Ultrasonic irrigation device for internal cleansing and sterilization

Abstract

A perfusion device for ultrasonically cleaning an internal element within a body cavity, the device comprising: at least one container sized and shaped to hold at least one liquid and at least one type of nanomaterial; at least one ultrasonic transducer configured to generate ultrasonic waves in the at least one liquid within the container; and at least one adhesive element coupled to the at least one container and configured to ensure a reversible sealed bond between the at least one container and the body cavity.

Description

[Technical Field]

[0001] The present disclosure relates generally to ultrasonic medical devices, and more particularly to ultrasonic medical devices that use nanoparticles to cleanse and sterilize body parts of a subject. [Background technology]

[0002] The approaches described in this section are approaches that could be pursued, but not necessarily approaches that have been previously conceived and pursued. Thus, unless otherwise expressly noted, it should not be assumed that any of the approaches described in this section qualify as prior art solely by virtue of their inclusion in this section. Likewise, it should not be assumed that problems identified with one or more approaches qualify as prior art by virtue of this section, unless otherwise expressly noted.

[0003] Arthroscopy (also known as arthroscopic surgery or keyhole surgery) is a minimally invasive surgical procedure on joints that uses an arthroscope, an endoscope that is inserted into the joint through a small incision, to examine the joint and sometimes treat injuries.

[0004] The most common cause of joint infection is septic arthritis, which occurs when an infection, such as a skin infection or urinary tract infection, spreads to the joint via the bloodstream. Less commonly, pathogenic bacteria can enter the joint cavity through a puncture wound, drug injection, or surgery in or near a joint. In all of these cases, the joint infection must be treated to prevent further infection within the body.

[0005] Before, during, and after arthroscopy, joint irrigation and aspiration may be performed to cleanse the area, remove foreign material, and / or improve visualization of the surgical field. In fact, they are used as part of the treatment of bacterial arthritis.

[0006] According to the American Academy of Orthopaedic Surgeons, one in 100 people who undergo hip or knee replacement surgery develop an infection. Treatment for knee replacement infections includes both nonsurgical and surgical procedures. Some knee replacement infections are superficial, meaning that the infection reaches the skin and tissues around the joint but does not affect the prosthesis itself. These infections can be treated with oral or intravenous antibiotics. If the knee replacement infection is deeper than the skin and tissues around the joint, surgical treatment involves debridement and surgical irrigation of the joint. All contaminated soft tissue is surgically removed, and the prosthesis is cleaned and sterilized. Staged surgery is also an option, involving a series of surgeries to remove and replace the prosthesis. This is necessary if an infection develops months or years after the initial knee replacement. After the prosthesis is removed, a joint irrigation is performed to remove the infected soft tissue within the joint.

[0007] The antibacterial, bactericidal, and disinfectant effects of various metal particles, such as silver, copper, zinc, and titanium, are well known in the art, and extensive literature exists on this topic. Nanoparticle carriers can be used with ultrasound-generated microbubbles as a means of drug delivery to improve their efficacy and enhance cell and vascular permeability, as described in Mullin et al., 2013 ("Nanoparticle delivery enhancement with acoustically activated microbubbles." IEEE Transactions on Ultrasonics, Ferroelectrics, and Frequency Control, 60, 1.). There is also literature in the art on the sonication of metal particles. Pradhan et al., 2016 ("Effect of sonication on particle dispersion, administered dose, and metal release of non-functionalized, non-inert metal nanoparticles") describes probe sonication of metal nanoparticles and shows that probe sonication is the preferred method for dispersion and metal release.

[0008] Similarly, infections of other permanent or temporary implanted devices such as pacemakers, valves, grafts, etc. pose a problem for the healthcare system. Therefore, improvements in medical devices and procedures can improve the level of patient care and have clear medical and economic benefits. Summary of the Invention

[0009] Certain embodiments disclosed herein include a method of ultrasonically cleaning an internal body part by providing a liquid within a body cavity of a living organism, providing nanoparticles within the body cavity, generating ultrasound waves in the liquid with the nanoparticles suspended therein for a predetermined period of time, and removing the liquid with the nanoparticles suspended therein from the body cavity after the predetermined period of time has elapsed.

[0010] Certain embodiments disclosed herein also include a perfusion system for ultrasonically cleaning an internal body part, the perfusion system comprising: an ultrasonic transducer, a perfusion chamber having an inlet and an outlet; at least one control unit including at least one power source configured to supply power to at least the control unit; a liquid supply configured to store a liquid; a nanoparticle supply configured to store one or more nanoparticle powders; a pump configured to supply liquid from the liquid supply to the body cavity via the perfusion chamber inlet and / or extract liquid from the body cavity; and a control circuit configured to couple to one or more of the power source, the liquid supply, the nanoparticle supply, and the pump, wherein the control circuit and the at least one power source are further configured to couple to the ultrasonic transducer, the ultrasonic transducer being configured to contact the liquid.

[0011] A non-limiting aspect of the present invention is Included is a method for ultrasonically cleaning and sterilizing an internal element within a body cavity, the method comprising: providing at least one device, the at least one device comprising at least one ultrasound transducer in communication with at least one container, the container in communication with at least one adhesive element adapted to ensure a reversible sealed connection between the container and the body cavity; reversibly sealingly coupling the container to the body cavity; providing at least one liquid and at least one type of nanoparticles in a container; generating ultrasonic waves for a predetermined period of time in a liquid in which the nanoparticles are suspended by at least one ultrasonic transducer; and removing the liquid in which the nanoparticles are suspended from the at least one container after the predetermined period of time has elapsed.

[0012] In some embodiments, the element is selected from the group consisting of an implant, a soft tissue, a hard tissue, and any combination thereof.

[0013] In some embodiments, the container is either flexible or rigid.

[0014] In some embodiments, the container includes a lid having at least one opening through which at least one ultrasound transducer is inserted.

[0015] In some embodiments, the step of generating ultrasound waves in the liquid in which the nanoparticles are suspended by at least one ultrasound transducer is applied simultaneously with the steps of providing at least one liquid and at least one type of nanoparticles into a container and removing the liquid in which the nanoparticles are suspended from the body cavity.

[0016] In some embodiments, the body cavity is a joint cavity.

[0017] In some embodiments, at least one ultrasound transducer comprises at least one coil at least partially circulating therethrough.

[0018] In some embodiments, the method further comprises providing an electromagnetic field with at least one coil.

[0019] In some embodiments, the step of removing the liquid in which the nanoparticles are suspended from the body cavity after a specified period of time has elapsed is accomplished by generating a magnetic field within the body cavity with at least one ultrasound transducer to collect at least one type of nanoparticles.

[0020] In some embodiments, providing at least one liquid and at least one type of nanoparticles in the container further comprises premixing the at least one liquid and the at least one type of nanoparticles.

[0021] In some embodiments, the pre-mixing is accomplished by means selected from at least one ultrasonic transducer, a mechanical mixer, a magnetic mixer, and any combination thereof.

[0022] In some embodiments, the steps of providing at least one liquid and at least one type of nanoparticles into the container are performed sequentially by (a) providing at least one liquid into the container, followed by (b) providing at least one type of nanoparticles.

[0023] In some embodiments, the step of providing at least one liquid and at least one type of nanoparticles in the container is performed after mixing the at least one liquid and the at least one type of nanoparticles.

[0024] In some embodiments, the perfusion device comprises at least one control unit adapted to regulate the supplying of at least one liquid and at least one type of nanoparticles into the container.

[0025] In some embodiments, the control unit is adapted to adjust at least one parameter selected from the group consisting of speed, flow rate, pressure, timing, frequency of at least one ultrasonic transducer, and any combination thereof, in the step of supplying at least one liquid and at least one type of nanoparticles into the container.

[0026] In some embodiments, the method further comprises providing at least one antibiotic composition.

[0027] In some embodiments, the antibiotic composition comprises one or more of vancomycin, gentamicin, or ciprofloxacin.

[0028] In some embodiments, the method for ultrasonic cleaning and sterilization is performed in minimally invasive arthroscopic knee surgery.

[0029] In some embodiments, the method further comprises the step of providing at least one coloring reagent selected from crystal violet stain into the body cavity.

[0030] In some embodiments, the step of generating ultrasound waves with at least one ultrasonic transducer in the liquid in which the nanoparticles are suspended for a defined period of time continues until all of the coloring reagent has been removed.

[0031] In some embodiments, an irrigation device for ultrasonically cleaning and sterilizing internal components within a body cavity is provided, the irrigation device comprising: at least one container sized and shaped to hold at least one liquid and at least one type of nanomaterial; at least one ultrasonic transducer in communication with the container; The at least one container is associated with at least one adhesive element, the at least one adhesive element adapted to ensure a reversible sealed connection between the at least one container and the body cavity.

[0032] In some embodiments, the internal element is selected from the group consisting of an implant, soft tissue, hard tissue, and any combination thereof.

[0033] In some embodiments, the container is either flexible or rigid.

[0034] In some embodiments, the container includes a lid having at least one opening through which at least one ultrasound transducer is inserted.

[0035] In some embodiments, the body cavity is a joint cavity.

[0036] In some embodiments, at least one ultrasound transducer comprises at least one coil at least partially circulating therethrough.

[0037] In some embodiments, the electromagnetic field is provided by at least one coil.

[0038] In some embodiments, the perfusion device is in communication with at least one premixing device adapted to mix at least one liquid with at least one type of nanoparticles.

[0039] In some embodiments, the pre-mixing is performed by means selected from at least one ultrasonic transducer, a mechanical mixer, a magnetic mixer, and any combination thereof.

[0040] In some embodiments, the perfusion device further comprises a control unit adapted to regulate at least one parameter selected from the group consisting of speed, flow rate, pressure, timing, frequency of at least one ultrasound transducer, and any combination thereof.

[0041] In some embodiments, the perfusion device is used in arthroscopic knee surgery.

[0042] In some embodiments, the perfusion device is in communication with at least one fluid supply configured to store a fluid.

[0043] In some embodiments, the perfusion device is in communication with at least one nanoparticle supply configured to store one or more nanoparticle powders.

[0044] In some embodiments, the perfusion device is in communication with at least one pump configured to deliver at least one fluid into the body cavity.

[0045] In some embodiments, the pump is adapted to extract at least one liquid from at least one container.

[0046] In some embodiments, the perfusion apparatus further comprises a waste unit configured to store fluid extracted from the body cavity.

[0047] In some embodiments, at least one of the liquids comprises an antibiotic composition.

[0048] In some embodiments, the antibiotic composition comprises one or more of vancomycin, gentamicin, ciprofloxacin, and any combination thereof.

[0049] In some embodiments, the at least one container comprises at least one inlet port for introducing at least one liquid into the at least one container.

[0050] In some embodiments, the at least one container comprises at least one outlet port for allowing at least one liquid to be expelled from the at least one container.

[0051] In some embodiments, a method for ultrasonically cleaning and sterilizing an internal element within a body cavity is provided, the method comprising: providing at least one liquid and at least one type of nanoparticles into a body cavity; generating ultrasonic waves by at least one ultrasonic transducer to fluidly contact at least one liquid and at least one type of nanoparticles suspended therein for a defined period of time, the at least one ultrasonic transducer including at least one coil at least partially circulating therethrough; and removing the liquid with the suspended nanoparticles from the body cavity after a predetermined period of time has elapsed; After a predetermined period of time, step (c) of removing the liquid in which the nanoparticles are suspended from the body cavity is performed by generating a magnetic field within the body cavity with at least one ultrasound transducer to collect at least one type of nanoparticles.

[0052] In some embodiments, the internal element is selected from the group consisting of an implant, soft tissue, hard tissue, and any combination thereof.

[0053] In some embodiments, the step of generating ultrasound waves in the liquid in which the nanoparticles are suspended by at least one ultrasound transducer is applied simultaneously with the steps of providing at least one liquid and at least one type of nanoparticles into a container and removing the liquid in which the nanoparticles are suspended from the body cavity.

[0054] In some embodiments, the method further comprises coupling at least one ultrasound transducer to the at least one container.

[0055] In some embodiments, the container is either flexible or rigid.

[0056] In some embodiments, the container is associated with at least one adhesive element adapted to ensure a reversible sealed connection between the container and the body cavity.

[0057] In some embodiments, the method further comprises reversibly sealingly coupling the container with the body cavity.

[0058] In some embodiments, the method further comprises providing at least one liquid and at least one type of nanoparticles in the container.

[0059] In some embodiments, the container comprises a lid, the lid comprising at least one opening through which the at least one ultrasound transducer is inserted.

[0060] In some embodiments, the body cavity is a joint cavity.

[0061] In some embodiments, providing at least one liquid and at least one type of nanoparticles further comprises premixing the at least one liquid and the at least one type of nanoparticles.

[0062] In some embodiments, the pre-mixing is provided by means selected from at least one ultrasonic transducer, a mechanical mixer, a magnetic mixer, and any combination thereof.

[0063] In some embodiments, the steps of providing at least one liquid and at least one type of nanoparticles are performed sequentially by (a) providing at least one liquid, followed by (b) providing at least one type of nanoparticles.

[0064] In some embodiments, the step of providing at least one liquid and at least one type of nanoparticles is performed after mixing the at least one liquid and the at least one type of nanoparticles.

[0065] In some embodiments, the method further comprises providing at least one control unit adapted to regulate the steps of supplying the at least one liquid and the at least one type of nanoparticles.

[0066] In some embodiments, the control unit is adapted to adjust at least one parameter selected from the group consisting of speed, flow rate, pressure, timing, frequency of at least one ultrasonic transducer, and any combination thereof, in the step of supplying at least one liquid and at least one type of nanoparticles into the container.

[0067] In some embodiments, the method further comprises providing at least one antibiotic composition.

[0068] In some embodiments, the antibiotic composition comprises one or more of vancomycin, gentamicin, or ciprofloxacin.

[0069] In some embodiments, the method for ultrasonic cleaning and sterilization is performed in minimally invasive arthroscopic knee surgery.

[0070] In some embodiments, the method further comprises the step of providing at least one coloring reagent selected from crystal violet stain into the body cavity.

[0071] In some embodiments, the step of generating ultrasound waves with at least one ultrasonic transducer in the liquid in which the nanoparticles are suspended for a defined period of time continues until all of the coloring reagent has been removed.

[0072] Particular embodiments of the present disclosure may include some, all, or none of the above advantages. One or more other technical advantages will be readily apparent to those skilled in the art from the drawings, descriptions, and claims contained herein. Additionally, while certain advantages have been enumerated above, various embodiments may include some, all, or none of the enumerated advantages. [Brief explanation of the drawings]

[0073] The foregoing and other objects, features and advantages will become apparent and will be more readily understood from the following detailed description taken in conjunction with the accompanying drawings. [Figure 1] FIG. 1 shows an exemplary, non-limiting schematic diagram of an ultrasonic internal cleaning and sterilization device according to one embodiment of the present invention. [Figure 2] FIG. 2 shows a schematic diagram of the above-described apparatus for use in sterilizing a body cavity via perfusion with ultrasound-transduced nanoparticles according to one embodiment of the present invention. [Figure 3] 3a and 3b show another embodiment of a perfusion device according to an embodiment of the present invention. [Figure 4] 4a-4f show another embodiment of a perfusion device according to an embodiment of the present invention. [Figure 5] 5a and 5b show one embodiment of an ultrasound transducer with a magnetic tip. [Figure 6] 6a and 6b show a specialized device for premixing perfusate with nanoparticles according to one embodiment of the present invention. [Figure 7] FIG. 7 illustrates a method of providing ultrasonic cleaning and sterilization to a body part, performed in accordance with one embodiment of the present invention. [Figure 8] FIG. 8 shows the results of experiments carried out to investigate the ability of the nanoparticles of the present invention to eliminate bacterial adhesion and biofilm formation compared to the current standard of care. [Figure 9] FIG. 9 shows the results of experiments carried out to investigate the ability of the nanoparticles of the present invention to eliminate bacterial adhesion and biofilm formation compared to the current standard of care. [Figure 10] FIG. 10 shows the results of experiments carried out to investigate the ability of the nanoparticles of the present invention to eliminate bacterial adhesion and biofilm formation compared to the current standard of care. [Figure 11] FIG. 11 shows the results of experiments carried out to investigate the ability of the nanoparticles of the present invention to eliminate bacterial adhesion and biofilm formation compared to the current standard of care. [Figure 12] FIG. 12 shows the results of experiments carried out to investigate the ability of the nanoparticles of the present invention to eliminate bacterial adhesion and biofilm formation compared to the current standard of care. [Figure 13] FIG. 13 shows the results of experiments carried out to investigate the ability of the nanoparticles of the present invention to eliminate bacterial adhesion and biofilm formation compared to the current standard of care. [Figure 14] FIG. 14 shows the results of experiments carried out to investigate the ability of the nanoparticles of the present invention to eliminate bacterial adhesion and biofilm formation compared to the current standard of care. [Figure 15] FIG. 15 shows the results of experiments carried out to investigate the ability of the nanoparticles of the present invention to eliminate bacterial adhesion and biofilm formation compared to the current standard of care. [Figure 16] FIG. 16 shows the results of experiments carried out to investigate the ability of the nanoparticles of the present invention to eliminate bacterial adhesion and biofilm formation compared to the current standard of care. [Figure 17]FIG. 17 shows the results of experiments carried out to investigate the ability of the nanoparticles of the present invention to eliminate bacterial adhesion and biofilm formation compared to the current standard of care. [Figure 18] FIG. 18 shows the results of experiments carried out to investigate the ability of the nanoparticles of the present invention to eliminate bacterial adhesion and biofilm formation compared to the current standard of care. DETAILED DESCRIPTION OF THE INVENTION

[0074] Hereinafter, exemplary embodiments will be described in detail with reference to the accompanying drawings so that those skilled in the art can easily realize them. The exemplary embodiments are not limited to the exemplary embodiments described herein, and may be implemented in various forms. Well-known parts will be omitted for clarity of description. Similar reference numerals refer to similar elements throughout.

[0075] It is important to note that the embodiments disclosed herein are merely examples of the many advantageous applications of the innovative teachings described herein. In general, statements in the specification of this application do not necessarily limit any of the various claims. Moreover, some statements may apply to some features but not to other features. In general, unless otherwise specified, singular elements may be in the plural and vice versa without loss of generality.

[0076] The nanoparticles are suspended in a fluid medium and activated using an ultrasound transducer in contact with the fluid medium. The fluid medium can include, but is not limited to, at least one of saline, water, chlorhexidine, or any suitable liquid having a pH level substantially similar to that of the area to be treated, without causing adverse reactions. In one embodiment, the fluid medium is introduced into a body cavity or directed to a body site at a pressure level sufficient to remove debris, biofilm, etc., or at or above that level. The fluid composition can also include other components that can assist in cleansing and disinfecting. The body cavity may contain hard surfaces such as bones, joints, artificial devices, or naturally occurring organic matter accumulations. In one embodiment, a perfusion device can be used before, during, or after a procedure, such as minimally invasive or open surgery, to cleanse, sanitize, or disinfect a body site via perfusion with an ultrasound-transduced fluid in which the nanoparticles are suspended. The perfusion device delivers fluid to the body cavity and can include an ultrasound transducer therein or can be applied using a separate device. In other embodiments, the cleaning and disinfection process can be performed as a treatment for infection in the absence of another surgical procedure. In one embodiment, the ultrasound transducer can be a unidirectional acoustic source that is redirected to become a hemispherical acoustic wave source.

[0077] Nanoparticles are suspended in a fluid medium and activated with an ultrasound transducer in contact with the fluid medium for the purpose of cleansing, purifying, disinfecting, and sterilizing body parts before, during, and / or after surgery. The fluid is intended to treat a body part, an implanted device, or a naturally occurring buildup of organic material, such as calcified tissue or a blood clot. In some embodiments, the body part is a joint, such as a knee, shoulder, or hip joint. In other embodiments, the area to be treated is a cavity, such as a subcutaneous cavity, in which an implanted device is placed. In certain embodiments, the surgery is a minimally invasive procedure, such as a knee arthroscopic procedure. In certain embodiments, the cleansing process itself is performed as a treatment for infection in the absence of another surgical procedure. In other embodiments, the process is used to cleanse and disinfect the interior of the body in open surgery. In yet other embodiments, the disclosed process can be used to cleanse the lumen of a blood vessel or a heart valve from thrombus formation or calcified lesions by inserting a miniature ultrasound transducer, for example, via catheterization, to generate cavitation near the source. In some embodiments, the fluid can further include an antibiotic composition to aid in sterilization. Although occasional references to the human body are made throughout this disclosure, this is not intended to be read as limiting in any way. Similar procedures using the devices disclosed herein can be performed mutatis mutandis on other animals as well. Likewise, it will be apparent that the disclosed methods and devices can be used in open surgery as well as in closed body cavities.

[0078] FIG. 1 is a cross-sectional schematic diagram of an ultrasonic cleaning and sterilization irrigator 100 for use in arthroscopic knee surgery, implemented according to one embodiment. The irrigator 100 includes a surgical probe 110 for insertion into the cavity of a joint 130. In one embodiment, the irrigator 100 may be rigid, while in other embodiments, the irrigator 100 may be flexible. The flexibility of the irrigator can be achieved through material selection or a mechanical design that allows for flexibility. The joint cavity may be the knee or hip joint, or any substantially enclosed structure. A multi-channel conduit 170 can connect the irrigator 100 to one or more controllers 240. The conduit 170 can include, for example, a first channel (not shown) for an electrical cable and a second channel (not shown) for allowing liquid flow. Alternatively, at least two conduits can be used. For example, one conduit can be used for routing electrical cables and another conduit can be used for fluid / liquid delivery. The one or more controllers 240 may include a nanoparticle supply 242, a power supply 244, a liquid supply 246, a control circuit 248, a pump 250, and a waste unit 252. In some embodiments, the nanoparticle supply 242 and the liquid supply 246 are a single supply, and the nanoparticles are already suspended in the liquid supplied into the perfusion device 100 via the conduit 170. Similarly, the liquid may further include an antibiotic. In some embodiments, multiple liquid and / or nanoparticle supplies may be utilized in any combination to produce various effects. For example, a first liquid may contain a first nanoparticle solution and a second liquid may contain an antibiotic solution. The liquids may be supplied alternately, sequentially, or in combination to achieve a desired effect. A valve 178 may be opened to allow the flow of liquid from the control unit 240 to the conduit 170. In certain embodiments, the valve may be one-way or two-way. The liquid may contain a suspension of insoluble nanoparticles. The nanoparticles may range in size from 1 to 900 nanometers. For purposes of this disclosure, micron-sized particles may also be used in certain embodiments, although they may not provide optimal results.Exemplary nanoparticles are metal oxides, metal nitrides, and metal carbides, such as silicon dioxide, aluminum dioxide, magnesium oxide, samarium oxide, titanium dioxide, and zinc oxide; metal nitrides, such as silicon nitride and titanium nitride; and metal carbides, such as silicon carbide, titanium carbide, and tungsten carbide. In certain embodiments, diamond nanoparticles can be used. In certain embodiments, the ratio between the liquid and the nanoparticles suspended therein is 1 part nanoparticle to 10,000 to 100,000 parts liquid medium. In other embodiments, the ratio may be 1 part nanoparticle to 1,000 to 1,000,000 parts liquid medium. In some embodiments, nanoparticles can have a morphology that includes one or more sharp shapes. For purposes of this disclosure, a sharp shape is one that includes surface features capable of penetrating or otherwise disrupting bacterial membranes (cell walls) or infectious agents (pathogens), such as virions. Examples of such shapes include rods (fibers) and stars. The power supply 244 provides power to various electronic and mechanical elements of the controller 240, such as a pump 250 used to regulate flow from the liquid supply 246 through the conduit 170 to the perfusion device 100, and the ultrasound transducer 125 of the perfusion device 100. A control circuit 248 can be used to regulate the power supply 244, or the nanoparticle supply 242 and the liquid supply 246, to vary the type of nanoparticles (if different particles are used), the ratio between the liquid and the nanoparticles, etc. The control circuit 248 can be coupled to actuators (not shown), sensors (not shown), etc. The controller 240 can use the pump 250 to supply a flow of liquid with suspended nanoparticles to the irrigation device 100. The controller 240 can then be configured to activate the ultrasound transducer, which generates cavitation bubbles that impinge, forming microjets in the liquid carrying the nanoparticles, resulting in the nanoparticles impinging on the surfaces of the joint cavity. This allows for the cleansing and disinfection of the joint cavity and structures within it by destroying pathogens through membrane disruption.Biofilms can develop and grow on implanted medical devices, such as pacemakers, artificial heart valves, artificial joints, and catheters, potentially causing infection. The microjet injects nanoparticles through the membrane's surface, destroying the microorganisms within. Disrupting the biofilm also allows for further penetration of additional antibiotics, if needed. The irrigator 100 can be used before, after, during, and to assist in surgical procedures requiring sterilization. Once the procedure is complete, fluids and debris are aspirated through the conduit 170 and stored in the waste unit 252. The position and location of the probe 110 within the body cavity (e.g., joint cavity) can be visualized by images generated by the endoscope 120. In some embodiments, fluid can be retained within the body cavity (e.g., joint cavity) during surgery to expand the cavity and create a more maneuverable surgical space for surgical tools, such as the surgical tool 140. In another embodiment, after the fluid has collected within the body cavity, ultrasonic transduction of the airborne nanoparticles can be performed via a separate transducer device. In some embodiments, the liquid can include one or more additional antibiotic compositions such as vancomycin, gentamicin, or a fluoroquinolone such as ciprofloxacin in a concentration range of 10 mg to 50 mg per liter, and any combination thereof.

[0079] It should be noted that the devices and methods of the present invention may also be utilized to clean and sterilize internal elements within a body cavity, such as implants, soft tissue, hard tissue (e.g., bone), and any combination thereof.

[0080] FIG. 2 is a schematic diagram of a perfusion device 100 implemented according to one embodiment. The device 100 includes a surgical probe 110, a transducer 125, a handle 116, and a valve 115. The transducer 125 is attached to a multi-channel conduit 170, which carries, for example, at least one electrical cable 176 and at least one fluid transfer tube 177. While a multi-channel conduit is described in this example, it will be readily apparent to one skilled in the art that one or more conduits may be utilized to provide one or more fluid supplies, nanoparticle supplies, or power supplies without departing from the scope of this disclosure. Power to the transducer 125 may be provided by a controller (described in more detail in connection with FIG. 1) or a separate power source. The valve 115 allows for manual control of fluid flow from the device 100 to the probe 110 and into a body cavity (e.g., a joint cavity), thereby allowing an operator of the device 100 to manually control the pressure of fluid exiting the device. A mechanical handle 116 allows for manual control of the device. For example, a visual image of the interior, obtained by an endoscope, can be used to guide the probe 110 to the correct location within the body cavity. In some embodiments, the transducer 125 can have a portion that extends through at least a portion of the probe 110 to activate the nanoparticles closer to the body cavity. In some embodiments, the perfusion device 100 can be operable to eject fluid from the probe 110 at high pressure to form a fluid jet having kinetic energy that can assist in debris removal by transferring kinetic energy from the fluid jet to the treatment surface upon impact. For example, the probe 110 can be tapered toward an outlet from which the fluid is ejected, creating a pressure differential. Such perfusion devices are described in more detail in PCT Publication WO 2020 / 141,533, which has at least one co-inventor, the contents of which are incorporated herein by reference.

[0081] Reference is now made to Figures 3a, 3b, 4a and 4b, which illustrate another embodiment of a perfusion device 300 of the present invention.

[0082] According to this embodiment, the device is reversibly attachable to a body cavity (e.g., the knee, as shown in Figures 4a and 4b) and serves as both an ultrasound transducer holder and a container for holding the perfusion fluid and nanoparticles.

[0083] It should be noted that the body cavity may be any cavity selected from among the knee, shoulder, elbow, wrist, ankle or hip joints.

[0084] According to this embodiment, the perfusion device 300 comprises at least one ultrasound transducer 301 and a flexible container 302 having a lid 303. The lid 303 comprises at least one opening through which the at least one ultrasound transducer 301 can pass. As described above, the at least one ultrasound transducer 301 is adapted to generate acoustic waves within the flexible container 302, which contains perfusion fluid and nanoparticles, as described below, that sterilize the implant by imparting kinetic energy to the nanoparticles and assist in removing debris from the treated surface (e.g., implant) upon impact.

[0085] The flexible container 302 is designed to hold a sufficient amount of liquid therein, including the nanoparticle suspension.

[0086] It should be noted that the liquid medium is mixed with the nanoparticles in the flexible container 302, for example, by manipulating one or more ultrasonic transducers 301. In another embodiment, the liquid and nanoparticles are premixed in separate devices, as shown in Figures 6a and 6b below. After mixing, the liquid and nanoparticles are delivered into the area to be treated.

[0087] In a further embodiment, the liquid and nanoparticles are delivered separately into the body cavity, with mixing occurring only within the body cavity.

[0088] As described above, the ultrasound transducer generates ultrasound waves to create disruptive cavitations in the fluid, causing nanoparticles to impact surfaces within the body cavity of the subject, sterilizing the subject (by disrupting the membranes of bacteria and pathogens that come into contact with the nanoparticles in the fluid, thereby inhibiting their function). It is also within the scope of the present invention for the perfusion fluid to contain one or more additional antibiotic compositions, such as vancomycin, gentamicin, or a fluoroquinolone, such as ciprofloxacin.

[0089] According to one embodiment, the perfusion device 300 includes a valve that allows for controlled flow of ultrasound-transducing nanoparticles suspended in a liquid into the body cavity.

[0090] According to another embodiment, the perfusion device 300 further includes at least one adhesive element 304 that connects to the bottom of the flexible container 302. The adhesive element 304 is adapted to provide a sealed connection with a body cavity (e.g., the knee 305, as shown in Figures 4a and 4b), thereby obtaining a tight connection between the flexible container 302 and the body cavity (i.e., no liquid leaks out).

[0091] According to one embodiment, adhesive elements 304 are bonded to the bottom of flexible container 302 at multiple locations. According to another embodiment, adhesive elements 304 are bonded to the entire periphery of the bottom of flexible container 302.

[0092] Referring now to Figures 4a and 4b, the perfusion device 300 and adhesive element 304 are shown in sealing connection with the knee when the knee is held in a bent position (Figure 4a) and a straight position (Figure 4b).

[0093] 4c-4e, another embodiment of the present invention is shown in which the transducer is disposed perpendicular to the flexible container 302. An adhesive element 304 is connected to the bottom of the flexible container 302. The adhesive element 304 is adapted to provide a sealed connection with a body cavity (e.g., the knee 305 as shown in FIGS. 4a and 4b), thereby providing a tight connection between the flexible container 302 and the body cavity (i.e., no liquid leaks out).

[0094] Also shown in this embodiment are perfusion inlet and outlet ports 306a and 306b, through which perfusion fluid is introduced into and withdrawn from the container 302.

[0095] According to another embodiment, the ultrasonic transducer 301 may be rigid. Alternatively, the ultrasonic transducer 301 may be flexible, for example as shown in Figure 4f.

[0096] Referring to Figures 5a and 5b, another optional embodiment of the ultrasonic transducer 301 is shown.

[0097] At the end of the procedure (after sterilization), aspiration of the liquid medium is performed, which requires extraction of the nanoparticles and debris. According to this embodiment, at least one of the ultrasonic transducers 301 comprises at least one magnetic coil 401 wound at least partially around the body 402 of the ultrasonic transducer 301.

[0098] That is, the ultrasound transducer 301 has a magnetic tip by generating an electromagnetic field with at least one coil 401. Such a magnetic tip is used to collect nanoparticles from within a body cavity.

[0099] The remaining liquid and debris can be sucked into the waste unit, for example by a pump.

[0100] It should be noted that the time frame for such extraction may vary depending on considerations such as the location of the body cavity, the type of infection and / or the level of infection.

[0101] Instead of applying a magnet, the nanoparticles can also be subjected to suction of the perfusate.

[0102] Referring to Figures 6a and 6b, a dedicated device is shown for pre-mixing the perfusate and nanoparticles to provide a liquid medium containing a nanoparticle suspension.

[0103] According to this embodiment, a dedicated device 500 comprises at least one ultrasound transducer 503 coupled to an electronic connector 501 via at least one electronic cable 502. Such ultrasound transducer 503 is in fluid communication with a container 504 for holding nanoparticles and perfusion fluid.

[0104] In operation, perfusion fluid is provided through at least one inlet 505. Once the nanoparticles are provided into the container 504, the ultrasonic transducer 503 is activated to generate ultrasonic waves, which impart kinetic energy to the nanoparticles, causing them to mix and resulting in a liquid fluid containing the nanoparticles as a suspension.

[0105] In some embodiments, a dedicated premixing device 500 can mix nanoparticles in multiple different liquids, or alternatively, a dedicated premixing device 500 can mix multiple different nanoparticles in one liquid.

[0106] In some embodiments, a dedicated premixing device 500 can dispense a first nanoparticle suspension into a first liquid and then dispense a second nanoparticle suspension into a second liquid. The first and second suspensions can differ in nanoparticle type, nanoparticle size, nanoparticle to liquid ratio, liquid type, liquid viscosity, or a combination thereof.

[0107] FIG. 7 is a flowchart of an exemplary method for providing ultrasonic cleaning and sterilization inside a body using an ultrasonic irrigation device, performed according to one embodiment.

[0108] In step S310, a surgical probe of the perfusion device is inserted into a particular body cavity through a keyhole incision. The body cavity may be the knee, shoulder, elbow, wrist, ankle, or hip cavity. Of course, other joints are possible, but these are listed here as the most common types undergoing arthroscopic surgery. In certain embodiments, other body cavities or organs, such as arteries, may also be treated. The body cavity must be capable of containing a certain amount of liquid containing the nanoparticle suspension. It will be appreciated that if the ultrasonic cleaning method is performed during open surgery, the insertion through the keyhole incision in step S310 is not required.

[0109] In optional step S320, a controller coupled to the perfusion device mixes the liquid medium with the nanoparticles, for example, by operating one or more actuators. In another embodiment, the liquid and nanoparticles are premixed in a prepared solution. In yet another embodiment, the liquid and nanoparticles are delivered into the treatment area.

[0110] In step S330, the liquid medium with the nanoparticles suspended therein is supplied to the perfusion device by a pump through a conduit. In one embodiment, the liquid and nanoparticles are supplied separately to the perfusion device and mixed within the perfusion device. In a further embodiment, the liquid and nanoparticles are supplied separately to the body cavity and mixed only within the body cavity. The pump can be manually turned on via a controller. In one embodiment, the liquid medium can move through the conduit only when a valve in the controller is in an open state.

[0111] In step S340, power is supplied to the ultrasound transducer by a controller or a separate power source. In some embodiments, the ultrasound transducer is housed in a perfusion device. The transducer generates ultrasound waves to create disruptive cavitations in the fluid, thereby impacting nanoparticles on surfaces within the body cavity to be treated. The perfusion device can include a valve to control the flow of ultrasound-converted nanoparticles suspended in the liquid into the body cavity. In another embodiment, the transducer can be a separate device inserted through a separate surgical incision (if the procedure is not an open surgery) to generate ultrasound waves from within the body cavity. The surfaces of the body cavity are impacted by nanoparticles suspended in the liquid. In certain embodiments, a liquid medium is supplied at a regulated flow rate and pressure. In some embodiments, the perfusion device can supply a first liquid containing a first nanoparticle suspension, followed by a second liquid containing a second nanoparticle suspension. The first and second suspensions can differ in nanoparticle type, nanoparticle size, nanoparticle to liquid ratio, liquid type, liquid viscosity, or combinations thereof. Bacteria and pathogens that come into contact with the nanoparticles in the fluid have their membranes disrupted, inhibiting their function, resulting in the disruptive cavitation and resulting nanoparticle impacts cleaning and disinfecting the joint. In some embodiments, the fluid can contain one or more additional antibiotic compositions, such as vancomycin, gentamicin, or a fluoroquinolone such as ciprofloxacin, or any combination thereof, in a concentration range of 10 mg to 50 mg per liter.

[0112] In step S350, after the liquid medium has been in contact with the body cavity for a period of time, aspiration of the liquid medium is performed. This time frame varies depending on considerations such as the location of the body cavity, the type and / or level of infection. Once irrigation is complete, the remaining liquid and any debris are aspirated from the probe into the irrigator and sent to a waste unit by a pump and power source. In another embodiment, the waste unit is separate from the control unit. In another embodiment, aspiration can be performed by a suction device separate from the irrigator. In some embodiments, the liquid remains in the joint during surgery to allow the joint to expand. In other embodiments, aspiration is performed after irrigation / sterilization, followed by the delivery of a second liquid with or without nanoparticles to expand the joint. The surgery is then performed, followed by additional activation and final aspiration to remove all of the nanoparticle-suspended liquid from the joint.

[0113] In optional step S360, a fluid medium containing one or more antibiotic compositions, such as vancomycin, gentamicin, or a fluoroquinolone such as ciprofloxacin, or any combination thereof, at a concentration ranging from 10 mg to 50 mg per liter, is delivered to the perfusion device via a conduit by a pump. The pump can be turned on manually via a controller. In some embodiments, steps S320-S350 can be performed multiple times. In certain embodiments, steps S320-S360 can also be performed multiple times, such that a first set of steps S320-S350 is performed and step S360 is performed before or after each set of steps.

[0114] According to another embodiment, ultrasound is provided simultaneously with irrigation. Unlike other methods, according to this embodiment, a fluid medium and nanoparticles suspended therein are delivered to and simultaneously expelled from a body cavity (e.g., knee 305) while ultrasound is applied to treat the body cavity (e.g., knee 305).

[0115] According to another embodiment, a coloring agent that adheres to biofilm is added to the body cavity to be treated. The treatment is continued until the color disappears. Such a coloring agent can be used to confirm that all biofilm has been removed and the treatment is complete. An example of such an agent is crystal violet stain.

[0116] Example 1 The incidence of surgical site infections after knee replacement surgery has been reported to be 1%–20%. These infections lead to increased patient morbidity and reduced hospital reimbursement. Hardware implanted during surgery is susceptible to colonization by bacterial biofilms, resulting in hardware loosening and infection. While the causes of colonization are still under investigation, it is likely a combination of bacteria introduced through the skin during the initial surgery, as well as systemic infection by bacteria normally present in the oral cavity and / or intestinal tract. Once formed, bacterial biofilms are difficult to treat and are typically treated clinically by physical removal during additional surgeries (Ehrlich et al., 2012; Stoodley et al., 2011; Stoodley et al., 2013).

[0117] The present inventors have expanded on a successful in vitro study to evaluate the effect of nanoparticle sonication on bacterial adhesion and biofilm formation in a whole-joint infection model. This study examined orthopedic implants and surrounding host tissue 21 days after implantation / inoculation. The objective was to investigate the ability of the nanoparticles of the present invention to eliminate bacterial adhesion and biofilm formation compared to the current standard of care.

[0118] method Animal Experiments: Animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of the Allegheny Health Network Research Institute (AHNRI) under Animal Experiment Protocol # 1096. Animals were housed in the AHNRI animal care facility. Animal model: Adult male Sprague-Dawley rats (approximately 350 g) Incubation period: 21 days

[0119] Bacterial inoculation: A clinical strain of methicillin-resistant Staphylococcus aureus was streaked onto blood agar plates and incubated overnight at 37°C and 5% CO2. A single colony was picked from the plate and incubated in 10 ml of brain heart infusion for 2 hours until logarithmic growth phase was reached. At this point, the culture contained approximately 1 x 10 6 The broth culture was diluted to a concentration of CFU / 2.5 μL and was divided into 1 ml aliquots for single use and transported to the animal facility on ice.

[0120] Surgical Procedure: Preoperative baseline measurements, including body weight and local temperature, were performed the day before surgery. On the day of surgery, rats were anesthetized with 1-3% isoflurane in 1 L / min of oxygen. Buprenorphine 0.05 mg / kg was administered IP 30 minutes before surgery. A 3-4 cm skin incision was made on the cranial lateral side of the right femoral surface, extending from the supracondylar region of the right leg to the tibia. An intercondylar canal was drilled in the tibia, and 10 μl of 2 × 107 Staphylococcus aureus was injected. A 1.5 mm diameter, 5.0 mm long titanium maxillofacial screw was then implanted. Control animals received saline instead of the inoculum. The wound was closed with 5-0 Vicryl deep dermal sutures. Rats received one postoperative dose of buprenorphine-SR and three doses of meloxicam at 24-h intervals.

[0121] On postoperative days 1, 3, 7, 14, and 21, the animals were monitored for changes in body weight, incision integrity, changes in gait, and static weight-bearing ability. On day 21, the animals were anesthetized, and the native joint (left knee) was incised as described above. The animals were then anesthetized and administered the nanoparticle treatment disclosed herein to the native knee (i.e., an Ag nanoparticle solution at a concentration of 12 mg nanoparticles / 300 mL saline was administered for 15 minutes). Removed screws: The implanted screws were removed, and the designated treatment (nanoparticles or standard treatment) was administered.

[0122] Treatment according to the present invention: A silicon container (as disclosed in this invention) was screwed and 20 ml of HBSS containing 12 mg / 300 mL of nanoparticles was added.

[0123] The samples were sonicated for 15 minutes. At the end of the sonication, the magnetic beads were removed from the solution. The samples were then serially diluted and plated onto BHI (brain heart infusion) medium. After 24 hours of incubation at 37°C, the colony-forming units (CFUs) were counted at the appropriate dilutions and the CFUs were calculated. Crystal violet staining was then performed.

[0124] SOC treatment: The screw was placed in a sterile polystyrene centrifuge tube containing 1 mL of HBSS. The sample was vortexed for 10 seconds for three cycles. The sample was then serially diluted and plated onto BHI (brain heart infusion) medium. After 24 hours of incubation at 37°C, colony-forming units (CFUs) were counted at appropriate dilutions and the CFU count was calculated. Crystal violet staining was then performed.

[0125] Crystal violet staining: After treatment, the screws were immersed in 1 mL of crystal violet solution and incubated at room temperature for 15 minutes. After incubation, the crystal violet was removed and the screws were transferred to a new tube containing 1 mL of deionized water. After 1 minute, the screws were transferred to a new tube containing 1 mL of water. This process was repeated three times. The screws were then placed in an empty tube and allowed to dry overnight. The next day, 100 μL of 100% ethanol was added to the tube containing the screws and incubated for 10 minutes. After incubation, the crystal violet-ethanol solution was transferred to a 96-well plate. The absorbance of the solution was measured at 570 nm using a Tecan plate reader.

[0126] Screw cavity: After removing the embedded screw, the screw cavity was filled with 20 μL of HBSS and collected. The collected solution was then serially diluted and plated onto BHI (brain heart infusion) medium. After 24 hours of incubation at 37°C, colony-forming units (CFU) were counted at appropriate dilutions, and CFU / mL was calculated.

[0127] Tissue CFU: At the time of collection, samples were placed in HBSS in a sterile 1 mL polystyrene centrifuge tube. Tissue sections were used for CFU calculations.

[0128] To calculate the number of bacteria removed using the technology of the present invention, a sample was weighed, placed in a silicone container, and 25 ml of HBSS containing 12 mg / 300 mL of nanoparticles was added. The sample was sonicated for 15 minutes using the device of the present invention. After sonication, the magnetic beads were removed from the solution. The sample was then serially diluted and plated onto BHI (brain heart infusion) medium. After 24 hours of incubation at 37°C, colony-forming units (CFUs) were counted at the appropriate dilutions, and the number of CFUs per mg of tissue was calculated.

[0129] To calculate the residual bacteria after using the present technique, tissue was placed in a new tube containing 1 mL of HBSS. This was sonicated for 5 minutes, vortexed for 10 seconds, sonicated for 5 minutes, and vortexed for 10 seconds. The sample was then serially diluted and plated onto BHI (brain heart infusion) medium. After 24 hours of incubation at 37°C, colony-forming units (CFU) were counted at the appropriate dilutions to calculate CFU / mL.

[0130] Histology: Upon collection, samples were placed in sterile 1 mL polystyrene centrifuge tubes. Tissues were fixed in FFPE, paraffin-embedded, sliced ​​into 10 μm sections using a microtome, and mounted on microscope slides. Tissues were stained with hematoxylin and eosin and imaged with EVOS FL. Tissue sections were subjected to histopathological examination by a board-certified clinical pathologist to confirm signs of focal cellular damage and the presence of nanoparticles.

[0131] result Summary of visual examination: On day 21, all animals showed various signs of inflammation, including redness, tenderness, and elevated temperature in the implanted joint. Upon opening the joint capsule, many animals showed cloudy serous fluid in the joint cavity and evidence of scar tissue within the knee joint. Upon screw removal, most animals showed bone defects around the implanted screws, indicating the presence of a slow-acting infection within the tissue.

[0132] Joint capsule swab: A total of 25 pockets were tested for the presence of planktonic bacteria, with a positive result indicating the presence of one or more colonies on the blood agar plate.

[0133] Joint capsule swab test summary: Mannitol salts are selective for S. aureus, as evidenced by a red-to-yellow color change observed upon growth on culture media. Capsular swabs tested positive for planktonic S. aureus with varying levels of bacterial growth in all 20 capsules (Table 1). This was reflected by visual inspection of the pockets at the time of material retrieval, which revealed cloudy fluid within the capsule cavity. TIFF2025537738000002.tif30170

[0134] Summary of CFU testing: Overall, nanoparticles eliminated more bacteria compared to SOC, and fewer bacteria remained after nanoparticle treatment.

[0135] Referring now to Figure 8, the CFU of the screws is shown compared between the two treatments. The nanoparticle bar shows the median CFU of 15 screws. The SOC bar shows the median CFU of 10 screws. A total of 25 screws were analyzed. The nanoparticle treatment (labeled "nanoparticle" in the figure) removed more bacteria compared to the SOC treatment.

[0136] Furthermore, the nanoparticles eliminated an average of 1 x 104 S. aureus per gram of tissue. Figure 9 shows the calculated CFU of tissue after nanoparticle treatment. The CFU / ml bar represents the average CFU of fluid after the tissue was subjected to the treatment of the present invention (labeled "nanoparticles" in the figure). The CFU / gram tissue bar represents the bacteria eliminated per gram of tissue after the treatment of the present invention. A total of 25 tissue samples were analyzed.

[0137] Crystal violet staining Overall, crystal violet readings using the technology of the present invention were reduced five-fold compared to standard of care. Figure 10 shows the trend for reduction in individual screws treated with the nanoparticles of the present invention, and Figure 11 shows the statistical significance between the two treatment groups.

[0138] Note that in Figure 10, each bar represents a crystal violet absorbance measurement from each screw after its assigned treatment, and in Figure 11, the nanoparticle bar represents the median crystal violet result for the 15 screws assigned the inventive treatment. The standard care bar represents the median crystal violet result for the 10 screws assigned the SOC treatment.

[0139] Statistical analysis of the CV absorbance measurements yielded a significant p-value of less than 0.5. TIFF2025537738000003.tif17170

[0140] Safety Results All animals were sonicated with the device of the present invention without visible signs of tissue damage. Animals tolerated the sonication under anesthesia with no signs of pain, distress, or adverse events reported.

[0141] Histology Summary: The histological staining results did not suggest any damage to the host tissue. No traces of the nanoparticles of the present invention were found, and intact cellular structures were observed. These findings were verified by a board-certified clinical pathologist.

[0142] Consideration The results of this study further validate in vitro studies that demonstrated the effectiveness of the present technology against methicillin-resistant Staphylococcus aureus (MRSA) biofilms on titanium screws and test specimens.

[0143] The rat whole-joint infection model was successfully performed, and all test animals reached the 21-day endpoint without any reported adverse events. At 21 days, all animals showed various signs of inflammation in the implanted joint, including redness, tenderness, and elevated temperature. Upon screw removal, most animals had bone defects around the implanted screw, and swabs of the joint capsule were positive for Staphylococcus aureus, indicating the presence of a slow-acting infection within the tissue, demonstrating the validity of this model for inducing chronic whole-joint infection.

[0144] The technology of the present invention has demonstrated its effectiveness in removing bacterial biofilm from implanted titanium screws. Evaluation of nanoparticles in the ultrasonic treatment process of the present invention showed that the nanoparticles were capable of removing approximately 1 x 104 Staphylococcus aureus per gram of tissue, more bacteria than the SOC process. Crystal violet measurements in the treatment using the nanoparticles of the present invention were five times lower than standard of care, with a statistically significant difference between the two treatment groups, based on a Mann-Whitney analysis with a p-value of 0.002335.

[0145] Furthermore, the nanoparticle sonication process of the present invention has been shown to be safe in an in vivo model. No adverse events were reported and no evidence of pain or distress was observed in the test animals when sonication was performed under anesthesia. Visual examination of the host tissue after sonication revealed no visible damage. Furthermore, pathological examination reported no evidence of nanoparticles and confirmed the presence of intact host tissue.

[0146] In conclusion, these results support our hypothesis that the nanoparticle sonication technique of the present invention can remove adherent bacterial biofilms from implanted orthopedic hardware and joint capsules without disrupting native tissue. Therefore, the technique of the present invention has the potential to provide a novel solution for patients with suspected infections after joint replacement surgery.

[0147] Example 2 As mentioned above, the incidence of surgical site infections after knee replacement surgery has been reported to be approximately 1.5%. These infections lead to increased patient morbidity and reduced hospital reimbursement. Hardware implanted during surgery is susceptible to colonization by bacterial biofilms, resulting in hardware loosening and infection. While the causes of colonization are still under investigation, it is likely to result from a combination of bacteria introduced through the skin during the initial surgery, as well as systemic infection by bacteria normally present in the oral cavity and / or intestines. Once formed, bacterial biofilms are difficult to treat and are typically treated clinically by physically removing them during additional surgery.

[0148] The following example demonstrates the optimization of nanoparticle concentrations. As will be shown later, USN (ultrasound and nanoparticles) is statistically significantly superior to US (ultrasound alone) by 1.45-2.36 times in both bacterial and artificial biofilms.

[0149] Research Overview Titanium elements (disks and screws) 1. Infect the animals with a bacterial biofilm of methicillin-resistant Staphylococcus aureus, a strain commonly isolated from orthopedic infections. 2. The membranes were coated with an artificial material, gellan-based hydrogel and methylcellulose, which mimicked the mechanical properties of biofilms.

[0150] Each specimen was treated with either USN (ultrasound and nanoparticles) or US (ultrasound alone) in which two types of nanoparticles were compared: Ti2O3 (30–50 Nm) and Ag (30–50 Nm).

[0151] Two types of titanium elements were used to best represent the surface of implants used in total knee replacements and the deep grooves that form between the implant and the bony elements of the knee.

[0152] Testing using biofilm simulants background Biofilm-forming microorganisms are ubiquitous, yet continuous cultivation of these microorganisms with predictable biofilm growth and structural characteristics remains challenging. The development of reliable simulated biofilms has been limited by a lack of information on the fluid-structure interactions and microbial subpopulations involved in biofilm formation and detachment under mechanical stress. We present gellan-based hydrogels as an alternative physicochemical biofilm mimic. The hydrogel's mechanical properties, storage modulus (G') and loss modulus (G''), can be tuned and adapted to mimic biofilms of various strengths by varying the concentrations of gellan and methylcellulose.

[0153] The storage modulus of the hydrogel ranges from 2 to 20 kPa, and the loss modulus ranges from 0.1 to 2.0 kPa. The material constants of the hydrogel and Pseudomonas putida KT2440 biofilm were experimentally determined by rheometric analysis. A simplified biofilm mimic based on highly hydrolyzed gellan hydrogel and methylcellulose was established using experimental design. This model system design was compared with actual biofilms and adapted to mimic the mechanical properties of biofilms, thereby obtaining biofilm-like viscoelastic behavior. The use of gellan-based hydrogels allows us to mimic biofilm behavior in the absence of growth effects, thereby simplifying the system. Biofilm characterization tools can be tested and validated before application to actual biofilms. In general, this method allows for more rapid and reliable testing of the mechanical properties of biofilms (Gunther S., Kwad A., A Biomimetic Gellan-Based Hydrogel as a Physicochemical Biofilm Model Journal of Biomaterials and Nanobiotechnology 05(02), 2014).

[0154] Testing was performed using a material that best represents the mechanical properties of biofilms naturally produced by bacteria. The material used was a mixture of 3% w / v hydrogel and 3% w / v methylcellulose. This mixture produced a layer (50-100 mg) substantial enough to mimic a biofilm layer produced in the human body over a long period of at least several months. This artificial biofilm could be produced in a relatively short time (1 day). Cultivating the same amount of biofilm in a laboratory could take several months.

[0155] method Using a small brush, the simulant (3% w / v hydrogel and 3% w / v methylcellulose) was applied three times to the titanium sample surface. The sample was then heated in an oven at 100°C for 15 minutes until completely dry. The total weight of the coating was 50–100 mg.

[0156] Two beakers containing 300 ml of water were inserted into an ultrasonic bath filled with 2400 ml of water, bringing the total water volume to 3 liters. The ultrasonic bath was operated at 150 watts (50 watts per liter).

[0157] One beaker contained only water, while the other contained nanoparticles at different concentrations. The samples were placed in the beakers and subjected to ultrasonic irradiation until complete cleaning was achieved. The cleaning process was determined to be complete when the sample weight differed by less than 5 mg between cleaning cycles and the total weight of the cleaned element was 5 mg different from the original element weight. Once complete, the samples were thoroughly dried in an oven and weighed.

[0158] Optimizing nanoparticle concentration Concentrations were measured as milligrams of Ti2O3 nanoparticles (30-50 nm) in 300 ml of water in a 450 ml beaker.

[0159] The ultrasound used was 50 watts per liter. The test concentrations used were 0 (sonication only, no nanoparticles), 3, 6, 12, and 20 mg per 300 ml.

[0160] Test Procedure Two beakers containing 300 ml of water were placed in an ultrasonic bath containing an additional 2400 ml of water. The bath was set at 150 watts (150 watts for a total of 3000 ml - 50 watts per liter). The bath was not preheated and reached a maximum temperature of 35°C during the test.

[0161] Fifteen disks were coated with the simulant. The disks were washed until they were clean. The effectiveness of the cleaning was determined by weighing the disks and comparing the weight of the washed disks with the original weight of the disks. This process was performed three times for each nanoparticle concentration.

[0162] The cleaning rate parameter was calculated as (contaminated disc weight - final weight after cleaning) / (time required for cleaning).

[0163] The effectiveness of the washout was expressed in mg / min and calculated for each concentration level tested.

[0164] result The table below shows the concentrations and cleaning rates. TIFF2025537738000004.tif30170

[0165] Please note that "nano concentration" refers to mg / 300 ml of water and "wash rate" refers to mg / min.

[0166] Figure 12 shows the cleaning rate results for different concentrations of Ti2O3 nanoparticles: 0, 3, 6, 12, and 20 mg per 300 ml.

[0167] The table below summarizes the nanoparticle concentrations and average cleaning rates. TIFF2025537738000005.tif22170

[0168] As can be seen, the 12 mg per 300 ml concentration resulted in the highest cleaning rate of 3.62 mg / min on average, compared to 1.94 mg / min with sonication alone (0 mg / 300 ml), a factor of 1.86. It can also be seen that the addition of nanoparticles increased the average cleaning rate by 51.3%.

[0169] Example 3 Testing titanium discs with Ti2O3 nanoparticles and biofilm simulants In the following example, superior cleaning effectiveness is demonstrated by adding a predetermined concentration of nanoparticles to a cleaning solution of 12 mg per 300 ml. This effect was demonstrated using titanium disks and biofilm simulants. The concentration was measured as milligrams of Ti2O3 nanoparticles in 300 ml of water placed in a 450 ml beaker.

[0170] Test parameters Test item - disk (diameter 20 mm) Nanoparticles - Ti2O3 (30-50nm) Ultrasonic - 50 watts / liter Concentrations - 0, 12, 20 mg per 300 ml (0 is for sonication only, no nanoparticles)

[0171] Test Procedure Two beakers containing 300 ml of water were placed in an ultrasonic bath containing an additional 2400 ml of water. The bath was set at 150 watts (150 watts for a total of 3000 ml - 50 watts per liter). The bath was not preheated and reached a maximum temperature of 35°C during the test.

[0172] Twelve disks were contaminated using the simulant. The disks were washed until clean. The cleaning effectiveness was determined by weighing the disks and comparing the washed disks with the original and contaminated disk weights. This process was performed four times for each nanoparticle concentration. The cleaning rate parameter was calculated as (contaminated disk weight - final disk weight after washing) / (time required for cleaning).

[0173] This provided a cleaning rate parameter to compare the cleaning efficiency of sonication with nanoparticles to sonication alone. The cleaning rate was measured in mg / min.

[0174] result The table below summarizes the nanoparticle concentrations (mg / 300 ml water) and washing rates (mg / min). TIFF2025537738000006.tif35170

[0175] Figure 14 shows the wash rates for three concentrations (four times each concentration).

[0176] The table below summarizes the average cleaning rates for each nanoparticle concentration. TIFF2025537738000007.tif26170

[0177] Figure 15 shows the average wash rates for three concentrations (four runs at each concentration).

[0178] The results show that cleaning rates for contaminated titanium disks using Ti2O3 nanoparticles at a concentration of 12 mg per 300 ml and ultrasound were, on average, 1.85 times more efficient than using ultrasound alone (0 mg per 300 ml). Cleaning results using USN were significantly better than US (based on a one-way ANOVA with Bonferroni post-hoc correction for three groups (p<0.05)).

[0179] Example 4 Testing titanium screws with Ti2O3 nanoparticles and biofilm simulants In this example, superior cleaning effectiveness is demonstrated by adding a predetermined concentration of nanoparticles, 12 mg per 300 ml, to a cleaning solution. This effect was demonstrated using a titanium screw and a biofilm simulant. The concentration was measured as milligrams of TiO nanoparticles (30-50 nm) in 300 ml of water in a 450 ml beaker.

[0180] Test parameters Test item - Screw (M6, length 20 mm) Nanoparticles - Ti2O3 (30-50nm) Ultrasonic - 50 watts / liter Concentrations - 0, 12, 20 mg per 300 ml (0 is for sonication only, no nanoparticles)

[0181] Test Procedure Two beakers containing 300 ml of water were placed in an ultrasonic bath containing an additional 2400 ml of water. The bath was set at 150 watts (150 watts for a total of 3000 ml - 50 watts per liter). The bath was not preheated and reached a maximum temperature of 35°C during the test.

[0182] The simulant was applied to 12 screws. The screws were washed until they were clean. The effectiveness of the cleaning was determined by weighing the screws and comparing the weight of the screw after cleaning with the original weight of the screw and the weight of the contaminated screw. This process was carried out four times for each nanoparticle concentration. The cleaning rate parameter was calculated as (contaminated screw weight - final screw weight after cleaning) / (time required for cleaning).

[0183] This provided a cleaning rate parameter to compare the cleaning efficiency of sonication with nanoparticles to sonication alone. The cleaning rate was measured in mg / min.

[0184] result The table below summarizes the nanoparticle concentrations (mg / 300 ml water) and their washing rates (mg / min). TIFF2025537738000008.tif36170

[0185] Figure 16 shows the wash rates for three concentrations (four times each concentration).

[0186] The table below summarizes the average cleaning speed for each concentration. TIFF2025537738000009.tif26170

[0187] conclusion The above example shows that cleaning rates for contaminated titanium screws using Ti2O3 nanoparticles at a concentration of 12 mg per 300 ml and ultrasound are, on average, 2.36 times more efficient than US (0 mg per 300 ml). This result is statistically significant based on a one-way ANOVA with Bonferroni correction (p<0.05) for three groups.

[0188] It can be seen that the effects demonstrated using the screw are much more pronounced than those demonstrated using the disc. The inventors believe this difference is related to the rough surface morphology of the screw compared to the smoothness of the polished disc. Therefore, in the real-life situation of TKA surgery, where biofilms may be present in many cracks and voids, the advantages of USN over US are expected to be much greater. The cleaning results using USN were significantly better.

[0189] Example 5 Testing titanium screws with Ag silver nanoparticles and biofilm simulants This example demonstrates superior cleaning effectiveness by adding a predetermined concentration of silver nanoparticles to a cleaning solution of 12 mg per 300 ml. This effect was demonstrated using a titanium screw and a biofilm simulant. The concentration was measured as milligrams of Ag nanoparticles in 300 ml of water in a 450 ml beaker.

[0190] Test parameters Test item - Screw (M6, length 20 mm) Nanoparticles - Ag (30-50nm) Ultrasonic - 50 watts / liter Concentrations - 0, 12, 20 mg per 300 ml (0 is for sonication only, no nanoparticles)

[0191] Test Procedure Two beakers containing 300 ml of water were placed in an ultrasonic bath containing an additional 2400 ml of water. The bath was set at 150 watts (150 watts for a total of 3000 ml - 50 watts per liter). The bath was not preheated and reached a maximum temperature of 35°C during the test.

[0192] Six screws were coated with the simulant. The screws were washed until they were clean. The effectiveness of the cleaning was determined by weighing the screws and comparing the weight of the cleaned screws with the original weight of the screws and the weight of the contaminated screws. This process was carried out twice for each nanoparticle concentration.

[0193] The cleaning rate parameter was calculated as (contaminated screw weight - final screw weight after cleaning) / (time required for cleaning).

[0194] result The table below summarizes the average cleaning rate (mg / min) for each nanoparticle concentration (mg / 300 ml water). TIFF2025537738000010.tif30170

[0195] Figure 17 shows the calculated wash rates for three concentrations (two runs at each concentration).

[0196] The table below shows the average cleaning rate for each nanoparticle concentration. TIFF2025537738000011.tif26170

[0197] conclusion These results show that the average cleaning rate of coated titanium screws using AG (silver) nanoparticles at a concentration of 12 mg per 300 ml and ultrasound is 1.28 times more efficient than using ultrasound alone (0 mg per 300 ml).

[0198] The effect demonstrated with the screw and Ag nanoparticles was less pronounced than that demonstrated with the screw and Ti2O3 nanoparticles, which was expected due to the larger weight of the Ag nanoparticles compared to the Ti2O3 nanoparticles and the lower velocity of the particles moving through the liquid when using the same amount of ultrasonic energy (50 watts per liter).

[0199] This resulted in a lower particle-surface collision velocity, reducing the mechanical effect of the system. A further mechanical effect was the hardness of the different nanoparticles. The hardness (Shore A) of Ti2O3 nanoparticles is much higher than that of Ag nanoparticles, so the mechanical cleaning effect based on the nanoscale sandblasting effect of Ti2O3 was much higher, as expected. Both the different nanoparticle velocity and hardness phenomena indicate that the primary cleaning mechanism using nanoparticle-enhanced ultrasound is mechanical and unrelated to the inherent bactericidal properties of titanium.

[0200] Example 6 Biofilm - Testing with bacterial cultures In this example, superior cleaning efficacy was demonstrated by adding a predetermined concentration of nanoparticles to a cleaning solution of 12 mg per 300 ml. This efficacy was demonstrated by culturing methicillin-resistant Staphylococcus aureus for 14 days to form a biofilm on a titanium screw. The concentration was measured as milligrams of TiO nanoparticles in 300 ml of water in a 450 ml beaker.

[0201] Test Method A clinical isolate of methicillin-resistant Staphylococcus aureus was grown and diluted to a concentration of 6 log10 CFU to form a robust bacterial biofilm on titanium screws. Eight screws per condition (24 screws total) were inoculated with 6 log10 CFU of S. aureus in Petri dishes. Bacteria were fed daily to promote biofilm growth. After 14 days of incubation at 37°C, the screws were sonicated. Eight screws were washed with USN and eight with US. An additional eight screws served as untreated controls. After 20 minutes of sonication, the titanium screws were fixed and immersed in crystal violet stain, which stains the bacteria present on the screws. Samples were analyzed using a plate reader, and absorbance was measured at 550 nm.

[0202] Test parameters Test item - Screw (M6, length 20 mm) Sonication time: 20 minutes Ultrasonic power: 50 watts / liter, liquid volume: 300 ml Nano Ti2O3 concentration - 0mg (sonication only), 12mg per 300ml

[0203] result The table below shows the biofilm growth (weight in grams) on days 7 and 14. Two screws from each group were measured. TIFF2025537738000012.tif22170

[0204] Crystal violet luminescence was used to measure the remaining biofilm / bacteria on the screws after cleaning in absorbance units, and the results (measured in absorbance units) are shown in the table below. TIFF2025537738000013.tif64170

[0205] Referring to Figure 18, crystal violet emission is shown.

[0206] Cleaning efficacy was measured on biofilms grown for 14 days. Crystal violet staining results after 20 min of sonication were significantly different (P<0.05) between sonication with nanoparticles and sonication alone (ANOVA + post-hoc Bonferroni correction), indicating better cleaning results (less crystal violet emission) with nanoparticles.

[0207] The results show that USN is statistically significantly 1.45- to 2.36-fold superior to US in both bacterial and artificial biofilms, highlighting the mechanistic mechanism of action of USN.

[0208] Therefore, the present technology demonstrates excellent biofilm cleansing efficacy in TKA patients, warranting the continuation of this project to the next level of using USN in a preclinical total knee arthroplasty model.

[0209] The results below show the mean USN compared to US for Examples 2-6. TIFF2025537738000014.tif56170

[0210] It should be understood that certain features of the disclosure, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosure that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or in any other described embodiment of the disclosure, as appropriate. A feature described in the context of an embodiment is not considered an essential feature of that embodiment, unless expressly stated otherwise.

[0211] Although the steps of some embodiments of the methods may be described in a particular order, the methods of the present disclosure may include some or all of the described steps performed in a different order. The methods of the present disclosure may include some or all of the described steps. No particular step in a disclosed method is considered an essential step of the method unless otherwise specified.

[0212] The phraseology and terminology used herein are for the purpose of description and should not be regarded as limiting. Citation or identification of a reference in this application should not be construed as an admission that such reference is available as prior art to the present disclosure. Section headings are used to facilitate understanding of this specification and should not be construed as necessarily limiting.

Claims

1. 1. An irrigation device for ultrasonically cleaning an internal element within a body cavity, comprising: at least one container sized and shaped to hold at least one liquid and at least one type of nanomaterial; at least one ultrasonic transducer configured to generate ultrasonic waves in at least one liquid within the container; and at least one adhesive element coupled to the at least one container and configured to ensure a reversible sealed connection between the at least one container and the body cavity.

2. 2. The perfusion device of claim 1, A perfusion device, wherein the at least one ultrasound transducer includes an electromagnetic element for generating a magnetic field.

3. 3. The perfusion device according to claim 1 or 2, The perfusion device is configured to communicate with at least one premixing device, and the at least one premixing device is configured to mix the at least one liquid with at least one type of nanoparticles.

4. The perfusion device according to any one of claims 1 to 3, The perfusion apparatus, further comprising a control unit configured to regulate the supply of one or more of the at least one liquid and the at least one type of nanomaterial into the at least one container.

5. 5. The perfusion device according to claim 4, The perfusion device, wherein the control unit is further configured to adjust at least one parameter selected from the group consisting of speed, flow rate, pressure, timing, frequency of the at least one ultrasonic transducer, and any combination thereof.

6. The perfusion device according to any one of claims 1 to 5, 10. A perfusion device, the device being configured to communicate with at least one fluid supply configured to store said at least one fluid.

7. The perfusion device according to any one of claims 1 to 6, The perfusion apparatus, further comprising at least one pump configured to supply the at least one fluid to the body cavity.

8. The perfusion device according to any one of claims 1 to 7, A perfusion device, characterized in that the at least one container is flexible.

9. The perfusion device according to any one of claims 1 to 8, The perfusion device, wherein the at least one type of nanomaterial comprises nanoparticles, the nanoparticles having surface features capable of penetrating or otherwise destroying infectious agents.

10. The perfusion device according to any one of claims 1 to 9, 10. The perfusion device according to claim 9, wherein the at least one container comprises a lid having at least one opening through which the at least one ultrasonic transducer passes.

11. The perfusion device according to any one of claims 1 to 10, 10. A perfusion device, wherein the at least one container comprises at least one inlet port configured to introduce the at least one liquid into the at least one container.

12. The perfusion device according to any one of claims 1 to 11, 10. A perfusion device, wherein the at least one container comprises at least one outlet port configured to remove the at least one liquid from the at least one container.

13. The perfusion device according to any one of claims 1 to 12, The perfusion device, wherein the internal element is selected from the group consisting of an implant, soft tissue, hard tissue, and combinations thereof.

14. The perfusion device according to any one of claims 1 to 13, A perfusion device wherein the body cavity includes a joint.

15. The perfusion device according to any one of claims 1 to 14, 10. The perfusion device according to claim 9, wherein the at least one liquid comprises a coloring agent that adheres to biofilm on the internal component.

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