Description:MICROFLUIDIC SENSING DEVICE FOR DISEASE MODELLING AND DRUG SCREENING STUDIES
FIELD OF THE DISCLOSURE
[0001] This invention generally relates to a field of a bio-sensing device, in particular relates to a microfluidic sensing device for disease modelling and drug screening studies and a method for operating the microfluidic sensing device for disease modelling and drug screening studies.

BACKGROUND
[0002] The subject matter discussed in the background section should not be assumed to be prior art merely as a result of its mention in the background section. Similarly, a problem mentioned in the background section or associated with the subject matter of the background section should not be assumed to have been previously recognized in the prior art. The subject matter in the background section merely represents different approaches, which in and of themselves may also correspond to implementations of the claimed technology.
[0003] Three-dimensional (3D) cell technology may revolutionized the disease modelling by providing a physiologically environment which may resemble complex microenvironments of tissues and organs in vitro. Unlike conventionally known two-dimensional (2D) cell cultures, 3D cell models may show better spatial organization, cell-cell interactions, and extracellular matrix architecture found in vivo, offering more accurate representations of disease processes. 3D models have become indispensable tools for studying various diseases, including cancer, neurodegenerative disorders, and infectious diseases, allowing researchers to explore disease mechanisms, screen drug candidates, and develop personalized treatment strategies. Further, the researcher may understand disease progression, identify novel therapeutic targets, and ultimately leading to improved patient outcomes by using the 3D model cell technology.
[0004] Further, the conventionally known 3D cell models may have some limitations in accurately representing the multicellular complexity of human skin, primarily due to several key factors. Firstly, the 3D cell models may lack vascular networks, which play a crucial role in supplying nutrients, oxygen, and signalling molecules to skin cells and facilitating waste removal. Without adequate vascularization, conventionally known 3D models may not fully capture the physiological responses and interactions of skin cells under normal or pathological conditions. Further, the traditional 3D models may exhibit poor barrier characteristics as compared to a native skin. Additionally, the conventionally known 3D models may lack skin appendages such as hair follicles, sweat glands, sebaceous glands, which is integral component of the skin microenvironment.
[0005] According to a patent application “US20080003142A1” titled “Microfluidic devices” discloses Microfluidic devices. The present invention provides novel microfluidic substrates and methods that are useful for performing biological, chemical and diagnostic assays. The substrates can include a plurality of electrically addressable, channel bearing fluidic modules integrally arranged such that a continuous channel is provided for flow of immiscible fluids.
[0006] According to another patent application “EP3546067A1” titled “Microfluidic chip” discloses a Microfluidic chip thereof. A microfluidic chip for conducting microbiological assays, comprises a substrate in which incubation segments, a sample reservoir and microfluidic channels connecting said sample reservoir with said incubation segments are arranged. Said microfluidic chip further comprise a non-aqueous fluid reservoir for containing non-aqueous liquid wherein said reservoir is connectable via a releasable airtight and liquid-tight valve with said microfluidic channels connecting said sample reservoir with said incubation segments.
[0007] However, the traditional 3D cell model may lack vascular structures capable of exchanging nutrient and oxygen supply crucial in living tissue. Further, the conventional 3D cell model may struggle to replicate the barrier function of skin. Further, these traditional 3D cell model may lack skin appendages which may hamper the cell model ability to resemble complex skin biology, drug responses and disease mechanisms.
OBJECTIVES OF THE INVENTION
[0008] The objective of invention is to provide a microfluidic sensing device for disease modelling and drug screening studies.
[0009] The objective of invention is to provide a method for operating the microfluidic sensing device for disease modelling and drug screening studies.
[0010] Furthermore, the objective of present invention is to provide the microfluidic sensing device for disease modelling and drug screening studies that is capable of sensing an essential amino acids utilized in wound healing mechanism.
[0011] Furthermore, the objective of present invention is to provide the microfluidic sensing device for disease modelling and drug screening studies utilizes a cost effective flexible electrode.
[0012] Furthermore, the objective of present invention is to provide the microfluidic sensing device for disease modelling and drug screening studies that is capable of improving a skin physiology in vivo and a high-throughput.
SUMMARY
[0013] According to an aspect, the present embodiments the microfluidic sensing device for disease modelling and drug screening studies antibiotics, the sensing device comprises a first layer. Further, the first layer comprising a culture chamber contained with a 3D (3-dimensional) cell culture. Further, a second layer having a microvascular channel. Further, a third layer having at least one membrane and a pair of pneumatic channels. Further, a sensing surface having a plurality of sensors is arranged on a first layer. Further, each of the pneumatic channels are configured to put the 3D cell culture through one or more strain cycles by providing a predefined amount of pneumatic pressure to the 3D cell culture. Further, the at least one membrane is configured to transfer a nutrient, an oxygen and one or more signalling molecules to the 3D cell culture during the one or more strain cycles of the 3D cell culture. Further, wherein the plurality of sensors is configured to detect one or more parameters, wherein the one or more parameters may correspond to healing of wound, formation of the 3D cell culture and drug administration during a physiological process and cellular responses in a real time.
[0014] In one embodiment, a method for operating the microfluidic sensing device for disease modelling and drug screening studies comprising providing, via a first layer, a culture chamber contained with a 3D (3-dimensional) cell culture. Further, the method comprises providing, via a second layer, a microvascular channel. Further, the method comprises providing, via a third layer, at least one membrane and a pair of pneumatic channels. Further, the method comprises providing, via each of the pair of pneumatic channels, a predefined amount of pneumatic pressure to the 3D cell culture. Further, the method comprises transferring, via the at least one membrane, a nutrient, an oxygen and one or more signalling molecules to the 3D cell culture during the one or more strain cycles of the 3D cell culture. Further, the method comprises detecting, via the plurality of sensors, one or more parameters may correspond to healing of wound, the 3D cell culture formation and drug administration during a physiological process and cellular responses in a real time.

BRIEF DESCRIPTION OF THE DRAWINGS
[0015] The accompanying drawings illustrate various embodiments of systems, methods, and embodiments of various other aspects of the disclosure. Any person with ordinary skills in the art will appreciate that the illustrated element boundaries (e.g. boxes, groups of boxes, or other shapes) in the figures represent one example of the boundaries. It may be that in some examples one element may be designed as multiple elements or that multiple elements may be designed as one element. In some examples, an element shown as an internal component of one element may be implemented as an external component in another, and vice versa. Furthermore, elements may not be drawn to scale. Non-limiting and non-exhaustive descriptions are described with reference to the following drawings. The components in the figures are not necessarily to scale, emphasis instead being placed upon illustrating principles.
[0016] FIG. 1 illustrates an isometric view of a microfluidic sensing device for disease modelling and drug screening studies, according to an embodiment of the present invention;
[0017] FIG. 2A illustrates a front view of the microfluidic sensing device for disease modelling and drug screening studies, according to an embodiment of the present invention;
[0018] FIG. 2B illustrates a rear view of the microfluidic sensing device for disease modelling and drug screening studies, according to an embodiment of the present invention;
[0019] FIG. 2C illustrates a left view of the microfluidic sensing device for disease modelling and drug screening studies, according to an embodiment of the present invention;
[0020] FIG. 2D illustrates a right view of the microfluidic sensing device for disease modelling and drug screening studies, according to an embodiment of the present invention;
[0021] FIG. 2E illustrates a top view of the microfluidic sensing device for disease modelling and drug screening studies, according to an embodiment of the present invention;
[0022] FIG. 2F illustrates a bottom view of the microfluidic sensing device for disease modelling and drug screening studies, according to an embodiment of the present invention;
[0023] FIG. 2G illustrates a perspective view of the microfluidic sensing device (100) for disease modelling and drug screening studies, according to an embodiment of the present invention;
[0024] FIG. 3 illustrates a measurement representation of a sensing surface, according to an embodiment of the present invention;
[0025] FIG. 4 illustrates a flowchart for the preparation of the flexible ITO, according to an embodiment of the present invention;
[0026] FIG. 5 illustrates a flow chart of the prepared flexible ITO for sensing arginine, according to an embodiment of the present invention;
[0027] FIG. 6 illustrates a graphical representation of XRD (X-ray diffraction) of a Pd/GO nanoparticles, according to an embodiment of the present invention;
[0028] FIG. 7A and 7B illustrates a graphical representation of a Fourier Transform Infrared spectroscopy (FT-IR) of GO and Pd/GO nanoparticles, according to an embodiment of the present invention;
[0029] FIG. 8 illustrates a graphical representation of an Electrodeposition and cycle optimization by using Pd/GO nanoparticles, according to an embodiment of the present invention;
[0030] FIG. 9 illustrates a graphical representation of a deposition of palladium and Graphene oxide, according to an embodiment of the present invention;
[0031] FIG. 10 illustrates a graphical representation of a Cyclic voltammetry of electrodeposited Pd/GO nanoparticles, according to an embodiment of the present invention;
[0032] FIG. 11A illustrates a graphical representation of a cyclic voltammetry of the electrodeposition of Palladium graphene oxide (Pd-GO) over flexible ITO, according to an embodiment of the present invention;
[0033] FIG. 11B illustrates a graphical representation of a cyclic voltammetry of the CV response of electrodeposited Pd-GO/ITO with prominent cathodic and anodic peak currents, according to an embodiment of the present invention;
[0034] FIG. 11C illustrates a graphical representation of a cyclic voltammetry of EIS response of electrodeposited Pd-GO/ITO, according to an embodiment of the present invention.
[0035] FIG. 12A illustrates a graphical representation of a cyclic voltammetry of sensing arginine at different concentrations, according to an embodiment of the present invention;
[0036] FIG. 12B illustrates a graphical representation of a cyclic voltammetry of sensing arginine at a concentration ranging from 1 µg/mL to 80 µg/mL, according to embodiment of the present invention;
[0037] FIG. 13 illustrates a graphical representation of the electrochemical impedance spectroscopy (EIS) of arginine at different concentration, according to an embodiment of the present invention;
[0038] FIG. 14 illustrates a graphical representation of an inference studies of the sensing device for different target analytes, according to an embodiment of the present invention;
[0039] FIG. 15 illustrates a graphical representation of a different pH of electrolyte buffer solution vs. current responses, according to an embodiment of the present invention; and
[0040] FIG. 16 illustrates a method for operating the microfluidic sensing device for disease modelling and drug screening studies, according to an embodiment of the present invention.

DETAILED DESCRIPTION
[0041] Some embodiments of this disclosure, illustrating all its features, will now be discussed in detail. The words “comprising,” “having,” “containing,” and “including,” and other forms thereof, are intended to be equivalent in meaning and be open ended in that an item or items following any one of these words is not meant to be an exhaustive listing of such item or items, or meant to be limited to only the listed item or items. It must also be noted that as used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural references unless the context clearly dictates otherwise.
[0042] Although any systems and methods similar or equivalent to those described herein can be used in the practice or testing of embodiments of the present disclosure, the preferred, systems and methods are now described. Embodiments of the present disclosure will be described more fully hereinafter with reference to the accompanying drawings in which like numerals represent like elements throughout the several figures, and in which example embodiments are shown. Embodiments of the claims may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. The examples set forth herein are non-limiting examples and are merely examples among other possible examples.
[0043] The present invention discloses about a microfluidic sensing device for disease modelling and drug screening studies and a method for operating the microfluidic sensing device for disease modelling and drug screening studies. The microfluidic sensing device is capable of monitoring a wound healing mechanism in a real time.
[0044] FIG. 1 illustrates an isometric view of the microfluidic sensing device (100) for disease modelling and drug screening studies, according to an embodiment of the present invention. FIG. 2A illustrates a front view (200) of the microfluidic sensing device (100) for disease modelling and drug screening studies, according to an embodiment of the present invention. FIG. 2B illustrates a rear view (200) of the microfluidic sensing device (100) for disease modelling and drug screening studies, according to an embodiment of the present invention. FIG. 2C illustrates a left view (200) of the microfluidic sensing device (100) for disease modelling and drug screening studies, according to an embodiment of the present invention. FIG. 2D illustrates a right view (200) of the microfluidic sensing device (100) for disease modelling and drug screening studies, according to an embodiment of the present invention. FIG. 2E illustrates a top view (200) of the microfluidic sensing device (100) for disease modelling and drug screening studies, according to an embodiment of the present invention. FIG. 2F illustrates a bottom view (200) of the microfluidic sensing device (100) for disease modelling and drug screening studies, according to an embodiment of the present invention. FIG. 2G illustrates a perspective view (200) of the microfluidic sensing device (100) for disease modelling and drug screening studies, according to an embodiment of the present invention. FIGS. 2A-2G is described in conjunction with FIG. 1.
[0045] In one embodiment, the microfluidic sensing device (100) may comprise a first layer (102), a second layer (104) and a third layer (106). Further, the first layer (102) may contain a culture chamber. Further, the second layer (104) may comprise a microvascular channel. Further, the third layer (106) having at least one membrane and a pair of pneumatic channels. Further, a sensing surface (108) is arranged on the first layer (102). Further, the sensing surface (108) may comprise a plurality of sensors. Further, the plurality of sensor may comprise an optical sensor, an electrochemical sensor and an electrical sensor. Further, the sensing surface (108) comprises at least one pair of connecting terminals (110) connected with a 3-Dimensional (3D) cell culture. Further, the sensing surface (108) comprises a plurality of ports (112).
[0046] In one embodiment, the first layer (102) of the sensing device (100) comprises the culture chamber having the 3D cell culture. Further, the 3D cell culture may resemble a natural architecture and microenvironment for the cells in vivo. Further, the culture chamber is configured to provide a controlled environment for growth, maintenance and interaction of cells in one or more dimensions. Further, the 3D cell culture may comprise various cells of different types. Further, each of the cells may include a primary cell, cell lines, and a stem cell. Further, the cells are arranged spatially to resemble a structure of a tissue and functioning of the tissue.
[0047] In one embodiment, the second layer (104) of the sensing device (100) comprises the microvascular channel. Further, the microvascular channel may resemble one or more properties of an intricate network of a plurality of blood vessels that are found in living tissues. Further, the microvascular channel is configured to provide a conduit for channelizing one or more constituents that may support the viability and functionality of the 3D cell culture contained in the culture chamber of the first layer (102). Further, the one or more constituents may comprise at least one of nutrients, oxygen, and one or more signalling molecules.
[0048] In one embodiment, the third layer (106) comprises the at least one membrane and the pair of pneumatic channels. Further, the third layer (106) corresponds to a centre chamber. Further, the third layer (106) is sandwiched between the first layer (102) and the second layer (104). Further, the at least one membrane may correspond to a porous membrane. Further, the porous membrane is a material having a predefined thickness. Further the porous member comprises a network of interconnected pores or voids within the membrane. Further, the porous membrane is configured to develop communication between the first layer (102) and the second layer (104). Further, the porous membrane is configured to transfer the one or more constituents (i.e. nutrients, oxygen and the one or more signalling molecules) from the microvascular channel to the 3D cell culture. Further, the porous nature of the porous membrane may enable a diffusion of the nutrients into the 3D cell culture. Further, the diffusion of the nutrients into the 3D cell culture is necessary for a cellular metabolism. Further, the nutrients may ensure a continuous supply of energy and provide building blocks required for cellular growth from the microvascular channel to the 3D cell culture.
[0049] Further, the porous membrane is configured to ensure a diffusion of oxygen into the 3D cell culture. Further, the diffusion of the oxygen into the 3D cell culture is a vital substrate for aerobic respiration. Further, the aerobic respiration is essential for generating cellular energy in the form of ATP (adenosine triphosphate). Further, the porous membrane may provide a passage of the one or more signalling molecules. Further, the one or more signalling molecules may comprise hormones, cytokines, or etc. which may regulate the various cellular process. Further, the cellular process may include proliferation, differentiation, and immune responses of the 3D cell culture.
[0050] Further, the pair of pneumatic channels is configured to perform a pneumatic operation. Further, the pair of pneumatic channels are configured to supply a pre-defined amount of pneumatic pressure to the 3D cell culture of the first layer (102). Further, the pair of pneumatic channels are configured to exert a controlled mechanical stress on the cellular environment of the 3D cell culture. Further, the pair of pneumatic channels is configured to provide a mechanical stimulus to the 3D cell culture. Further, the mechanical stress exerting on the 3D cell culture via the pair of pneumatic channels may induce one or more strain cycles. Further, the pair of pneumatic channels is configured to provide the pre-defined amount of pneumatic pressure to the 3D cell culture at a pre-defined time interval for stimulate the physiological conditions experienced by the cells in vivo. Further, the one or more strain cycles are configured to modulate a cellular behaviour by stimulating a cell process. Further, the cell process may comprise cell morphology, proliferation, differentiation and a gene expression.
[0051] In one embodiment, the sensing surface (108) having the plurality of sensors is arranged on the first layer (102). Further, the sensing surface (108) is configured to detect one or more biomolecules. Further, the one or more biomolecules may correspond to arginine, tyrosine, cysteine, uric acid, ascorbic acid, or alike. Further, the plurality of sensors may comprise the optical sensor, the electrochemical sensor and the electrical sensor. For example, the optical sensor works on different optical properties. Further, the optical properties may comprise fluoresce, absorbance, luminescence, or other optical properties. Further, during wound healing process of the cell of the 3D cell culture, 3D cell culture formation, and a drug administration, the optical sensor may continuously collect one or more signals emitted by the cells or molecules of the 3D cell culture. For example, if the fluorescence is present within the 3D cell culture, the optical sensor may receive the one or more signals corresponding to an emitted light having a specific wavelength.
[0052] For example, the electrochemical sensor is configured to detect the drug administration in the 3D cell culture in a real time. Further, the electrochemical sensor is configured to continuously monitor an analyte concentration of a drug or any other molecules. Further, the drug administration within the 3D cell culture may cause change in analyte concentration of the 3D cell culture. Further, changes in the analyte concentration may induce changes in current, voltage, impedance, or capacitance. Further, the changes in the current, the voltage, the impedance, or capacitance may represent in the form of the one or more signals.
[0053] For example, the electrical senor is configured to detect changes in one or more electrical properties of the 3D cell culture. Further, the changes in the electric properties within the 3D cell culture is due to variations in a cell density, morphology and the presence of analyte in the 3D cell culture during a healing process of the cells. Further, the changes in the electrical properties of the 3D cell culture may represent in the form of the one or more signals. Further, the one or more signals obtained from the plurality of sensors are in the form of amplitudes, frequencies, slopes, or etc. Further the one or more signals are stored in a digital format within a database associated with the device (100).
[0054] In one embodiment, the sensing surface (108) may comprise the at least one pair of connecting terminals (110) integrated within the 3D cell culture. Further, the at least one pair of connecting terminals (110) is configured to enable a connection of the sensing device (100) with one or more external devices. Further, the one or more external devices may comprise a computing unit, a microscope, microelectrode arrays, and etc. Further, the at least one pair of connecting terminals (110) are configured to provide an interface between the sensing device (100) and the one or more external devices, enabling the exchange of the one or more signals, data, or etc.
[0055] In an example embodiment, each of the at least one pair of connecting terminals (110) is coated with a flexible Indium tin oxide (ITO). Further, the flexible ITO is configured to sense essential amino acids that are used for would healing mechanism and cell proliferation mechanism of the cells of the 3D cell culture. Further, an electrode is integrated within each of the at least one pair of connection terminals (110) may detect and quantify the amino acids within the 3D cell culture. Further, the amino acids are essential for a protein synthesis, cell proliferation, tissue repair in the wound healing mechanism. Further, a selective binding of the amino acids with an electrode surface causes changes in the one or more electric signals in a real time. Further, the one or more signals are transmitted to the one or more external devices. Further, the one or more signals may allow researchers to track dynamic processes, such as cellular responses, drug diffusion, or chemical reactions, wound healing mechanism in the real-time.
[0056] In one embodiment, the sensing surface (108) may comprise the plurality of ports (112). Further, the plurality of ports (112) is configured to provide controlled introduction of liquids, gases, or particles within the 3D cell culture. Further. an influx and efflux of a plurality of substances through the plurality of ports (112) may allow the researchers to regulate several factors such as nutrient availability, oxygen tension, and the presence of one or more signalling molecules, thereby influencing cellular behaviour and responses. Further, the plurality of ports (112) is configured to administrate drugs, growth factors, or other bioactive compounds, as well as a removal of waste products or metabolic by-products from the 3D cell culture.
[0057] In an example embodiment, an anti-microbial study is conducted using a carbon nanomaterial through an agar well diffusion and broth micro-dilution procedures. Further, the anti-microbial activity of the 3D cell culture is carried out by measuring an inhibition zones formed around wells containing tested materials in agar plates. Further, the anti-microbial activity of the 3D cell culture is determined by of a minimum inhibitory concentration (MIC) required to inhibit microbial growth in a liquid culture. Further, a carbon nanomaterial is selected as a bio nanomaterial. Further, the carbon nanomaterial is integrated within the sensing device (100). Further, the anti-microbial activity of the 3D cell culture is evaluated by using the plurality of sensors.
[0058] In one embodiment, a structure directing agent is incorporated into the culture chamber, or the at least one membrane. The structure directing agent is selected to guide the formation and arrangement of cells within the 3D cell culture. The structure-directing agent is a substance used in microfluidic sensing device (100) to guide the formation, arrangement, and growth of cells or materials within a specific environment. The structure-directing agents is configured to create structured three-dimensional environments to mimic natural biological tissues by controlling the organization of cells and interactions of the cells with the extracellular matrix (ECM). The structure-directing agent is configured to ensure the proper spatial distribution and organization of cells, which is crucial for developing 3-dimesnional models that replicate the structure and function of in vivo tissues.
[0059] In some embodiments, the structure-directing agent is used periodically to maintain the desired environment within the culture chamber. The periodical use of the structure-directing agent may include using the structure-guiding agent as a part of materials used to fabricate the sensing device (100).
[0060] Furthermore, the structure directing agent may include arginine, thermresponsive polymers, block copolymers, and so on. In an example embodiment, the structure guiding agent is arginine.
[0061] FIG. 3 illustrates a measurement representation (300) of a sensing surface (108), according to an embodiment of the present invention.
[0062] In an example embodiment, the sensing surface (108) may comprise a middle portion. Further, the middle portion may define a length L mm. Further, the length L may define as 12.0766 mm. Further, the middle portion may define a width W mm. Further, the width W may define as 2.9333mm. Further, the distance between the plurality of ports (112) and the middle portion of the sensing surface (108) may correspond to D. Further, the D may define as 4.233mm. Further, the width of a channel of the sensing surface (108) may correspond to d. Further, d may define as 0.9334 mm.
[0063] FIG. 4 illustrates a flowchart (400) for the preparation of the flexible ITO, according to an embodiment of the present invention. FIG. 5 illustrates a flow chart (500) of the prepared flexible ITO for sensing arginine, according to an embodiment of the present invention. FIG. 5 is described in conjunction with FIG. 4.
[0064] In an example embodiment, the method for the preparation of the flexible ITO comprises of following steps which include: dispersing of a graphene oxide (GO) in a solvent. Further, the solvent may comprise a water or an ethanol. Further, the dispersion process is achieved by a process of sonication. Further, adding the palladium chloride (PdCl2) into the GO dispersion. Further, adding a reducing solution (NaBH4) in the solution while stirring. Further, the reducing agent may reduce Pd(II) ions to Pd(0), resulting in the formation of palladium nanoparticles on the surface of GO.
[0065] Further, electrodeposition of Pd/GO nanoparticles to form the flexible ITO. Electrodeposition of Pd/GO nanoparticles to form flexible ITO (Indium Tin Oxide) electrodes provides a good approach for fabricating transparent, conductive coatings suitable for flexible electronics. The process integrates the benefits of Pd nanoparticles for enhanced conductivity and graphene oxide (GO) for mechanical flexibility and stability. Further, a potential difference is applied between the working electrode and a counter electrode immersed in the electrolyte solution. Further, the working electrode may correspond to Indium Tin oxide (ITO) coated substrate on the working electrode. Further, the potential is choosing to facilitate the reduction of Pd ions and the deposition of Pd nanoparticles onto the electrode surface. During the process of electrodeposition, the palladium complex is catalysed. Further, the glycals is configured to modify arginine guanidine groups in one step with high functional group tolerance at ambient temperature.
[0066] In one embodiment, after optimizing the electrodeposition of Pd/GO nanoparticles, a potential range is choosing for the process ranged from a positive potential of +0.5 V to a negative potential of -1.5 V. The scan rate was maintained at 0.05 V/s, and the number of scans was optimized for 5 cycles. Further, the potential range dictates the electrochemical conditions during electrodeposition. Starting from a positive potential of +0.5 V facilitates the reduction of Pd ions, leading to the formation of Pd nanoparticles on the electrode surface. Further, the scanning to negative potentials as low as -1.5 V ensures efficient deposition of Pd nanoparticles and promotes the interaction with graphene oxide (GO) to form the Pd/GO nanocomposite.
[0067] Further, the scan rate of 0.05 V/s indicates the speed at which the potential is swept across the chosen range. A moderate scan rate is typically preferred to control the electrodeposition process and ensure uniform coating deposition on the substrate surface. Further, optimizing the number of scans for 5 cycles suggests that the electrodeposition process undergoes multiple cycles to achieve the desired coating thickness and morphology.
[0068] In an example embodiment, the sensing surface on the flexible ITO electrode interacts selectively with arginine through specific binding interactions, such as hydrogen bonding, electrostatic interactions, or ligand-receptor interactions, depending on the nature of the recognition element upon exposure of the 3D cell culture containing arginine. Further, the interaction leads to changes in the electrical properties of the sensing surface, such as conductivity, capacitance, or impedance that is measured using electrochemical or impedance spectroscopy techniques. Further, the changes in the electrical properties of the sensing surface induced by arginine binding are transduced into measurable signals, such as current or voltage, using appropriate readout techniques.
[0069] FIG. 6 illustrates a graphical representation (600) of XRD (X-ray diffraction) of a Pd/GO nanoparticles, according to an embodiment of the present invention.
[0070] In one embodiment, the diffraction peak corresponding to the (001) plane of graphene oxide typically appears at a low 2? angle (around 10-15 degrees) due to the interlayer spacing of the graphene oxide sheets. The peak indicates the presence of the layered structure of GO, with the diffraction arising from the periodic stacking of graphene sheets. Further, the diffraction peaks corresponding to crystallographic planes of palladium appear at higher 2? angles informs about a crystalline nature of the Pd nanoparticles in the nanocomposite. The (111) peak is the highest peaks in the XRD pattern, indicating the preferential orientation of Pd crystallites along the (111) plane. Further, (200), (220), (311) peaks may correspond to other crystallographic planes of Pd and provide additional information about the crystal structure and orientation of the Pd nanoparticles. Further, the (222) peak indicates the presence of high-quality Pd crystallites with a face-centered cubic (FCC) crystal structure. The XRD representation confirms the successful formation of the Pd-GO nanocomposite.
[0071] FIG. 7A and 7B illustrates a graphical representation (700) of a Fourier Transform Infrared spectroscopy (FT-IR) of GO and Pd/GO nanoparticles, according to an embodiment of the present invention.
[0072] In one embodiment, the FTIR of the GO and Pd/GO particles may indicate the presence of C-H stretching, C=O stretching and O-H stretching. The presence of these functional groups is essential for the chemical properties and reactivity of both graphene oxide and the Pd-GO complex. Further, the FT-IR indicates the chemical structure of graphene oxide which is preserved even after the incorporation of palladium nanoparticles. These functional groups also play a significant role in potential applications such as catalysis, sensing, and electronics.
[0073] FIG. 8 illustrates a graphical representation (800) of an Electrodeposition and cycle optimization by using Pd/GO nanoparticles, according to an embodiment of the present invention.
[0074] In one embodiment, the cycle optimization plot is plotted the performance metric (e.g., conductivity, electrochemical activity) of the Pd/GO electrode as a function of the number of electrodeposition cycles. Further, the graph visualizes the optimization process, showing how the performance improves or stabilizes with increasing cycles before reaching a plateau.
[0075] FIG. 9 illustrates a graphical representation (900) of a deposition of palladium and Graphene oxide, according to an embodiment of the present invention. FIG. 10 illustrates a graphical representation (1000) of a Cyclic voltammetry of electrodeposited Pd/GO nanoparticles, according to an embodiment of the present invention. FIG. 10 is described in conjunction with FIG. 9.
[0076] In one embodiment, the CV plot shows the redox behavior of the electrodeposited Pd/GO nanoparticles during successive potential scans. The CV plot typically consists of multiple overlapped curves, each representing a single potential scan. The shape and features of these curves provide information about the electrochemical processes occurring at the electrode surface. Peaks observed in the CV plot correspond to specific electrochemical reactions, such as the oxidation or reduction of Pd species or functional groups on the graphene oxide surface. Anodic peaks correspond to oxidation processes, while cathodic peaks correspond to reduction processes.
[0077] FIG. 11A illustrates a graphical representation (1100) of a cyclic voltammetry of the electrodeposition of Palladium graphene oxide (Pd-GO) over flexible ITO, according to an embodiment of the present invention.
[0078] In one embodiment, the electrodeposition process of Pd-GO nanoparticles onto a flexible Indium Tin Oxide (ITO) substrate. The graphical representation depicts the setup of the electrochemical cell, with the flexible ITO electrode as the working electrode, a counter electrode, and a reference electrode immersed in an electrolyte solution containing Pd precursor and graphene oxide. The process of electrodeposition is shown, with Pd ions being reduced and deposited onto the surface of the ITO electrode, forming a Pd-GO nanocomposite.
[0079] FIG. 11B illustrates a graphical representation (1100) of a cyclic voltammetry of the CV response of electrodeposited Pd-GO/ITO with prominent cathodic and anodic peak currents, according to an embodiment of the present invention.
[0080] In one embodiment, the cyclic voltammetry (CV) response of the electrodeposited Pd-GO/ITO electrode. The graph shows the current (y-axis) as a function of applied potential (x-axis) during successive potential scans. Prominent cathodic and anodic peak currents are observed in the CV curve, indicating redox reactions associated with the Pd-GO nanoparticles. The position, shape, and intensity of these peaks provide information about the electrochemical behavior and catalytic activity of the Pd-GO/ITO electrode.
[0081] FIG. 11C illustrates a graphical representation (1100) of a cyclic voltammetry of EIS response of electrodeposited Pd-GO/ITO, according to an embodiment of the present invention.
[0082] In one embodiment, the electrochemical impedance spectroscopy (EIS) response of the electrodeposited Pd-GO/ITO electrode. The Nyquist plot shows the impedance (imaginary component) versus frequency (real component) of the electrode system. The EIS response provides insights into the electrical properties, interfacial capacitance, and charge transfer kinetics of the Pd-GO/ITO electrode. The shape and characteristics of the Nyquist plot is analyzed to evaluate the performance and stability of the electrode for various applications, such as sensing or catalysis.
[0083] FIG. 12A illustrates a graphical representation (1200) of a cyclic voltammetry of sensing arginine at different concentrations, according to an embodiment of the present invention. FIG. 12B illustrates a graphical representation of a cyclic voltammetry of sensing arginine at a concentration ranging from 1 µg/mL to 80 µg/mL, according to embodiment of the present invention.
[0084] In one embodiment, the graphical representation comprises a CV curves showing a characteristic peaks corresponding to the redox reactions associated with the recognition element's interaction of the Pd/GO nanoparticles with arginine. Further, the increase in the arginine concentration led to changes in the peak currents or peak areas, demonstrating the sensing surface response to varying analyte concentrations.
[0085] FIG. 13 illustrates a graphical representation (1300) of the electrochemical impedance spectroscopy (EIS) of arginine at different concentration, according to an embodiment of the present invention.
[0086] In one embodiment, the impedance spectra illustrate characteristic features, such as semicircles in the high-frequency region corresponding to charge transfer processes and linear segments in the low-frequency region associated with diffusion processes. Further, the changes in the impedance spectra are observed with increasing arginine concentration, reflecting alterations in the electrode interface caused by the interaction between the recognition element of the flexible ITO and arginine molecules.
[0087] FIG. 14 illustrates a graphical representation (1400) of an inference studies of the sensing device (100) for different target analytes, according to an embodiment of the present invention.
[0088] In an example embodiment, the selectivity analysis reveals the sensing device (100) ability to specifically detect the target analyte (arginine) in the presence of interferents. Minimal interference from interferents illustrated by low cross-reactivity and negligible changes in sensing device (100) response, demonstrates the high selectivity of the sensing device (100).
[0089] FIG. 15 illustrates a graphical representation (1500) of a different pH of electrolyte buffer solution vs. current responses, according to an embodiment of the present invention.
[0090] In an example embodiment, the graphical representation revealed that pH 7 of the electrolyte buffer solution resulted in the highest current response for the sensing device (100). Further, the optimal pH condition is crucial for achieving maximum sensitivity, accuracy, and reliability of the sensing device (100) for practical applications.
[0091] FIG. 16 illustrates a method (1600) for operating microfluidic sensing device (100) for disease modelling and drug screening studies, according to an embodiment of the present invention.
[0092] At operation 1602, providing, via the first layer (102), the culture chamber contained with the 3D (3-dimensional) cell culture. Further, the 3D cell culture may resemble the natural architecture and microenvironment for the cells in vivo. Further, the culture chamber is configured to provide the controlled environment for growth, maintenance and interaction of cells in three dimensions.
[0093] At operation 1604, providing, via the second layer (104), the microvascular channel. Further, the microvascular channel may resemble the intricate network of the plurality of blood vessels that are found in living tissues. Further, the microvascular channel is configured to provide the conduit for channelizing the nutrients, oxygen and the one or more signals molecules to support the physiological behaviour of the 3D cell culture.
[0094] At operation 1606, providing, via the third layer (106), at least one membrane and the pair of pneumatic channels. Further, the third layer (106) is sandwiched between the first layer (102) and the second layer (104). Further, the at least one membrane may correspond to the porous membrane. Further, the porous membrane is the thin material having the network of interconnected pores or voids within the membrane.
[0095] At operation 1608, providing, via each of the pair of pneumatic channels, the predefined amount of pneumatic pressure to the 3D cell culture. Further, the pair of pneumatic channels are configured to deliver the pre-defined amount of pneumatic pressure to the 3D cell culture of the first layer (102). Further, the pair of pneumatic channels are configured to exert the controlled mechanical force on the cellular environment of the 3D cell culture. Further, the pair of pneumatic channels is configured to provide the mechanical stimulus to the 3D cell culture. Further, the mechanical force exerting on the 3D cell culture via the pair of pneumatic channels may induce the one or more strain cycles.
[0096] At operation 1610, transferring, via the at least one membrane, the nutrient, the oxygen and the one or more signalling molecules to the 3D cell culture during the one or more strain cycles of the 3D cell culture. Further, the porous membrane is configured to transfer the nutrient, the oxygen and the one or more signalling molecules from the microvascular channel to the 3D cell culture. Further, the one or more strain cycles are configured to modulate the cellular behaviour by stimulating cell process. Further, the cell process may comprise cell morphology, proliferation, differentiation and a gene expression.
[0097] At operation 1612, detecting, via the plurality of sensors, the one or more parameters may correspond to healing of wound, the 3D cell culture formation and drug administration during the physiological process and cellular responses in the real time. Further, the plurality of sensor may comprise the optical sensor, the electrochemical sensor and the electrical sensor. Further, the one or more from the optical signals corresponding to the emitted light at the specific wavelength due to the presence of cell and molecules in the 3D cell culture.
[0098] Further, the electrochemical sensor is configured to detect the drug administration in the 3D cell culture in the real time. Further, the administration of drug within the 3D cell culture may cause change in analyte concentration of the 3D cell culture. Further, the change in the current, the voltage, the impedance, or capacitance may represent in the form of the one or more signals.
[0099] Further, the electrical senor is configured to detect the changes in the electrical properties of the 3D cell culture. For example, the changes in the electric properties within the 3D cell culture is due to the variations in a cell density, morphology and the presence of analyte in the 3D cell culture during the healing process of the cells. Further, the one or more signals obtained from the plurality of sensors are in the form of amplitudes, frequencies, slopes, or etc. Further the one or more signals are stored in a digital format within a database associated with the device (100) for analysis.
[00100] It has thus been seen that the microfluidic sensing device (100) for disease modelling and drug screening studies and the method (1600) for operating microfluidic sensing device (100) for disease modelling and drug screening studies, as described. The microfluidic sensing device (100) for disease modelling and drug screening studies and the method (1600) for operating the microfluidic sensing device (100) for disease modelling and drug screening studies in any case could undergo numerous modifications and variants, all of which are covered by the same innovative concept; moreover, all of the details can be replaced by technically equivalent elements. In practice, the components used, as well as the numbers, shapes, and sizes of the components can be whatever according to the technical requirements. The scope of protection of the invention is therefore defined by the attached claims. , C , Claims:CLAIMS
We Claim:
1. A microfluidic sensing device (100) for disease modelling and drug screening studies, the sensing device (100) comprising:
a first layer (102) comprising:
a culture chamber contained with a 3D (3-dimensional) cell culture;
a second layer (104) having a microvascular channel;
a third layer (106) having at least one membrane and a pair of pneumatic channels; and
a sensing surface (108) having a flexible ITO and a plurality of sensors is arranged on the first layer (102).
wherein each of the pneumatic channels are configured to put the 3D cell culture through one or more strain cycles by providing a predefined amount of pneumatic pressure to the 3D cell culture,
wherein the at least one membrane is configured to transfer a nutrient, an oxygen and one or more signalling molecules to the 3D cell culture during the one or more strain cycles of the 3D cell culture; and
wherein the flexible ITO and the plurality of sensors is configured to detect one or more parameters, wherein the one or more parameters may correspond to healing of wound, formation of the 3D cell culture and drug administration during a physiological process and cellular responses in a real time.
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2. The microfluidic sensing device (100) as claimed in claim 1, further the sensing surface (108) comprising at least one pair of connecting terminals (110) connected with the 3D cell culture, wherein the at least one pair of connecting terminals (110) are configured to enable connection of one or more external device (100) with the sensing surface (108).
3. The microfluidic sensing device (100) as claimed in claim 1, wherein the third layer (106) is sandwiched between the first layer (102) and the second layer (104) and the third layer (106) correspond to a centre chamber
4. The microfluidic sensing device (100) as claimed in claim 1, wherein the microvascular channel corresponds to a vascular network found in a human body.
5. The microfluidic sensing device (100) as claimed in claim 1, wherein the sensing surface (108) further comprises a plurality of ports (112) that are configured to provide controlled introduction of liquids, gases, or particles within the 3D cell culture.
6. The microfluidic sensing device (100) as claimed in claim 1, wherein the plurality of sensors may correspond to an optical sensor, an electrochemical sensor and an electrical sensor.
7. The microfluidic sensing device (100) as claimed in claim 1, wherein a structure directing agent is incorporated into the culture chamber, or the at least one membrane, wherein the structure directing agent is selected to guide the formation and arrangement of cells within the 3D cell culture.
8. The microfluidic sensing device (100) as claimed in claim 8, wherein the structure directing agent may include arginine, thermresponsive polymers, block copolymers, and so on.
9. The microfluidic sensing device (100) as claimed in claim 1, wherein the sensing surface (108) is configured to detect one or more biomolecules, wherein the one or more biomolecules may correspond to arginine, tyrosine, cysteine, uric acid, ascorbic acid, or alike.
10. A method (1600) comprising:
providing, via a first layer (102), a culture chamber contained with a 3D (3-dimensional) cell culture, at step 1602;
providing, via a second layer (104), a microvascular channel, at step 1604;
providing, via a third layer (106), at least one membrane and a pair of pneumatic channels, at step 1608;
providing, via each of the pair of pneumatic channels, a predefined amount of pneumatic pressure to the 3D cell culture, at step 1608;
transferring, via the at least one membrane, a nutrient, an oxygen and one or more signalling molecules to the 3D cell culture during the one or more strain cycles of the 3D cell culture, at step 1610; and
detecting, via the plurality of sensors, one or more parameters may correspond to healing of wound, the 3D cell culture formation and drug administration during a physiological process and cellular responses in a real time, at step 1612.