics, e.g., an external electrical system such as a computer or a transmitter (for instance, a radio transmitter, a wireless transmitter, etc.), e.g., as discussed herein. The interfacing may occur at any suitable time, e.g., before or after exposure to cells, before or after a biological structure (e.g., an organoid or an organism) has formed, before or after sale to a user, or the like.

[0121] For instance, in some cases, electronic testing of the flexible scaffold may be performed. The flexible scaffold, or a portion thereof, can be connected to an external electrical circuit, e.g., to electronically interrogate or otherwise determine the electronic state of the flexible scaffold. For example, the flexible scaffold may comprise one or more nanoscale wires, or other nanoelectronic components, that can be used as sensors. Such determinations may be performed quantitatively and / or qualitatively, depending on the application, and can involve all, or only a portion, of the flexible scaffold, e.g., as discussed herein.

[0122] Thus, as mentioned, in some aspects, the flexible scaffold can comprise one or more nanoscale wires. For instance, one or more nodes may contain nanoscale wires, and / or nanoscale wires may be contained within interconnects, or the like. In some cases, the flexible scaffold within the organoids, organs, or organisms may include one or more sensors or stimulators, interconnected with stretchable mesh interconnects, to form a network, e.g., as is shown in Figs. 7 and 8. The sensors or stimulators may, in some embodiments, comprise nanoscale wires, such as those described herein. Such sensors may be monitored, e.g., individually or collectively.

[0123] Non-limiting examples of suitable nanoscale wires include carbon nanotubes, nanorods, nanowires, organic and inorganic conductive and semiconducting polymers, metal nanoscale wires, semiconductor nanoscale wires (for example, formed from silicon), and the like. If carbon nanotubes are used, they may be single-walled and / or multi-walled, and may be metallic and / or semiconducting in nature. Other conductive or semiconducting elements that may not be nanoscale wires, but are of various small nanoscopic-scale dimension, also can be used within the flexible scaffold.

[0124] In general, a “nanoscale wire” (also known herein as a “nanoscopic-scale wire” or “nanoscopic wire”) generally is a wire or other nanoscale object, that at any point along its length, has at least one cross-sectional dimension and, in some embodiments, two orthogonal

[0125] 20

[0126] 12713828.1 cross-sectional dimensions (e.g., a diameter) of less than 1 micrometer, less than about 500 nm, less than about 200 nm, less than about 150 nm, less than about 100 nm, less than about 70, less than about 50 nm, less than about 20 nm, less than about 10 nm, less than about 5 nm, than about 2 nm, or less than about 1 nm. In some embodiments, the nanoscale wire is generally cylindrical. In other embodiments, however, other shapes are possible; for example, the nanoscale wire can be faceted, i.e., the nanoscale wire may have a polygonal cross-section. The cross-section of a nanoscale wire can be of any arbitrary shape, including, but not limited to, circular, square, rectangular, annular, polygonal, or elliptical, and may be a regular or an irregular shape. The nanoscale wire can also be solid or hollow.

[0127] In some cases, the nanoscale wire has one dimension that is substantially longer than the other dimensions of the nanoscale wire. For example, the nanoscale wire may have a longest dimension that is at least about 1 micrometer, at least about 3 micrometers, at least about 5 micrometers, or at least about 10 micrometers or about 20 micrometers in length, and / or the nanoscale wire may have an aspect ratio (longest dimension to shortest orthogonal dimension) of greater than about 2:1, greater than about 3:1, greater than about 4:1, greater than about 5:1, greater than about 10:1, greater than about 25:1, greater than about 50:1, greater than about 75:1, greater than about 100:1, greater than about 150:1, greater than about 250:1, greater than about 500:1, greater than about 750:1, or greater than about 1000:1 or more in some cases.

[0128] In some embodiments, a nanoscale wire is substantially uniform, or the nanowire may have a variation in average diameter of the nanoscale wire of less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, or less than about 5%. For example, the nanoscale wires may be grown from substantially uniform nanoclusters or particles, e.g., colloid particles. See, e.g., U.S. Patent No. 7,301,199, issued November 27, 2007, entitled “Nanoscale Wires and Related Devices,” by Lieber, et al., incorporated herein by reference in its entirety. In some cases, the nanoscale wire may be one of a population of nanoscale wires having an average variation in diameter, of the population of nanowires, of less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, or less than about 5%.

[0129] In some embodiments, a nanoscale wire has a conductivity of or of similar magnitude to any semiconductor or any metal. The nanoscale wire can be formed of suitable materials, e.g., semiconductors, metals, etc., as well as any suitable combinations thereof. In some cases, the nanoscale wire will have the ability to pass electrical charge, for example, being electrically conductive. For example, the nanoscale wire may have a relatively low

[0130] 21

[0131] 12713828.1 resistivity, e.g., less than about 10'3Ohm m, less than about 10'4Ohm m, less than about 10'6Ohm m, or less than about 10'7Ohm m. The nanoscale wire can, in some embodiments, have a conductance of at least about 1 microsiemens, at least about 3 microsiemens, at least about 10 microsiemens, at least about 30 microsiemens, or at least about 100 microsiemens.

[0132] The nanoscale wire can be solid or hollow, in various embodiments. As used herein, a “nanotube” is a nanoscale wire that is hollow, or that has a hollowed-out core, including those nanotubes known to those of ordinary skill in the art. As another example, a nanotube may be created by creating a core / shell nanowire, then etching away at least a portion of the core to leave behind a hollow shell. Accordingly, in one set of embodiments, the nanoscale wire is a non-carbon nanotube. In contrast, a “nanowire” is a nanoscale wire that is typically solid (i.e., not hollow). Thus, in one set of embodiments, the nanoscale wire may be a semiconductor nanowire, such as a silicon nanowire.

[0133] For example, in one embodiment, a nanoscale wire may comprise or consist essentially of a metal. Non-limiting examples of potentially suitable metals include aluminum, gold, silver, copper, molybdenum, tantalum, titanium, nickel, tungsten, chromium, or palladium. In another set of embodiments, a nanoscale wire comprises or consists essentially of a semiconductor. Typically, a semiconductor is an element having semiconductive or semi-metallic properties (i.e., between metallic and non-metallic properties). An example of a semiconductor is silicon. Other non-limiting examples include elemental semiconductors, such as gallium, germanium, diamond (carbon), tin, selenium, tellurium, boron, or phosphorous. In other embodiments, more than one element may be present in the nanoscale wire as the semiconductor, for example, gallium arsenide, gallium nitride, indium phosphide, cadmium selenide, etc. Still other examples include a Group II- VI material (which includes at least one member from Group II of the Periodic Table and at least one member from Group VI, for example, ZnS, ZnSe, ZnSSe, ZnCdS, CdS, or CdSe), or a Group III-V material (which includes at least one member from Group III and at least one member from Group V, for example GaAs, GaP, GaAsP, InAs, InP, AlGaAs, or InAsP).

[0134] In certain embodiments, the semiconductor can be undoped or doped (e.g., p-type or n-type). For example, in one set of embodiments, a nanoscale wire may be a p-type semiconductor nanoscale wire or an n-typc semiconductor nanoscale wire, and can be used as a component of a transistor such as a field effect transistor (“FET”). For instance, the nanoscale wire may act as the “gate” of a source-gate-drain arrangement of a FET, while metal leads or other conductive pathways (as discussed herein) are used as the source and drain electrodes.

[0135] 22

[0136] 12713828.1 In some embodiments, a dopant or a semiconductor may include mixtures of Group IV elements, for example, a mixture of silicon and carbon, or a mixture of silicon and germanium. In other embodiments, the dopant or the semiconductor may include a mixture of a Group III and a Group V element, for example, BN, BP, BAs, AIN, A1P, AlAs, AlSb, GaN, GaP, GaAs, GaSb, InN, InP, InAs, or InSb. Mixtures of these may also be used, for example, a mixture of BN / BP / BAs, or BN / A1P. In other embodiments, the dopants may include alloys of Group III and Group V elements. For example, the alloys may include a mixture of AlGaN, GaPAs, InPAs, GalnN, AlGalnN, GalnAsP, or the like. In other embodiments, the dopants may also include a mixture of Group II and Group VI semiconductors. For example, the semiconductor may include ZnO, ZnS, ZnSe, ZnTe, CdS, CdSe, CdTe, HgS, HgSe, HgTe, BeS, BeSe, BeTe, MgS, MgSe, or the like. Alloys or mixtures of these dopants are also be possible, for example, (ZnCd)Se, or Zn(SSe), or the like. Additionally, alloys of different groups of semiconductors may also be possible, for example, a combination of a Group II-Group VI and a Group III-Group V semiconductor, for example, (GaAs)x(ZnS)i-x. Other examples of dopants may include combinations of Group IV and Group VI elemnts, such as GeS, GeSe, GeTe, SnS, SnSe, SnTe, PbO, PbS, PbSe, or PbTe. Other semiconductor mixtures may include a combination of a Group I and a Group VII, such as CuF, CuCl, CuBr, Cui, AgF, AgCl, AgBr, Agl, or the like. Other dopant compounds may include different mixtures of these elements, such as BeSiN2, CaCN2, ZnGeP2, CdSnAs2, ZnSnSb2, CuGeP3, CuSi2P3, Si3N4, Ge3N4, A12O3, (Al, Ga, In)2(S, Se, Te)3, AI2CO, (Cu, Ag)(Al, Ga, In, Tl, Fe)(S, Se, Te)2 and the like.

[0137] The doping of the semiconductor to produce a p-type or n-typc semiconductor may be achieved via bulk-doping in certain embodiments, although in other embodiments, other doping techniques (such as ion implantation) can be used. Many such doping techniques that can be used will be familiar to those of ordinary skill in the art, including both bulk doping and surface doping techniques. A bulk-doped article (e.g. an article, or a section or region of an article) is an article for which a dopant is incorporated substantially throughout the crystalline lattice of the article, as opposed to an article in which a dopant is only incorporated in particular regions of the crystal lattice at the atomic scale, for example, only on the surface or exterior. For example, some articles are typically doped after the base material is grown, and thus the dopant only extends a finite distance from the surface or exterior into the interior of the crystalline lattice. It should be understood that “bulk-doped” does not define or reflect a concentration or amount of doping in a semiconductor, nor does it necessarily indicate that the doping is uniform. “Heavily doped” and “lightly doped” are

[0138] 23

[0139] 12713828.1 terms the meanings of which are clearly understood by those of ordinary skill in the art. In some embodiments, one or more regions comprise a single monolayer of atoms (“deltadoping”). In certain cases, the region may be less than a single monolayer thick (for example, if some of the atoms within the monolayer are absent). As a specific example, the regions may be arranged in a layered structure within the nanoscale wire, and one or more of the regions can be delta-doped or partially delta-doped.

[0140] Accordingly, in one set of embodiments, the nanoscale wires may include a heterojunction, e.g., of two regions with dissimilar materials or elements, and / or the same materials or elements but at different ratios or concentrations. The regions of the nanoscale wire may be distinct from each other with minimal cross-contamination, or the composition of the nanoscale wire can vary gradually from one region to the next. The regions may be both longitudinally arranged relative to each other, or radially arranged (e.g., as in a core / shell arrangement) on the nanoscale wire. Each region may be of any size or shape within the wire. The junctions may be, for example, a p / n junction, a p / p junction, an n / n junction, a p / i junction (where i refers to an intrinsic semiconductor), an n / i junction, an i / i junction, or the like. The junction can also be a Schottky junction in some embodiments. The junction may also be, for example, a semiconductor / semiconductor junction, a semiconductor / metal junction, a semiconductor / insulator junction, a metal / metal junction, a metal / insulator junction, an insulator / insulator junction, or the like. The junction may also be a junction of two materials, a doped semiconductor to a doped or an undoped semiconductor, or a junction between regions having different dopant concentrations. The junction can also be a defected region to a perfect single crystal, an amorphous region to a crystal, a crystal to another crystal, an amorphous region to another amorphous region, a defected region to another defected region, an amorphous region to a defected region, or the like. More than two regions may be present, and these regions may have unique compositions or may comprise the same compositions. As one example, a wire can have a first region having a first composition, a second region having a second composition, and a third region having a third composition or the same composition as the first composition. Non-limiting examples of nanoscale wires comprising heterojunctions (including core / shell heterojunctions, longitudinal heterojunctions, etc., as well as combinations thereof) are discussed in U.S. Patent No. 7,301,199, issued November 27, 2007, entitled “Nanoscale Wires and Related Devices,” by Lieber, et al., incorporated herein by reference in its entirety.

[0141] In some embodiments, a nanoscale wire is a bent or a kinked nanoscale wire. A kink is typically a relatively sharp transition or turning between a first substantially straight

[0142] 24

[0143] 12713828.1 portion of a wire and a second substantially straight portion of a wire. For example, a nanoscale wire may have 1, 2, 3, 4, or 5 or more kinks. In some cases, the nanoscale wire is formed from a single crystal and / or comprises or consists essentially of a single crystallographic orientation, for example, a <110> crystallographic orientation, a <112> crystallographic orientation, or a <1 120> crystallographic orientation. It should be noted that the kinked region need not have the same crystallographic orientation as the rest of the semiconductor nanoscale wire. In some embodiments, a kink in the semiconductor nanoscale wire may be at an angle of about 120° or a multiple thereof. The kinks can be intentionally positioned along the nanoscale wire in some cases. For example, a nanoscale wire may be grown from a catalyst particle by exposing the catalyst particle to various gaseous reactants to cause the formation of one or more kinks within the nanoscale wire. Non-limiting examples of kinked nanoscale wires, and suitable techniques for making such wires, are disclosed in International Patent Application No. PCT / US2010 / 050199, filed September 24, 2010, entitled “Bent Nanowires and Related Probing of Species,” by Tian, et al., published as WO 2011 / 038228 on March 31, 2011, incorporated herein by reference in its entirety.

[0144] In one set of embodiments, the nanoscale wire is formed from a single crystal, for example, a single crystal nanoscale wire comprising a semiconductor. A single crystal item may be formed via covalent bonding, ionic bonding, or the like, and / or combinations thereof. While such a single crystal item may include defects in the crystal in some cases, the single crystal item is distinguished from an item that includes one or more crystals, not ionically or covalently bonded, but merely in close proximity to one another.

[0145] In some embodiments, the nanoscale wires used herein are individual or free-standing nanoscale wires. For example, an “individual” or a “free-standing” nanoscale wire may, at some point in its life, not be attached to another article, for example, with another nanoscale wire, or the free-standing nanoscale wire may be in solution. This is in contrast to nanoscale features etched onto the surface of a substrate, e.g., a silicon wafer, in which the nanoscale features are never removed from the surface of the substrate as a free-standing article. This is also in contrast to conductive portions of articles which differ from surrounding material only by having been altered chemically or physically, in situ, i.e., where a portion of a uniform article is made different from its surroundings by selective doping, etching, etc. An “individual” or a “free-standing” nanoscale wire is one that can be (but need not be) removed from the location where it is made, as an individual article, and transported to a different location and combined with different components to make a functional device such as those

[0146] 25

[0147] 12713828.1 described herein and those that would be contemplated by those of ordinary skill in the art upon reading this disclosure.

[0148] In various embodiments, more than one nanoscale wire may be present within the flexible scaffold. The nanoscale wires may each independently be the same or different. For example, the flexible scaffold can comprise at least 5 nanoscale wires, at least about 10 nanoscale wires, at least about 30 nanoscale wires, at least about 50 nanoscale wires, at least about 100 nanoscale wires, at least about 300 nanoscale wires, at least about 1000 nanoscale wires, etc. The nanoscale wires may be distributed uniformly or non-uniformly throughout the flexible scaffold. In some cases, the nanoscale wires may be distributed at an average density of at least about 10 nanoscale wires / mm3, at least about 30 nanoscale wires / mm3, at least about 50 nanoscale wires / mm3, at least about 75 nanoscale wires / mm3, or at least about 100 nanoscale wires / mm3. In certain embodiments, the nanoscale wires are distributed within the flexible scaffold such that the average separation between a nanoscale wire and its nearest neighboring nanoscale wire is less than about 2 mm, less than about 1 mm, less than about 500 micrometers, less than about 300 micrometers, less than about 100 micrometers, less than about 50 micrometers, less than about 30 micrometers, or less than about 10 micrometers.

[0149] Within the flexible scaffold, some or all of the nanoscale wires may be individually electronically addressable. For instance, in some cases, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or substantially all of the nanoscale wires within the flexible scaffold may be individually electronically addressable. In some embodiments, an electrical property of a nanoscale wire can be individually determinable (e.g., being partially or fully resolvable without also including the electrical properties of other nanoscale wires), and / or such that the electrical property of a nanoscale wire may be individually controlled (e.g., by applying a desired voltage or current to the nanoscale wire, for instance, without simultaneously applying the voltage or current to other nanoscale wires). In other embodiments, however, at least some of the nanoscale wires can be controlled within the same electronic circuit (e.g., by incorporating the nanoscale wires in series and / or in parallel), such that the nanoscale wires can still be electronically controlled and / or determined.

[0150] The nanoscale wire, in some embodiments, may be responsive to a property external of the nanoscale wire, e.g., a chemical property, an electrical property, a physical property, etc. Such determination may be qualitative and / or quantitative. For example, in one set of embodiments, the nanoscale wire may be responsive to voltage. For instance, the nanoscale

[0151] 26

[0152] 12713828.1 wire may exhibits a voltage sensitivity of at least about 5 microsiemens / V; by determining the conductivity of a nanoscale wire, the voltage surrounding the nanoscale wire may thus be determined. In other embodiments, the voltage sensitivity can be at least about 10 microsiemens / V, at least about 30 microsiemens / V, at least about 50 microsiemens / V, or at least about 100 microsiemens / V. Other examples of electrical properties that can be determined include resistance, resistivity, conductance, conductivity, impendence, or the like.

[0153] As another example, a nanoscale wire may be responsive to a chemical property of the environment surrounding the nanoscale wire. For example, an electrical property of the nanoscale wire can be affected by a chemical environment surrounding the nanoscale wire, and the electrical property can be thereby determined to determine the chemical environment surrounding the nanoscale wire. As a specific non-limiting example, the nanoscale wires may be sensitive to pH or hydrogen ions. Further non-limiting examples of such nanoscale wires are discussed in U.S. Patent No. 7,129,554, filed October 31, 2006, entitled “Nanosensors,” by Lieber, et al., incorporated herein by reference in its entirety.

[0154] As an example, the nano scale wire may have the ability to bind to an analyte indicative of a chemical property of the environment surrounding the nanoscale wire (e.g., hydrogen ions for pH, or concentration for an analyte of interest), and / or the nanoscale wire may be partially or fully functionalized, i.e. comprising surface functional moieties, to which an analyte is able to bind, thereby causing a determinable property change to the nanoscale wire, e.g., a change to the resistivity or impedance of the nanoscale wire. The binding of the analyte can be specific or non-specific. Functional moieties may include simple groups, selected from the groups including, but not limited to, -OH, -CHO, -COOH, -SO3H, -CN, - NH2, -SH, -COSH, -COOR, halide; biomolecular entities including, but not limited to, amino acids, proteins, sugars, DNA, antibodies, antigens, and enzymes; grafted polymer chains with chain length less than the diameter of the nanowire core, selected from a group of polymers including, but not limited to, polyamide, polyester, polyimide, polyacrylic; a shell of material comprising, for example, metals, semiconductors, and insulators, which may be a metallic element, an oxide, an sulfide, a nitride, a selenide, a polymer and a polymer gel.

[0155] In some embodiments, a reaction entity may be bound to a surface of the nanoscale wire, and / or positioned in relation to the nanoscale wire such that the analyte can be determined by determining a change in a property of the nanoscale wire. The “determination” may be quantitative and / or qualitative, depending on the application. The term “reaction entity” refers to any entity that can interact with an analyte in such a manner to cause a detectable change in a property (such as an electrical property) of a nanoscale wire.

[0156] 27

[0157] 12713828.1 The reaction entity may enhance the interaction between the nanowire and the analyte, or generate a new chemical species that has a higher affinity to the nanowire, or to enrich the analyte around the nanowire. The reaction entity can comprise a binding partner to which the analyte binds. The reaction entity, when a binding partner, can comprise a specific binding partner of the analyte. For example, the reaction entity may be a nucleic acid, an antibody, a sugar, a carbohydrate or a protein. Alternatively, the reaction entity may be a polymer, catalyst, or a quantum dot. A reaction entity that is a catalyst can catalyze a reaction involving the analyte, resulting in a product that causes a detectable change in the nanowire, e.g. via binding to an auxiliary binding partner of the product electrically coupled to the nanowire. Another exemplary reaction entity is a reactant that reacts with the analyte, producing a product that can cause a detectable change in the nanowire. The reaction entity can comprise a shell on the nanowire, e.g. a shell of a polymer that recognizes molecules in, e.g., a gaseous sample, causing a change in conductivity of the polymer which, in turn, causes a detectable change in the nanowire.

[0158] The term “binding partner” refers to a molecule that can undergo binding with a particular analyte, or “binding partner” thereof, and includes specific, semi-specific, and nonspecific binding partners as known to those of ordinary skill in the art. The term “specifically binds,” when referring to a binding partner (e.g., protein, nucleic acid, antibody, etc.), refers to a reaction that is determinative of the presence and / or identity of one or other member of the binding pair in a mixture of heterogeneous molecules (e.g., proteins and other biologies). Thus, for example, in the case of a receptor / ligand binding pair the ligand would specifically and / or preferentially select its receptor from a complex mixture of molecules, or vice versa. An enzyme would specifically bind to its substrate, a nucleic acid would specifically bind to its complement, an antibody would specifically bind to its antigen. Other examples include, nucleic acids that specifically bind (hybridize) to their complement, antibodies specifically bind to their antigen, and the like. The binding may be by one or more of a variety of mechanisms including, but not limited to ionic interactions, and / or covalent interactions, and / or hydrophobic interactions, and / or van der Waals interactions, etc.

[0159] Additionally, as discussed, a flexible scaffold in some aspects may include a photoresist, such as a soft photoresist. For example, in some embodiments, the photoresist may comprise a polymer formed by photo-curing a fluorinated monomer including crosslinkable function groups using a photoinitiator. This may, for example, facilitate stretchability of the flexible scaffold. One example of such a polymer is perfluoropolyether dimethacrylate (PFPE-DMA). In addition, in some cases, the photoresist may be a photo-

[0160] 28

[0161] 12713828.1 curable composition. In some embodiments, a photo-curable composition includes: a fluorinated monomer including cross -linkable functional groups; and a photoinitiator. Additional non-limiting examples of photoresist may be found in Int. Pat. Apl. Pub. No. WO 2019 / 084498, incorporated herein by reference in its entirety.

[0162] Some embodiments of this disclosure are directed to a photo-curable composition that can be cured to form an elastomer exhibiting high stretchability and that is chemically orthogonal to various development solvents used in photolithography and, hence, compatible with photolithography. Further, the elastomer can be patterned with fine feature resolution, and can be used as a photoresist for patterning various materials, including electrically (or electronically) active materials. Examples of applications of such photo-pattemable composition include forming stretchable and transparent substrates, stretchable and transparent dielectric / passivation / encapsulation films or layers for elastic or stretchable microelectronics, and photoresists for patterning of materials, such as in the context of implantable medical devices, wearable electronic devices, and soft electronic devices; other biomedical devices; cosmetics; prosthetics; and other applications involving an interface with a human body, an animal body, or other biological tissue where matching of mechanical properties with the biological tissue is desired.

[0163] In some embodiments, a kit may be provided, e.g., comprising a flexible scaffold as is discussed herein. Cells may or may not be provided with the kit. The kit may include a package or an assembly including the flexible scaffold, and optionally other components associated with the flexible scaffold, such as cells. Examples of other components include, but are not limited to, solvents, surfactants, diluents, salts, buffers, emulsifiers, chelating agents, fillers, antioxidants, binding agents, bulking agents, preservatives, drying agents, antimicrobials, needles, syringes, packaging materials, tubes, bottles, flasks, beakers, dishes, frits, filters, rings, clamps, wraps, patches, containers, and the like, for example, for using, administering, modifying, assembling, storing, packaging, preparing, mixing, diluting, and / or preserving the flexible scaffold.

[0164] A kit may include instructions in any form that are provided in connection with the components of the kit in such a manner that one of ordinary skill in the art would recognize that the instructions are to be associated with those components. For instance, the instructions may include instructions for the use, modification, mixing, diluting, preserving, administering, assembly, storage, packaging, and / or preparation of the flexible scaffold. The instructions may be provided in any form recognizable by one of ordinary skill in the art as a suitable vehicle for containing such instructions, for example, written or published, verbal,

[0165] 29

[0166] 12713828.1 audible (e.g., telephonic), digital, optical, visual (e.g., videotape, DVD, etc.) or electronic communications (including Internet or web-based communications), provided in any manner.

[0167] In some cases, a material of a device can be chosen to be one that is readily introduced into the device, e.g., using techniques compatible with lithographic techniques. For example, in one set of embodiments, lithographic techniques such as e-beam lithography, photolithography, X-ray lithography, extreme ultraviolet lithography, ion projection lithography, etc. may be used to layer or deposit one or more metals on a substrate. Additional processing steps can also be used to define or register the pathways in some cases.

[0168] In some embodiments, more than one metal can be used within a pathway. For example, two, three, or more metals may be used within a pathway. The metals may be deposited in different regions or alloyed together, or in some cases, the metals may be layered on top of each other, e.g., layered on top of each other using various lithographic techniques. If dissimilar metals are layered on top of each other, they may be layered in some embodiments in a “stressed” configuration (although in other embodiments, they may not necessarily be stressed). For example, a chromium / palladium / chromium deposition process, in some embodiments, may form a pre- stressed arrangement that is able to spontaneously form a 3-dimensional structure after release from the substrate. See, e.g., U.S. Pat. Nos. 9,457,128 or 9,786,850, each incorporated herein by reference in its entirety.

[0169] In some embodiments, the conductive pathway may be relatively narrow. For example, the conductive pathway may have a smallest dimension or a largest cross-sectional dimension of less than about 5 micrometers, less than about 4 micrometers, less than about 3 micrometers, less than about 2 micrometers, less than about 1 micrometer, less than about 700 nm, less than about 600 nm, less than about 500 nm, less than about 300 nm, less than about 200 nm, less than about 100 nm, less than about 80 nm, less than about 50 nm, less than about 30 nm, less than about 10 nm, less than about 5 nm, less than about 2 nm, etc. The conductive pathway may have any suitable cross-sectional shape, e.g., circular, square, rectangular, polygonal, elliptical, regular, irregular, etc. As is discussed in detail below, such conductive pathways may be achieved using lithographic or other techniques.

[0170] In some cases, the conductive pathways may define an electrical circuit that is internally contained within the device, and / or that extends externally of the device, e.g., such that the electrical circuit is in electrical communication with an external electrical system, such as a computer or a transmitter (for instance, a radio transmitter, a wireless transmitter, an Internet connection, etc.). The device, in some embodiments, may contain components such as nanoelectric components. Non-limiting examples of such components include nanoscale

[0171] 30

[0172] 12713828.1 wires, sensors such as nanosensors, transistors such as field effect transistors, resistors, capacitors, inductors, diodes, integrated circuits, batteries, power sources, RFID tags, antennae, transmitter, or the like, which may be present in one or more electrical circuit within the device.

[0173] In certain embodiments, a conductive pathway may define an electrical circuit that is interfaceable or connectable with an external electrical device, such as a computer, using a suitable connector. For example, the flexible scaffold may be directly connected to an external device (for instance, using an interface such as described in U.S. Pat. Apl. Pub. No. 2018 / 0328884, incorporated herein by reference in its entirety). In some cases, a suitable connector, such as a cable, may be used to make such a connection between an electrical circuit within a flexible scaffold and the external device. Non-limiting examples of cables include those commercially available, such as ribbon cables, flexible flat cable, 8-pin cables, 16-pin cables, etc., or other electrical cables. However, an external connection is not always required, and in some cases, the scaffold may be a self-contained electrical circuit. For example, the circuit may be able to transmit information wirelessly to an external device, store information for later access (e.g., after sacrificing the organoids, organs, or organisms), or the like.

[0174] In addition, in certain embodiments, the device may be able to communicate with an external device using wireless communications, e.g., in addition to and / or instead of an electrical connection. For example, the device may contain a transmitter (for instance, a radio transmitter, a wireless transmitter, an Internet connection, etc.) and / or a receiver, e.g., which may be in communication with a transmitter and / or a receiver on an external device.

[0175] In some embodiments, more than one electrical circuit and / or more than one conductive pathway may be used within a device. For example, multiple conductive pathways or circuits can be used such that some or all of the nodes may be individually electronically addressable within the flexible scaffold. However, in other embodiments, more than one node may be addressable by a particular conductive pathway.

[0176] In some cases, the device may be connected to a computer system, e.g., a general or a special purpose computer system. The system may be used in some embodiments to apply electrical stimulation to one or more portions of the device, e.g., to be applied to one or more cells within the scaffold. For instance, one or more portions of the device may be individually addressable, e.g., via the computer system. In addition, in some embodiments, the system may be used to determine a property of the device, e.g., when it is embedded within a biological structure or system, such as described elsewhere herein. For instance, a

[0177] 31

[0178] 12713828.1 property, such as a chemical property, an electrical property, a mechanical property, or the like may be determined and recorded by the computer system. In some cases, one or more electrodes may be used. In certain embodiments, the computer system may be used to apply electrical stimulation to the scaffold, determine one or more properties of the flexible scaffold, and predict electrical stimulation to be applied based on properties of the flexible scaffold.

[0179] A non-limiting illustrative implementation of a special purpose computer system 300 that may be specially programmed to be used in connection with any of the embodiments of the disclosure provided herein is shown in Fig. 4. The computer system 300 may include one or more processors 310 and one or more articles of manufacture that comprise non-transitory computer-readable storage media (e.g., memory 320 and one or more non-volatile storage media 330). The processor 310 may control writing data to and reading data from the memory 320 and the non-volatile storage device 330 in any suitable manner. To perform any of the functionality described herein (e.g., build predictive models, provide inputs to the predictive models, apply electrical stimuli to one or more portions of the scaffold, etc.), the processor 310 may execute one or more processor-executable instructions stored in one or more non-transitory computer-readable storage media (e.g., the memory 320), which may serve as non-transitory computer-readable storage media storing processor-executable instructions for execution by the processor 310.

[0180] The terms “program” or “software” or “app” are used herein in a generic sense to refer to any type of computer code or set of processor-executable instructions that can be employed to program a computer or other processor to implement various aspects of embodiments as discussed above. Additionally, it should be appreciated that according to one aspect, one or more computer programs that when executed perform methods of the disclosure provided herein need not reside on a single computer or processor, but may be distributed in a modular fashion among different computers or processors to implement various aspects of the disclosure provided herein.

[0181] Processor-executable instructions may be in many forms, such as program modules, executed by one or more computers or other devices. Generally, program modules include routines, programs, objects, components, data structures, etc. that perform particular tasks or implement particular abstract data types. Typically, the functionality of the program modules may be combined or distributed as desired in various embodiments.

[0182] Also, data structures may be stored in one or more non-transitory computer-readable storage media in any suitable form. For simplicity of illustration, data structures may be

[0183] 32

[0184] 12713828.1 shown to have fields that are related through location in the data structure. Such relationships may likewise be achieved by assigning storage for the fields with locations in a non-transitory computer-readable medium that convey relationships between the fields. However, any suitable mechanism may be used to establish relationships among information in fields of a data structure, including through the use of pointers, tags or other mechanisms that establish relationships among data elements.

[0185] Also, various concepts may be embodied as one or more processes, of which examples have been provided herein. The acts performed as part of each process may be ordered in any suitable way. Accordingly, embodiments may be constructed in which acts are performed in an order different than illustrated, which may include performing some acts simultaneously, even though shown as sequential acts in illustrative embodiments.

[0186] As noted above and elsewhere herein, a computer-readable medium may be associated with a biological system to receive and / or transmit electrical signals. In some such embodiments, the computer-readable medium is configured to process a first signal based on a first current having a first peak shape to determine a second current having a second peak shape, different from the first peak shape, to apply to the biological system.

[0187] In some embodiments, the computer-readable medium is configured to use machine learning to determine a set of subsequent outputs. For example, a biological system may provide a series of inputs (e.g., electrical impedance measurements), and the computer- readable medium is configured to determine a set of subsequent outputs. In some embodiments, the computer-readable medium is configured to train using a predictive model and at least one of a voltage supplied during electrical stimulation, a duration of electrical stimulation, a volume or density of the portions of the tissue undergoing electrical stimulation, type of cells comprising tissue undergoing electrical stimulation, an intensity or pattern of electrical stimulation, a frequency of electrical stimulation, periodicity of electrical stimulation, and, optionally, further trained on a resulting tissue response.

[0188] The following documents are incorporated herein by reference in their entireties: U.S. Provisional Patent Application Serial No. 62 / 865,648, filed June 24, 2019, entitled “Organoids Containing Electronics, and Methods Thereof,” by Liu, et al. U.S. Provisional Patent Application Serial No. 62 / 872,031, filed July 9, 2019, entitled “Organoids Containing Electronics, and Methods Thereof,” by Liu, et al. ; U.S. Patent No. 7,211,464, issued May 1, 2007, entitled “Doped Elongated Semiconductors, Growing Such Semiconductors, Devices Including Such Semiconductors, and Fabricating Such Devices”; U.S. Patent No. 7,301,199, issued November 27, 2007, entitled “Nanoscale Wires and Related Devices”; and

[0189] 33

[0190] 12713828.1 International Patent Application No. PCT / US2010 / 050199, filed September 24, 2010, entitled “Bent Nanowires and Related Probing of Species,” published as WO 2011 / 038228 on March 31, 2011. In addition, the following are each incorporated herein by reference in their entireties: U.S. Pat. Nos. 9,786,850 and 9,457,128; U.S. Pat. Apl. Pub. Nos. 2017 / 0069858, 2014 / 0073063, 2017 / 0072109, and 2014 / 0074253; and Int. Pat. Apl. Pub. No. WO 2019 / 084498. Also incorporated herein by reference in their entireties are U.S. Pat. Apl. Pub. No. 2022-0213425 and Int. Pat. Apl. Pub. No. WO 2020 / 263772. In addition, U.S. Pat. Apl. Ser. Nos. 63 / 681,576 and 63 / 681,627, and Int. Pat. Apl. Ser. No. PCT / US25 / 20504 are each incorporated herein by reference in its entirety.

[0191] The following examples are intended to illustrate certain embodiments of the present invention, but do not exemplify the full scope of the invention.

[0192] EXAMPLE 1

[0193] In this example, nanoelectronic mesh recordings as well as initial stimulation data on two healthy rats was performed. Briefly, healthy CD rats received 5% isoflurane by inhalation and 0.3mg / kg buprenorphine before the chest cavity was opened, and 1000U heparin solution was injected to prevent coagulation and occlusion. The heart was excised, and the aorta was cannulated for retrograde ex vivo perfusion of warmed and oxygenated Tyrode’s solution using the Langendorff Hanging Heart System. A nanoelectronic mesh electrode was placed on the left atrial appendage for detection of irregular cardiac conduction, and a reference electrode was placed on the right atrial appendage. After taking healthy atrial recordings at baseline, stimulation was applied for the induction of irregular atrial electrophysiology. The initial stimulation protocol applied current at a frequency of 20Hz and amplitude of 0.5mA for a duration of 1 millisecond and a period of 2 seconds. The pace current was applied from Stimulus A and / or B, using 1, 2, or 3 channels (Table 1.)

[0194] Table 1

[0195] 34

[0196] 12713828.1

[0197] EXAMPLE 2

[0198] In this example, electrophysiological recordings were performed on cardiac organoids, which typically formed in ~48 hours (Figs. 2A-2B). Fig. 2C shows the voltage trace of 14-channel (out of 16 channels) electrophysiological recording on cardiac organoid at day 35 of differentiation. The zoom-in plot of single spikes (Fig. 2D) showed non-uniform electrophysiological behaviors of the cells distributed across the organoid, as well as a time latency revealing tissue- wide propagation of local field potentials (LFP). Chronic tracing of LFP at millisecond temporal resolution during organogenesis (Fig. 2E) revealed changes in the spike dynamics from initially slow waveform through the emergence of repolarization to fast depolarization. The averaged amplitude of the fast component associated with depolarization remains undetectable until day 31 of differentiation, and then increased monotonically (Fig. 2F). The field potential duration (FPD) was found to remain relatively steady between 0.7 and 0.8 s (Fig. 2G) with a slight increase. The activation mapping confirmed the gradually synchronized LFP propagation across the cardiac organoid from day 26 to day 35 of differentiation (Figs. 2H, 21, and 3). These recording data suggested that the functional maturation of a cardiac organoid was marked by the synchronization of the bursting phase, instead of bursting frequency or FPD.

[0199] Fig. 2 shows chronic, multiplex, tissue-wide electrophysiological mapping of cardiac organoid during organogenesis. Fig. 2A is a schematic that shows the set-up that connects cyborg cardiac organoids to external recording equipment for multiplexing electrophysiology during organoid development. In Fig. 2B, optical images show representative processes (0, 24, and 48 hours) of 3D organization of cardiac cyborg organoids in culture chamber connected for electrophysiological recording. In Fig. 2C, 14-channel voltage traces recorded from the cardiac cyborg organoid at day 35 of differentiation. In Fig. 2D, zoom-in views of the box highlights in Fig. 2C show single-spiked field potential recording. Fig. 2E shows zoom-in views of the dashed box highlighted channel in Fig. 2C on three different culturing days (day 26, 31, and 35 of differentiation). Figs. 2F-2I show amplitude of fast peak (Fig. 2F) and field potential duration (Fig. 2G) defined in Fig. 2E as a function of differentiation

[0200] 35

[0201] 12713828.1 time. Gray lines show individual channels. Black line shows the averaged results from 14 channels. (Value = mean + / - s.e.m., n= 14.) In Figs. 2H and 21, isochronal mappings at day 26 (Fig. 2H) and day 35 (Fig. 21) of differentiation show delays of activation time (max. - dV / df) for three consecutive spiking events labeled in Figs. 3A-3B.

[0202] Fig. 3 shows electrophysiology data for 14 channels during day 26 (Fig. 3A) and 35 (Fig. 3B) of culture shows transition from irregular activity patterns to synchronous beating. This dataset was used to build the activation maps in Fig. 2H, for three consecutive spiking events inside the dashed red rectangles. Dashed segments at day 26 represent the activation time for spike #1 for each channel.

[0203] EXAMPLE 3

[0204] Following are various materials and method used in the above examples.

[0205] Fabrications of soft, stretchable mesh nanoelectronics. Important steps in the fabrication of the ultra-thin, stretchable mesh nanoelectronics, illustrated in Fig. 1, included:

[0206] (1) Cleaning a silicon wafer grown with thermal oxide (500-nm thickness) with acetone, isopropyl alcohol, and water.

[0207] (2) Depositing 100-nm-thick nickel (Ni) using electron-beam evaporator as a sacrificial layer.

[0208] (3) Spin-coating SU-8 precursor (SU-8 2000.5, MicroChem) at 4000 rpm, which was pre-baked at (65 °C, 95 °C) for 2 min each, exposed to 365 nm ultra-violet (UV) for 200 mJ / cm2, post-baked at (65 °C, 95 °C) for 2 min each, developed using SU-8 developer (MicroChem) for 60 s, and hard-baked at 180 °C for 40 min to define mesh SU-8 patterns (400-nm thickness) for bottom encapsulation. For fluorescence imaging, 0.004 wt%© of Rhodamin 6G powder (Sigma- Aldrich) was added into SU-8 precursor. Note that other materials may be used besides SU-8, including those described herein, and those in Int. Pat. Apl. Pub. No. WO 2019 / 084498, incorporated herein by reference in its entirety.

[0209] (4) Spin-coating LOR3A photoresist (MicroChem) at 4000 rpm, followed by prebaking at 180 °C for 5 min; spin-coating SI 805 photoresist (MicroChem) at 4000 rpm, followed by pre-backing at 115 °C for 1 min; the sample was then exposed to 405 nm UV for 40 mJ / cm2, and developed using CD-26 developer (Micropost) for 70 s to define interconnects patterns.

[0210] (5) Depositing 5 / 40 / 5-nm-thick chromium / gold / chromium (Cr / Au / Cr) by electronbeam evaporator, followed by a standard lift-off procedure in remover PG (MicroChem) overnight to define the Au interconnects.

[0211] 36

[0212] 12713828.1 (6) Repeating Step (4) to define electrode array patterns in LOR3A / S1805 bilayer photoresists;

[0213] (7) Depositing 5 / 50-nm-thick chromium / platinum (Cr / Pt) by electron-beam evaporator, followed by a standard lift-off procedure in remover PG (MicroChem) for 10 min to define the electrode array;

[0214] (8) Repeating Step (3) for top SU-8 encapsulation (400-nm thickness).

[0215] (9) Electrochemically polymerizing poly (3, 4-ethylenedioxy thiophene) (PEDOT) on the Pt electrode array using a precursor consisting of 14 mM 3, 4-ethylenedioxy thiophene (EDOT, Sigma- Aldrich) in lx phosphate-buffered saline (PBS) solution. The precursor was drop-casted onto the device, followed by passage of a 1 V DC voltage (Dr. Meter DC power supply PS-305DM) for 200 s using device electrodes as the anode and an external Pt wire as the cathode. The device was then rinsed with DI water for 30 s and dried.

[0216] (10) Soldering a 16-channel flexible flat cable (Molex) onto the input / output pads using a flip-chip bonder (Finetech Fineplacer).

[0217] (11) Gluing a chamber onto the substrate wafer to completely enclose the mesh part of the device using a bio-compatible adhesive (Kwik-Sil, WPI).

[0218] (12) Treating surface of the device with light oxygen plasma (Anatech 106 oxygen plasma barrel asher), followed by adding 3 mL of Ni etchant (type TFB, Transene) into the chamber for 2 to 4 hours to completely release the mesh electronics from the substrate wafer. The device was then ready for subsequent sterilization steps before cell culture.

[0219] Organoid culture. Materials for cell culture. Human induced pluripotent stem cells hiPSCs-(IMR90)-l were purchased from WiCell Research Institute (Madison, WI, USA). Authentication and test for the free of mycoplasma were performed by WiCell Research Institute. A cell bank was established after the cells were received. For experiments, each aliquot from the cell bank was used for less than 5 additional passages. Human mesenchymal stem cells (hMSCs; PT-2501) were obtained from Lonza (Walkersville, MD, USA) and used for less than 5 additional passages. Reagents and their supplies: Essential 8™ (E8) medium (cat. no. A1517001, Life Technologies); MSCGM BulletKit (cat. no. PT-3238 & PT-4105, Lonza); RPMI 1640 Medium (cat. no. 11875093, life technologies); B-27 Supplement (cat. no. 17504044, life technologies); B-27™ Supplement, minus insulin (cat. no. A1895601, Life Technologies); CHIR 99021, (cat. no. 44-231-0, Tocris Bioscienc ™); IWR-1 (cat. no. CAYM-13659-5, Cayman Chemical); Normal donkey serum (cat. no. 017-000-121, Jackson ImmunoResearch); Accutase (cat. no. Al 110501, Life Technologies); Trypsin-EDTA 0.05% (cat. no. 25-300-054, Invitrogen); EDTA 0.5M pH 8.0 (cat. no. AM9260G, Invitrogen); Y-

[0220] 37

[0221] 12713828.1 27632 (cat. no. 129830-38-2, Sigma); Matrigel (cat. no. 08-774-552, Corning); DAPI (cat. no. D9542, Sigma). Phalloidin-iFluor 647 (cat. no. abl76759, Abeam); poly-D-lysine hydrobromide (cat. no. P7280-5MG, Sigma); photoinitiator 2,20-azobis[2-(2-imidazolin-2-yl) propane] dihydrochloride (cat. no. VA-044, Wako Chemicals). Antibodies used in these examples: Nanog (Santa Cruz, 1:100); CD44 (Abeam, 1:100); Cardiac troponin T (TNT) (Invitrogen, 1:100); a-actinin (Sigma, 1:100); NKX2.5 (Abeam, 1:100); Alexa 594 donkey anti-mouse (Jackson ImmunoResearch, 1:500); Alexa 647 Donkey anti-rabbit (Invitrogen, 1:500).

[0222] Two dimensional (2D) cells culture. hiPSCs-(IMR90)-l were maintained in 6-well plate coated with Matrigel in Essential 8™ medium. Steps were as follows:

[0223] (1) 6-well plate was coated with Matrigel at 37 °C at least one hour.

[0224] (2) Matrigel from the well was aspirated and replaced with 2 mL E8 medium.

[0225] (3) hiPSCs were incubated with 0.5 mM EDTA at room temperature for 5 minutes.

[0226] (4) EDTA solution was aspirated and replaced with 1 mL E8 medium. Cells were pipetted into clusters.

[0227] (5) 100-200 microliters cells were seeded in the Matrigel coated 6 well-plate.

[0228] (6) Medium was changed daily. Cells were passaged every 4 days. hMSCs were cultured in T-75 flasks in MSCGM medium. Steps were as follows:

[0229] (1) 5000-6000 cells per cm2were seeded in T-75 flask with about 30 ml MSCGM medium.

[0230] (2) Medium was changed every 3-4 days.

[0231] (3) When hMSCs reached ~ 90% confluence, cells were dissociated with Trypsin- EDTA 0.05%. The cells were used by passage 5.

[0232] 2D hiPSC-derived cardiomyocytes differentiation. Cardiomyocytes differentiation steps were as follows:

[0233] (1) hiPSCs were cultured in 6-well plate with E8 medium for 3-4 days.

[0234] (2) E8 medium was removed. RPMI 1640 medium plus 1% B27-insulin and 12 micromolar CHIR99021 were added for differentiation. This day was defined as Day 0.

[0235] (3) On day 1, the medium was changed to RPMI 1640 medium plus 1% B27-insulin.

[0236] (4) On day 3, the medium was changed to RPMI 1640 medium plus 1% B27-insulin and 5 micromolar IWR1.

[0237] (5) On day 5, the medium was changed to RPMI 1640 medium plus 1% B27-insulin.

[0238] (6) On day 7, the medium was changed to RPMI 1640 medium plus 1% B27.

[0239] 38

[0240] 12713828.1 (7) The medium was then replaced with fresh RPMI 1640 medium plus 1% B27 every other day. The cells started beating from day 8 or day 10.

[0241] 3D culture of cyborg organoids. The integration of stretchable mesh nanoelectronics with cells to create cyborg organoids followed immediately from fabrications and release of the device in the chamber, which involved the following steps:

[0242] (1) The released device was sterilized by 1) DI water rinse for three times, and 2) 70% ethanol for at least 15 minutes.

[0243] (2) The device was washed with PBS three times followed by incubating with Poly- D-lysine hydrobromide (0.01% w / v) overnight.

[0244] (3) Poly-D-lysine hydrobromide solution was removed, and 300 microliters liquid Matrigel (10 mg / mL) was added to the chamber from the device-free side on ice. The device was incubated for at least 30 minutes at 37 °C to solidify the Matrigel layer.

[0245] (4) hiPSCs or hiPSC-derived cardiac progenitor cells (hiPSC-CPCs) (2~3xl06cells) and hMSCs (2~4xl05cells) were suspended in a mixture of E8 or RPMI 1640 (supplemented with 1% B27-insulin or 1% B27) and MSCGM medium, and then transferred onto the cured Matrigel in the chamber and cultured at 37 °C, 5% CO2.

[0246] (5) For electrophysiological measurement, hiPSC-CPCs at day 24 and hMSCs mixture were transferred onto the Matrigel and cultured at 37 °C, 5% CO2. hiPSCs-derived cyborg cardiac organoid was formed within 48-72 hours. Electrical activity from the organoid was recorded from 48 hours of its formation, which corresponded to day 26 of cardiac differentiation in Fig. 2.

[0247] Characterization was performed as follows.

[0248] Organoids clearing, section, immunostaining and imaging. For whole organoids imaging, procedures were adapted from tissue clearing techniques CLARITY and passive clarity technique (PACT). The organoids were fixed with 4% PFA at 4 °C overnight and incubated with hydrogel solution (0.25% (w / v) VA-044 and 4% (w / v) acrylamide in PBS) at 4 °C for 24 hours. The samples were placed in X-CLARITY hydrogel polymerization device for 3-4 hours at 37 °C with -90 kPa vacuum, followed by wash in PBS overnight before electrophoretic lipid extraction for 24 hours in the X-CLARITY electrophoretic tissue clearing (ETC) chamber. Then immunostaining was performed by staining the primary antibodies for 3-5 days and the secondary antibodies for 2-4 days, respectively. The samples were submerged in optical clearing solution overnight and embedded in 2% agarose gel before imaging using Zeiss 2-photon time-lapse confocal microscopy at Harvard Center for Biological Imaging (HCBI). For characterization of cyborg cardiac organoids, the organoids

[0249] 39

[0250] 12713828.1 at different stages were fixed with 4% PFA at 4 °C overnight and immersed in 30% sucrose for at least 12 hours. Then the samples were embedded in optimal cutting temperature (OCT) compound and cryostat section of 30 micrometer-thick slices. Cardiac organoids without device integrating were used as control. For staining, the first antibodies were incubated at 4 °C overnight and the secondary antibodies were stained at RT for 3-4 hours. For 2D culture, cells were fixed with 4% paraformaldehyde (PFA) at room temperature (RT) for 15 minutes. Cells were incubated with primary antibodies at 4 °C overnight and the secondary antibodies were stained at RT for 1-2 hours. Finally, 4’,6-diamidino-2-phenylindole (DAPI) were stained for 10 minutes. Cells were washed with PBS three times before imaging. All samples were imaged by Zeiss 880 confocal microscopy at HCBI. Imaging was analysis by Zen (Blue edition) and Fiji. Fluorescence intensity was calculated by Fiji. Data analysis and statistical tests were performed by Graphpad Prism.

[0251] Device impedance characterization. A three-electrodes setup was used to measure the electrochemical impedance spectrum of the electrodes from each device. Platinum wire (300 micrometers in diameter, 1.5 cm in length immersed) and a standard silver / silver chloride electrode were used as counter electrode and reference electrode, respectively. The device was immersed in IxPBS solution during measurement. The SP-150 potentiostat (Bio-logic) along with its commercial software EC-lab was used to perform the measurements. For each measurement, at least three frequency sweeps were measured from 1 MHz down to 1Hz to obtain statistical results. A sinusoidal voltage of 100 mV peak-to-peak was applied. For each data point, the response to 10 consecutive sinusoids (spaced out by 10% of the period duration) was accumulated and averaged.

[0252] Electrophysiological measurement and data analysis. The Blackrock CerePlex Direct voltage amplifier along with a 32 channels Blackrock p (mu) digital headstage connected to the device was used to record electrical activity from organoids. The headstage-to-device connector (16 channels only) was homemade. The organoid culture media was grounded to earth and a reference electrode was also inserted in the media, far from the device (distance above 1 cm). Platinum wires were used as ground and reference electrodes. A sampling rate of 2000 samples per second was used. A 0.3 to 1000 Hz band-pass filter (Butterworth, 4th order) was applied to reduce high frequency noise. Matlab codes provided by Blackrock were used to convert raw data files into accessible format. Data were then transferred to Graphpad Prism for post-processing.

[0253] EQUIVALENTSAND SCOPE

[0254] 40

[0255] 12713828.1 While several embodiments of the present disclosure have been described and illustrated herein, those of ordinary skill in the art will readily envision a variety of other means and / or structures for performing the functions and / or obtaining the results and / or one or more of the advantages described herein, and each of such variations and / or modifications is deemed to be within the scope of the present disclosure. More generally, those skilled in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are meant to be exemplary and that the actual parameters, dimensions, materials, and / or configurations will depend upon the specific application or applications for which the teachings of the present disclosure is / are used. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the disclosure described herein. It is, therefore, to be understood that the foregoing embodiments are presented by way of example only and that, within the scope of the appended claims and equivalents thereto, the disclosure may be practiced otherwise than as specifically described and claimed. The present disclosure is directed to each individual feature, system, article, material, kit, and / or method described herein. In addition, any combination of two or more such features, systems, articles, materials, kits, and / or methods, if such features, systems, articles, materials, kits, and / or methods are not mutually inconsistent, is included within the scope of the present disclosure.

[0256] In cases where the present specification and a document incorporated by reference include conflicting and / or inconsistent disclosure, the present specification shall control. If two or more documents incorporated by reference include conflicting and / or inconsistent disclosure with respect to each other, then the document having the later effective date shall control.

[0257] All definitions, as defined and used herein, should be understood to control over dictionary definitions, definitions in documents incorporated by reference, and / or ordinary meanings of the defined terms.

[0258] The indefinite articles “a” and “an,” as used herein in the specification and in the claims, unless clearly indicated to the contrary, should be understood to mean “at least one.”

[0259] The phrase “and / or,” as used herein in the specification and in the claims, should be understood to mean “either or both” of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Multiple elements listed with “and / or” should be construed in the same fashion, i.e., “one or more” of the elements so conjoined. Other elements may optionally be present other than the elements specifically identified by the “and / or” clause, whether related or unrelated to those elements

[0260] 41

[0261] 12713828.1 specifically identified. Thus, as a non-limiting example, a reference to “A and / or B”, when used in conjunction with open-ended language such as “comprising” can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc.

[0262] As used herein in the specification and in the claims, “or” should be understood to have the same meaning as “and / or” as defined above. For example, when separating items in a list, “or” or “and / or” shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly indicated to the contrary, such as “only one of’ or “exactly one of,” or, when used in the claims, “consisting of,” will refer to the inclusion of exactly one element of a number or list of elements. In general, the term “or” as used herein shall only be interpreted as indicating exclusive alternatives (i.e. “one or the other but not both”) when preceded by terms of exclusivity, such as “either,” “one of,” “only one of,” or “exactly one of.”

[0263] As used herein in the specification and in the claims, the phrase “at least one,” in reference to a list of one or more elements, should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. This definition also allows that elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase “at least one” refers, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, “at least one of A and B” (or, equivalently, “at least one of A or B,” or, equivalently “at least one of A and / or B”) can refer, in one embodiment, to at least one, optionally including more than one, A, with no B present (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements); etc.

[0264] When the word “about” is used herein in reference to a number, it should be understood that still another embodiment of the disclosure includes that number not modified by the presence of the word “about.”

[0265] 42

[0266] 12713828.1 It should also be understood that, unless clearly indicated to the contrary, in any methods claimed herein that include more than one step or act, the order of the steps or acts of the method is not necessarily limited to the order in which the steps or acts of the method are recited. In the claims, as well as in the specification above, all transitional phrases such as

[0267] “comprising,” “including,” “carrying,” “having,” “containing,” “involving,” “holding,” “composed of,” and the like are to be understood to be open-ended, i.e., to mean including but not limited to. Only the transitional phrases “consisting of’ and “consisting essentially of’ shall be closed or semi-closed transitional phrases, respectively, as set forth in the United States Patent Office Manual of Patent Examining Procedures, Section 2111.03.

[0268] What is claimed is:

[0269] 43

[0270] 12713828.1

Claims

1. CLAIMS1. A method for treating an arrythmia, the method comprising: administering, to the heart of a subject having or at risk of arrythmia, a device comprising a mesh of nanoelectrodes.

2. A method for treating an arrythmia, the method comprising: applying electrical stimulation to the heart of a subject having or at risk of having arrythmia using a device comprising a mesh of nanoelectrodes.

3. A method for treating an arrythmia, the method comprising: determining an electrical signal of the heart of a subject having or at risk of arrythmia using a device comprising a mesh of nanoelectrodes; and applying an electrical stimulation to the heart using the mesh of nanoelectrodes, wherein the electrical stimulation alters the heartbeat to treat the arrythmia.

4. A device, comprising: a thoracoscope; and a device comprising a mesh of nanoelectrodes positionable using the thoracoscope.

5. A method for treating an arrythmia , the method comprising: placing a flexible scaffold comprising a nanoelectronic mesh on a tissue at a target location in a heart; and applying electrical stimuli to portions of the tissue at the target location using one or more of electrodes within the nanoelectronic mesh of the flexible scaffold, wherein the electrical stimuli are generated using a predictive model programmed by electrically stimulating the tissue at the target location using one or more of the electrodes and determining a tissue response based on the electrical stimulation.

6. The method of claim 5, wherein the arrythmia is an atrial fibrillation.4412713828.

17. The method of claim 6, wherein the atrial fibrillation originates at the pulmonary veins, coronary sinus, or a posterior wall of the heart.

8. The method of any one of claims 5-7, where the target location in the heart is selected from the pulmonary veins, coronary sinus, the posterior wall of the heart or a left atrial appendage.

9. The method of any one of claims 5-8, wherein applying the electrical stimuli places the heart into a normal sinus rhythm.

10. The method of any one of claims 1-9, wherein the nanoelectronic mesh comprises at least a portion of an electrical circuit.

11. The method of claim 10, wherein at least a portion of the electrical circuit is electrically connected to an electrical device external of the target location.

12. The method of any one of claims 1-11, wherein the nanoelectronic mesh is a rectangular mesh or an irregular mesh.

13. The method of any one of claims 1-12, wherein the nanoelectronic mesh comprises plurality of nodes, at least some of which nodes are connected by interconnects to form the mesh.

14. The method of claim 13, wherein at least some of the interconnects are serpentine.

15. The method of claim 13 or 14, wherein at least some of the interconnects are stretchable.

16. The method of any one of claims 13-15, wherein at least some of the interconnects comprise at least one inflection point.

17. A method for treating atrial fibrillation , the method comprising: placing a flexible scaffold defining at least a portion of an electrical circuit and comprising a plurality of electrodes on a tissue at one or more pulmonary vein-left4512713828.1atrial junctions in a heart; and applying electrical stimuli to portions of the tissue at one or more pulmonary vein-left atrial junctions using one or more of the electrodes within the flexible scaffold, wherein the electrical stimuli are generated using a predictive model programmed by electrically stimulating the tissue at the target location using one or more of the electrodes and determining a tissue response based on the electrical stimulation.

18. The method of claim 17, further comprising placing the plurality of electrodes at a coronary sinus, a posterior wall of the heart or a left atrial appendage of the heart.

19. A device comprising : a flexible scaffold comprising a nanoelectronic mesh and a catheter for delivering said flexible scaffold, wherein the flexible scaffold is disposed within at least a portion of the catheter prior to delivery, and wherein the flexible scaffold is configured to selfassemble into a mesh structure after release from the catheter.4612713828.1

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