DRESSING APPLICATION SYSTEMS AND DEVICES

JOSEPH DEL ROSSI, TROY PALUSZCYK

FIELD

[001] Biologic tissues and cells are affected by electrical stimulus. Accordingly, apparatuses and techniques for applying electric stimulus to biological tissue and cells have been developed to address a number of medical issues. The present specification relates to bioelectric devices and methods of storage, application, manufacture, and use thereof.

SUMMARY

[002] Embodiments disclosed herein include systems, devices, and methods for storing and applying dressings, for example, dressings for treating tissues. These tissues may have sustained injury and/or wounds (including surgical incisions), or could benefit from treatment for skin-related conditions (for example, acne, rosacea, rash, or the like), or could benefit from treatment or pre-treatment to minimize risk of injury (for example, muscle damage). Disclosed systems, devices, and methods can comprise a multi-array matrix of biocompatible microcells and provide a treatment site with a localized voltage and/or microcurrent.

[003] Disclosed systems and devices can comprise a multi-array matrix on a substrate or base layer that is reversibly attached to, for example, a backing layer or card, for example a cardboard backing layer. The substrate can comprise an adhesive to reversibly attach the substrate to the backing layer and then to the treatment site.

[004] In embodiments, the substrate can comprise a "tab" to allow the user to remove the dressing from the backing layer or card. In embodiments, the tab can be reversibly attached to both the substrate as well as the backing layer, and used to remove the substrate from the backing layer. During application of the dressing to a treatment area, the tab can be removed.

[005] In embodiments, the backing layer covers the adhesive to maintain the adhesive's effectiveness prior to use and provide for more efficient storage. For example, an irregularly- shaped bandage can be associated via adhesive with a square or rectangular backing layer to provide a more efficiently-stored system.

[006] In embodiments the backing layer comprises a port or void exposing the multi-array matrix. The port or void can provide access to the multi-array matrix, for example to hydrate the matrix or apply a hydrogel or active agent or the like.

[007] Disclosed systems and devices can retain the ability to produce voltage and/or microcurrent at a treatment site for a longer period of time than conventional devices, for example through the use of a hydrogel. For example, in an embodiment the system or device comprises a dehydrated hydrogel, which can provide a conductive environment upon rehydration or reconstitution. Further, in certain embodiments the hydrogel helps to maintain a moist, conductive environment.

[008] Other embodiments provide methods of treating tissue, for example, the skin, a wound, or a muscle. Embodiments disclosed herein include treatment of a muscle or muscle group (for example a muscle group surrounding a joint), either before, during, or after athletic activity or exercise. For example, a method of treatment disclosed herein can comprise applying an embodiment disclosed herein to the area where treatment is desired.

[009] Embodiments disclosed herein can be used to treat irregular surfaces of the body, including the face, the shoulder, the elbow, the wrist, the finger joints, the hip, the knee, the ankle, the toe joints, etc. Additional embodiments disclosed herein can be used in areas where tissue is prone to movement, for example the eyelid, the ear, the lips, the nose, the shoulders, the back, etc.

[010] Still other embodiments provide methods for manufacturing application and storage systems comprising systems and devices capable of providing a low level micro-current to a treatment area. Disclosed methods can comprise applying a hydrogel to an array of microcells associated with or attached or dried to or bonded to a substrate and dehydrating the hydrogel.

[011] Disclosed embodiments can activate enzymes, increase glucose uptake, drive redox signaling, increase H ₂ 0 ₂ production, increase cellular protein sulfhydryl levels, and increase (IGF)-1 R phosphorylation. Embodiments can also up-regulate integrin production and accumulation in treatment areas.

BRIEF DESCRIPTION OF THE DRAWINGS

[012] FIG. 1 is a detailed plan view of a substrate comprising a multi-array matrix of an embodiment disclosed herein.

[013] FIG. 2 is a detailed plan view of a substrate comprising a pattern of applied electrical conductors in accordance with an embodiment disclosed herein.

[014] FIG. 3 is an embodiment using the applied pattern of FIG. 2.

[015] FIG. 4 is a cross-section of FIG. 3 through line 3-3.

[016] FIG. 5 is a detailed plan view of an alternate embodiment disclosed herein which includes fine lines of conductive metal solution connecting electrodes upon the base layer.

[017] FIG. 6 is a detailed plan view of another alternate embodiment having a line pattern and dot pattern.

[018] FIG. 7 is a detailed plan view of yet another alternate embodiment having two line patterns. [019] FIGs. 8A-8E depict alternate embodiments showing the location of discontinuous regions as well as anchor regions of the system.

[020] FIG. 9 depicts a detailed plan view of a substrate layer electrode pattern disclosed herein.

[021] FIG. 10 depicts a detailed plan view of a substrate layer electrode pattern disclosed herein.

[022] FIG. 1 1 depicts a detailed plan view of a substrate layer electrode pattern disclosed herein.

[023] FIG. 12 depicts an exploded view of a disclosed embodiment.

[024] FIG. 13 depicts an embodiment in use, with the user removing the substrate from the backing layer.

[025] FIG. 14 depicts void regions in a backing layer with a multi-array matrix visible.

DETAILED DESCRIPTION

[026] Described herein are systems, devices, and methods for storing and applying dressings for treating tissues, for example, organs such as skin or muscles, including skin conditions, wounds, and the like. For example, embodiments comprise a dressing system comprising a substrate comprising a multi-array matrix, and a backing layer or card with which the substrate is associated reversibly. In embodiments the backing layer or card can comprise a void or "cut-out."

[027] Embodiments disclosed herein comprise methods, systems and devices that can provide a low level electric field (LLEF) to a tissue or organism (thus a "LLEF system") or, when brought into contact with an electrically conducting material, can provide a low level electric micro-current (LLEC) to a tissue or organism (thus a "LLEC system"). Thus, in embodiments a LLEC system is a LLEF system that is in contact with an electrically conducting material, for example a liquid material. In certain embodiments, the microcurrent or electric field can be modulated, for example, to alter the duration, size, shape, field depth, duration, current, polarity, or voltage of the system. For example, it can be desirable to employ an electric field of greater strength or depth in an area to achieve optimal treatment. I n embodiments the watt-density of the system can be modulated. Embodiments can comprise a gel, for example a hydrogel.

[028] Definitions

[029] "Activation agent" as used herein means a composition useful for maintaining a moist environment within and about the skin. Activation agents can be in the form of gels (for example a hydrogel) or liquids. Activation agents can be conductive. Activation gels can also be antibacterial. In one embodiment, an activation agent can be a liquid such as sweat or topical substance such as petroleum jelly (for example with a conductive component added).

[030] "Affixing" as used herein can mean contacting a patient or tissue with a device or system disclosed herein. In embodiments "affixing" can comprise the use of straps, elastic, etc.

[031] "Antimicrobial agent" as used herein refers to an agent that kills or inhibits the growth of microorganisms. One type of antimicrobial agent can be an antibacterial agent. "Antibacterial agent" or "antibacterial" as used herein refers to an agent that interferes with the growth and reproduction of bacteria. Antibacterial agents are used to disinfect surfaces and eliminate potentially harmful bacteria. Unlike antibiotics, they are not used as medicines for humans or animals, but are found in products such as soaps, detergents, health and skincare products and household cleaners. Antibacterial agents may be divided into two groups according to their speed of action and residue production: The first group contains those that act rapidly to destroy bacteria, but quickly disappear (by evaporation or breakdown) and leave no active residue behind (referred to as non-residue-producing). Examples of this type are the alcohols, chlorine, peroxides, and aldehydes. The second group consists mostly of compounds that leave long-acting residues on the surface to be disinfected and thus have a prolonged action (referred to as residue-producing). Common examples of this group are triclosan, triclocarban, and benzalkonium chloride. Another type of antimicrobial agent can be an anti-fungal agent that can be used with the devices described herein.

[032] "Applied" or "apply" as used herein refers to contacting a substrate or surface with a conductive material, for example printing, painting, or spraying a conductive ink on a surface. Alternatively, "applying" can mean contacting a patient or tissue or organism with a device or system disclosed herein.

[033] "Backing layer" or "card" as used herein refers to a layer with which the substrate comprising the multi-array matrix is associated, for example reversibly associated using an adhesive. A backing layer can include a port or void area of an appropriate shape, for example, a square, a circle, a slit, etc.

[034] "Conductive material" as used herein refers to an object or type of material which permits the flow of electric charges. Conductive materials can comprise solids such as metals or carbon, or liquids such as conductive metal solutions and conductive gels. Conductive materials can be applied to form at least one matrix. Conductive liquids can dry, cure, or harden after application to form a solid material.

[035] "Discontinuous region" as used herein refers to a "void" in a substrate material such as a hole, slot, or the like. The term can mean any void in the material though typically the void is of a regular shape. A void in the material can be entirely within the perimeter of a material or it can extend to the perimeter of a material.

[036] "Dots" as used herein refers to discrete deposits of dissimilar reservoirs that can function as at least one battery cell. The term can refer to a deposit of any suitable size or shape, such as squares, circles, triangles, lines, etc. The term can be used synonymously with "electrodes," "microcells," "microspheres," etc. "Microspheres" refers to small spherical particles, with diameters in the micrometer range (typically 1 μηι to 3000 μηι (3 mm)). Microspheres are sometimes referred to as microparticles. Microspheres can be manufactured from various natural and synthetic materials. The term can be used synonymously with "micro-balloons," "beads," "particles," etc.

[037] "Electrode" refers to similar or dissimilar conductive materials. In embodiments utilizing an external power source the electrodes can comprise similar conductive materials. In embodiments that do not use an external power source, the electrodes can comprise dissimilar conductive materials that can define an anode and a cathode.

[038] "Expandable" as used herein refers to the ability to stretch while retaining structural integrity and not tearing. The term can refer to solid regions as well as discontinuous or void regions; solid regions as well as void regions can stretch or expand.

[039] "Matrix" or "matrices" or "array" or "arrays" as used herein refer to a pattern or patterns, such as those formed by electrodes on a surface, such as a fabric or a fiber, or the like. Matrices can also comprise a pattern or patterns within a solid or liquid material or a three dimensional object. Matrices can be designed to vary the electric field or electric current or microcurrent generated. For example, the strength and shape of the field or current or microcurrent can be altered, or the matrices can be designed to produce an electric field(s) or current or microcurrent of a desired strength or shape. "Matrices" can also refer to the random distribution of electrodes in a gel, such as a hydrogel.

[040] "Sheets" as used herein refer to substrate, typically in bulk quantities. As such, "sheets" can refer to a continuous roll or unit of substrate.

[041] "Stretchable" as used herein refers to the ability of embodiments that stretch without losing their structural integrity. That is, embodiments can stretch to accommodate irregular skin surfaces or surfaces wherein one portion of the surface can move relative to another portion.

[042] "Tab" as used herein refers to an area of the dressing or backing layer or substrate that provides the user means to remove the substrate from the backing layer. The tab can comprise a "tear-away" such that it is removable.

[043] "Treatment" as used herein can include the use of disclosed embodiments on tissue to prevent, reduce, or repair damage. Treatment can include use on an injury, for example a wound. [044] "Viscosity" as used herein refers to a measurement of a fluid's resistance to gradual deformation by shear stress or tensile stress.

[045] LLEC / LLEF Systems, Devices, and Methods of Manufacture

[046] In embodiments, disclosed methods, systems, and devices can comprise a backing layer or card with which the substrate comprising the multi-array matrix is associated. The substrate can be reversibly associated with the backing layer via, for example, an adhesive layer. The backing layer or card can comprise a void region or port. In embodiments the port can expose the multi-array matrix. This port can provide access to the multi-array matrix, for example to hydrate the matrix, to apply an active agent to the matrix, to apply a hydrogel to the matrix, or the like.

[047] In embodiments, the backing layer or card can be shaped to follow the outline of the dressing or substrate. For example, in embodiments the backing layer or card can be circular when used with round dressings. Alternatively, the backing layer or card can comprise a shape contrasting with that of the dressing or substrate. For example, in embodiments the backing layer or card can be square or rectangular when used with round dressings. In embodiments, the system is provided as a single card associated with a single substrate or dressing. In further embodiments, the system is provided as a single card associated with multiple substrates or dressings.

[048] The adhesive layer can, in embodiments, allow the substrate to be reversibly associated with an area where treatment is desired, for example a tissue, or the like. The adhesive layer can maintain the association between the substrate and the backing layer prior to application of the substrate to a treatment area, for example during storage periods.

[049] The backing layer or substrate can comprise at least one "tab" to allow the user to remove the dressing comprising the substrate from the backing layer or card.

[050] In embodiments, disclosed methods, systems, and devices can retain their ability to provide localized voltage and/or amperage at a treatment site for a sustained period of time. In embodiments, this sustained period of time can be achieved by including a hydrogel in or with the multi-array matrix of biocompatible microcells and dehydrating the hydrogel. Once dehydrated, the device can be stored without losing its ability to later deliver a localized voltage and/or amperage. The localized voltage and/or amperage can be triggered or activated by rehydrating the hydrogel as described herein.

[051] In embodiments, the herein-described methods, systems, and devices provide a multi-array matrix of biocompatible microcells coated or otherwise impregnated with a hydrogel, which can then be dried to remove the water in the hydrogel.

[052] The methods, systems, and devices described herein can comprise a multi-array matrix of biocompatible microcells that can produce a localized treatment voltage or microcurrent or both at a treatment site. In some embodiments, the voltage can be a low level electric field (LLEF). This electric filed can be delivered to a tissue or organism (thus a "LLEF system") or, when brought into contact with an electrically conducting material, can provide a low level electric micro-current (LLEC) to a tissue or organism (thus a "LLEC system"). Thus, in embodiments a LLEC system is a LLEF system that is in contact with an electrically conducting material, for example a liquid material. In certain embodiments, the micro-current or electric field can be modulated, for example, to alter the duration, size, shape, field depth, duration, current, polarity, or voltage of the system. In embodiments, the field is very short, for example in the range of physiologic electric fields. In some embodiments, the direction of the electric field produced by devices disclosed herein is omnidirectional over the surface of the wound and more in line with the physiologic electric fields.

[053] In some embodiments, the multi-array matrix of biocompatible microcells can comprise a first array comprising a pattern of microcells formed of a conductive material and a second array comprising a pattern of microcells formed from a second conductive material. The first conductive material can be formed from, for example, a first conductive solution and the second conductive material can be formed from, for example, a second conductive solution. The first and/or second conductive solutions can include a metal species such as a metal species capable of defining at least one voltaic cell for spontaneously generating at least one electrical current with the metal species of the first array when said first and second arrays are introduced to an electrolytic solution and said first and second arrays are not in physical contact with each other. Certain embodiments utilize an external power source such as AC or DC power, or pulsed RF, or pulsed current, such as high voltage pulsed current. In one embodiment, the electrical energy is derived from the dissimilar metals creating a battery at each electrode/electrode interface, whereas those embodiments with an external power source can employ conductive electrodes in a spaced configuration to predetermine the electric field shape and strength.

[054] In certain embodiments, for example treatment methods, it can be preferable to utilize AC or DC current. For example, embodiments disclosed herein can employ phased array, pulsed, square wave, sinusoidal, or other wave forms, combinations, or the like. Certain embodiments utilize a controller to produce and control power production and/or distribution to the device.

[055] Embodiments of the LLEC or LLEF methods, systems, and devices disclosed herein can comprise electrodes or microcells. Electrodes or microcells can comprise discrete deposits of dissimilar reservoirs that can function as at least one battery cell. The deposits can be of any suitable size or shape, such as squares, circles, triangles, lines, etc. In some embodiments, "dots" can be used synonymously with, "microcells," and the like. Each electrode or microcell can be or comprise a conductive material, for example, a metal. In embodiments, the electrodes or microcells can comprise any electrically-conductive material, for example, an electrically conductive hydrogel, metals, electrolytes, superconductors, semiconductors, plasmas, and nonmetallic conductors such as graphite and conductive polymers. Electrically conductive metals can include, for example, silver, copper, gold, aluminum, molybdenum, zinc, lithium, tungsten, brass, carbon, nickel, iron, palladium, platinum, tin, bronze, carbon steel, lead, titanium, stainless steel, mercury, Fe/Cr alloys, and the like. The electrode can be coated or plated with a different metal such as aluminum, gold, platinum or silver.

[056] Turning to the figures, in FIG. 1 , the dissimilar first electrode 6 and second electrode 10 are applied onto base layer or substrate 2 of an article 4, for example a fabric. In an embodiment a primary surface is a surface of a LLEC or LLEF system that comes into direct contact with an area to be treated, for example a skin surface.

[057] In various embodiments the difference of the standard potentials of the electrodes or dots or reservoirs can be in a range from about 0.05 V to approximately about 5.0 V. For example, the standard potential can be about 0.05 V, about 0.06 V, about 0.07 V, about 0.08 V, about 0.09 V, about 0.1 V, about 0.2 V, about 0.3 V, about 0.4 V, about 0.5 V, about 0.6 V, about 0.7 V, about 0.8 V, about 0.9 V, about 1 .0 V, about 1 .1 V, about 1 .2 V, about 1 .3 V, about 1 .4 V, about 1 .5 V, about 1 .6 V, about 1 .7 V, about 1 .8 V, about 1 .9 V, about 2.0 V, about 2.1 V, about 2.2 V, about 2.3 V, about 2.4 V, about 2.5 V, about 2.6 V, about 2.7 V, about 2.8 V, about 2.9 V, about 3.0 V, about 3.1 V, about 3.2 V, about 3.3 V, about 3.4 V, about 3.5 V, about 3.6 V, about 3.7 V, about 3.8 V, about 3.9 V, about 4.0 V, about 4.1 V, about 4.2 V, about 4.3 V, about 4.4 V, about 4.5 V, about 4.6 V, about 4.7 V, about 4.8 V, about 4.9 V, about 5.0 V, about 5.1 V, about 5.2 V, about 5.3 V, about 5.4 V, about 5.5 V, about 5.6 V, about 5.7 V, about 5.8 V, about 5.9 V, about 6.0 V, about 6.1 V, about 6.2 V, about 6.3 V, about 6.4 V, about 6.5 V, about 6.6 V, about 6.7 V, about 6.8 V, about 6.9 V, about 7.0 V, about 7.1 V, about 7.2 V, about 7.3 V, about 7.4 V, about 7.5 V, about 7.6 V, about 7.7 V, about 7.8 V, about 7.9 V, about 8.0 V, about 8.1 V, about 8.2 V, about 8.3 V, about 8.4 V, about 8.5 V, about 8.6 V, about 8.7 V, about 8.8 V, about 8.9 V, about 9.0 V, or the like.

[058] In other embodiments the difference of the standard potentials of electrodes or dots or reservoirs can be at least 0.05 V, at least 0.06 V, at least 0.07 V, at least 0.08 V, at least 0.09 V, at least 0.1 V, at least 0.2 V, at least 0.3 V, at least 0.4 V, at least 0.5 V, at least 0.6 V, at least 0.7 V, at least 0.8 V, at least 0.9 V, at least 1 .0 V, at least 1 .1 V, at least 1 .2 V, at least 1 .3 V, at least 1 .4 V, at least 1 .5 V, at least 1 .6 V, at least 1 .7 V, at least 1 .8 V, at least 1.9 V, at least 2.0 V, at least 2.1 V, at least 2.2 V, at least 2.3 V, at least 2.4 V, at least 2.5 V, at least 2.6 V, at least 2.7 V, at least 2.8 V, at least 2.9 V, at least 3.0 V, at least 3.1 V, at least 3.2 V, at least 3.3 V, at least 3.4 V, at least 3.5 V, at least 3.6 V, at least 3.7 V, at least 3.8 V, at least 3.9 V, at least 4.0 V, at least 4.1 V, at least 4.2 V, at least 4.3 V, at least 4.4 V, at least 4.5 V, at least 4.6 V, at least 4.7 V, at least 4.8 V, at least 4.9 V, at least 5.0 V, or the like, once the device or system is rehydrated for use from a dehydrated state.

[059] In still other embodiments, the difference of the standard potentials of electrodes or dots or reservoirs can be less than 0.05 V, less than 0.06 V, less than 0.07 V, less than 0.08 V, less than 0.09 V, less than 0.1 V, less than 0.2 V, less than 0.3 V, less than 0.4 V, less than 0.5 V, less than 0.6 V, less than 0.7 V, less than 0.8 V, less than 0.9 V, less than 1 .0 V, less than 1 .1 V, less than 1.2 V, less than 1 .3 V, less than 1 .4 V, less than 1 .5 V, less than

1.6 V, less than 1 .7 V, less than 1.8 V, less than 1 .9 V, less than 2.0 V, less than 2.1 V, less than 2.2 V, less than 2.3 V, less than 2.4 V, less than 2.5 V, less than 2.6 V, less than 2.7 V, less than 2.8 V, less than 2.9 V, less than 3.0 V, less than 3.1 V, less than 3.2 V, less than 3.3 V, less than 3.4 V, less than 3.5 V, less than 3.6 V, less than 3.7 V, less than 3.8 V, less than 3.9 V, less than 4.0 V, less than 4.1 V, less than 4.2 V, less than 4.3 V, less than 4.4 V, less than 4.5 V, less than 4.6 V, less e than 4.7 V, less than 4.8 V, less than 4.9 V, less than 5.0 V, or the like, once the device or system is rehydrated for use from a dehydrated state.

[060] In embodiments, systems and devices disclosed herein can produce a LLEC of between for example about 1 and about 200 micro-amperes, between about 10 and about 190 micro-amperes, between about 20 and about 180 micro-amperes, between about 30 and about 170 micro-amperes, between about 40 and about 160 micro-amperes, between about 50 and about 150 micro-amperes, between about 60 and about 140 micro-amperes, between about 70 and about 130 micro-amperes, between about 80 and about 120 microamperes, between about 90 and about 100 micro-amperes, between about 100 and about 150 micro-amperes, between about 150 and about 200 micro-amperes, between about 200 and about 250 micro-amperes, between about 250 and about 300 micro-amperes, between about 300 and about 350 micro-amperes, between about 350 and about 400 micro-amperes, between about 400 and about 450 micro-amperes, between about 450 and about 500 microamperes, between about 500 and about 550 micro-amperes, between about 550 and about 600 micro-amperes, between about 600 and about 650 micro-amperes, between about 650 and about 700 micro-amperes, between about 700 and about 750 micro-amperes, between about 750 and about 800 micro-amperes, between about 800 and about 850 micro-amperes, between about 850 and about 900 micro-amperes, between about 900 and about 950 microamperes, between about 950 and about 1000 micro-amperes (1 milli-amp [mA]), between about 1 .0 and about 1 .1 mA, between about 1 .1 and about 1 .2mA, between about 1 .2 and about 1 .3mA, between about 1.3 and about 1 .4mA, between about 1 .4 and about 1.5mA, between about 1 .5 and about 1 .6mA, between about 1 .6 and about 1 .7mA, between about

1.7 and about 1 .8mA, between about 1 .8 and about 1 .9mA, between about 1 .9 and about 2.0mA, between about 2.0 and about 2.1 mA, between about 2.1 and about 2.2mA, between about 2.2 and about 2.3mA, between about 2.3 and about 2.4mA, between about 2.4 and about 2.5mA, between about 2.5 and about 2.6mA, between about 2.6 and about 2.7mA, between about 2.7 and about 2.8mA, between about 2.8 and about 2.9mA, between about 2.9 and about 3.0mA, between about 3.0 and about 3.1 mA, between about 3.1 and about 3.2mA, between about 3.2 and about 3.3mA, between about 3.3 and about 3.4mA, between about 3.4 and about 3.5mA, between about 3.5 and about 3.6mA, between about 3.6 and about 3.7mA, between about 3.7 and about 3.8mA, between about 3.8 and about 3.9mA, between about 3.9 and about 4.0mA, between about 4.0 and about 4.1 mA, between about 4.1 and about 4.2mA, between about 4.2 and about 4.3mA, between about 4.3 and about 4.4mA, between about 4.4 and about 4.5mA, between about 4.5 and about 5.0mA, between about 5.0 and about 5.5mA, between about 5.5 and about 6.0mA, between about 6.0 and about 6.5mA, between about 6.5 and about 7.0mA, between about 7.5 and about 8.0mA, between about 8.0 and about 8.5mA, between about 8.5 and about 9.0mA, between about 9.0 and about 9.5mA, between about 9.5 and about 10.0mA, between about 10.0 and about 10.5mA, between about 10.5 and about 1 1 .0mA, between about 1 1 .0 and about 1 1 .5mA, between about 1 1 .5 and about 12.0mA, between about 12.0 and about 12.5mA, between about 12.5 and about 13.0mA, between about 13.0 and about 13.5mA, between about 13.5 and about 14.0mA, between about 14.0 and about 14.5mA, between about 14.5 and about 15.0mA, or the like.

[061] In embodiments, systems and devices disclosed herein can produce a LLEC of between for example about 1 and about 400 micro-amperes, between about 20 and about 380 micro-amperes, between about 40 and about 360 micro-amperes, between about 60 and about 340 micro-amperes, between about 80 and about 320 micro-amperes, between about 100 and about 300 micro-amperes, between about 120 and about 280 micro-amperes, between about 140 and about 260 micro-amperes, between about 160 and about 240 microamperes, between about 180 and about 220 micro-amperes, or the like.

[062] In embodiments, systems and devices disclosed herein can produce a LLEC of between for example about 1 micro-ampere and about 1 milli-ampere, between about 50 and about 800 micro-amperes, between about 200 and about 600 micro-amperes, between about 400 and about 500 micro-amperes, or the like.

[063] In embodiments, systems and devices disclosed herein can produce a LLEC of about 10 micro-amperes, about 20 micro-amperes, about 30 micro-amperes, about 40 microamperes, about 50 micro-amperes, about 60 micro-amperes, about 70 micro-amperes, about 80 micro-amperes, about 90 micro-amperes, about 100 micro-amperes, about 1 10 micro-amperes, about 120 micro-amperes, about 130 micro-amperes, about 140 microamperes, about 150 micro-amperes, about 160 micro-amperes, about 170 micro-amperes, about 180 micro-amperes, about 190 micro-amperes, about 200 micro-amperes, about 210 micro-amperes, about 220 micro-amperes, about 240 micro-amperes, about 260 microamperes, about 280 micro-amperes, about 300 micro-amperes, about 320 micro-amperes, about 340 micro-amperes, about 360 micro-amperes, about 380 micro-amperes, about 400 micro-amperes, about 450 micro-amperes, about 500 micro-amperes, about 550 microamperes, about 600 micro-amperes, about 650 micro-amperes, about 700 micro-amperes, about 750 micro-amperes, about 800 micro-amperes, about 850 micro-amperes, about 900 micro-amperes, about 950 micro-amperes, about 1 milli-ampere (mA), about 1 .1 mA, about 1.2mA, about 1 .3mA, about 1 .4mA, about 1 .5mA, about 1 .6mA, about 1 .7mA, about 1 .8mA, about 1 .9mA, about 2.0mA, about 2.1 mA, about 2.2mA, about 2.3mA, about 2.4mA, about 2.5mA, about 2.6mA, about 2.7mA, about 2.8mA, about 2.9mA, about 3.0mA, about 3.1 mA, about 3.2mA, about 3.3mA, about 3.4mA, about 3.5mA, about 3.6mA, about 3.7mA, about 3.8mA, about 3.9mA, about 4.0mA, about 4.1 mA, about 4.2mA, about 4.3mA, about 4.4mA, about 4.5mA, about 4.6mA, about 4.7mA, about 4.8mA, about 4.9mA, about 5.0mA, about 5.1 mA, about 5.2mA, about 5.3mA, about 5.4mA, about 5.5mA, about 5.6mA, about 5.7mA, about 5.8mA, about 5.9mA, about 6.0mA, about 6.1 mA, about 4.2mA, about 6.3mA, about 6.4mA, about 6.5mA, about 6.6mA, about 6.7mA, about 6.8mA, about 6.9mA, about 7.0mA, about 7.1 mA, about 7.2mA, about 7.3mA, about 7.4mA, about 7.5mA, about 7.6mA, about 7.7mA, about 7.8mA, about 7.9mA, about 8.0mA, about 8.1 mA, about 8.2mA, about 8.3mA, about 8.4mA, about 8.5mA, about 8.6mA, about 8.7mA, about 8.8mA, about 8.9mA, about 9.0mA, about 9.1 mA, about 9.2mA, about 9.3mA, about 9.4mA, about 9.5mA, about 9.6mA, about 9.7mA, about 9.8mA, about 9.9mA, about 10.0mA, about 10.1 mA, about 10.2mA, about 10.3mA, about 10.4mA, about 10.5mA, about 10.6mA, about 10.7mA, about 10.8mA, about 10.9mA, about 1 1 .0mA, about 1 1 .1 mA, about 1 1 .2mA, about 1 1.3mA, about 1 1 .4mA, about 1 1 .5mA, about 1 1 .6mA, about 1 1 .7mA, about 1 1 .8mA, about 1 1.9mA, about 12.0mA, about 12.1 mA, about 12.2mA, about 12.3mA, about 12.4mA, about 12.5mA, about 12.6mA, about 12.7mA, about 12.8mA, about 12.9mA, about 13.0mA, about 13.1 mA, about 13.2mA, about 13.3mA, about 13.4mA, about 13.5mA, about 13.6mA, about 13.7mA, about 13.8mA, about 13.9mA, about 14.0mA, about 14.1 mA, about 14.2mA, about 14.3mA, about 14.4mA, about 14.5mA, about 14.6mA, about 14.7mA, about 14.8mA, about 14.9mA, about 15.0mA, about 15.1 mA, about 15.2mA, about 15.3mA, about 15.4mA, about 15.5mA, about 15.6mA, about 15.7mA, about 15.8mA, or the like.

[064] In embodiments, the disclosed systems and devices can produce a LLEC of not more than about 10 micro-amperes, or not more than about 20 micro-amperes, not more than about 30 micro-amperes, not more than about 40 micro-amperes, not more than about 50 micro-amperes, not more than about 60 micro-amperes, not more than about 70 microamperes, not more than about 80 micro-amperes, not more than about 90 micro-amperes, not more than about 100 micro-amperes, not more than about 1 10 micro-amperes, not more than about 120 micro-amperes, not more than about 130 micro-amperes, not more than about 140 micro-amperes, not more than about 150 micro-amperes, not more than about 160 micro-amperes, not more than about 170 micro-amperes, not more than about 180 micro-amperes, not more than about 190 micro-amperes, not more than about 200 microamperes, not more than about 210 micro-amperes, not more than about 220 micro-amperes, not more than about 230 micro-amperes, not more than about 240 micro-amperes, not more than about 250 micro-amperes, not more than about 260 micro-amperes, not more than about 270 micro-amperes, not more than about 280 micro-amperes, not more than about 290 micro-amperes, not more than about 300 micro-amperes, not more than about 310 micro-amperes, not more than about 320 micro-amperes, not more than about 340 microamperes, not more than about 360 micro-amperes, not more than about 380 micro-amperes, not more than about 400 micro-amperes, not more than about 420 micro-amperes, not more than about 440 micro-amperes, not more than about 460 micro-amperes, not more than about 480 micro-amperes, not more than about 500 micro-amperes, not more than about 520 micro-amperes, not more than about 540 micro-amperes, not more than about 560 micro-amperes, not more than about 580 micro-amperes, not more than about 600 microamperes, not more than about 620 micro-amperes, not more than about 640 micro-amperes, not more than about 660 micro-amperes, not more than about 680 micro-amperes, not more than about 700 micro-amperes, not more than about 720 micro-amperes, not more than about 740 micro-amperes, not more than about 760 micro-amperes, not more than about 780 micro-amperes, not more than about 800 micro-amperes, not more than about 820 micro-amperes, not more than about 840 micro-amperes, not more than about 860 microamperes, not more than about 880 micro-amperes, not more than about 900 micro-amperes, not more than about 920 micro-amperes, not more than about 940 micro-amperes, not more than about 960 micro-amperes, not more than about 980 micro-amperes, not more than about 1 milli-ampere (mA), not more than about 1.1 mA, not more than about 1.2mA, not more than about 1.3mA, not more than about 1.4mA, not more than about 1.5mA, not more than about 1.6mA, not more than about 1.7mA, not more than about 1.8mA, not more than about 1.9mA, not more than about 2.0mA, not more than about 2.1 mA, not more than about 2.2mA, not more than about 2.3mA, not more than about 2.4mA, not more than about 2.5mA, not more than about 2.6mA, not more than about 2.7mA, not more than about 2.8mA, not more than about 2.9mA, not more than about 3.0mA, not more than about 3.1 mA, not more than about 3.2mA, not more than about 3.3mA, not more than about 3.4mA, not more than about 3.5mA, not more than about 3.6mA, not more than about 3.7mA, not more than about 3.8mA, not more than about 3.9mA, not more than about 4.0mA, not more than about 4.1 mA, not more than about 4.2mA, not more than about 4.3mA, not more than about 4.4mA, not more than about 4.5mA, not more than about 4.6mA, not more han about 4.7mA, not more han about 4.8mA, not more than about 4.9mA, not more han about 5.0mA, not more han about 5.1 mA, not more than about 5.2mA, not more han about 5.3mA, not more han about 5.4mA, not more than about 5.5mA, not more han about 5.6mA, not more han about 5.7mA, not more than about 5.8mA, not more han about 5.9mA, not more han about 6.0mA, not more than about 6.1 mA, not more han about 4.2mA, not more han about 6.3mA, not more than about 6.4mA, not more han about 6.5mA, not more han about 6.6mA, not more than about 6.7mA, not more han about 6.8mA, not more han about 6.9mA, not more than about 7.0mA, not more han about 7.1 mA, not more han about 7.2mA, not more than about 7.3mA, not more han about 7.4mA, not more han about 7.5mA, not more than about 7.6mA, not more han about 7.7mA, not more han about 7.8mA, not more than about 7.9mA, not more han about 8.0mA, not more han about 8.1 mA, not more than about 8.2mA, not more han about 8.3mA, not more han about 8.4mA, not more than about 8.5mA, not more han about 8.6mA, not more han about 8.7mA, not more than about 8.8mA, not more han about 8.9mA, not more han about 9.0mA, not more than about 9.1 mA, not more han about 9.2mA, not more han about 9.3mA, not more than about 9.4mA, not more han about 9.5mA, not more han about 9.6mA, not more than about 9.7mA, not more han about 9.8mA, not more han about 9.9mA, not more than about 10.0mA, not more than about 10.1 mA not more than about 10.2mA, not more than about 10.3mA, not more than about 10.4mA not more than about 10.5mA, not more than about 10.6mA, not more than about 10.7mA not more than about 10.8mA, not more than about 10.9mA, not more than about 1 1.0mA not more than about 1 1.1 mA, not more than about 11.2mA, not more than about 1 1.3mA not more than about 1 1.4mA, not more than about 11.5mA, not more than about 1 1.6mA not more than about 1 1.7mA, not more than about 11.8mA, not more than about 1 1.9mA not more than about 12.0mA, not more than about 12.1 mA, not more than about 12.2mA not more than about 12.3mA, not more than about 12.4mA, not more than about 12.5mA not more than about 12.6mA, not more than about 12.7mA, not more than about 12.8mA not more than about 12.9mA, not more than about 13.0mA, not more than about 13.1 mA not more than about 13.2mA, not more than about 13.3mA, not more than about 13.4mA not more than about 13.5mA, not more than about 13.6mA, not more than about 13.7mA not more than about 13.8mA, not more than about 13.9mA, not more than about 14.0mA not more than about 14.1 mA, not more than about 14.2mA, not more than about 14.3mA not more than about 14.4mA, not more than about 14.5mA, not more than about 14.6mA not more than about 14.7mA, not more than about 14.8mA, not more than about 14.9mA not more than about 15.0mA, not more than about 15.1 mA, not more than about 15.2mA not more than about 15.3mA, not more than about 15.4mA, not more than about 15.5mA, not more than about 15.6mA, not more than about 15.7mA, not more than about 15.8mA, and the like.

[065] In embodiments, systems and devices disclosed herein can produce a LLEC of not less than 10 micro-amperes, not less than 20 micro-amperes, not less than 30 microamperes, not less than 40 micro-amperes, not less than 50 micro-amperes, not less than 60 micro-amperes, not less than 70 micro-amperes, not less than 80 micro-amperes, not less than 90 micro-amperes, not less than 100 micro-amperes, not less than 1 10 micro-amperes, not less than 120 micro-amperes, not less than 130 micro-amperes, not less than 140 microamperes, not less than 150 micro-amperes, not less than 160 micro-amperes, not less than 170 micro-amperes, not less than 180 micro-amperes, not less than 190 micro-amperes, not less than 200 micro-amperes, not less than 210 micro-amperes, not less than 220 microamperes, not less than 230 micro-amperes, not less than 240 micro-amperes, not less than 250 micro-amperes, not less than 260 micro-amperes, not less than 270 micro-amperes, not less than 280 micro-amperes, not less than 290 micro-amperes, not less than 300 microamperes, not less than 310 micro-amperes, not less than 320 micro-amperes, not less than 330 micro-amperes, not less than 340 micro-amperes, not less than 350 micro-amperes, not less than 360 micro-amperes, not less than 370 micro-amperes, not less than 380 microamperes, not less than 390 micro-amperes, not less than 400 micro-amperes, not less than about 420 micro-amperes, not less than about 440 micro-amperes, not less than about 460 micro-amperes, not less than about 480 micro-amperes, not less than about 500 microamperes, not less than about 520 micro-amperes, not less than about 540 micro-amperes, not less than about 560 micro-amperes, not less than about 580 micro-amperes, not less than about 600 micro-amperes, not less than about 620 micro-amperes, not less than about 640 micro-amperes, not less than about 660 micro-amperes, not less than about 680 microamperes, not less than about 700 micro-amperes, not less than about 720 micro-amperes, not less than about 740 micro-amperes, not less than about 760 micro-amperes, not less than about 780 micro-amperes, not less than about 800 micro-amperes, not less than about 820 micro-amperes, not less than about 840 micro-amperes, not less than about 860 microamperes, not less than about 880 micro-amperes, not less than about 900 micro-amperes, not less than about 920 micro-amperes, not less than about 940 micro-amperes, not less than about 960 micro-amperes, not less than about 980 micro-amperes, not less than about 1 milli-ampere (mA), not less than about 1.1 mA, not less than about 1 .2mA, not less than about 1 .3mA, not less than about 1 .4mA, not less than about 1 .5mA, not less than about 1.6mA, not less than about 1 .7mA, not less than about 1 .8mA, not less than about 1.9mA, not less than about 2.0mA, not less than about 2.1 mA, not less than about 2.2mA, not less than about 2.3mA, not less than about 2.4mA, not less than about 2.5mA, not less than about 2.6mA, not less than about 2.7mA, not less than about 2.8mA, not less than about 2.9mA, not less than about 3.0mA, not less than about 3.1 mA, not less than about 3.2mA, not less than about 3.3mA, not less than about 3.4mA, not less than about 3.5mA, not less than about 3.6mA, not less than about 3.7mA, not less than about 3.8mA, not less than about 3.9mA, not less than about 4.0mA, not less than about 4.1 mA, not less than about 4.2mA, not less than about 4.3mA, not less than about 4.4mA, not less than about 4.5mA, not less than about 4.6mA, not less than about 4.7mA, not less than about 4.8mA, not less than about 4.9mA, not less than about 5.0mA, not less than about 5.1 mA, not less than about 5.2mA, not less than about 5.3mA, not less than about 5.4mA, not less than about 5.5mA, not less than about 5.6mA, not less than about 5.7mA, not less than about 5.8mA, not less than about 5.9mA, not less than about 6.0mA, not less than about 6.1 mA, not less than about 4.2mA, not less than about 6.3mA, not less than about 6.4mA, not less than about 6.5mA, not less than about 6.6mA, not less than about 6.7mA, not less than about 6.8mA, not less than about 6.9mA, not less than about 7.0mA, not less than about 7.1 mA, not less than about 7.2mA, not less than about 7.3mA, not less than about 7.4mA, not less than about 7.5mA, not less than about 7.6mA, not less than about 7.7mA, not less than about 7.8mA, not less than about 7.9mA, not less than about 8.0mA, not less than about 8.1 mA, not less than about 8.2mA, not less than about 8.3mA, not less than about 8.4mA, not less than about 8.5mA, not less than about 8.6mA, not less than about 8.7mA, not less than about 8.8mA, not less than about 8.9mA, not less than about 9.0mA, not less than about 9.1 mA, not less than about 9.2mA, not less than about 9.3mA, not less than about 9.4mA, not less than about 9.5mA, not less than about 9.6mA, not less than about 9.7mA, not less than about 9.8mA, not less than about 9.9mA, not less than about 10.0mA, not less than about 10.1 mA, not less than about 10.2mA, not less than about 10.3mA, not less than about 10.4mA, not less than about 10.5mA, not less than about 10.6mA, not less than about

10.7mA, not less than about 10.8mA, not less than about 10.9mA, not less than about

11.0mA, not less than about 1 1.1 mA, not less than about 1 1.2mA, not less than about

11.3mA, not less than about 1 1.4mA, not less than about 1 1.5mA, not less than about

11.6mA, not less than about 1 1.7mA, not less than about 1 1.8mA, not less than about

11.9mA, not less than about 12.0mA, not less than about 12.1 mA, not less than about

12.2mA, not less than about 12.3mA, not less than about 12.4mA, not less than about

12.5mA, not less than about 12.6mA, not less than about 12.7mA, not less than about

12.8mA, not less than about 12.9mA, not less than about 13.0mA, not less than about

13.1 mA, not less than about 13.2mA, not less than about 13.3mA, not less than about

13.4mA, not less than about 13.5mA, not less than about 13.6mA, not less than about

13.7mA, not less than about 13.8mA, not less than about 13.9mA, not less than about

14.0mA, not less than about 14.1 mA, not less than about 14.2mA, not less than about

14.3mA, not less than about 14.4mA, not less than about 14.5mA, not less than about 14.6mA, not less than about 14.7mA, not less than about 14.8mA, not less than about 14.9mA, not less than about 15.0mA, not less than about 15.1 mA, not less than about 15.2mA, not less than about 15.3mA, not less than about 15.4mA, not less than about 15.5mA, not less than about 15.6mA, not less than about 15.7mA, not less than about 15.8mA, and the like.

[066] In some embodiments the electrodes or microcells can comprise a clear conductive material. For example, in certain embodiments indium tin oxide (ITO) can be used. In other embodiments other transparent conductive oxides (TCOs), conductive polymers, metal grids, carbon nanotubes, graphene, and nanowire thin films can be employed.

[067] In certain embodiments, array, reservoir or electrode geometry can comprise shapes including circles, polygons, lines, zigzags, ovals, stars, or any suitable variety. This provides the ability to design/customize surface electric field shapes as well as depth of penetration. For example, in embodiments it can be desirable to employ an electric field of greater strength or depth to achieve optimal treatment.

[068] Reservoir or electrode or dot sizes and concentrations can vary, as these variations can allow for changes in the properties of the electric field created by embodiments of the invention. Certain embodiments provide an electric field at about, for example, 0.5-5.0 V at the device surface under normal tissue loads with resistance of 100 to 100K ohms.

[069] In embodiments, disclosed devices can provide an electric field of greater than physiological strength, for example to a depth of, for example, at least 1 mm, at least 2 mm, at least 3 mm, at least 4 mm, at least 5 mm, at least 6 mm, at least 7 mm, at least 8 mm, at least 9 mm, at least 10 mm, at least 1 1 mm, at least 12 mm, at least 13 mm, at least 14 mm, at least 15 mm, at least 16 mm, at least 17 mm, at least 18 mm, at least 19 mm, at least 20 mm, at least 21 mm, at least 22 mm, at least 23 mm, at least 24 mm, at least 25 mm, at least 26 mm, at least 27 mm, at least 28 mm, at least 29 mm, at least 30 mm, at least 31 mm, at least 32 mm, at least 33 mm, at least 34 mm, at least 35 mm, at least 36 mm, at least 37 mm, at least 38 mm, at least 39 mm, at least 40 mm, or more.

[070] In various embodiments dissimilar metals can be used to create a customized electric field with a desired voltage or microcurrent. In certain embodiments the pattern of reservoirs can control the watt density and shape of the electric field. For example. In embodiments it can be desirable to employ an electric field of greater strength or depth in an area, for example where skin is thicker to achieve optimal treatment.

[071] In embodiments devices disclosed herein the electric field or current or both applied to a tissue can be designed or produced or adjusted based upon feedback from the tissue or upon an algorithm within sensors operably connected to the embodiment and a control module. The electric field, electric current, or both can be stronger in one zone and weaker in another. The electric field, electric current, or both can change with time and be modulated based on treatment goals or feedback from the tissue or patient. The control module can monitor and adjust the size, strength, density, shape, or duration of electric field or electric current based on tissue parameters. For example, embodiments disclosed herein can produce and maintain localized electrical events. For example, embodiments disclosed herein can produce specific values for the electric field duration, electric field size, electric field shape, field depth, current, polarity, and/or voltage of the device or system.

[072] As described, the disclosed systems and devices can include a base layer. The base layer can be useful in reducing the amount of motion between tissue and device and/or can be a substrate for the multi-array matrix of biocompatible microcells. In some embodiments, the base layer can be elastic. In other embodiments, the base layer or the substrate can include components such as straps to maintain or help maintain its position. In some embodiments, the base layer or substrate can comprise a strap on either end of the long axis, or a strap linking on end of the long axis to the other. The straps can comprise velcro, snaps, or a similar fastening system. In further embodiments the strap can comprise a conductive material, for example a wire to electrically link the device with other components, such as monitoring equipment or a power source.

[073] In further embodiments the strap can comprise a conductive material, for example a wire, to electrically link the device with other components, such as monitoring equipment or a power source. In embodiments the device can be wirelessly linked to monitoring or data collection equipment, for example linked via Bluetooth to a cell phone or computer that collects data from the device. In certain embodiments the device can comprise data collection means, such as temperature, pH, pressure, or conductivity data collection means. In certain embodiments, disclosed devices and systems can comprise data collection means, such as temperature, pH, pressure, or conductivity data collection means. Embodiments can comprise a display, for example to visually present, for example, the temperature, pH, pressure, or conductivity data to a user. Embodiments can include, for example, tracking equipment so as to track and/or quantify a user's movements or performance. Embodiments can include, for example, an accelerometer, so as to measure acceleration or impact forces on a user.

[074] If an elastic material is used in the base layer or substrate, it can comprise an elastic film with elasticity, for example, similar to that of skin, or greater than that of skin, or less than that of skin. In embodiments, the LLEC or LLEF system can comprise a laminate where layers of the laminate can be of varying elasticities. For example, an outer layer may be highly elastic and an inner layer in-elastic or less elastic. In embodiments, a layer can be made to stretch by placing stress relieving discontinuous regions or slits through the thickness of the material so there is a mechanical displacement rather than stress that would break the fabric weave before stretching would occur. In embodiments the slits can extend completely through a layer or the system or can be placed where expansion is required. In embodiments of the system the slits do not extend all the way through the system or a portion of the system such as the substrate. In embodiments the discontinuous regions can pass halfway through the long axis of the substrate.

[075] In embodiments the substrate can be shaped to fit an area of desired use or treatment. For example, in embodiments the device can be shaped to fit the area around the eye or the eye itself, to treat, for example, a corneal injury. In embodiments the device can be shaped to fit the area around the eye to be used prior to or following surgery, for example blepharoplasty.

[076] In some embodiments, the substrate can be a bandage. When provided as a bandage, the bandage can include any or all of the features described herein.

[077] In some embodiments, the substrate can comprise a fabric, a fiber, or the like. In embodiments the substrate can be pliable, for example to better follow the contours of an area to be treated, such as the face or back. In embodiments the substrate can comprise a gauze or mesh or plastic. Suitable types of pliable substrates for use in embodiments disclosed herein can be absorbent or non-absorbent textiles, low-adhesives, vapor permeable films, hydrocolloids, alginates, foams, foam-based materials, cellulose-based materials including Kettenbach fibers, hollow tubes, fibrous materials, such as those impregnated with anhydrous/hygroscopic materials, beads and the like, or any suitable material as known in the art.

[078] In some embodiments, the substrate can comprise "anchor" regions or "arms" or straps to affix the system securely. For example, a system or device as described herein can be secured to or around a curved surface, and anchor regions of the substrate can extend to areas of minimal stress or movement to securely affix the system in place. In embodiments, the backing layer can be designed to accommodate substrates of a particular shape or size.

[079] In some embodiments, the backing layer or card can comprise, for example, cardboard, a fabric, a fiber, or the like. In embodiments the backing layer or card can be pliable. Suitable types of pliable backing layers or cards for use in embodiments disclosed herein can be absorbent or non-absorbent textiles, low-adhesives, vapor permeable films, hydrocolloids, alginates, foams, foam-based materials, cellulose-based materials including Kettenbach fibers, hollow tubes, fibrous materials, such as those impregnated with anhydrous/hygroscopic materials, beads and the like, or any suitable material as known in the art.

[080] As described, the devices described herein can comprise at least one hydrogel that coats or otherwise impregnates the multi-array matrix of biocompatible microcells of the device. A hydrogel as described herein can include any hydrogel known in the art that can provide rehydration characteristics that allow bioelectric devices as described herein to function as if the hydrogel were not present or substantially as if the hydrogel were not present, yet keep the microcell batteries activated for an extended time as if an amorphous hydrogel were applied at time of use. In embodiments, the hydrogel can coat the matrix present on the base layer or substrate. In further embodiments, the hydrogel can comprise the matrix.

[081] Further, the hydrogel can function to retain or "lock" the eventual rehydration voltages and/or amperages that provide localized treatment.

[082] Suitable hydrogels can include, but are not limited to polyvinyl alcohol, sodium polyacrylate, acrylate based polymers, glycolated polymers, cellulose, glycerol, sugars, agarose, methylcellulose, hyaluronan, other naturally derived polymers, and combinations thereof.

[083] A hydrogel can be configured in a variety of viscosities. Viscosity is a measurement of a fluid or material's resistance to gradual deformation by shear stress or tensile stress. In embodiments the electrical field can be extended through a semi-liquid hydrogel with a low viscosity. In other embodiments the electrical field can be extended through a solid hydrogel with a high viscosity. In some embodiments, the hydrogel(s) described herein may be configured to have a viscosity of between about 0.5 Pa s and greater than about 10 ¹² Pa s. In embodiments, the viscosity of a hydrogel can be, for example, between 0.5 and 10 ¹² Pa s, between 1 Pa s and 10 ⁶ Pa s, between 5 and 10 ³ Pa s, between 10 and 100 Pa s, between 15 and 90 Pa s, between 20 and 80 Pa s, between 25 and 70 Pa s, between 30 and 60 Pa s, or the like when applied to a device.

[084] The hydrogel can be supplied in a device as described herein as an amount of hydrogel per square foot of system or device. In some embodiments, about 1 g, about 5 g, about 10 g, about 15 g, about 20 g, about 25 g, about 30 g, about 35 g, about 40 g, about 45 g, about 50 g, at least about 1 g, at least about 5 g, at least about 10 g, at least about 20 g, between about 1 g and about 20 g, between about 10 g and about 20 g or between about 15 g and about 25 g of a hydrogel per square foot of device can be sufficient to provide the herein desired results.

[085] Embodiments disclosed herein can comprise active agents or cosmetic agents or drugs, for example applied prior to applying the dressing to the treatment area, or applied to the substrate. Suitable active agents con comprise, for example, hypoallergenic agents, drugs, biologies, stem cells, growth factors, skin substitutes, cosmetic products, combinations, or combinations thereof, or the like. Stem cells can include, for example, embryonic stem cells, bone-marrow stem cells, adipose stem cells, and the like.

[086] A growth factor is a naturally-occurring substance capable of stimulating cellular growth, proliferation, healing, and cellular differentiation, often a protein or a steroid hormone. Growth factors are important for regulating a variety of cellular processes. Growth factors typically act as signaling molecules between cells. Examples are cytokines and hormones that bind to specific receptors on the surface of their target cells. They often promote cell differentiation and maturation, which varies between growth factors. For example, bone morphogenetic proteins stimulate bone cell differentiation, while fibroblast growth factors and vascular endothelial growth factors stimulate blood vessel differentiation.

[087] Growth factors can include, for example, Adrenomedullin (AM), Angiopoietin (Ang), Autocrine motility factor, Bone morphogenetic proteins (BMPs), Brain-derived neurotrophic factor (BDNF), Epidermal growth factor (EGF), Erythropoietin (EPO), Fibroblast growth factor 1 or 2(FGF-1 or -2), Fetal Bovine Somatotrophin (FBS), Glial cell line-derived neurotrophic factor (GDNF), Granulocyte colony-stimulating factor (G-CSF), Granulocyte macrophage colony-stimulating factor (GM-CSF), Growth differentiation factor-9 (GDF9), Hepatocyte growth factor (HGF), Hepatoma-derived growth factor (HDGF), Insulin-like growth factor (IGF), Keratinocyte growth factor (KGF), Migration-stimulating factor (MSF), Myostatin (GDF-8), Nerve growth factor (NGF) and other neurotrophins, Platelet-derived growth factor (PDGF), Thrombopoietin (TPO), T-cell growth factor (TCGF), Transforming growth factor alpha (TGF-a), Transforming growth factor beta (TGF-β), Tumor necrosis factor-alpha (TNF-a), Vascular endothelial growth factor (VEGF), Wnt Signaling Pathway, Placental growth factor (PGF), IL-1 , IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, Renalase, or combinations thereof. Active agents can include alpha granules.

[088] Cosmetic products can include, for example, moisturizers, exfoliants, antioxidants, sunscreens, and the like.

[089] Drugs can include but are not limited to, for example, anti-inflammatories, painkillers, antibiotics, antivirals, and wound treatment compositions.

[090] In embodiments, active agents or cosmetic agents or drugs can be mixed with a hydrogel prior to application to a multi-array matrix of biocompatible microcells, or can be otherwise attached to the hydrogel when the hydrogel is already part of a device, such as by chemical substitution or through the use of intermolecular forces. In embodiments the active agent can be applied to an area of treatment prior to contacting the area with a system or device disclosed herein.

[091] In embodiments, dissimilar conductive metals used to make a LLEC or LLEF system disclosed herein can be silver and zinc, and the electrolytic solution can include sodium chloride in water. In certain embodiments the electrodes are applied onto a non-conductive surface to create a pattern, most preferably an array or multi-array of voltaic cells that do not spontaneously react until they contact an electrolytic solution. Sections of this description use the terms "printing" with "ink," but it is to be understood that the patterns may also be "painted" with "paints." The use of any suitable means for applying a conductive material is contemplated. In embodiments, "ink" or "paint" can comprise any material such as a solution suitable for forming an electrode on a surface such as a conductive material including a conductive metal solution. In embodiments "printing" or "painted" can comprise any method of applying a solution to a material upon which a matrix is desired, for example a transparent or translucent material.

[092] The applied electrodes or reservoirs or dots can adhere or bond to the primary surface or substrate because a biocompatible binder is mixed, in embodiments into separate mixtures, with each of the dissimilar metals that will create the pattern of voltaic cells, in embodiments. Most inks are simply a carrier, and a binder mixed with pigment. Similarly, conductive metal solutions can be a binder mixed with a conductive element. The resulting conductive metal solutions can be used with an application method such as screen printing to apply the electrodes to the primary surface in predetermined patterns. Once the conductive metal solutions dry and/or cure, the patterns of spaced electrodes can substantially maintain their relative position, even on a flexible material such as that used for a LLEC or LLEF system. The conductive metal solution can be allowed to dry before being applied to a surface so that the conductive materials do not mix, which could interrupt the array and cause direct reactions that will release the elements.

[093] In certain embodiments that utilize a poly-cellulose binder, the binder itself can have a beneficial effect such as reducing the local concentration of matrix metallo-proteases through an iontophoretic process that drives the cellulose into the surrounding tissue. This process can be used to electronically drive other components such as drugs, active agents, or cosmetic agents, into the surrounding tissue.

[094] The binder can comprise any biocompatible liquid material that can be mixed with a conductive element (for example, metallic crystals of silver or zinc) to create a conductive solution. One suitable binder is a solvent reducible polymer, such as the polyacrylic nontoxic silk-screen ink manufactured by COLORCON ^® Inc., a division of Berwind Pharmaceutical Services, Inc. (see COLORCON ^® NO-TOX ^® product line, part number NT28). In an embodiment the binder is mixed with high purity (at least 99.99%, in an embodiment) metallic silver crystals to make the silver conductive solution. The silver crystals, which can be made by grinding silver into a powder, are preferably smaller than 100 microns in size or about as fine as flour. In an embodiment, the size of the crystals is about 325 mesh, which is typically about 40 microns in size or a little smaller. The binder is separately mixed with high purity (at least 99.99%, in an embodiment) metallic zinc powder which has also preferably been sifted through standard 325 mesh screen, to make the zinc conductive solution.

[095] Other powders of metal can be used to make other conductive metal solutions in the same way as described in other embodiments. [096] In embodiments, the size of the metal crystals, the availability of the surface to the conductive fluid and the ratio of metal to binder can affect the release rate of the metal from the mixture. For example, when COLORCON ^® polyacrylic ink is used as the binder, about 10 to 40 percent of the mixture should be metal for a long term bandage (for example, one that stays on for about 10 days). If the same binder is used, but the percentage of the mixture that is metal is increased to 60 percent or higher, a typical system or device will be effective for longer. For example, for a longer term device, the percent of the mixture that should be metal can be 40 percent, or 42 percent, 44 percent, 46 percent, 48 percent, 50 percent, 52 percent, 54 percent, 56 percent, 58 percent, 60 percent, 62 percent, 64 percent, 66 percent, 68 percent, 70 percent, 72 percent, 74 percent, 76 percent, 78 percent, 80 percent, 82 percent, 84 percent, 86 percent, 88 percent, 90 percent, or the like.

[097] For LLEC or LLEF systems comprising a pliable substrate it can be desired to decrease the percentage of metal down to, for example, 20 percent, 18 percent, 16 percent, 14 percent, 12 percent, 10 percent, 5 percent, or less, or to use a binder that causes the crystals to be more deeply embedded, so that the primary surface will be antimicrobial for a very long period of time and will not wear prematurely. Other binders can dissolve or otherwise break down faster or slower than a polyacrylic ink, so adjustments can be made to achieve the desired rate of spontaneous reactions from the voltaic cells.

[098] To maximize the number of voltaic cells, in various embodiments, a pattern of alternating silver masses or electrodes or reservoirs and zinc masses or electrodes or reservoirs can create an array of electrical currents across the primary surface or base layer. A basic pattern, shown in FIG. 1 , has each mass of silver equally spaced from four masses of zinc, and has each mass of zinc equally spaced from four masses of silver, according to an embodiment. The first electrode 6 is separated from the second electrode 10 by a spacing 8. The designs of first electrode 6 and second electrode 10 are simply round dots, and in an embodiment, are repeated. Numerous repetitions 12 of the designs result in a pattern. For an exemplary device comprising silver and zinc, each silver design preferably has about twice as much mass as each zinc design, in an embodiment. For the pattern in FIG. 1 , the silver designs are most preferably about a millimeter from each of the closest four zinc designs, and vice-versa. The resulting pattern of dissimilar metal masses defines an array of voltaic cells when introduced to an electrolytic solution. Further disclosure relating to methods of producing micro-arrays can be found in U.S. Patent No. 7,813,806 entitled CURRENT PRODUCING SURFACE FOR TREATING BIOLOGIC TISSUE issued October 12, 2010, which is incorporated by reference in its entirety.

[099] A dot pattern of masses like the alternating round dots of FIG. 1 can be preferred when applying conductive material onto a flexible base layer, as the dots won't significantly affect the flexibility of the material. To maximize the density of electrical current over a primary surface the pattern of FIG. 2 can be used. The first electrode 6 in FIG. 2 is a large hexagonally shaped dot, and the second electrode 10 is a pair of smaller hexagonally shaped dots that are spaced from each other. The spacing 8 that is between the first electrode 6 and the second electrode 10 maintains a relatively consistent distance between adjacent sides of the designs. Numerous repetitions 12 of the designs result in a pattern 14 that can be described as at least one of the first design being surrounded by six hexagonally shaped dots of the second design.

[0100] FIGS. 3 and 4 show how the pattern of FIG . 2 can be used to make an embodiment disclosed herein. The pattern shown in detail in FIG. 2 is applied to the primary surface 2 of an embodiment. The back 20 of the printed material is fixed to a substrate layer 22. This layer is adhesively fixed to a pliable layer 16.

[0101] FIG. 5 shows an additional feature, which can be added between designs, that can initiate the flow of current in a poor electrolytic environment. A fine line 24 is printed using one of the conductive metal solutions along a current path of each voltaic cell. The fine line will initially have a direct reaction but will be depleted until the distance between the electrodes increases to where maximum voltage is realized. The initial current produced is intended to help control edema so that the LLEC system will be effective. If the electrolytic solution is highly conductive when the system is initially applied the fine line can be quickly depleted and the device will function as though the fine line had never existed.

[0102] FIGS. 6 and 7 show alternative patterns that use at least one line design. The first electrode 6 of FIG. 6 is a round dot similar to the first design used in FIG. 1 . The second electrode 10 of FIG . 6 is a line. When the designs are repeated, they define a pattern of parallel lines that are separated by numerous spaced dots. FIG . 7 uses only line designs. The first electrode 6 can be thicker or wider than the second electrode 10 if the oxidation- reduction reaction requires more metal from the first conductive element (mixed into the first design's conductive metal solution) than the second conductive element (mixed into the second design's conductive metal solution). The lines can be dashed. Another pattern can be silver grid lines that have zinc masses in the center of each of the cells of the grid. The pattern can be letters printed from alternating conductive materials so that a message can be printed onto the primary surface-perhaps a brand name or identifying information such as patient blood type.

[0103] Because the spontaneous oxidation-reduction reaction of an embodiment utilizing silver and zinc uses a ratio of approximately two silver atoms to one zinc atom, in an embodiment the silver design can contain about twice as much mass as the zinc design in an embodiment. At a spacing of about 1 mm between the closest dissimilar metals (closest edge to closest edge) each voltaic cell that contacts a conductive fluid such as a cosmetic cream can create approximately 1 volt of potential that will penetrate substantially through its surrounding surfaces. Closer spacing of the dots can decrease the resistance, providing less potential, and the current will not penetrate as deeply. Therefore, spacing between the closest conductive materials on the base layer or substrate can be, for example, about 1 μηι, about 2 μηι, about 3 μηι, about 4 μηι, about 5 μηι, about 6 μηι, about 7 μηι, about 8 μηι, about 9 μηι, about 10 μηι, about 11 μηι, about 12 μηι, about 13 μηι, about 14 μηι, about 15 μηι, about 16 μηι, about 17 μηι, about 18 μηι, about 19 μηι, about 20 μηι, about 21 μηι, about 22 μηι, about 23 μηι, about 24 μm, about 25 μηι, about 26 μm, about 27 μηι, about 28 μηι, about 29 μm, about 30 μηι, about 31 μm, about 32 μηι, about 33 μηι, about 34 μm, about 35 μηι, about 36 μm, about 37 μηι, about 38 μηι, about 39 μm, about 40 μηι, about 41 μm, about 42 μηι, about 43 μηι, about 44 μm, about 45 μηι, about 46 μm, about 47 μηι, about 48 μηι, about 49 μm, about 50 μηι, about 51 μm, about 52 μηι, about 53 μηι, about 54 μm, about 55 μηι, about 56 μm, about 57 μηι, about 58 μηι, about 59 μm, about 60 μηι, about 61 μm, about 62 μηι, about 63 μηι, about 64 μm, about 65 μηι, about 66 μm, about 67 μηι, about 68 μηι, about 69 μm, about 70 μηι, about 71 μm, about 72 μηι, about 73 μηι, about 74 μm, about 75 μηι, about 76 μm, about 77 μηι, about 78 μηι, about 79 μm, about 80 μηι, about 81 μm, about 82 μηι, about 83 μηι, about 84 μm, about 85 μηι, about 86 μm, about 87 μηι, about 88 μηι, about 89 μm, about 90 μηι, about 91 μm, about 92 μηι, about 93 μηι, about 94 μm, about 95 μηι, about 96 μm, about 97 μηι, about 98 μηι, about 99 μm, about 0.1 mm, about 0.2 mm, about 0.3 mm, about 0.4 mm, about 0.5 mm, about 0.6 mm, about 0.7 mm, about 0.8 mm, about 0.9 mm, about 1 mm, about 1.1 mm, about 1.2 mm, about 1.3 mm, about 1.4 mm, about 1.5 mm, about 1.6 mm, about 1.7 mm, about 1.8 mm, about 1.9 mm, about 2 mm, about 2.1 mm, about 2.2 mm, about 2.3 mm, about 2.4 mm, about 2.5 mm, about 2.6 mm, about 2.7 mm, about 2.8 mm, about 2.9 mm, about 3 mm, about 3.1 mm, about 3.2 mm, about 3.3 mm, about 3.4 mm, about 3.5 mm, about 3.6 mm, about 3.7 mm, about 3.8 mm, about 3.9 mm, about 4 mm, about 4.1 mm, about 4.2 mm, about 4.3 mm, about 4.4 mm, about 4.5 mm, about 4.6 mm, about 4.7 mm, about 4.8 mm, about 4.9 mm, about 5 mm, about 5.1 mm, about 5.2 mm, about 5.3 mm, about 5.4 mm, about 5.5 mm, about 5.6 mm, about 5.7 mm, about 5.8 mm, about 5.9 mm, about 6 mm, or the like.

[0104] In certain embodiments the closest spacing between conductive materials on the base layer or substrate can be not more than 0.1 mm, not more than 0.2 mm, not more than 0.3 mm, not more than 0.4 mm, not more than 0.5 mm, not more than 0.6 mm, not more than 0.7 mm, not more than 0.8 mm, not more than 0.9 mm, not more than 1 mm, not more than 1.1 mm, not more than 1.2 mm, not more than 1.3 mm, not more than 1.4 mm, not more than 1.5 mm, not more than 1.6 mm, not more than 1.7 mm, not more than 1.8 mm, not more than 1.9 mm, not more than 2 mm, not more than 2.1 mm, not more than 2.2 mm, not more than 2.3 mm, not more than 2.4 mm, not more than 2.5 mm, not more than 2.6 mm, not more than 2.7 mm, not more than 2.8 mm, not more than 2.9 mm, not more than 3 mm, not more than 3.1 mm, not more than 3.2 mm, not more than 3.3 mm, not more than 3.4 mm, not more than 3.5 mm, not more than 3.6 mm, not more than 3.7 mm, not more than 3.8 mm, not more than 3.9 mm, not more than 4 mm, not more than 4.1 mm, not more than 4.2 mm, not more than 4.3 mm, not more than 4.4 mm, not more than 4.5 mm, not more than 4.6 mm, not more than 4.7 mm, not more than 4.8 mm, not more than 4.9 mm, not more than 5 mm, not more than 5.1 mm, not more than 5.2 mm, not more than 5.3 mm, not more than 5.4 mm, not more than 5.5 mm, not more than 5.6 mm, not more than 5.7 mm, not more than 5.8 mm, not more than 5.9 mm, not more than 6 mm, or the like.

[0105] In certain embodiments closest spacing between the conductive materials on the base layer or substrate can be not less than 0.1 mm, or not less than 0.2 mm, not less than 0.3 mm, not less than 0.4 mm, not less than 0.5 mm, not less than 0.6 mm, not less than 0.7 mm, not less than 0.8 mm, not less than 0.9 mm, not less than 1 mm, not less than 1 .1 mm, not less than 1 .2 mm, not less than 1 .3 mm, not less than 1 .4 mm, not less than 1 .5 mm, not less than 1 .6 mm, not less than 1 .7 mm, not less than 1 .8 mm, not less than 1 .9 mm, not less than 2 mm, not less than 2.1 mm, not less than 2.2 mm, not less than 2.3 mm, not less than 2.4 mm, not less than 2.5 mm, not less than 2.6 mm, not less than 2.7 mm, not less than 2.8 mm, not less than 2.9 mm, not less than 3mm, not less than 3.1 mm, not less than 3.2 mm, not less than 3.3 mm, not less than 3.4 mm, not less than 3.5 mm, not less than 3.6 mm, not less than 3.7 mm, not less than 3.8 mm, not less than 3.9 mm, not less than 4 mm, not less than 4.1 mm, not less than 4.2 mm, not less than 4.3 mm, not less than 4.4 mm, not less than 4.5 mm, not less than 4.6 mm, not less than 4.7 mm, not less than 4.8 mm, not less than 4.9 mm, not less than 5 mm, not less than 5.1 mm, not less than 5.2 mm, not less than 5.3 mm, not less than 5.4 mm, not less than 5.5 mm, not less than 5.6 mm, not less than 5.7 mm, not less than 5.8 mm, not less than 5.9 mm, not less than 6 mm, or the like.

[0106] Disclosures of the present specification include LLEC or LLEF systems comprising a primary surface of a material wherein the material is adapted to be applied to an area of tissue such as a muscle; a first electrode design formed from a first conductive liquid that includes a mixture of a polymer and a first element, the first conductive liquid being applied into a position of contact with the primary surface, the first element including a metal species, and the first electrode design including at least one dot or reservoir, wherein at least one of the at least one dot or reservoir has approximately a 1 .5 mm +/- 1 mm mean diameter; a second electrode design formed from a second conductive liquid that includes a mixture of a polymer and a second element, the second element including a different metal species than the first element, the second conductive liquid being printed into a position of contact with the primary surface, and the second electrode design including at least one other dot or reservoir, wherein at least one of the at least one other dot or reservoir has approximately a 2.5 mm +/- 2 mm mean diameter; a spacing on the primary surface that is between the first electrode design and the second electrode design such that the first electrode design does not physically contact the second electrode design, wherein the spacing is approximately 1 .5 mm +/- 1 mm, and at least one repetition of the first electrode design and the second electrode design, the at least one repetition of the first electrode design being substantially adjacent the second electrode design, wherein the at least one repetition of the first electrode design and the second electrode design, in conjunction with the spacing between the first electrode design and the second electrode design, defines at least one pattern of at least one voltaic cell for spontaneously generating at least one electrical current when introduced to an electrolytic solution. Therefore, electrodes, dots or reservoirs can have a mean diameter of, for example, about 0.2 mm, about 0.3 mm, about 0.4 mm, about 0.5 mm, about 0.6 mm, about 0.7 mm, about 0.8 mm, about 0.9 mm, about 1 .0 mm, about 1 .1 mm, about 1 .2 mm, about 1 .3 mm, about 1 .4 mm, about 1 .5 mm, about 1 .6 mm, about 1 .7 mm, about 1 .8 mm, about 1 .9 mm, about 2.0 mm, about 2.1 mm, about 2.2 mm, about 2.3 mm, about 2.4 mm, about 2.5 mm, about 2.6 mm, about 2.7 mm, about 2.8 mm, about 2.9 mm, about 3.0 mm, about 3.1 mm, about 3.2 mm, about 3.3 mm, about 3.4 mm, about 3.5 mm, about 3.6 mm, about 3.7 mm, about 3.8 mm, about 3.9 mm, about 4.0 mm, about 4.1 mm, about 4.2 mm, about 4.3 mm, about 4.4 mm, about 4.5 mm, about 4.6 mm, about 4.7 mm, about 4.8 mm, about 4.9 mm, about 5.0 mm, or the like.

[0107] In further embodiments, electrodes, dots or reservoirs can have a mean diameter of not less than 0.2 mm, not less than 0.3 mm, not less than 0.4 mm, not less than 0.5 mm, not less than 0.6 mm, not less than 0.7 mm, not less than 0.8 mm, not less than 0.9 mm, not less than 1 .0 mm, not less than 1 .1 mm, not less than 1 .2 mm, not less than 1 .3 mm, not less than 1 .4 mm, not less than 1 .5 mm, not less than 1 .6 mm, not less than 1 .7 mm, not less than 1 .8 mm, not less than 1 .9 mm, not less than 2.0 mm, not less than 2.1 mm, not less than 2.2 mm, not less than 2.3 mm, not less than 2.4 mm, not less than 2.5 mm, not less than 2.6 mm, not less than 2.7 mm, not less than 2.8 mm, not less than 2.9 mm, not less than 3.0 mm, not less than 3.1 mm, not less than 3.2 mm, not less than 3.3 mm, not less than 3.4 mm, not less than 3.5 mm, not less than 3.6 mm, not less than 3.7 mm, not less than 3.8 mm, not less than 3.9 mm, not less than 4.0 mm, not less than 4.1 mm, not less than 4.2 mm, not less than 4.3 mm, not less than 4.4 mm, not less than 4.5 mm, not less than 4.6 mm, not less than 4.7 mm, not less than 4.8 mm, not less than 4.9 mm, not less than 5.0 mm, or the like.

[0108] In further embodiments, electrodes, dots or reservoirs can have a mean diameter of not more than 0.2 mm, not more than 0.3 mm, not more than 0.4 mm, not more than 0.5 mm, not more than 0.6 mm, not more than 0.7 mm, not more than 0.8 mm, not more than 0.9 mm, not more than 1 .0 mm, not more than 1 .1 mm, not more than 1 .2 mm, not more than 1 .3 mm, not more than 1 .4 mm, not more than 1 .5 mm, not more than 1 .6 mm, not more than 1 .7 mm, not more than 1 .8 mm, not more than 1 .9 mm, not more than 2.0 mm, not more than 2.1 mm, not more than 2.2 mm, not more than 2.3 mm, not more than 2.4 mm, not more than 2.5 mm, not more than 2.6 mm, not more than 2.7 mm, not more than 2.8 mm, not more than 2.9 mm, not more than 3.0 mm, not more than 3.1 mm, not more than 3.2 mm, not more than 3.3 mm, not more than 3.4 mm, not more than 3.5 mm, not more than 3.6 mm, not more than 3.7 mm, not more than 3.8 mm, not more than 3.9 mm, not more than 4.0 mm, not more than 4.1 mm, not more than 4.2 mm, not more than 4.3 mm, not more than 4.4 mm, not more than 4.5 mm, not more than 4.6 mm, not more than 4.7 mm, not more than 4.8 mm, not more than 4.9 mm, not more than 5.0 mm, or the like.

[0109] FIG. 9 shows an embodiment utilizing two electrodes (one positive and one negative) . Upper arms 140 and 145 can be, for example, about 1 , 2, 3, or 4 mm in width. Lower arm 147 and serpentine 149 can be, for example, about 1 , 2, 3, or 4 mm in width. The electrodes can be, for example, 1 , 2, or 3 mm in depth.

[0110] FIG. 10 shows an embodiment utilizing two electrodes (one positive and one negative) . Upper arms 150 and 155 can be, for example, about 1 , 2, 3, or 4 mm in width. The extensions protruding from the lower arm 156 can be, for example, about 1 , 1 .5, 2, 2.5, 3, 3.5, or 4 mm in width . The extensions protruding from the comb 158 can be, for example, about 1 , 2, 3, 4, 5, 6, or 7 mm in width. The electrodes can be, for example, about 1 , 2, or 3 mm in depth .

[0111] FIG. 1 1 shows an embodiment utilizing two electrodes (one positive and one negative) . Upper arms 160 and 165 can be, for example, about 1 , 2, 3, or 4 mm in width. Lower block 167 can be, for example, about 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, or 54 mm along its shorter axis. Lower block 167 can be, for example, about 60, 65, 70, 75, 80, 85, 90, 95, or 100 mm along its longer axis. The electrodes can be, for example, about 1 , 2, or 3 mm in depth.

[0112] In embodiments such as those in FIG.s 9-1 1 , the width and depth of the various areas of the electrode can be designed to produce a particular electric field, or, when both electrodes are in contact with a conductive material, a particular electric current. For example, the width of the various areas of the electrode can be, for example, about 0.1 mm, about 0.2 mm, about 0.3 mm, about 0.4 mm, about 0.5 mm, about 0.6 mm, about 0.7 mm, about 0.8 mm, about 0.9 mm, about 1 mm, about 1 .1 mm, about 1 .2 mm, about 1 .3 mm, about 1 .4 mm, about 1 .5 mm, about 1 .6 mm, about 1 .7 mm, about 1 .8 mm, about 1 .9 mm, about 2 mm, about 2.1 mm, about 2.2 mm, about 2.3 mm, about 2.4 mm, about 2.5 mm, about 2.6 mm, about 2.7 mm, about 2.8 mm, about 2.9 mm, about 3 mm, about 3.1 mm, about 3.2 mm, about 3.3 mm, about 3.4 mm, about 3.5 mm, about 3.6 mm, about 3.7 mm, about 3.8 mm, about 3.9 mm, about 4 mm, about 4.1 mm, about 4.2 mm, about 4.3 mm, about 4.4 mm, about 4.5 mm, about 4.6 mm, about 4.7 mm, about 4.8 mm, about 4.9 mm, about 5 mm, about 5.1 mm, about 5.2 mm, about 5.3 mm, about 5.4 mm, about 5.5 mm, about 5.6 mm, about 5.7 mm, about 5.8 mm, about 5.9 mm, about 6 mm, about 7 mm, about 8 mm, about 9 mm, about 10 mm, about 11 mm, or the like.

[0113] In embodiments such as those in FIG.s 9-1 1 , the depth or thickness of the various areas of the electrode can be, for example, about 0.1 mm, about 0.2 mm, about 0.3 mm, about 0.4 mm, about 0.5 mm, about 0.6 mm, about 0.7 mm, about 0.8 mm, about 0.9 mm, about 1 mm, about 1.1 mm, about 1.2 mm, about 1.3 mm, about 1.4 mm, about 1.5 mm, about 1.6 mm, about 1.7 mm, about 1.8 mm, about 1.9 mm, about 2 mm, about 2.1 mm, about 2.2 mm, about 2.3 mm, about 2.4 mm, about 2.5 mm, about 2.6 mm, about 2.7 mm, about 2.8 mm, about 2.9 mm, about 3 mm, about 3.1 mm, about 3.2 mm, about 3.3 mm, about 3.4 mm, about 3.5 mm, about 3.6 mm, v3.7 mm, about 3.8 mm, about 3.9 mm, about 4 mm, about 4.1 mm, about 4.2 mm, about 4.3 mm, about 4.4 mm, about 4.5 mm, about 4.6 mm, about 4.7 mm, about 4.8 mm, about 4.9 mm, about 5 mm, about 5.1 mm, about 5.2 mm, about 5.3 mm, about 5.4 mm, about 5.5 mm, about 5.6 mm, about 5.7 mm, about 5.8 mm, about 5.9 mm, about 6 mm, or the like.

[0114] In embodiments such as those in FIG.s 9-1 1 , the shortest distance between the two electrodes in an embodiment can be, for example, about 0.1 mm, or about 0.2 mm, about 0.3 mm, about 0.4 mm, about 0.5 mm, about 0.6 mm, about 0.7 mm, about 0.8 mm, about 0.9 mm, about 1 mm, about 1.1 mm, about 1.2 mm, about 1.3 mm, about 1.4 mm, about 1.5 mm, about 1.6 mm, about 1.7 mm, about 1.8 mm, about 1.9 mm, about 2 mm, about 2.1 mm, about 2.2 mm, about 2.3 mm, about 2.4 mm, about 2.5 mm, about 2.6 mm, about 2.7 mm, about 2.8 mm, about 2.9 mm, about 3 mm, about 3.1 mm, about 3.2 mm, about 3.3 mm, about 3.4 mm, about 3.5 mm, about 3.6 mm, about 3.7 mm, about 3.8 mm, about 3.9 mm, about 4 mm, about 4.1 mm, about 4.2 mm, about 4.3 mm, about 4.4 mm, about 4.5 mm, about 4.6 mm, about 4.7 mm, about 4.8 mm, about 4.9 mm, about 5 mm, about 5.1 mm, about 5.2 mm, about 5.3 mm, about 5.4 mm, about 5.5 mm, about 5.6 mm, about 5.7 mm, about 5.8 mm, about 5.9 mm, about 6 mm, about 7 mm, about 8 mm, about 9 mm, about 10 mm, about 11 mm, about 12 mm, about 13 mm, about 14 mm, about 15 mm, about 16 mm, about 17 mm, about 18 mm, about 19 mm, about 20 mm, about 21 mm, about 22 mm, about 23 mm, about 24 mm, about 25 mm, about 26 mm, about 27 mm, about 28 mm, about 29 mm, about 30 mm, about 31 mm, about 32 mm, about 33 mm, about 34 mm, or the like.

[0115] In embodiments such as those in FIG.s 9-1 1 , the length of the long axis of the electrode can be, for example, about 2 mm, about 2.1 mm, about 2.2 mm, about 2.3 mm, about 2.4 mm, about 2.5 mm, about 2.6 mm, about 2.7 mm, about 2.8 mm, about 2.9 mm, about 3 mm, about 3.1 mm, about 3.2 mm, about 3.3 mm, about 3.4 mm, about 3.5 mm, about 3.6 mm, about 3.7 mm, about 3.8 mm, about 3.9 mm, about 4 mm, about 4.1 mm, about 4.2 mm, about 4.3 mm, about 4.4 mm, about 4.5 mm, about 4.6 mm, about 4.7 mm, about 4.8 mm, about 4.9 mm, about 5 mm, about 5.1 mm, about 5.2 mm, about 5.3 mm, about 5.4 mm, about 5.5 mm, about 5.6 mm, about 5.7 mm, about 5.8 mm, about 5.9 mm, about 6 mm, about 7 mm, about 8 mm, about 9 mm, about 10 mm, about 11 mm, about 12 mm, about 13 mm, about 14 mm, about 15 mm, about 16 mm, about 17 mm, about 18 mm, about 19 mm, about 20 mm, about 21 mm, about 22 mm, about 23 mm, about 24 mm, about 25 mm, about 26 mm, about 27 mm, about 28 mm, about 29 mm, about 30 mm, about 31 mm, about 32 mm, about 33 mm, about 34 mm, about 35 mm, about 36 mm, about 37 mm, about 38 mm, about 39 mm, about 40 mm, about 41 mm, about 42 mm, about 43 mm, about 44 mm, about 45 mm, about 46 mm, about 47 mm, about 48 mm, about 49 mm, about 50 mm, about 75 mm, about 100 mm, about 150 mm, about 200 mm, about 250 mm, about 300 mm, about 350 mm, about 400 mm, about 450 mm, about 500 mm, about 600 mm, about 700 mm, about 800 mm, about 900 mm, about 1000 mm, or more, or the like.

[0116] In embodiments such as those in FIG.s 9-1 1 , the length of the short axis of the electrode can be, for example, about 2 mm, about 2.1 mm, about 2.2 mm, about 2.3 mm, about 2.4 mm, about 2.5 mm, about 2.6 mm, about 2.7 mm, about 2.8 mm, about 2.9 mm, about 3 mm, about 3.1 mm, about 3.2 mm, about 3.3 mm, about 3.4 mm, about 3.5 mm, about 3.6 mm, about 3.7 mm, about 3.8 mm, about 3.9 mm, about 4 mm, about 4.1 mm, about 4.2 mm, about 4.3 mm, about 4.4 mm, about 4.5 mm, about 4.6 mm, about 4.7 mm, about 4.8 mm, about 4.9 mm, about 5 mm, about 5.1 mm, about 5.2 mm, about 5.3 mm, about 5.4 mm, about 5.5 mm, about 5.6 mm, about 5.7 mm, about 5.8 mm, about 5.9 mm, about 6 mm, about 7 mm, about 8 mm, about 9 mm, about 10 mm, about 11 mm, about 12 mm, about 13 mm, about 14 mm, about 15 mm, about 16 mm, about 17 mm, about 18 mm, about 19 mm, about 20 mm, about 21 mm, about 22 mm, about 23 mm, about 24 mm, about 25 mm, about 26 mm, about 27 mm, about 28 mm, about 29 mm, about 30 mm, about 31 mm, about 32 mm, about 33 mm, about 34 mm, about 35 mm, about 36 mm, about 37 mm, about 38 mm, about 39 mm, about 40 mm, about 41 mm, about 42 mm, about 43 mm, about 44 mm, about 45 mm, about 46 mm, about 47 mm, about 48 mm, about 49 mm, about 50 mm, about 75 mm, about 100 mm, or more, or the like.

[0117] FIG. 12 depicts a backing layer 123, substrate 124, tear tab 125, and void region 120.

[0118] FIG. 13 depicts an embodiment in use, with a user removing the substrate layer 130 from the backing layer 135.

[0119] FIG. 14 depicts void regions 140 in a backing layer with visible multi-array matrix 145. [0120] The material concentrations or quantities within and/or the relative sizes (e.g., dimensions or surface area) of the first and second reservoirs can be selected deliberately to achieve various characteristics of the systems' behavior. For example, the quantities of material within a first and second reservoir can be selected to provide an apparatus having an operational behavior that depletes at approximately a desired rate and/or that "dies" after an approximate period of time after activation. In an embodiment the one or more first reservoirs and the one or more second reservoirs are configured to sustain one or more currents for an approximate pre-determined period of time, after activation. It is to be understood that the amount of time that currents are sustained can depend on external conditions and factors (e.g., the quantity and type of activation material), and currents can occur intermittently depending on the presence or absence of activation material. Further disclosure relating to producing reservoirs that are configured to sustain one or more currents for an approximate pre-determined period of time can be found in U.S. Patent No. 7,904, 147 entitled SUBSTANTIALLY PLANAR ARTICLE AND METHODS OF MANUFACTURE issued March 8, 201 1 , which is incorporated by reference herein in its entirety.

[0121] In one embodiment, a coating of a hydrogel can be manually spread on the multi- array matrix of biocompatible microcells. This process can be accomplished using a coating system similar to one used in silkscreening.

[0122] In some embodiments, the hydrogel can be thinned by adding additional water to the hydrogel before application. When the viscosity of the hydrogel has been reduced sufficiently, the thinned hydrogel can be applied. A reduced viscosity hydrogel can be used for dip coating.

[0123] In another embodiment, a hydrogel can be sprayed onto a multi-array matrix of biocompatible microcells in a manner similar to spray painting. In some embodiments, a thinned hydrogel can be used for spraying.

[0124] LLEC / LLEF Systems, Devices; Methods of Use

[0125] Disclosed methods can comprise hydrating the multi-array matrix, removing the dressing from the backing layer using the "tab" (as seen in FIG. 13), then applying the dressing to an area where treatment is desired.

[0126] Disclosed methods of use comprise application of a system or device described herein to a tissue, for example skin (such as around the eyes), a joint, a muscle, or a muscle group. In embodiments, the application can be performed prior to, during, or after use of the muscle or muscle group to be treated. For example, a shoulder can be treated prior to engaging in an athletic activity, for example pitching a baseball. Disclosed embodiments can increase glucose uptake, drive redox signaling, increase H ₂ 0 ₂ production, increase cellular protein sulfhydryl levels, and increase (IGF)-1 R phosphorylation. [0127] Disclosed embodiments include devices and methods for increasing capillary density.

[0128] Further aspects include a method of directing cell migration using a device disclosed herein. These aspects include methods of improving re-epithelialization.

[0129] Further aspects include methods of increasing cellular thiol levels. Additional aspects include a method of energizing mitochondria.

[0130] Further aspects include a method of stimulating cellular protein expression.

[0131] Further aspects include a method of stimulating cellular DNA synthesis.

[0132] Further aspects include a method of stimulating cellular Ca ²⁺ uptake.

[0133] Embodiments include devices and methods for increasing transcutaneous partial pressure of oxygen. Further embodiments include methods and devices for treating or preventing pressure ulcers.

[0134] In embodiments, these systems, devices, and methods can increase ATP production, and angiogenesis, thus accelerating the healing process. Disclosed systems, devices, and methods can also reduce bacterial population and/or proliferation, for example, in and around injuries or wounds.

[0135] Additional aspects include methods of preventing bacterial biofilm formation. Aspects also include a method of reducing microbial or bacterial proliferation, killing microbes or bacteria, killing bacteria through a biofilm layer, or preventing the formation of a biofilm. Embodiments include methods using devices disclosed herein in combination with antibiotics for reducing microbial or bacterial proliferation, killing microbes or bacteria, killing bacteria through a biofilm layer, or preventing the formation of a biofilm.

[0136] Further aspects include methods of treating diseases related to metabolic deficiencies, such as diabetes, or other diseases wherein the patient exhibits a compromised metabolic status.

[0137] Embodiments disclosed herein include LLEC and LLEF systems that can promote and/or accelerate the muscle recovery process and optimize muscle performance.

[0138] Further, embodiments disclosed herein can increase or decrease cell migration.

[0139] Further embodiments can increase cellular protein sulfhydryl levels and cellular glucose uptake. Increased glucose uptake can result in greater mitochondrial activity and thus increased glucose utilization.

[0140] Disclosed embodiments can produce an electrical stimulus and/or can electro- motivate, electro-conduct, electro-induct, electro-transport, and/or electrophorese one or more therapeutic materials in areas of target tissue (e.g., iontophoresis), and/or can cause one or more biologic or other materials in proximity to, on or within target tissue to be rejuvenated. Further disclosure relating to materials that can produce an electrical stimulus can be found in U.S. Patent No. 7,662, 176 entitled FOOTWEAR APPARATUS AND METHODS OF MANUFACTURE AND USE issued February 16, 2010, which is incorporated herein by reference in its entirety.

[0141] Methods disclosed herein can include applying a disclosed embodiment to an area to be treated. Embodiments can include selecting or identifying a patient in need of treatment. In embodiments, methods disclosed herein can include application of a device disclosed herein to an area to be treated. For example, in an embodiment, a user can remove a substrate comprising a multi-array matrix from the backing layer using the "tear" tab. In embodiments the tear tab can be detachable from the backing layer.

[0142] In embodiments, disclosed methods include application to the treatment area or the device of an antibacterial. In embodiments the antibacterial can be, for example, alcohols, aldehydes, halogen-releasing compounds, peroxides, anilides, biguanides, bisphenols, halophenols, heavy metals, phenols and cresols, quaternary ammonium compounds, and the like. In embodiments the antibacterial agent can comprise, for example, ethanol, isopropanol, glutaraldehyde, formaldehyde, chlorine compounds, iodine compounds, hydrogen peroxide, ozone, peracetic acid, formaldehyde, ethylene oxide, triclocarban, chlorhexidine, alexidine, polymeric biguanides, triclosan, hexachlorophene, PCMX (p-chloro- m-xylenol), silver compounds, mercury compounds, phenol, cresol, cetrimide, benzalkonium chloride, cetylpyridinium chloride, ceftolozane/tazobactam, ceftazidime/avibactam, ceftaroline/avibactam, imipenem/MK-7655, plazomicin, eravacycline, brilacidin, and the like.

[0143] In embodiments, compounds that modify resistance to common antibacterials can be employed. For example, some resistance-modifying agents may inhibit multidrug resistance mechanisms, such as drug efflux from the cell, thus increasing the susceptibility of bacteria to an antibacterial. In embodiments, these compounds can include Phe-Arg-β- naphthylamide, or β-lactamase inhibitors such as clavulanic acid and sulbactam.

[0144] In embodiments, the system can also be used for preventative treatment of tissue injuries. Preventative treatment can include preventing the reoccurrence of previous muscle injuries. For example, an embodiment can be shaped to fit a patient's shoulder to prevent recurrence of a deltoid injury.

EXAMPLES

[0145] The following non-limiting examples are provided for illustrative purposes only in order to facilitate a more complete understanding of representative embodiments. These examples should not be construed to limit any of the embodiments described in the present specification.

Example 1

Cell Migration Assay

[0146] The in vitro scratch assay is a well-developed method to measure cell migration in vitro. The basic steps involve creating a "scratch" in a cell monolayer, capturing images at the beginning and at regular intervals during cell migration to close the scratch, and comparing the images to quantify the migration rate of the cells. Compared to other methods, the in vitro scratch assay is particularly suitable for studies on the effects of cell- matrix and cell-cell interactions on cell migration, mimic cell migration during wound healing in vivo and are compatible with imaging of live cells during migration to monitor intracellular events if desired. In addition to monitoring migration of homogenous cell populations, this method has also been adopted to measure migration of individual cells in the leading edge of the scratch.

[0147] Human keratinocytes were plated under plated under placebo or a LLEC system as described herein (labeled "PROCELLERA ^® "). Cells were also plated under silver-only or zinc-only dressings. After 24 hours, the scratch assay was performed. Cells plated under the PROCELLERA ^® device displayed increased migration into the "scratched" area as compared to any of the zinc, silver, or placebo dressings. After 9 hours, the cells plated under the PROCELLERA ^® device had almost "closed" the scratch. This demonstrates the importance of electrical activity to cell migration and infiltration.

[0148] In addition to the scratch test, genetic expression was tested. Increased insulin growth factor (IGF)-1 R phosphorylation was demonstrated by the cells plated under the PROCELLERA ^® device as compared to cells plated under insulin growth factor alone.

[0149] Integrin accumulation also affects cell migration. An increase in integrin accumulation was achieved with the LLEC system. Integrin is necessary for cell migration, and is found on the leading edge of migrating cell.

[0150] Thus, the tested LLEC system enhanced cellular migration and IGF-1 R / integrin involvement. This involvement demonstrates the effect that the LLEC system had upon cell receptors involved with the wound healing process.

Example 2

Wound Care Study

[0151] The medical histories of patients who received "standard-of-care" wound treatment ("SOC"; n = 20), or treatment with a LLEC device as disclosed herein (n = 18), were reviewed. The wound care device used in the present study consisted of a discrete matrix of silver and zinc dots. A sustained voltage of approximately 0.8 V was generated between the dots. The electric field generated at the device surface was measured to be 0.2-1 .0 V, 10-50 μΑ.

[0152] Wounds were assessed until closed or healed. The number of days to wound closure and the rate of wound volume reduction were compared. Patients treated with LLEC received one application of the device each week, or more frequently in the presence of excessive wound exudate, in conjunction with appropriate wound care management. The LLEC was kept moist by saturating with normal saline or conductive hydrogel. Adjunctive therapies (such as negative pressure wound therapy [NPWT], etc.) were administered with SOC or with the use of LLEC unless contraindicated. The SOC group received the standard of care appropriate to the wound, for example antimicrobial dressings, barrier creams, alginates, silver dressings, absorptive foam dressings, hydrogel, enzymatic debridement ointment, NPWT, etc. Etiology-specific care was administered on a case-by-case basis. Dressings were applied at weekly intervals or more. The SOC and LLEC groups did not differ significantly in gender, age, wound types or the length, width, and area of their wounds.

[0153] Wound dimensions were recorded at the beginning of the treatment, as well as interim and final patient visits. Wound dimensions, including length (L), width (W) and depth (D) were measured, with depth measured at the deepest point. Wound closure progression was also documented through digital photography. Determining the area of the wound was performed using the length and width measurements of the wound surface area.

[0154] Closure was defined as 100% epithelialization with visible effacement of the wound. Wounds were assessed 1 week post-closure to ensure continued progress toward healing during its maturation and remodeling phase.

[0155] Wound types included in this study were diverse in etiology and dimensions, thus the time to heal for wounds was distributed over a wide range (9- 124 days for SOC, and 3-44 days for the LLEC group). Additionally, the patients often had multiple co-morbidities, including diabetes, renal disease, and hypertension. The average number of days to wound closure was 36.25 (SD = 28.89) for the SOC group and 19.78 (SD = 14.45) for the LLEC group, p = 0.036. On average, the wounds in the LLEC treatment group attained closure 45.43% earlier than those in the SOC group.

[0156] Based on the volume calculated, some wounds improved persistently while others first increased in size before improving. The SOC and the LLEC groups were compared to each other in terms of the number of instances when the dimensions of the patient wounds increased (i.e. , wound treatment outcome degraded) . In the SOC group, 10 wounds (50% for n = 20) became larger during at least one measurement interval, whereas 3 wounds (16.7% for n = 18) became larger in the LLEC group (p = 0.018). Overall, wounds in both groups responded positively. Response to treatment was observed to be slower during the initial phase, but was observed to improve as time progressed.

[0157] The LLEC wound treatment group demonstrated on average a 45.4% faster closure rate as compared to the SOC group. Wounds receiving SOC were more likely to follow a "waxing-and-waning" progression in wound closure compared to wounds in the LLEC treatment group. [0158] Compared to localized SOC treatments for wounds, the LLEC (1) reduces wound closure time, (2) has a steeper wound closure trajectory, and (3) has a more robust wound healing trend with fewer incidence of increased wound dimensions during the course of healing.

Example 3

LLEC Influence on Human Keratinocyte Migration

[0159] An LLEC-generated electrical field was mapped, leading to the observation that LLEC generates hydrogen peroxide, known to drive redox signaling. LLEC-induced phosphorylation of redox-sensitive IGF-1 R was directly implicated in cell migration. The LLEC also increased keratinocyte mitochondrial membrane potential.

[0160] The LLEC was made of polyester printed with dissimilar elemental metals as described herein. It comprises alternating circular regions of silver and zinc dots, along with a proprietary, biocompatible binder added to lock the electrodes to the surface of a flexible substrate in a pattern of discrete reservoirs. When the LLEC contacts an aqueous solution, the silver positive electrode (cathode) is reduced while the zinc negative electrode (anode) is oxidized. The LLEC used herein consisted of metals placed in proximity of about 1 mm to each other thus forming a redox couple and generating an ideal potential on the order of 1 Volt. The calculated values of the electric field from the LLEC were consistent with the magnitudes that are typically applied (1 -10 V/cm) in classical electrotaxis experiments, suggesting that cell migration observed with the bioelectric dressing is likely due to electrotaxis.

[0161] Measurement of the potential difference between adjacent zinc and silver dots when the LLEC is in contact with de-ionized water yielded a value of about 0.2 Volts. Though the potential difference between zinc and silver dots can be measured, non-intrusive measurement of the electric field arising from contact between the LLEC and liquid medium was difficult. Keratinocyte migration was accelerated by exposure to an Ag/Zn LLEC. Replacing the Ag/Zn redox couple with Ag or Zn alone did not reproduce the effect of keratinocyte acceleration.

[0162] Exposing keratinocytes to an LLEC for 24h significantly increased green fluorescence in the dichlorofluorescein (DCF) assay indicating generation of reactive oxygen species under the effect of the LLEC. To determine whether H ₂ 0 ₂ is generated specifically, keratinocytes were cultured with a LLEC or placebo for 24h and then loaded with PF6-AM (Peroxyfluor-6 acetoxymethyl ester; an indicator of endogenous H ₂ 0 ₂ ). Greater intracellular fluorescence was observed in the LLEC keratinocytes compared to the cells grown with placebo. Over-expression of catalase (an enzyme that breaks down H ₂ 0 ₂ ) attenuated the increased migration triggered by the LLEC. Treating keratinocytes with N-Acetyl Cysteine (which blocks oxidant-induced signaling) also failed to reproduce the increased migration observed with LLEC. Thus, H ₂ 0 ₂ signaling mediated the increase of keratinocyte migration under the effect of the electrical stimulus.

[0163] External electrical stimulus can up-regulate the TCA (tricarboxylic acid) cycle. The stimulated TCA cycle is then expected to generate more NADH and FADH ₂ to enter into the electron transport chain and elevate the mitochondrial membrane potential (Am). Fluorescent dyes JC-1 and TMRM were used to measure mitochondrial membrane potential. JC- 1 is a lipophilic dye which produces a red fluorescence with high Am and green fluorescence when Am is low. TMRM produces a red fluorescence proportional to Am. Treatment of keratinocytes with LLEC for 24h demonstrated significantly high red fluorescence with both JC-1 and TMRM , indicating an increase in mitochondrial membrane potential and energized mitochondria under the effect of the LLEC. As a potential consequence of a stimulated TCA cycle, available pyruvate (the primary substrate for the TCA cycle) is depleted resulting in an enhanced rate of glycolysis. This can lead to an increase in glucose uptake in order to push the glycolytic pathway forward. The rate of glucose uptake in HaCaT cells treated with LLEC was examined next. More than two fold enhancement of basal glucose uptake was observed after treatment with LLEC for 24h as compared to placebo control.

[0164] Keratinocyte migration is known to involve phosphorylation of a number of receptor tyrosine kinases (RTKs) . To determine which RTKs are activated as a result of LLEC, scratch assay was performed on keratinocytes treated with LLEC or placebo for 24h. Samples were collected after 3h and an antibody array that allows simultaneous assessment of the phosphorylation status of 42 RTKs was used to quantify RTK phosphorylation. It was determined that LLEC significantly induces IGF-1 R phosphorylation. Sandwich ELISA using an antibody against phospho-IGF- 1 R and total IGF-1 R verified this determination. As observed with the RTK array screening, potent induction in phosphorylation of IGF-1 R was observed 3h post scratch under the influence of LLEC. IGF- 1 R inhibitor attenuated the increased keratinocyte migration observed with LLEC treatment.

[0165] MBB (monobromobimane) alkylates thiol groups, displacing the bromine and adding a fluoresce nt tag (lamda emission = 478 nm) . MCB (monochlorobimane) reacts with only low molecular weight thiols such as glutathione. Fluorescence emission from UV laser- excited keratinocytes loaded with either MBB or MCB was determined for 30 min. Mean fluorescence collected from 10,000 cells showed a significant shift of MBB fluorescence emission from cells. No significant change in MCB fluorescence was observed, indicating a change in total protein thiol but not glutathione. HaCaT cells were treated with LLEC for 24 h followed by a scratch assay. Integrin expression was observed by immuno-cytochemistry at different time points. Higher integrin expression was observed 6h post scratch at the migrating edge.

[0166] Consistent with evidence that cell migration requires H ₂ 0 ₂ sensing, we determined that by blocking H ₂ 0 ₂ signaling by decomposition of H ₂ 0 ₂ by catalase or ROS scavenger, N- acetyl cysteine, the increase in LLEC -driven cell migration is prevented. The observation that the LLEC increases H ₂ 0 ₂ production is significant because in addition to cell migration, hydrogen peroxide generated in the wound margin tissue is required to recruit neutrophils and other leukocytes to the wound, regulates monocyte function, and VEGF signaling pathway and tissue vascularization. Therefore, external electrical stimulation can be used as an effective strategy to deliver low levels of hydrogen peroxide over time to mimic the environment of the healing wound and thus should help improve wound outcomes. Another phenomenon observed during re-epithelialization is increased expression of the integrin subunit alpha-v. There is evidence that integrin, a major extracellular matrix receptor, polarizes in response to applied ES and thus controls directional cell migration. It may be noted that there are a number of integrin subunits, however we chose integrin aV because of evidence of association of alpha-v integrin with IGF-1 R, modulation of IGF-1 receptor signaling, and of driving keratinocyte locomotion. Additionally, integrin alpha v has been reported to contain vicinal thiols that provide site for redox activation of function of these integrins and therefore the increase in protein thiols that we observe under the effect of ES may be the driving force behind increased integrin mediated cell migration. Other possible integrins which may be playing a role in LLEC -induced IGF-1 R mediated keratinocyte migration are a5 integrin and a6 integrin.

[0167] MATERIALS AND METHODS

[0168] Cell culture— Immortalized HaCaT human keratinocytes were grown in Dulbecco's low-glucose modified Eagle's medium (Life Technologies, Gaithersburg, MD, U.S.A.) supplemented with 10% fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin. The cells were maintained in a standard culture incubator with humidified air containing 5% C02 at 37°C.

[0169] Scratch assay— A cell migration assay was performed using culture inserts (IBIDI®, Verona, Wl) according to the manufacturer's instructions. Cell migration was measured using time-lapse phase-contrast microscopy following withdrawal of the insert. Images were analyzed using the AxioVision Rel 4.8 software.

[0170] N-Acetyl Cysteine Treatment— Cells were pretreated with 5mM of the thiol antioxidant N-acetylcysteine (Sigma) for 1 h before start of the scratch assay.

[0171] IGF-1 R inhibition— When applicable, cells were preincubated with 50nM IGF-1 R inhibitor, picropodophyllin (Calbiochem, MA) just prior to the Scratch Assay. [0172] Cellular H ₂ 0 ₂ Analysis - To determine intracellular H ₂ 0 ₂ levels, HaCaT cells were incubated with 5 pM PF6-AM in PBS for 20 min at room temperature. After loading, cells were washed twice to remove excess dye and visualized using a Zeiss Axiovert 200M microscope.

[0173] Catalase gene delivery— HaCaT cells were transfected with 2.3 x 107 pfu

AdCatalase or with the empty vector as control in 750 μΙ of media. Subsequently, 750 μΙ of additional media was added 4 h later and the cells were incubated for 72 h.

[0174] RTK Phosphorylation Assay— Human Phospho-Receptor Tyrosine Kinase phosphorylation was measured using Phospho-RTK Array kit (R & D Systems).

[0175] ELISA— Phosphorylated and total IGF-1 R were measured using a DuoSet IC ELISA kit from R&D Systems.

[0176] Determination of Mitochondrial Membrane Potential— Mitochondrial membrane potential was measured in HaCaT cells exposed to the LLEC or placebo using TMRM or JC- 1 (MitoProbe JC-1 Assay Kit for Flow Cytometry, Life Technologies), per manufacturer's instructions for flow cytometry.

[0177] Integrin alpha V Expression— Human HaCaT cells were grown under the MCD or placebo and harvested 6h after removing the IBIDI® insert. Staining was done using antibody against integrin aV (Abeam, Cambridge, MA).

Example 4

Generation of Superoxide

[0178] A LLEC system was tested to determine the effects on superoxide levels which can activate signal pathways. PROCELLERA ^® LLEC system increased cellular protein sulfhydryl levels. Further, the PROCELLERA ^® system increased cellular glucose uptake in human keratinocytes. Increased glucose uptake can result in greater mitochondrial activity and thus increased glucose utilization, providing more energy for cellular migration and proliferation. This can "prime" the wound healing process before a surgical incision is made and thus speed incision healing.

Example 5

Effect on Propionibacterium acnes

[0179] Bacterial Strains and Culture

[0180] The main bacterial strain used in this study is Propionibacterium acnes and multiple antibiotics-resistant P. acnes isolates are to be evaluated.

[0181] ATCC medium (7 Actinomyces broth) (BD) and/or ATCC medium (593 chopped meat medium) is used for culturing P. acnes under an anaerobic condition at 37°C. All experiments are performed under anaerobic conditions. [0182] Culture

[0183] LNA (Leeming-Notman agar) medium is prepared and cultured at 34°C for 14 days.

[0184] Planktonic cells

[0185] P. acnes is a relatively slow-growing, typically aero-tolerant anaerobic, Gram-positive bacterium (rod) . P. acnes is cultured under anaerobic condition to determine for efficacy of an embodiment disclosed herein (PROCELLERA®). Overnight bacterial cultures are diluted with fresh culture medium supplemented with 0.1 % sodium thioglycolate in PBS to10 ⁵ colony forming units (CFUs). Next, the bacterial suspensions (0.5 mL of about 105) are applied directly on PROCELLERA® (2" x 2") and control fabrics in Petri-dishes under anaerobic conditions. After 0 h and 24 h post treatments at 37°C, portions of the sample fabrics are placed into anaerobic diluents and vigorously shaken by vortexing for 2 min. The suspensions are diluted serially and plated onto anaerobic plates under an anaerobic condition. After 24 h incubation, the surviving colonies are counted. The LLEC limits bacterial proliferation.

Example 6

Availability of Cellular Energy and Lactate Threshold

[0186] The lactate threshold, also known as lactate inflection point or anaerobic threshold, is the exercise intensity at which lactate (more specifically, lactic acid) starts to accumulate in the blood stream. The reason for the acidification of the blood at high exercise intensities is two-fold: the high rates of ATP hydrolysis in the muscle release hydrogen ions, as they are co-transported out of the muscle into the blood via the monocarboxylate transporter, and also bicarbonate stores in the blood begin to be used up. This happens when lactate is produced faster than it can be removed (metabolized) in the muscle. When exercising at or below the lactate threshold, lactate produced by the muscles is removed by the body without it building up (e.g. , aerobic respiration) . When exercising at or above the lactate threshold (e.g. anaerobic respiration) , excess lactate can build up in tissue causing a lower pH and soreness, called acidosis. This excess lactate build-up decreases athletic ability during exercise as well tissue recovery after exercise and can be a primary source of post-exercise muscle stiffness/pain.

[0187] Prior to exercise or activity the patient applies a disclosed embodiment to a muscle involved in the activity. The embodiment increases cellular glucose uptake. Increased glucose uptake can result in greater mitochondrial activity and thus increased glucose utilization, providing more energy for cellular activity to remove lactic acid from muscle tissue. It has been shown that increased cellular glucose utilization can also sustain anaerobic respiration for a longer period of time during exercise, thus increasing a person's lactate threshold. An increased lactate threshold prevents lactate from building-up in muscle tissue, thus reducing or preventing muscle damage and/or pain. Example 7

Treatment of Lateral Epicondylitis

[0188] A 29 year-old tennis player reports pain on the outside of her elbow. Her doctor performs arthroscopic surgery to correct the damaged tissue. Following surgery, an embodiment as disclosed herein is applied to the patient's elbow to stimulate healing and prevent post-surgical infection. The doctor hydrates the matrix through the void in the backing layer, removes the substrate/matrix from the backing layer, and applies the substrate/matrix to the treatment area.

Example 8

Treatment of Medial Epicondylitis

[0189] A 42 year-old golfer reports pain on the inside of his elbow. His doctor performs arthroscopic surgery to correct the damaged tissue. Following surgery, an embodiment as disclosed herein is applied to the patient's elbow to stimulate healing and prevent postsurgical infection. The doctor hydrates the matrix, removes the substrate/matrix from the backing layer and applies the substrate/matrix to the treatment area.

Example 9

Treatment Following Blepharoplasty

[0190] A patient underwent a blepharoplasty procedure. Following the procedure, a device disclosed herein was applied to the incision sites above the eye. The incisions heal faster as compared to incision sites not treated with the device.

[0191] In closing, it is to be understood that although aspects of the present specification are highlighted by referring to specific embodiments, one skilled in the art will readily appreciate that these disclosed embodiments are only illustrative of the principles of the subject matter disclosed herein. Therefore, it should be understood that the disclosed subject matter is in no way limited to a particular methodology, protocol, and/or reagent, etc., described herein. As such, various modifications or changes to or alternative configurations of the disclosed subject matter can be made in accordance with the teachings herein without departing from the spirit of the present specification. Lastly, the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present disclosure, which is defined solely by the claims. Accordingly, embodiments of the present disclosure are not limited to those precisely as shown and described.

[0192] Certain embodiments are described herein, including the best mode known to the inventor for carrying out the methods and devices described herein. Of course, variations on these described embodiments will become apparent to those of ordinary skill in the art upon reading the foregoing description. Accordingly, this disclosure includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described embodiments in all possible variations thereof is encompassed by the disclosure unless otherwise indicated herein or otherwise clearly contradicted by context.

[0193] Groupings of alternative embodiments, elements, or steps of the present disclosure are not to be construed as limitations. Each group member may be referred to and claimed individually or in any combination with other group members disclosed herein. It is anticipated that one or more members of a group may be included in, or deleted from, a group for reasons of convenience and/or patentability. When any such inclusion or deletion occurs, the specification is deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.

[0194] Unless otherwise indicated, all numbers expressing a characteristic, item, quantity, parameter, property, term, and so forth used in the present specification and claims are to be understood as being modified in all instances by the term "about." As used herein, the term "about" means that the characteristic, item, quantity, parameter, property, or term so qualified encompasses a range of plus or minus ten percent above and below the value of the stated characteristic, item, quantity, parameter, property, or term. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and attached claims are approximations that may vary. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical indication should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and values setting forth the broad scope of the disclosure are approximations, the numerical ranges and values set forth in the specific examples are reported as precisely as possible. Any numerical range or value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements. Recitation of numerical ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate numerical value falling within the range. Unless otherwise indicated herein, each individual value of a numerical range is incorporated into the present specification as if it were individually recited herein.

[0195] The terms "a," "an," "the" and similar referents used in the context of describing the disclosure (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., "such as") provided herein is intended merely to better illuminate the disclosure and does not pose a limitation on the scope otherwise claimed. No language in the present specification should be construed as indicating any non-claimed element essential to the practice of embodiments disclosed herein.

[0196] Specific embodiments disclosed herein may be further limited in the claims using consisting of or consisting essentially of language. When used in the claims, whether as filed or added per amendment, the transition term "consisting of excludes any element, step, or ingredient not specified in the claims. The transition term "consisting essentially of limits the scope of a claim to the specified materials or steps and those that do not materially affect the basic and novel characteristic(s). Embodiments of the present disclosure so claimed are inherently or expressly described and enabled herein.