Patent Application
20220039737
The present disclosure generally relates to cancer diagnosis, and particularly, to diagnosis of cancer involved lymph nodes by measuring lipid contents in a lymph node, which may be suspected to be cancerous, utilizing an electrochemical impedance spectroscopy (EIS) response recorded from the suspected lymph node.
Lymph nodes (LNs) are parts of lymphatic system responsible for body immunity. There are hundreds of lymph nodes spread all over body linked together by lymphatic vessels (LV) and lymphatic fluid (LF). Continuous circulation of LF through LV makes LNs susceptible to be invaded at beginning of cancer invasion, and each involved LN may work as a site for tumor spreading. Hence, early detection of involved LNs, e.g., involved sentinel LNs (SLNs) is vital for managing therapeutic protocols.
On the other hand, most of the current diagnostic clinical approaches for detecting cancer involved LNs are based on dissecting a suspicious lymph node (LN) and evaluating the dissected LN. However, LN dissection has many side effects, such as causing body inflation and causing immune system defects because LN dissection may cause removing healthy LNs from a person's body. Lymph nodes play a key role in immune system; basically, lymph nodes filter foreign and undesirable substances such as infections, dead cells, cancer cells, etc. Hence, removing more lymph nodes causes greater damage to immune system. So, in-vivo approaches that do not require LN dissection from a patient's body are preferred to avoid dissection of cancer free LNs from body.
Hence, there is a need for a highly accurate device and method for diagnosing cancer involved LNs in early stages of cancer progression. Additionally, there is a need for a device and method for intraoperative in-vivo detection as well as in-vitro diagnosis applications of cancerous LNs that should be highly precise, fast, simple, and biocompatible with human body without causing any side effects.
This summary is intended to provide an overview of the subject matter of the present disclosure, and is not intended to identify essential elements or key elements of the subject matter, nor is it intended to be used to determine the scope of the claimed implementations. The proper scope of the present disclosure may be ascertained from the claims set forth below in view of the detailed description below and the drawings.
In one general aspect, the present disclosure describes an exemplary method for detecting cancerous status of a suspected lymph node (LN) to be cancerous. The method may include forming a lipid detection probe (LDP) by forming three lipid sensitive parts at three respective distal ends of three hollow needle electrodes by laser-assisted welding of a layer of carbon nanotubes (CNTs) onto surface of the respective distal ends of three platinum hollow needles, connecting a proximal end of each respective hollow needle electrode to an electrochemical stimulator-analyzer device, inserting the three lipid sensitive parts into the suspected LN, injecting a biocompatible electrolyte solution into the LN through a respective proximal end of at least one hollow needle electrode of the three hollow needle electrodes, measuring fatty acid oxidation (FAO) in the suspected LN by measuring a charge transfer resistance (RCT) associated with the suspected LN, and detecting, utilizing one or more processors, a cancerous status of the suspected LN.
In an exemplary implementation, measuring FAO in the suspected LN by measuring the RCT associated with the suspected LN may include recording an electrochemical impedance spectroscopy (EIS) from the suspected LN utilizing the LDP and the electrochemical stimulator-analyzer device where the recorded EIS may include a pseudo-semicircular curve and calculating RCT of the recorded EIS.
In an exemplary implementation, calculating the RCT of the recorded EIS may include forming a semicircle curve by complementing the pseudo-semicircular curve and measuring a diameter of the formed semicircle curve. In an exemplary implementation, calculating the RCT of the recorded EIS may be done utilizing one or more processors.
In an exemplary implementation, detecting the cancerous status of the suspected LN may include detecting the suspected LN being cancer involved responsive to the measured RCT being less than 110 kΩ, or detecting the suspected LN being healthy responsive to the measured RCT being more than 110 kΩ.
In another general aspect, the present disclosure describes an exemplary method for detecting cancerous status of a suspected LN to be cancerous. The method may include measuring FAO in the suspected LN by measuring a RCT associated with the suspected LN and detecting, utilizing one or more processors, a cancerous status of the suspected LN. In an exemplary implementation, detecting the cancerous status of the suspected LN may include one of detecting the suspected LN being cancer involved responsive to the measured RCT being less than a reference RCT value and detecting the suspected LN being healthy responsive to the measured RCT being more than the reference RCT value. In an exemplary implementation, measuring FAO in the suspected LN and detecting the cancerous status of the suspected LN may be done in a time period of less than one minute.
In an exemplary implementation, detecting the cancerous status of the suspected LN may include comparing, utilizing one or more processors, the measured RCT with a reference RCT value of 110 kΩ, and detecting, utilizing one or more processors, one of situations in which the suspected LN is cancer involved if the measured RCT is less than 110 kΩ or the suspected LN is healthy if the measured RCT is less than 110 kΩ.
In an exemplary implementation, the method may further include generating the reference RCT value. In an exemplary implementation, generating the reference RCT value may include measuring a first set of RCT values associated with a plurality of healthy lymph nodes (LNs), measuring a second set of RCT values associated with a plurality of cancer involved LNs, and determining the reference RCT value by determining a RCT value at a border line magnitude between the first set of RCT values and the second set of RCT values.
In an exemplary implementation, measuring each of RCT values associated with each LN of the plurality of cancer involved LNs, the plurality of healthy LNs, and the suspected LN may include inserting three lipid sensitive parts of three respective electrodes of a lipid detection probe (LDP) into a LN, increasing electrical conductivity inside the LN by injecting a biocompatible electrolyte solution into the LN, recording an electrochemical impedance spectroscopy (EIS) including a pseudo-semicircular curve from the LN utilizing the LDP, and calculating RCT of the recorded EIS by measuring a diameter of a semicircle associated with the pseudo-semicircular curve.
In an exemplary embodiment, the LN may include one of the suspected LN, a LN of the plurality of cancer involved LNs, and a LN of the plurality of healthy LNs. In an exemplary embodiment, each respective electrode ma include a hollow needle electrode and each lipid sensitive part may include a distal end of each respective hollow needle electrode coated with a layer of lipophilic electrically conductive nanostructures. In an exemplary embodiment, the layer of lipophilic electrically conductive nanostructures may include a layer of carbon nanotubes (CNTs).
In an exemplary implementation, inserting three lipid sensitive parts of three respective electrodes of the LDP into the LN may include inserting each respective distal end of each electrode of the three electrodes into at least one of a LN in a living body and a dissected LN from a living body.
In an exemplary implementation, recording the EIS from the LN may include connecting the LDP to an electrochemical stimulator-analyzer device, applying an AC voltage between 5 mV and 10 mV by sweeping a frequency range including a plurality of frequency values between 0.01 Hz and 100 kHz, measuring a set of electrical impedance of the LN respective to the swept frequency range, and plotting a respective set of imaginary part of impedance (Z″ (Ω)) of the set of electrical impedance versus a respective set of real part of impedance (Z′ (Ω)) of the set of electrical impedance.
In an exemplary implementation, calculating the RCT of the recorded EIS may include measuring a first intersection point of the pseudo-semicircular-shaped curve with Z′ (Ω) axis, generating a second intersection point between the pseudo-semicircular-shaped curve and Z′ (Ω) axis by adding a complementary sector to the pseudo-semicircular-shaped curve to form a semicircle, and measuring a distance between the first intersection point and the second intersection point.
In an exemplary implementation, injecting the biocompatible electrolyte solution into the LN may include injecting the biocompatible electrolyte solution through at least one of the three electrodes of the LDP by injecting the biocompatible electrolyte solution into a respective proximal end of the at least one of the three electrodes utilizing a syringe. In an exemplary implementation, injecting the biocompatible electrolyte solution into the LN may include injecting a biocompatible and electrically conductive solution of metal ions into the LN. In an exemplary implementation, injecting the biocompatible electrolyte solution into the LN may include injecting a solution of iron ions into the LN, where the solution of iron ions may include a colloidal solution of ferric carboxymaltose complex.
In an exemplary implementation, the method may further include fabricating the LDP. In an exemplary implementation, fabricating the LDP may include forming the three hollow needle electrodes with the three respective lipid sensitive parts, connecting respective first ends of three electrical connector lines to respective proximal ends of the three hollow needle electrodes, placing the three hollow needle electrodes with the respective electrical connector lines inside a handle, and forming an opening at a location of the handle above a location of respective proximal ends of the three hollow needle electrodes.
In an exemplary embodiment, a respective second end of each electrical connector line may be configured to be connected to an electrochemical stimulator-analyzer device. In an exemplary embodiment, the handle may include a hollow cylinder with a bottom surface at a first end of the hollow cylinder. In an exemplary embodiment, the handle may be configured to facilitate inserting the three hollow needle electrodes into the LN. In an exemplary embodiment, the respective distal end of each hollow needle electrode may be placed outside the bottom surface. In an exemplary embodiment, the opening may be configured to inject the biocompatible electrolyte solution there through into at least one of the three hollow needle electrodes.
In an exemplary implementation, forming the three hollow needle electrodes with the three respective lipid sensitive parts may include forming a bevel-shaped tip at a respective distal end of each of the three hollow needle electrodes and forming a layer of CNTs on the respective distal end of each of the three hollow needle electrodes. In an exemplary embodiment, the bevel-shaped tip may be configured to facilitate a non-invasive insertion of each of the three hollow needle electrodes into a LN. In an exemplary embodiment, the three hollow needle electrodes may include a set of electrochemical electrodes including a working electrode, a counter electrode, and a reference electrode.
In an exemplary implementation, forming the layer of CNTs on the respective distal end of each of the three hollow needle electrodes may include growing a layer of CNTs on each respective distal end of each of the three hollow needle electrodes and welding the grown layer of CNTs to the respective distal end of each of the three hollow needle electrodes utilizing a laser welding process. In an exemplary implementation, growing the layer of
CNTs on each respective distal end of each of the three hollow needle electrodes may include preparing a solution by dispersing CNTs in a mixture of ethanol and deionized water, immersing respective distal end of each of the three hollow needle electrodes in the solution of dispersed CNTs, and sonicating the solution of dispersed CNTs. In an exemplary implementation, welding the grown layer of CNTs to the respective distal end of each of the three hollow needle electrodes may include placing the three hollow needle electrodes with grown CNTs on the respective distal ends in a sealed container, filling the sealed container with a noble gas, and irradiating continuous-wave laser with a wavelength of 1024 nm to the three hollow needle electrodes with grown CNTs on the respective distal ends in the presence of the noble gas.
In an exemplary implementation, forming the three hollow needle electrodes with the three respective lipid sensitive parts may further include electrically insulating parts of surface of each hollow needle electrode by covering a layer of an electrical insulating material around the surface of each hollow needle electrode except a surface of the respective distal end of each hollow needle electrode.
In an exemplary implementation, placing the three hollow needle electrodes with the respective electrical connector lines inside the handle may include forming three openings with a triangular pattern at the bottom surface and placing the three hollow needle electrodes with the respective electrical connector lines inside the handle by passing each of the three hollow needle electrodes through a respective opening of the formed three openings. In an exemplary embodiment, three respective lipid sensitive parts may be placed outside the bottom surface. In an exemplary embodiment, each two openings may be apart from each other by a distance between 1 mm and 5 mm.
In an exemplary embodiment, the respective proximal end of each of the three hollow needle electrodes may be placed inside the handle. In an exemplary embodiment, a second end of each respective electrical connector line of the three electrical connector lines may be placed outside a second end of the handle.
The drawing figures depict one or more implementations in accord with the present teachings, by way of example only, not by way of limitation. In the figures, like reference numerals refer to the same or similar elements.
In the following detailed description, numerous specific details are set forth by way of examples in order to provide a thorough understanding of the relevant teachings. However, it should be apparent that the present teachings may be practiced without such details. In other instances, well known methods, procedures, components, and/or circuitry have been described at a relatively high-level, without detail, in order to avoid unnecessarily obscuring aspects of the present teachings. The following detailed description is presented to enable a person skilled in the art to make and use the methods and devices disclosed in exemplary embodiments of the present disclosure. For purposes of explanation, specific nomenclature is set forth to provide a thorough understanding of the present disclosure. However, it will be apparent to one skilled in the art that these specific details are not required to practice the disclosed exemplary embodiments. Descriptions of specific exemplary embodiments are provided only as representative examples. Various modifications to the exemplary implementations will be readily apparent to one skilled in the art, and the general principles defined herein may be applied to other implementations and applications without departing from the scope of the present disclosure. The present disclosure is not intended to be limited to the implementations shown, but is to be accorded the widest possible scope consistent with the principles and features disclosed herein.
In case of spreading a metastatic cancer throughout a patient' body, cancer cells may invade lymph nodes (LNs), for example, sentinel lymph nodes (SLNs). Cancer cells may invade LNs by migrating from a cancerous organ or tissue to a LN through circulatory system and/or lymphatic system. Herein, a “cancer involved lymph node (LN)” may refer to a LN which may be invaded by cancer cells by migrating cancer cells to the LN. Invasion of cancer cells into LNs causes changes in metabolism of cancer cells within LNs respective to cancer cells' metabolism in other organs/tissues. Specifically, cancer cells' metabolism may be hypoxia glycolysis in a cancerous tissue/organ, whereas cancer cells' metabolism may change to fatty acid oxidation (FAO) when they invade LNs since LNs are resources of lipids. Such metabolism change may cause consumption of lipid contents within an exemplary cancer involved LN. In other words, levels of fatty acid (FA) in normal LNs may be higher than those involved by malignant cells. Specifically, a metabolism change of cancer cells migrated to LNs from glycolysis toward FAO may cause lipid consumption by cancer cells within LNs, resulting in a depletion of LNs' tissue from lipid. Hence, herein an exemplary probe, method, and system based on tracing lipidic content variations in LNs is disclosed for cancer diagnosis within LNs and detection of cancer involved LNs.
Herein, an exemplary device and methods are disclosed for diagnosing cancer involved LNs via tracing fatty acid oxidation as a distinct metabolism of malignant cells that have invaded lymph nodes (LNs). Exemplary device and method are disclosed for intraoperative in-vivo detection as well as in-vitro diagnosis of cancer involved LNs; therefore, exemplary device and method may be capable of detecting a suspicious LN is either a cancer involved LN or a cancer free LN via a real-time, simple, and biocompatible with human body approach based on monitoring and/or measuring a lipid content within an exemplary lymph node.
In an exemplary embodiment, an exemplary method for distinguishing cancer involved LNs from normal (healthy) LNs is disclosed. In an exemplary embodiment, an exemplary method may include a bio-sensing method based on measuring levels of fatty acid content in LNs utilizing analysis of an electrochemical impedance spectroscopy (EIS) recorded response of an exemplary LN suspected to be cancerous utilizing an exemplary lipid detection probe (LDP). In an exemplary embodiment, an exemplary lipid detection probe (LDP) may be designed and fabricated for in-vivo and in-vitro FAO tracing in an exemplary LN via recording EIS of an exemplary LN utilizing exemplary fabricated LDP.
In an exemplary implementation, exemplary methods 100 and 100A may be conducted in-vitro for detecting cancerous status of a suspected LN dissected from a living body. In an exemplary implementation, exemplary methods 100 and 100A may be conducted in-vivo for detecting cancerous status of a suspected LN within a living body. In an exemplary implementation, measuring FAO in an exemplary suspected LN (step 110) and detecting a cancerous status of the suspected LN (step 120) may be done in a time period of less than one minute, for example, in 10 seconds.
In detail, step 102 may include fabricating an exemplary lipid detection probe (LDP) that may be utilized via exemplary method 100 for detecting cancerous status of a suspected lymph node LN to be cancerous.
In an exemplary embodiment, exemplary handle 304 may include a hollow cylinder with bottom surface 308 at exemplary first end 310 of the hollow cylinder. In an exemplary embodiment, exemplary handle 304 may be configured to facilitate inserting three hollow needle electrodes 302 into an exemplary LN. In an exemplary embodiment, three hollow needle electrodes 302 may be placed inside exemplary handle 304, so that an exemplary lipid sensitive part 306 of each hollow needle electrodes of hollow needle electrodes 302 may be placed outside bottom surface 308.
In an exemplary embodiment, each lipid sensitive part 306 may include a distal portion of each respective hollow needle electrode of three hollow needle electrodes 302. Herein, an exemplary “distal portion” may refer to a length of about 5 mm to about 15 mm of total length of each of hollow needle electrodes 302a, 302b, and 302c from a respective distal end of each of hollow needle electrodes 302a, 302b, and 302c. In an exemplary embodiment, each lipid sensitive part 306 may include a distal portion of each respective hollow needle electrodes 302a, 302b, and 302c coated with a layer of a lipophilic electrically conductive material. In an exemplary embodiment, each lipid sensitive part 306 may include a distal portion of each respective hollow needle electrode coated with a layer of lipophilic electrically conductive nanostructures, for example, a layer of carbon nanotubes (CNTs). In an exemplary embodiment, the layer of CNTs may include a layer of at least one of single wall carbon nanotubes (SWCNTs), multi-walled carbon nanotubes (MWCNTs), vertically aligned multi-walled carbon nanotubes (VAMWCNTs), and combinations thereof. In an exemplary embodiment, the layer of CNTs, due to their hydrophobic nature and metallic conductivity, may be configured to provide an appropriate interfacial contact between lipids within an exemplary LN and hollow needle electrodes 302 while maintaining high electrical conductivity for impedance recording from exemplary LN to avoid parasitic resistance.
In an exemplary embodiment, each hollow needle electrode 302a, 302b, or 302c may include a biocompatible electrically conductive hollow needle. In an exemplary embodiment, each hollow needle electrode 302a, 302b, or 302c may include a needle made of at least one of gold, stainless steel, platinum, and combinations thereof In an exemplary embodiment, each hollow needle electrode 302a, 302b, or 302c may include a needle with a thickness between about 0.2 mm and about 1 cm. In an exemplary embodiment, each hollow needle electrode 302a, 302b, or 302c may include a needle with a thickness of about 0.5 mm.
In an exemplary implementation, step 402 may include forming a bevel-shaped tip 312 at a respective distal end of each hollow needle electrode of three hollow needle electrodes 302; allowing for inserting and penetrating lipid sensitive part 306 into a LN without causing a rupture in the LN and surrounding tissue. In an exemplary implementation, forming bevel-shaped tip 312 at a respective distal end of each hollow needle electrode 302a, 302b, or 302c may further include thinning, utilizing wiring or cylinder formation methods, three cylindrical needles or three hollow wires to reach a thickness in a range between about 0.2 mm and 1 mm. In an exemplary implementation, forming bevel-shaped tip 312 at a respective distal end of each hollow needle electrode 302a, 302b, or 302c may include thinning three cylindrical needles or three hollow wires made of at least one of gold, stainless steel, platinum, and combinations thereof and forming a bevel-shaped tip 312 at a respective distal end of each of the three cylindrical needles or three hollow wires. In an exemplary implementation, each of the three cylindrical needles or three hollow wires may be thinned to reach a thickness size (or a diameter) at which deforming, displacing, bending, or breaking of hollow needle electrodes 302 may be prevented during penetration. In an exemplary implementation, each of the three cylindrical needles or three hollow wires may be thinned to reach a diameter of about 0.5 mm; allowing for preventing deforming, displacing, bending, or breaking of hollow needle electrodes 302 during penetration into an exemplary tissue or LN.
In an exemplary implementation, forming a bevel-shaped tip 312 at a respective distal end of each hollow needle electrode of three hollow needle electrodes 302 may be done via a mechanochemical method. In an exemplary implementation, forming bevel-shaped tip 312 at a respective distal end of each hollow needle electrode via an exemplary mechanochemical method may include forming bevel-shaped tip 312 via a process including both mechanical processes and chemical reactions. In an exemplary implementation, forming bevel-shaped tip 312 may include forming bevel-shaped tip 312 at a respective distal end of each hollow needle electrode 302a, 302b, or 302c using a mechanical process, including at least one of wedge metal cutting, metal shaping, metal polishing, and combinations thereof. Furthermore, forming bevel-shaped tip 312 may further include smoothing formed bevel-shaped tip 312 and removing microscale metal shavings from formed bevel-shaped tip 312 by applying a chemical process to formed bevel-shaped tip 312. In an exemplary implementation, an exemplary chemical process may include acid treatment, for example, using Nitric acid. In an exemplary implementation, a respective distal end of each hollow needle electrode 302a, 302b, or 302c may be treated by an acidic solution to form smooth and well sharped edges at the respective distal end.
Moreover, step 404 may include forming a layer of lipophilic electrically conductive nanostructures on a respective surface of each of hollow needle electrodes 302a, 302b, and 302c. In an exemplary implementation, forming the layer of lipophilic electrically conductive nanostructures on the respective surface of each of hollow needle electrodes 302a, 302b, and 302c may include forming a layer of CNTs on the respective surface of each of hollow needle electrodes 302a, 302b, and 302c. In an exemplary implementation, forming the layer of CNTs on the respective surface of each of hollow needle electrodes 302a, 302b, and 302c may include forming a layer of at least one of SWCNTs, MWCNTs, VAMWCNTs, and combinations thereof on the respective surface of each of hollow needle electrodes 302a, 302b, and 302c. In an exemplary embodiment, the layer of lipophilic electrically conductive nanostructures may include an array of VAMWCNTs with a length between about 2 μm and about 12 and a diameter between about 20 nm and about 75 nm for each VAMWCNT of the array of VAMWCNTs. In an exemplary embodiment, CNTs, for example, MWCNTs may have superhydrophobic properties as well as high electrical conductivity, so that the layer of CNTs may be configured to enhance an interaction of hollow needle electrodes 302a, 302b, and 302c with lipidic contents of an exemplary LN and increase electrical conductivity of hollow needle electrodes 302a, 302b, and 302c for signal transduction. For example, MWCNTs' contact resistance may be less than about 100Ω and MWCNTs' electrical conductivity may be about 40 S/cm to about 50 S/cm. Hydrophobicity of CNTs may be dependent on growth procedure and functionalization. However, hydrophobicity of CNTs may be measured by measuring a contact angle of CNTs with a non-lipidic solution. An exemplary measured contact angle of MWCNTs may be more than about 120° which shows a high hydrophobicity for MWCNTs.
In an exemplary implementation, forming the layer of CNTs on the respective surface of each of hollow needle electrodes 302a, 302b, and 302c may include forming the layer of CNTs on a portion of the respective surface of each of hollow needle electrodes 302a, 302b, and 302c. In an exemplary implementation, forming the layer of CNTs on the portion of the respective surface of each of hollow needle electrodes 302a, 302b, and 302c may include forming the layer of CNTs on a surface of a respective distal portion of each of hollow needle electrodes 302a, 302b, and 302c. In an exemplary implementation, forming the layer of CNTs on the portion of the respective surface of each of hollow needle electrodes 302a, 302b, and 302c may include forming the layer of CNTs on a portion surface of each of hollow needle electrodes 302a, 302b, and 302c corresponding to a length of a respective lipid sensitive part 306. In an exemplary implementation, forming the layer of CNTs on the portion of the respective surface of each of hollow needle electrodes 302a, 302b, and 302c may include forming the layer of CNTs on surface of respective bevel-shaped tip 312 of each of hollow needle electrodes 302a, 302b, and 302c.
In an exemplary implementation, forming the layer of CNTs on the respective surface of each of hollow needle electrodes 302a, 302b, and 302c may include growing a plurality of CNTs on the respective surface of each of hollow needle electrodes 302a, 302b, and 302c and welding the grown plurality of CNTs to the respective surface of each of hollow needle electrodes 302a, 302b, and 302c.
In an exemplary implementation, growing the plurality of CNTs on the respective surface of each of hollow needle electrodes 302a, 302b, and 302c may include exposing hollow needle electrodes 302a, 302b, and 302c to oxygen plasma and immersing whole or parts of hollow needle electrodes 302a, 302b, and 302c in a solution including dispersed CNTs. In an exemplary implementation, immersing whole or parts of hollow needle electrodes 302a, 302b, and 302c in the solution including dispersed CNTs may further include sonicating the solution to form a highly dispersed solution of CNTs. In an exemplary implementation, immersing whole or parts of hollow needle electrodes 302a, 302b, and 302c in the solution including dispersed CNTs may further include preparing the solution by dispersing CNTs in a mixture of ethanol and deionized (DI) water. In an exemplary implementation, sonicating the solution may include sonicating the solution utilizing at least one of a sonicator horn and a sonication bath. In an exemplary implementation, sonicating the solution may be done at a sonication power in a range between 100 W and 150 W. In an exemplary implementation, growing the plurality of CNTs on the respective surface of each of hollow needle electrodes 302a, 302b, and 302c may include adhesion of CNTs to hollow needle electrodes 302a, 302b, and 302c right after immersion of hollow needle electrodes 302a, 302b, and 302c in the solution.
In an exemplary implementation, forming the layer of CNTs on the respective surface of each of hollow needle electrodes 302a, 302b, and 302c may further include forming a firm attachment between the grown CNTs and the respective surface of each of hollow needle electrodes 302a, 302b, and 302c by welding or soldering the grown CNTs to the respective surface of each of hollow needle electrodes 302a, 302b, and 302c. In an exemplary implementation, welding the grown plurality of CNTs to the respective surface of each of hollow needle electrodes 302a, 302b, and 302c may include welding the grown plurality of CNTs to the respective surface of each of hollow needle electrodes 302a, 302b, and 302c by laser-assisted welding (nano-welding) of CNTs to an outer respective surface of each of hollow needle electrodes 302a, 302b, and 302c utilizing a laser-assisted welding (nano-welding) process. In an exemplary implementation, laser-assisted nano-welding of CNTs to the respective surface of each of hollow needle electrodes 302a, 302b, and 302c may include breaking C-C bonds of CNTs and forming carbon-metal (C-M) bonds; thereby, resulting in decreasing electrical contact resistance between CNTs and metallic surface of hollow needle electrodes 302, for example, platinum hollow needle electrodes.
In an exemplary implementation, laser-assisted welding of CNTs on the respective surface of each of three hollow needle electrodes 302a, 302b, and 302c may include placing hollow needle electrodes 302a, 302b, and 302c with grown CNTs thereon in a sealed container, filling the sealed container with a noble gas, for example, Argon (Ar), and irradiating laser to hollow needle electrodes 302a, 302b, and 302c with grown CNTs thereon. Irradiating the laser leads to rise temperature within the sealed container up to several thousand degrees Celsius (° C.), for example, more than about 1000° C. In an exemplary embodiment, the noble gas may be configured to be a neutralizer agent preventing oxidization of CNTs and/or hollow needle electrodes 302a, 302b, and 302c due to high temperatures caused by laser irradiation inside the sealed container. In an exemplary implementation, irradiating laser to hollow needle electrodes 302a, 302b, and 302c with grown CNTs thereon may include irradiating a continuous-wave laser with a wavelength of about 1064 nm. In an exemplary implementation, irradiating laser to hollow needle electrodes 302a, 302b, and 302c with grown CNTs thereon may be done within a time period in a range between 5 seconds and 10 seconds utilizing a laser irradiator device.
Moreover, step 406 may include electrically insulating the respective surface of each of hollow needle electrodes 302a, 302b, and 302c except a respective distal portion of each of hollow needle electrodes 302a, 302b, and 302c. In an exemplary implementation, electrically insulating the respective surface of each of hollow needle electrodes 302a, 302b, and 302c except the respective distal portion of each of hollow needle electrodes 302a, 302b, and 302c may include coating a layer of an electrical insulating material, for example, a layer of plastic, around parts of surface of each of hollow needle electrodes 302a, 302b, and 302c except a respective length of each of hollow needle electrodes 302a, 302b, and 302c configured to form exemplary lipid sensitive part 306 for each hollow needle electrode. In an exemplary embodiment, a respective length of each of hollow needle electrodes 302a, 302b, and 302c configured to form exemplary lipid sensitive part 306 may include a length of about 5 mm to 15 mm of respective distal portion of each of hollow needle electrodes 302a, 302b, and 302c.
In an exemplary implementation, three hollow needle electrodes 302a, 302b, and 302c with respective exemplary lipid sensitive part 306 for each hollow needle electrode may be formed by conducting steps 402, 404, and 406 of exemplary process 400 described hereinabove. Additionally, exemplary process 400 may further include connecting a respective first end of each electrical connector line of three electrical connector lines 316 to a respective proximal end of each of hollow needle electrodes 302a, 302b, and 302c (step 408). In an exemplary implementation, three electrical connector lines 316 may be configured to be connected to an electrochemical stimulator-analyzer device; thereby, connecting three hollow needle electrodes 302 to the electrochemical stimulator-analyzer device. In an exemplary embodiment, a respective second end of each electrical connector line of three electrical connector lines 316 may be configured to be connected to the electrochemical stimulator-analyzer device.
Furthermore, step 410 may include placing three hollow needle electrodes 302 with respective electrical connector lines 316 inside exemplary handle 304. In an exemplary implementation, placing three hollow needle electrodes 302 with respective electrical connector lines 316 inside exemplary handle 304 may include forming three holes at exemplary locations 324, 326, and 328 of exemplary bottom surface 308 and passing hollow needle electrodes 302a, 302b, and 302c through respective three holes at locations 324, 326, and 328, so that exemplary lipid sensitive part 306 of each hollow needle electrode of three hollow needle electrodes 302 may be placed outside of bottom surface 308. In an exemplary implementation, passing hollow needle electrodes 302a, 302b, and 302c through respective three holes at locations 324, 326, and 328 may include attaching a respective cross outer surface of hollow needle electrodes 302a, 302b, and 302c to a respective internal cross surface of three holes at locations 324, 326, and 328.
In an exemplary embodiment, handle 304 may be configured to facilitate inserting a respective lipid sensitive part 306 of each of three hollow needle electrodes 302 into an exemplary LN. In an exemplary embodiment, respective proximal ends of three hollow needle electrodes 302 may be placed inside handle 304. In an exemplary embodiment, the respective second end of each electrical connector line of three electrical connector lines 316 may be placed outside an exemplary second end 322 of handle 304.
In an exemplary implementation, forming three holes at exemplary locations 324, 326, and 328 of exemplary bottom surface 308 may include forming three circular openings with a respective diameter of each opening equal to a diameter of each hollow needle electrode of three hollow needle electrodes 302. In an exemplary embodiment, a diameter of each circular opening may be in a range between about 0.1 mm and 1 mm. In an exemplary embodiment, a diameter of each circular opening may be about 0.5 mm.
In an exemplary implementation, forming three holes at exemplary locations 324, 326, and 328 of exemplary bottom surface 308 may be done taking into account an average size of an exemplary LN, since all three respective lipid sensitive parts of three hollow needle electrodes 302 should be inserted into an exemplary LN to conduct an electrical measurement, for example, recording an electrochemical impedance spectroscopy (EIS) of lipid content in an exemplary LN. For example, an average size of adult female standard breast (sentinel or axillary) LN is less than about 1 cm in diameter, and a diameter of cancer involved LNs are mostly less than about 5 cm. Accordingly, an exemplary distance between each two respective electrode of three hollow needle electrodes 302 may be designed to lead to an optimum current density between three hollow needle electrodes 302 being generated. In an exemplary implementation, forming three holes at exemplary locations 324, 326, and 328 of exemplary bottom surface 308 may include forming three openings at exemplary locations 324, 326, and 328 with a triangular pattern at exemplary bottom surface 308. In an exemplary embodiment, each two openings at exemplary locations 324, 326, and 328 may be formed apart from each other by a distance between about 1 mm and about 5 mm. In an exemplary embodiment, an exemplary distance between each two respective electrode of three hollow needle electrodes 302 may be set equal to about 3 mm resulting in a current density of about 0.1 μA/mm2 to about 10 μA/mm2 in different frequencies of EIS recording, which may be an optimum situation for a dielectric spectroscopy approach.
Moreover, step 412 may include forming an opening 318 at a location of body of handle 304 above a location of respective proximal ends of three hollow needle electrodes 302. In an exemplary embodiment, exemplary opening 318 may be configured to inject a solution, for example, a biocompatible electrolyte solution through opening 318 into at least one hollow needle electrode of three hollow needle electrodes 302.
Referring to
In detail, step 104 may include generating a reference RCT value. In an exemplary implementation, generating the reference RCT value may include measuring a first set of RCT values associated with a plurality of healthy lymph nodes (LNs), measuring a second set of RCT values associated with a plurality of cancer involved LNs, and determining the reference RCT value by determining a RCT value at a border line between the first set of RCT values and the second set of RCT values. In an exemplary implementation, measuring the first set of RCT values associated with the plurality of healthy lymph nodes (LNs) and measuring the second set of RCT values associated with the plurality of cancer involved LNs may be done utilizing exemplary LDP 300.
In an exemplary implementation, determining the reference RCT value by determining a RCT value at a border line between the first set of RCT values and the second set of RCT values may include assigning a cut-off value between the first set of RCT values and the second set of RCT values to the reference RCT value. In an exemplary embodiment, the reference RCT value may be more than a maximum value of RCT values associated with the plurality of cancer involved LNs while being less than a minimum of the first set of RCT values associated with the plurality of healthy LNs. In an exemplary embodiment, the reference RCT value may be determined equal to 110 kΩ.
Furthermore, step 110 may include measuring FAO in a suspected LN by measuring a RCT value associated with the LN utilizing exemplary LDP 300. In an exemplary implementation, each step of measuring each of RCT values associated with each LN of the plurality of cancer involved LNs, the plurality of healthy LNs, and the suspected LN for exemplary steps similar to steps 104 and/or 110 may be done via an exemplary method 200 shown in
In an exemplary implementation, exemplary LDP 300 may be utilized for conducting method 200. In an exemplary implementation referring to
In detail, step 202 may include inserting exemplary three lipid sensitive parts 306 of three respective electrodes 302 of LDP 300 into an exemplary LN. In an exemplary implementation, inserting exemplary three lipid sensitive parts 306 of three respective electrodes 302 of LDP 300 into an exemplary LN may include inserting each respective distal end 312 of each of hollow needle electrodes 302a, 302b, and 302c into at least one of a LN in a living body in case of an in-vivo implementation and a dissected LN from a living body in case of an in-vitro implementation.
Furthermore, step 204 may include injecting a biocompatible electrolyte solution into an exemplary LN, for example, LN 502 shown in
In an exemplary embodiment, the biocompatible electrolyte solution may include a biocompatible and electrically conductive solution containing one or more metal ions. In an exemplary embodiment, the biocompatible electrolyte solution may include an injectable solution of iron ions. In an exemplary embodiment, the solution of iron ions may include a colloidal solution of ferric carboxymaltose complex. In an exemplary embodiment, the colloidal solution of ferric carboxymaltose complex may include a colloidal solution of ferric carboxymaltose complex with a concentration in a range between about 1 mg/ml and 50 mg/ml.
Moreover, step 206 may include recording an EIS from an exemplary LN, for example, LN 502, utilizing LDP 300 and an electrochemical stimulator-analyzer device. In an exemplary implementation, recording the EIS from exemplary LN 502 may include connecting exemplary LDP 300 to an electrochemical stimulator-analyzer device via three electrical connector lines 316, applying an AC voltage between about 5 mV and about 10 mV by sweeping a frequency range between about 0.01 Hz and about 100 kHz utilizing the electrochemical stimulator-analyzer device connected to LDP 300, measuring a set of electrical impedance of LN 502 respective to the swept frequency range utilizing the electrochemical stimulator-analyzer device connected to LDP 300, and plotting a respective set of imaginary part of impedance (Z″ (Ω)) of the set of electrical impedance versus a respective set of real part of impedance (Z′ (Ω)) of the set of electrical impedance.
In an exemplary embodiment, the frequency range may include a plurality of frequency values between about 0.01 Hz and about 100 kHz. In an exemplary embodiment, the electrochemical stimulator-analyzer device may include a potentiostat device.
In an exemplary implementation, plotting the respective set of imaginary part of impedance (Z″ (Ω)) of the set of electrical impedance versus the respective set of real part of impedance (Z′ (Ω)) of the set of electrical impedance may be conducted utilizing one or more processors. In an exemplary implementation, plotting the respective set of imaginary part of impedance (Z″ (Ω)) of the set of electrical impedance versus the respective set of real part of impedance (Z′ (Ω)) of the set of electrical impedance may include plotting a Nyquist plot with a pseudo-semicircular curve shape from an exemplary LN 502. In an exemplary embodiment, an exemplary plot of the set of electrical impedance (an exemplary EIS plot) may be a Nyquist plot which may be analyzed by a corresponding equivalent circuit (named a Randles circuit).
Furthermore, step 208 may include calculating RCT of the recorded EIS by measuring a diameter of a semicircle curve associated with the pseudo-semicircular curve. In an exemplary implementation, calculating RCT of the recorded EIS may be done utilizing one or more processors. Referring to
Referring again to
In an exemplary implementation, detecting the cancerous status of the suspected LN may further include detecting the suspected LN is a cancer involved LN if the measured RCT is less than the reference RCT value or detecting the suspected LN is a healthy LN if the measured RCT is more than the reference RCT value. In an exemplary implementation, detecting the cancerous status of the suspected LN may include comparing, utilizing one or more processors, the measured RCT with a reference RCT value of about 110 kΩ and detecting, utilizing one or more processors, the suspected LN is a cancer involved LN if the measured RCT is less than about 110 kΩ or detecting, utilizing one or more processors, the suspected LN is a healthy LN if the measured RCT is less than about 110 kΩ.
In this example, an exemplary LDP similar to LDP 300 was prepared for utilizing in exemplary methods 100 and 200 for diagnosis of cancer involved LNs. Three platinum, 0.5 mm thickness, hollow needle electrodes were formed thin with a bevel-shaped tip using wiring and cylinder formation methods, and mechanochemical method. Platinum needles were exposed to oxygen plasma and immersed in a dispersed CNT solution to weld CNTs on surface of needles. The dispersed CNT solution was prepared by dispersing 0.1 mg of multi-walled carbon nanotubes (MWCNTs) in ethanol:deionized (DI) water (3:1, V:V). To obtain well-dispersed CNTs, the solution was sonicated using a sonicator horn at a power of 100 W for about 30 minutes in a cycle of 7:3 seconds (On:Off).
Afterwards, CNT-Pt needles were placed in a sealed container purged by noble gas (Ar) and laser irradiation. The laser irradiation was conducted using a 1064 nm wavelength laser.
Furthermore, wettability of prepared electrodes was investigated through contact angle measurement. Droplet of fresh Dulbecco's Modified Eagle's medium (DMEM) cell culture media (a solution with no lipid) has been used to measure the contact angle.
To compare welded CNT electrodes (named as welded CNT Pt electrode (WCPE)) with electrodes that CNTs had been only grown thereon utilizing a direct-current plasma enhanced chemical vapor deposition (DC-PECVD) technique in a DC-PECVD reactor (named as grown CNT on Pt electrode (GCPE)), exemplary electrodes were penetrated into a euthanized animal tissue and a footprint image was recorded.
For analyzing effect of exemplary MWCNTs welded on exemplary platinum needle electrodes, a comparative graph of EIS results between exemplary bare Pt electrodes and Pt electrodes decorated by welded CNTs in Dulbecco's Modified Eagle Medium (DMEM) with 10% Fetal Bovine Serum (FBS), as a lipidic solution, was recorded.
A length of each of three exemplary MWCNT welded Pt electrodes, fabricated similar to exemplary hollow needle electrodes 302a, 302b, and 302c, were insulated except a distal end portion which remained non-insulated for insertion into an exemplary LN. A plastic material was covered around surface of each of three exemplary MWCNT welded Pt electrodes except a respective distal end portion. Three exemplary MWCNT welded Pt electrodes were attached and fixed at the end of a handle with a distance of 3 mm from each other. Three respective electrodes include a set of a working electrode (WE), a reference electrode (RE), and a counter electrode (CE) for that may be utilized in EIS recording and measurements.
Animal model procedures were accomplished in-vivo via an implementation of exemplar method 100 utilizing a miniaturized fabricated LDP similar to exemplary LDP 300. A proper animal model may be critical for mimicking human disease conditions. Hence, adult white strains of New Zealand rabbits with a weight of 2.5 kg to 3 kg were used in this trial. Anesthesia was performed using ketamine 50 mg/kg and xylazine 10 mg/kg intravenous and intramuscular, respectively. Rabbit LNs were used to clarify effect of lipid content in EIS response of LN ambient, and EIS was recorded before and after injection of Tumescent (as a lipolyzing agent) into an exemplary LN of rabbit to simulate lipid consumption by a cancer involved LN. Tumescent is a lipolysis solution that includes lidocaine and epinephrine, which increases phosphorylation status of perilipin in adipocytes and subjects them to lipolysis. Tumescent also has a local anesthesia effect on skin and its subcutaneous tissue. Tumescent solution contains 500 mg of lidocaine, 0.5-1 mg of epinephrine, and 10 mEq of sodium bicarbonate in 1 L of 0.9% normal saline. Before EIS recording of lymph nodes, 250 μl of a solution of ferric carboxymaltose complex (2 mg/ml) as an electrically conductive carrier solution had been directly injected into LNs of an exemplary animal model.
Additionally,
As exemplary EIS approach in animal model (described hereinabove in Example 2) showed meaningful results on differentiating LNs with various contents of lipids, exemplary method 100, utilizing exemplary LDP 300, was applied on dissected LNs of 41 patients, immediately after dissection. In this regard, exemplary LDP 300 was applied to 122 LNs (including sentinel and non-sentinel) from breast cancer patients, which had been dissected through standard guidelines. Patients undergoing breast cancer surgery were recruited to this analysis. During a breast cancer surgery, manual lymph node dissection was performed. Right after lymph node dissection, lymph nodes have been tested by injecting carriers and penetrating exemplary fabricated electrodes of exemplary fabricated LDP to them. Moreover, LN dissected samples were sent for frozen pathology. Obtained results of permanent pathology were conducted to compare with results obtained from EIS recording according to exemplary method 100. EIS recording procedure was carried out in less than 1 minute. To carry out electrochemical spectroscopy of dielectric components of dissected LNs, 250 μl of a solution of ferric carboxymaltose complex (2 mg/ml) was directly injected into LNs right after dissection. Exemplary EIS responses were recorded using a portable potentiostat by conducting EIS at 10 mV with a frequency range of 100 kHz to 0.01 Hz at 10 points. In order to ensure about Pt electrodes calibration, they tested and calibrated using potassium ferricyanide (K3Fe(CN)6) before each sterilization step. In each sterilization step, exemplary fabricated electrodes were sterilized by autoclave. The autoclaved electrodes were transferred to a biological safety container and sealed in a medical-grade sterilized package, including paraformaldehyde tablets. Sterilization step was renewed after 2 weeks of storing.
Table 1 shows measured RCT values of Patients' LNs using an exemplary LDP, responses of EIS measurements by LDP, frozen section, permanent pathology section, and EIS result evaluation in comparison to permanent histopathological diagnosis in 45 SLNs and 77 non-sentinel LNs. A border of normal and cancer involved LNs was detected at about 110 kΩ. True-positive and false-positive results have been specified by comparing EIS results with permanent pathology. Each test was repeated for 5 times with a standard deviation (STD) of about ±5%.
Results presented in Table 1 show a meaningful correlation between measured RCT of recorded EIS responses and pathological diagnosis about cancer cell involvement of LNs. Pathological diagnosis was done by H&E and Pan Cytokeratin, PCK, which are based immune histochemistry assays. Significantly, lower RCT was recorded for cancerous LNs in comparison with normal ones.
As presented in Table 1, only the first lymph node of patient No. 15 was falsely scored positive by EIS analysis. This case had undergone neoadjuvant chemotherapy with complete remission, but she lived with cancer for a while. Hence, cancer cells in invaded LNs by cancer cells had enough time to deplete LN lipids for their metabolism. So, LNs depleted from lipid due to cancer cells FAO metabolism before conducting a chemotherapy. As a consequence, this result might be due to a depletion of lipids that occurred from cancerous cells before chemotherapy.
It should be noted that the only way to quantify invading tumor cells in LNs is pathologists' diagnosis. Therefore, it will be very practitioner-dependent. Also, some other variables affect the pathological results. For instance, number of slides provided from tissue sections, evaluated area of the slides, and even number of slides that evaluated could directly affect a pathologists' diagnosis. Diagnosis of pathologists for defining a cancer involvement percentage were conducted for dissected LNs, and a mean value of their diagnosis was set as an involvement percentage to cancer. By plotting RCT versus involvement percentage, a relative relation between cancer involvement percentage and RCT magnitude was extrapolated.
By EIS investigation of diagnosed LNs (according to pathology gold standard), a primary warning factor for probability of lymph node involvement based on RCT was defined. EIS values obtained for clear (normal or healthy) and cancer involved LNs may be helpful to have an early estimation from the cancerous state of a LN before its dissection and sending to frozen pathology during surgery. The obtained results show real-time ability of such nano electro biochemical methodology based on a strong biological pathway, FAO, in in-vivo findings of LN cancer involvement before a LN dissection.
While the foregoing has described what are considered to be the best mode and/or other examples, it is understood that various modifications may be made therein and that the subject matter disclosed herein may be implemented in various forms and examples, and that the teachings may be applied in numerous applications, only some of which have been described herein. It is intended by the following claims to claim any and all applications, modifications and variations that fall within the true scope of the present teachings.
Unless otherwise stated, all measurements, values, ratings, positions, magnitudes, sizes, and other specifications that are set forth in this specification, including in the claims that follow, are approximate, not exact. They are intended to have a reasonable range that is consistent with the functions to which they relate and with what is customary in the art to which they pertain.
The scope of protection is limited solely by the claims that now follow. That scope is intended and should be interpreted to be as broad as is consistent with the ordinary meaning of the language that is used in the claims when interpreted in light of this specification and the prosecution history that follows and to encompass all structural and functional equivalents. Notwithstanding, none of the claims are intended to embrace subject matter that fails to satisfy the requirement of Sections 101, 102, or 103 of the Patent Act, nor should they be interpreted in such a way. Any unintended embracement of such subject matter is hereby disclaimed.
Except as stated immediately above, nothing that has been stated or illustrated is intended or should be interpreted to cause a dedication of any component, step, feature, object, benefit, advantage, or equivalent to the public, regardless of whether it is or is not recited in the claims.
It will be understood that the terms and expressions used herein have the ordinary meaning as is accorded to such terms and expressions with respect to their corresponding respective areas of inquiry and study except where specific meanings have otherwise been set forth herein. Relational terms such as first and second and the like may be used solely to distinguish one entity or action from another without necessarily requiring or implying any actual such relationship or order between such entities or actions. The terms “comprises,” “comprising,” or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. An element proceeded by “a” or “an” does not, without further constraints, preclude the existence of additional identical elements in the process, method, article, or apparatus that comprises the element.
The Abstract of the Disclosure is provided to allow the reader to quickly ascertain the nature of the technical disclosure. It is submitted with the understanding that it will not be used to interpret or limit the scope or meaning of the claims. In addition, in the foregoing Detailed Description, it can be seen that various features are grouped together in various implementations. This is for purposes of streamlining the disclosure, and is not to be interpreted as reflecting an intention that the claimed implementations require more features than are expressly recited in each claim. Rather, as the following claims reflect, inventive subject matter lies in less than all features of a single disclosed implementation. Thus, the following claims are hereby incorporated into the Detailed Description, with each claim standing on its own as a separately claimed subject matter.
While various implementations have been described, the description is intended to be exemplary, rather than limiting and it will be apparent to those of ordinary skill in the art that many more implementations and implementations are possible that are within the scope of the implementations. Although many possible combinations of features are shown in the accompanying figures and discussed in this detailed description, many other combinations of the disclosed features are possible. Any feature of any implementation may be used in combination with or substituted for any other feature or element in any other implementation unless specifically restricted. Therefore, it will be understood that any of the features shown and/or discussed in the present disclosure may be implemented together in any suitable combination. Accordingly, the implementations are not to be restricted except in light of the attached claims and their equivalents. Also, various modifications and changes may be made within the scope of the attached claims.
This application claims the benefit of priority from pending U.S. Provisional Patent Application Ser. No. 63/105,218 filed on Oct. 24, 2020, and entitled “A REAL-TIME INTRAOPERATIVE APPROACH TO DIAGNOSE METASTATIC LYMPH NODES”, which is incorporated herein by reference in its entirety.
| Number | Date | Country | |
|---|---|---|---|
| 63105218 | Oct 2020 | US |
