A microfluidic device for assessing fibrosis in material implantation and cancer

Abstract

Fibrosis diagnostic systems, subsystems, and components thereof are provided. The fibrosis diagnostic system may include a microfluidic device. The fibrosis diagnostic system may be preloaded with one or more components. For example, the fibrosis diagnostic system may include one or more reagents and a target object, such as a tissue, or a medical device, or both, intended for implantation. The fibrosis diagnostic system may include one or more additional components and subsystems, such as a detector including a sensor configured to detect fibrosis in the target object, a thermal subsystem, a fluidic subsystem, a processor, or a user interface, or any combination thereof. Kits are provided that include one or more components of the fibrosis diagnostic system. Screening methods for identifying therapeutic agents utilizing the fibrosis diagnostic system or components thereof are provided. Non-transitory computer readable media for storing programs are further provided.

Description

[Technical field]

[0001] (CROSS REFERENCE TO RELATED APPLICATIONS) This application is an international application under the Patent Cooperation Treaty, claiming priority to U.S. Provisional Patent Application No. 63 / 335,527, filed April 27, 2022, the contents of which are incorporated herein by reference in their entirety.

[0002] The present disclosure relates to systems, devices, and methods for analyzing foreign body-associated fibrosis, and therapeutic approaches based on that analysis. [Background technology]

[0003] Foreign body reactions lead to fibrosis of the implant and cancer tumor, which can cause multiple adverse effects such as hypertrophic scarring, excessive inflammation, and reduced efficacy of cancer therapeutics. During the fibrosis formation process, crosstalk mainly between macrophages and fibroblasts leads to the detachment of the biomaterial from the native tissue. Traditionally, these interactions have been evaluated by in vivo placement of foreign bodies in mouse or rat models. Available in vitro models attempt to mimic scarring, including associated fibrosis. Although these models can provide relevant data, they cannot provide a sufficient model of the interactions that take place in the human body and, as a result, can only have limited predictive value.

[0004] Solutions to these technical problems are provided by the present disclosure. Summary of the Invention

[0005] Fibrosis diagnostic systems, subsystems, and components thereof are provided by the present disclosure. The fibrosis diagnostic system can include a microfluidic device. The microfluidic device can include a device housing comprising a device interior, a plurality of channels within the device interior, a plurality of septa separating the channels from one another, and a plurality of channel ports within the device housing in fluid communication with the plurality of channels. The plurality of channels can include, for example, a first, second, and third channel. The first channel can be a central channel, and the second and third channels can be lateral channels on opposing sides of the central channel. The first channel can be configured to receive a target object, a substrate, and a first biological composition configured to promote fibrosis of the target object. The second channel can be configured to receive a second biological composition configured to promote fibrosis of the target object. The first septum can separate the first and second channels. The third channel can be configured to receive a third biological composition configured to promote fibrosis of the target object. The second septum can separate the first and third channels. The plurality of channel ports may include any number of channel ports having a minimum of two channel ports. The plurality of channel ports may include one or more sets of channel ports. The fibrosis diagnostic system may be configured to form a gradient of at least one type of molecule across the first channel, the second channel, or the third channel, or any combination thereof. The fibrosis diagnostic system may be preloaded with one or more components. For example, the fibrosis diagnostic system may include a substrate, a first biological composition, a second biological composition, or a third biological composition, or any combination thereof. The fibrosis diagnostic system may include a target object. The fibrosis diagnostic system may include one or more additional components and subsystems, such as a detector including a sensor configured to detect fibrosis in the target object, a thermal subsystem, a fluidic subsystem, a processor, or a user interface, or any combination thereof.Kits are provided that include one or more components of the fibrosis diagnostic system. Screening methods are provided that use the fibrosis diagnostic system or its components. Screening methods include, for example, methods of screening for anti-fibrotic or anti-cancer agents. Further provided are non-transitory computer readable media that store a program that includes instructions for performing the methods of the disclosure or for controlling the systems of the disclosure. [Brief description of the drawings]

[0006] For a fuller understanding of the nature, objects, and advantages of the present disclosure, reference should be made to the following detailed description which should be read in conjunction with the following drawings, in which like reference numerals indicate like elements and in which:

[0007] [Figure 1A] FIG. 1A is a perspective view of a microfluidic device according to the present disclosure. [Figure 1B] FIG. 1B is a top view of the microfluidic device shown in FIG. 1A. [Figure 2A] FIG. 2A is a perspective view of a fibrosis diagnostic system showing a microfluidic device disengaged from a detector according to the present disclosure. [Figure 2B] FIG. 2B is a perspective view of the fibrosis diagnostic system of FIG. 2A showing a microfluidic device engaged with a detector according to the present disclosure. [Diagram 3] FIG. 3 is a top view of a microfluidic device according to the present disclosure. [Figure 4] FIG. 4 is a top view of a microfluidic device according to the present disclosure. [Diagram 5] FIG. 5 is a top view of a microfluidic device according to the present disclosure. [Figure 6A] FIG. 6A is a top perspective view of a microfluidic device according to the present disclosure. [Figure 6B] FIG. 6B is a front perspective view of the microfluidic device shown in FIG. 6A. [Figure 6C] FIG. 6C is an enlarged perspective view of a portion of the microfluidic device shown in FIG. 6A. [Figure 6D]FIG. 6D is an enlarged plan view of a portion of the microfluidic device shown in FIG. 6A. [Figure 6E] FIG. 6E is a further enlarged plan view of a portion of the enlarged view shown in FIG. 6D. [Figure 7A] FIG. 7A is a schematic cross-sectional view of a loaded microfluidic device at a first point in time according to the present disclosure. [Figure 7B] FIG. 7B is a schematic cross-sectional view of the loaded microfluidic device shown in FIG. 7A at a second time point. [Figure 8] FIG. 8 is a schematic diagram of a fibrosis diagnostic system according to the present disclosure. [Figure 9] FIG. 9 is a schematic diagram of a fibrosis diagnostic system according to the present disclosure. [Figure 10] FIG. 10 is a schematic diagram of a fibrosis diagnostic system according to the present disclosure. [Figure 11] Subsections A, B, and C of FIG. 11 show magnified images of cells stained with picrosirius red according to the present disclosure. [Figure 12] FIG. 12 shows magnified fluorescent images of cells grown in uncoated or fibronectin-coated 96-well plates with L-ascorbic acid, with or without TGF-β1, and fixed with 4% paraformaldehyde (PFA) in accordance with the present disclosure. [Figure 13] FIG. 13 shows magnified fluorescent images of cells grown in uncoated or fibronectin-coated 96-well plates with L-ascorbic acid, with or without TGF-β1, and fixed with methanol in accordance with the present disclosure. [Figure 14] FIG. 14 shows a graph of quantified fluorescence intensity at 4× and 10× magnification of the collagen I and α-SMA channels of cells grown according to the present disclosure. [Figure 15] FIG. 15 shows a graph of quantified fluorescence intensity at 20× and 40× magnification of the collagen I and α-SMA channels of cells grown according to the present disclosure. [Figure 16]FIG. 16 shows magnified fluorescent images of cells grown in uncoated or fibronectin-coated 8-chamber glass slides with L-ascorbic acid, with or without TGF-β1, and fixed with 4% paraformaldehyde (PFA) in accordance with the present disclosure. [Figure 17] FIG. 17 shows magnified fluorescent images of cells grown in uncoated or fibronectin-coated 8-chamber glass slides with L-ascorbic acid, with or without TGF-β1, and fixed with methanol in accordance with the present disclosure. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0008] A fibrosis diagnostic system is provided by the present disclosure. The fibrosis diagnostic system can include a microfluidic device. The microfluidic device can include a device housing including a device interior, a plurality of channels within the device interior, a plurality of septa separating the channels from each other, and a plurality of channel ports within the device housing in fluid communication with the plurality of channels. The plurality of channels can include any number of channels with a minimum of two channels. The plurality of channels can be, for example, two, three, four, five, six, seven, eight, nine, or ten channels, or more than ten channels. The number of septa can be, for example, one less than the number of channels. For example, the plurality of channels can include a first, second, and third channel with a first and second septa. The number of septa can be, for example, one, two, three, four, five, six, seven, eight, or nine, or more than nine septa. The first channel can be a central channel, and the second and third channels can be lateral channels on opposite sides of the central channel (first channel). The first channel may be configured to receive a target object, a substrate, and a first biological composition configured to promote fibrosis of the target object. The second channel may be configured to receive a second biological composition configured to promote fibrosis of the target object. A first septum may separate the first and second channels. The first septum may include a first plurality of openings providing fluid communication between the first channel and the second channel. The third channel may be configured to receive a third biological composition configured to promote fibrosis of the target object. A second septum may separate the first and third channels. The second septum may include a second plurality of openings providing fluid communication between the first channel and the third channel. The multiple channel ports in the device housing in fluid communication with the multiple channels may comprise any number of channel ports having a minimum of two channel ports. The multiple channel ports may include one or more sets of channel ports having a set belonging to a particular channel.For example, the plurality of channel ports can include a first set of channel ports in fluid communication with a first channel, a second set of channel ports in fluid communication with a second channel, and a third set of channel ports in fluid communication with a third channel, each set of channel ports including one or more channel ports.

[0009] The fibrosis diagnostic system may be configured to form a gradient of at least one type of molecule across the first channel, the second channel, or the third channel, or any combination thereof. For example, the at least one type of molecule may include a chemotactic molecule. The chemotactic molecule may promote the migration of at least one cell type present in the first biological composition, the second biological composition, or the third biological composition, or any combination thereof. The at least one type of molecule may include a growth factor. The growth factor may promote the growth of a cell population, thereby forming a gradient that includes the cell population. The first biological composition, the second biological composition, or the third biological composition, or any combination thereof may include at least one type of molecule.

[0010] The plurality of channel ports may include any number of channel ports with a minimum of two channel ports. A channel may share one or more channel ports. A channel may have one or more exclusive channel ports. The plurality of channel ports may comprise at least one channel port for each channel. The plurality of channel ports may comprise at least two channel ports for at least one channel. The plurality of channel ports may include one or more sets of channel ports with the set of channel ports belonging to a particular channel. For example, the second and third sets of channel ports each include two or more channel ports. Each set of channel ports may include any desired number of channel ports, e.g., at least one channel port, two or more channel ports, three or more channel ports, or four or more channel ports. The second and third sets of channel ports each include a first channel port and a second channel port, e.g., a first channel port proximate a first end of its respective channel and a second channel port proximate a second end of its respective channel. Each of the sets of channel ports can include a first channel port and a second channel port, e.g., a first channel port proximal to a first end of its respective channel and a second channel proximal to a second end of its respective channel. The first channel can include a central channel port configured to receive a target object into the first channel. The central channel port can be between a first end of the first channel and a second end of the second channel. For example, the central channel port can be equidistant between the first end and the second end (the end of the first channel). One or more of the plurality of channel ports can include a seal, a valve, or a closure, or any combination thereof, configured for movement between an open state and a closed state. One or more of the plurality of channel ports can be configured to connect to a fluidic subsystem for moving one or more components into or out of the microfluidic device, or both.A channel port can be configured as an inlet, or an outlet, or both.

[0011] The one or more retaining elements can be located in one or more channels of the multi-fluidic device. For example, the one or more retaining elements can be located in one or more of the first, second, and third channels. The retaining elements can be configured to retain at least one component that is insertable into one or more of the channels of the microfluidic device, e.g., the first, second, and third channels. The retaining elements can be of any desired design or combination of designs. The retaining elements can be permanent or removable. The retaining elements can be movable within the microfluidic device or can be moved in and out of the microfluidic device. The retaining elements can be retractable. The retaining elements can include, for example, a modified channel surface. The modification can include, for example, a biomolecule. The biomolecule can include, for example, a polypeptide, a polysaccharide, a lipid, or a polynucleotide, or any combination thereof. The modification can include, for example, a polymer, a metal, or a ceramic, or any combination thereof. The device housing can include, for example, a polymer, a metal, or a ceramic, or any combination thereof. For example, the device housing can include a first polymer and the modification can include a second polymer. The first and second polymers may differ from each other in chemical composition. The modification may include, for example, a physical structure. The physical structure and the device housing may include a common material. The modification may include a container for receiving a target object. The container may include, for example, a recess in the channel surface. The container, recess, or other physical structure may have any desired geometric shape, or size, or both. For example, the container, recess, or physical structure may be circular, elliptical, triangular, square, rectangular, pentagonal, hexagonal, heptagonal, octagonal, or trapezoidal, or any combination thereof.The width, diameter, height, or depth of the vessel, depression, or other physical structure, or any combination thereof, can be less than about 1.0 μm, between about 1.0 μm and about 1.0 cm, between about 10 μm and about 100 mm, between about 50 μm and about 50 mm, between about 50 μm and about 5.0 mm, between about 50 μm and about 9.5 mm, between about 100 μm and about 1.0 mm, or greater than 1.0 cm. The physical structure can include multiple physical structures. For example, the multiple physical structures can include multiple recesses in the channel surface, or multiple protrusions from the channel surface, or both.

[0012] The microfluidic device and any of its components may have any desired geometry, shape, size, dimension, or relative scale, or any combination thereof. For example, the length, or width, or both, of the microfluidic device may be less than 1.0 mm, about 1.0 mm to about 10 cm, about 2.5 mm to about 5.0 cm, about 5.0 mm to about 2.5 cm, about 7.5 mm to about 1.0 cm, or more than 10 cm. The thickness of the microfluidic device may be less than about 0.001 mm, about 0.001 mm to about 1.0 cm, about 0.01 mm to about 0.5 cm, about 0.1 mm to about 0.1 cm, about 1.0 mm to about 0.5 cm, about 10 mm to about 100 mm, or more than 1.0 cm. The first channel, the second channel, or the third channel, or any combination thereof, may include a curved cross-section, or a straight cross-section, or both. The curved cross-section may be circular or elliptical. The first channel, the second channel, or the third channel, or any combination thereof, may include a constant width, or a constant diameter, or both. The first channel, the second channel, or the third channel, or any combination thereof, may include a variable width, or a constant diameter, or both. The width, diameter, or height of the channel, or any combination thereof, may be less than about 1.0 μm, about 1.0 μm to about 1.0 cm, about 10 μm to about 0.5 cm, about 50 μm to about 1.0 cm, about 100 μm to about 0.1 cm, about 1.0 mm to about 500 mm, about 10 mm to about 100 mm, or more than 1.0 cm, or any value therebetween, or any intervening range. Two or more of the channels may have a common length. The channel length, or width, or both, can be less than 1.0 mm, about 1.0 mm to about 10 cm, about 2.5 mm to about 5.0 cm, about 5.0 mm to about 2.5 cm, about 7.5 mm to about 1.0 cm, or greater than 10 cm, or any value therebetween, or any intervening range. The width of the channel, or device, or both, can be, for example, about 5.0 mm, about 6.0 mm, about 7.0 mm, about 8.0 mm, about 9.0 mm, or about 10 mm, or any value therebetween, or any intervening range.Two or more of the plurality of channels are parallel to one another. The first channel can be between the second channel and the third channel. Two or more of the first, second, and third channels can differ from one another in terms of volume, surface area, cross-sectional area, length, width, or diameter, or any combination thereof. For example, the width, diameter, or both of the first channel can be greater than the width, diameter, or both of the second channel, or the third channel, or both. The cross-sectional area, width, diameter, or any combination thereof of one or more of the first, second, and third channels can differ along their length.

[0013] Any channel may include any number of segments. Any two segments may be continuous or non-contiguous. Any two segments may be adjacent or non-adjacent. Any two adjacent segments may be open, closed, partially closed, or variably connected, or any combination thereof. For example, two adjacent segments, or two adjacent channels, or both, may be separated by a valve. Any type of valve may be used. The valve may be a one-way valve or a two-way valve. A channel may be divided, for example, into three segments. For example, one or more of the first, second, and third channels may include a first peripheral segment proximate a first end of the channel, a second peripheral segment proximate a second end of the channel, and a central segment between the first and second peripheral segments. A channel may have a central longitudinal axis, and each segment may have a respective central longitudinal axis. These axes may serve as a reference for the relationship, e.g., orientation, between each segment of a given channel, as well as between segments of different, e.g., adjacent, channels. The central peripheral segment may include a cross-sectional area, width, or diameter, or any combination thereof, that is greater than the cross-sectional area, width, or diameter, or any combination thereof, of one or both of the first and second peripheral segments. The central peripheral segment of the first channel may include a cross-sectional area, width, or diameter, or any combination thereof, that is greater than the cross-sectional area, width, or diameter, or any combination thereof, of one or both of the first and second peripheral segments of the first channel. One or both of the first and second peripheral segments of the channel may be collinear with the central segment with respect to their respective central longitudinal axes. One or both of the first and second peripheral segments may form an angle other than 180° with the central segment with respect to their respective central longitudinal axes. For example, the angle may include a right angle, an acute angle, or an obtuse angle, or any combination thereof.Two or more channels of the plurality of channels may lie in a common plane, for example, a first, second, and third channel of the plurality of channels may lie in a common plane.

[0014] The microfluidic device can include any desired number of septa. The number of septa can be, for example, one less than the number of channels. For example, the plurality of channels can include a first, second, and third channel having a first and second septa. Any two septa can vary or be identical with respect to a particular parameter, for example, a geometry, a material, or a design, or any combination thereof. For example, the length of the first septum can be shorter than the length of the first channel, or the length of the second channel, or both. The length of the second septum can be shorter than the length of the first channel, or the length of the third channel, or both. The septa can include a membrane. For example, the first septum, the second septum, or both can include a membrane. The membrane can include a porous membrane. The pore width, or diameter, or both, can be less than about 1.0 nm, about 1.0 nm to about 1.0 cm, about 10 nm to about 1.0 mm, about 10 nm to about 500 μm, about 50 nm to about 100 μm, about 100 nm to about 10 μm, or greater than 1.0 cm, or any value therebetween, or any intervening range. The porous membrane can include, for example, polydimethylsiloxane (PDMS), polytetrafluoroethylene (PTFE), polyethylene terephthalate (PET), and the like. The membrane can include a semi-permeable membrane. The membrane can include a biological membrane, or a synthetic membrane, or both.

[0015] The first septum can include a first plurality of rows separated by a first plurality of openings, and the second septum can include a second plurality of rows (pillars, micropillars, etc.) separated by a second plurality of openings, or both. Any desired number, size, shape, and spacing of columns can be used. For example, the first plurality of rows, or the second plurality of rows, or both, can have straight cross-sections, curved cross-sections, or both. For example, the first plurality of rows, or the second plurality of rows, or both, can be circular, elliptical, polygonal, square, triangular, rectangular, pentagonal, hexagonal, heptagonal, octagonal, or trapezoidal, or any combination thereof in terms of general shape or cross-sectional shape, or both. The width, diameter, height, or depth of the column, or any combination thereof, can be less than 1.0 μm, about 1.0 μm to about 1.0 mm, about 5.0 μm to about 300 μm, about 5.0 μm to about 500 μm, about 5.0 to about 700 μm, about 10 μm to about 250 μm, about 25 μm to about 100 μm, or greater than 1.0 mm, or any value therebetween, or any intervening range. A column can be an extension of any surface, or combination of surfaces of a channel or adjacent channels. A column can extend from a lateral channel wall, a channel ceiling, or a channel floor, or any combination thereof. A column may span partially or completely the channel width, or the channel diameter, or both. The number, arrangement, size, and shape of the columns can affect flow patterns, such as vortex, laminar, and turbulent flow.

[0016] The rows can be evenly or irregularly spaced to form a plurality of openings therebetween. The average, e.g., mean, distance between adjacent columns can be less than 1.0 μm, between about 1.0 μm and about 1.0 mm, between about 5.0 μm and about 300 μm, between about 5.0 and about 500 μm, between about 5.0 μm and about 700 μm, between about 10 μm and about 250 μm, between about 25 μm and about 100 μm, or greater than 1.0 mm, or any value therebetween, or any intervening range. The first plurality of openings can differ in number, size, shape, or spacing, or any combination thereof, or the openings of the second plurality of openings can differ in number, size, shape, or spacing, or any combination thereof, or both. The openings of the first plurality of apertures may vary in number, size, shape, or spacing, or any combination thereof, along one or more dimensions of the first septum, or the openings of the second plurality of apertures may vary in number, size, shape, or spacing, or any combination thereof, along one or more dimensions of the second septum, or both. The first plurality of apertures and the second plurality of apertures may be different from one another.

[0017] The device housing may include a substrate, and the plurality of channels and the plurality of channel ports are formed in the substrate. The device housing may include a polymer, a metal, or a ceramic, or any combination thereof. The polymer may be, for example, a silicone. The silicone may be polydimethylsiloxane (PDMS) or the like, for example, as described in Campbell et al., “Beyond Polydimethylsiloxane: Alternative Materials for Fabrication of Organ-on-a-Chip Devices and Microphysiological Systems,” ACS Biomater. Sci. Eng., 7, 7, 2880-2899 (2021), which is incorporated herein by reference in its entirety. The device housing may be made of polydimethylsiloxane (PDMS), polymethylmethacrylate (PMMA), agarose, alginate, hyaluronic acid, gelatin, other polymeric materials, glass, or quartz crystal, or other optically transparent materials, or any combination thereof. The device housing may comprise a flexible material, or a rigid material, or both. The device housing may comprise a conductor, or an insulator, or both.

[0018] Any surface of the microfluidic device or its contents may be coated with one or more materials. The interior, exterior, or ports of the microfluidic device, or any combination thereof, may be coated. The substrate on which the microfluidic device may comprise a first material and one or more channel surfaces may comprise a second material different from the first material. The second material may be different from the contents of the one or more channels in which it resides. The second material may be a coating on the first material. One or more channel surfaces may be coated with a plurality of different materials. One or more channel surfaces may be coated with a plurality of coatings. At least two of the plurality of coatings may differ in material composition. The substrate may comprise a first material, and the plurality of coatings may comprise a first coating on one or more channel surfaces comprising a second material adjacent to the substrate, and a second coating comprising a third material covering the first coating. The first and second materials may be different from each other, and the second material and the third material may be different from each other. The first, second, and third materials are different from each other. The plurality of coatings may comprise at least two layers that differ from each other in average thickness. The coating may have one, two, three, four, five, or ten or more layers. The thickness, or patterning, or a combination thereof, of the coating on one or more channel surfaces may vary along the length of one or more channels. Two or more of the channels may differ in the material composition of their surface coating. A first channel may include a first coating, a second channel may include a second coating, and a third channel may include a third coating, with the first, second, and third coatings differing in material composition, thickness, or patterning, or a combination thereof. The thickness of the coating or layer can be one atom, one molecule, less than 1.0 nm, from about 1.0 nm to about 1.0 mm, from about 10 nm to about 100 nm, from about 50 nm to about 5.0 μm, from about 0.5 μm to about 50 μm, from about 1.0 μm to about 10 μm, from about 10 μm to about 100 μm, from about 0.5 mm to about 1.0 mm, or greater than 1.0 mm, or any intervening thickness, or any range therebetween.The patterning can be linear, straight, curvilinear, diagonal, axial, radial, spiral, cross-hatched, spot, diffuse, random, lateral interior of the channel, bottom interior of the channel, bottom interior half of the channel, top interior of the channel, top interior half of the channel, and so on.

[0019] The first, second, and third channels may be coated with the same material. The one or more coatings may include organic or inorganic materials, or both. The one or more coatings may include a polymer. The one or more coatings may include a biomolecule. The one or more coatings may include a protein, or a nucleic acid, a lipid, or a carbohydrate, or any combination thereof. The one or more coatings include fibronectin, an extracellular matrix-derived composition, a basement membrane extract, a decellularized extracellular matrix, a solubilized basement membrane preparation, a mixture of extracellular matrix proteins, a tumor-derived composition, Matrigel, or hyaluronic acid, or any combination thereof. The substrate may include silicon, and the one or more channel surfaces may be coated with a coating that includes a silane. The one or more channel surfaces may be subjected to silanization. The one or more channel surfaces may be treated with a plasma. The plasma may include oxygen. The one or more coatings may be applied by emersion, spraying, brushing, pouring, precipitation, polymerization, or deposition, or any combination thereof. The one or more channel surfaces may include a laminate. The one or more channel surfaces may include an adhesive.

[0020] One or more surfaces may be etched, peened, polished, or smoothed, or combinations thereof, or modified, for example, to form a pattern such as those described herein for a coating, ridge, groove, ridge, or hole, or other surface feature, or any combination thereof. The physical surface features may have dimensions as described herein for a coating. One or more coatings may combine one or more physical surface features. The coatings and physical surface features may be hydrophilic, or hydrophobic, or both. The coatings and physical surface features may affect flow within one or more channels of the microfluidic device. The coatings and physical surface features may be applied to other surfaces of the microfluidic device in addition to or instead of the channel surfaces.

[0021] The device housing can include transparent materials, translucent materials, colored materials, colored materials, polarizing materials, materials that selectively transmit wavelengths of electromagnetic radiation, materials that selectively refract wavelengths of electromagnetic radiation, materials that selectively reflect wavelengths of electromagnetic radiation, or opaque materials, or any combination thereof. The device housing can include a backing material. The backing material can include any material that allows the transmission of light, for example, for fluorescence and optical microscopy. The device housing can include a window that allows a view into the device interior. The view can be of one or more of the multiple channels, or its components, or its contents, or any combination thereof. For example, the view can include a view of the first channel, or the target object, or both. The device housing can include multiple windows that allow multiple views into the device interior. The microfluidic device can be fabricated using any technique or combination of techniques. For example, the microfluidic device can be fabricated using molding, three-dimensional printing, or etching, or any combination thereof. Three-dimensional printing can include, for example, selective laser sintering (SLS), condensation deposition modeling (FDM), or photopolymerization, or any combination thereof.

[0022] The target object may include organic materials, or inorganic materials, or both. The target object may include cells, tissues, or medical devices, or any combination thereof. The cells or tissues may be human cells. The cells or tissues may be non-human mammals, such as rats, mice, hamsters, monkeys, apes, dogs, cats, sheep, cows, horses, or pigs. The target object may include spheroids, organoids, alginate-based biomaterials, scaffold-supported cell structures, or bioprinted structures, or any combination thereof. The spheroids may be generated using any suitable method, such as hanging drops, hydrogels, rolling cell cultures, cell suspensions with nanofibers, magnetic levitation, microfluidic technology, or bioprinting, or any combination thereof. The target object may include single cell suspensions, suspensions of spheroids, or suspensions of organoids, or any combination thereof.

[0023] The target object may include neoplastic cells, malignant cells, or benign cells, or any combination thereof. The target object may include cancer cells, or cancer tissue, or both. For example, cancer may be a pre-malignant growth, a malignant growth, or a tumor caused by abnormal and uncontrolled cell division, which may be metastatic or non-metastatic. The cancer may be, for example, breast cancer, prostate cancer, lung cancer, colon cancer, rectal cancer, bladder cancer, non-Hodgkin's lymphoma, lymphoma, melanoma, kidney cancer, pancreatic cancer, oral cancer, pharyngeal cancer, ovarian cancer, thyroid cancer, stomach cancer, brain cancer, glioma, multiple myeloma, esophageal cancer, liver cancer, cervical cancer, laryngeal cancer, intrahepatic bile duct cancer, acute myeloid leukemia, soft tissue sarcoma, small intestine cancer, testicular cancer, chronic lymphocytic leukemia, Hodgkin's lymphoma, chronic myeloid leukemia, acute lymphocytic leukemia, anal cancer, vulvar cancer, cervical cancer, gallbladder cancer, bladder cancer, malignant mesothelioma, bone cancer, joint cancer, hypopharyngeal cancer, eye cancer, nasal cancer, nasal cavity cancer, middle ear cancer, nasopharyngeal cancer, ureteral cancer, peritoneal cancer, omental cancer, mesenteric cancer, or gastrointestinal carcinoid, or a combination thereof. The target object can include alginate, alginate derivatives, polyethylene glycol (PEG), poly(lactic-co-glycolic acid) (PLGA), poly(ethylene glycol diacrylate) (PEGDA), collagen, or hyaluronic acid, or any combination thereof. The target object can include, for example, nanoparticles, nanomaterials, polymeric biomaterials, ceramic biomaterials, metallic biomaterials, or surface treatments, or any combination thereof. The target object can include a surgical device or a portion thereof, for example, a suture. The target object can include polypropylene, silk, polyester, polydioxanone, polyglucaprone, polyglactin, or nylon, or any combination thereof. The target object can be inserted into the microfluidic device using manual techniques, automated techniques, or both. For example, the target object can be inserted using pipetting, injection, or the use of one or more surgical tools.

[0024] The target may include one or more coatings. The one or more coatings of the target may be different from the substrate. The one or more coatings of the target may differ in terms of material composition, thickness, or patterning, or a combination thereof. Coatings and other surface modification properties described herein for the channel surfaces can be applied to the target, and vice versa. The one or more coatings of the target may be different or the same as the one or more coatings of the one or more channel surfaces in terms of material composition, thickness, or patterning, or a combination thereof. The target may be coated with multiple different materials. The target may be coated with multiple coatings. The multiple coatings may differ in material composition, thickness, or patterning, or a combination thereof. The multiple coatings may include a first coating comprising a first material adjacent to the target and a second coating comprising a second material covering the first coating. The target may be coated before, during, or after placement in the microfluidic device. The coating of the target may be partial or complete. At least 10%, at least 25%, at least 40%, at least 50%, at least 60%, at least 75%, at least 90%, at least 95%, or at least 99% of the target surface, or any intervening percentage, or any percentage range therebetween, can be coated or otherwise modified.

[0025] The matrix can include a hydrogel. The matrix can include a decellularized extracellular matrix, a solubilized basement membrane preparation, a mixture of extracellular matrix proteins, or a tumor-derived composition, or any combination thereof. For example, the matrix can include a solubilized basement membrane matrix secreted by Engelbreth-Holm-Swarm mouse sarcoma cells, and the matrix can include MATRIGEL (Corning). The matrix can include a natural polymer, an artificial polymer, or a cross-linked polymer, or any combination thereof. The matrix can include a polysaccharide, a polypeptide, a lipid, a polynucleotide, hyaluronic acid, hyaluronic acid methacrylate, hyaluronic acid diacrylate, fibronectin, fibrinogen, collagen, or collagen methacrylate, or any combination thereof.

[0026] Components described as being present in one biological composition can also be present in another or other biological composition. The first biological composition can include, for example, fibroblasts, or endothelial cells, or both. Cells that can be used include, for example, B16-F10 cells, MDA-MB-231 cells, MCF-7 cells, HEPG2 cells, etc., or any combination thereof. The cells can include immune cells, RAW264.7 macrophages, L929 fibroblasts, primary cells, skin fibroblasts, peripheral blood mononuclear cells, pathogenic or non-pathogenic bacterial cells, or any combination thereof. The components can include virus particles. The cells can be introduced into the microfluidic device, for example, in the form of a suspension of cells in cell culture medium, a buffer solution, a three-dimensional gel, or a pre-gel solution, or any combination thereof. Pharmaceutical agents or therapeutic materials (small molecule drugs, large molecule drugs, antibodies, proteins, nanoparticles, or lipids) may be introduced into the microfluidic device or incorporated into biomaterials or hydrogels disposed within the microfluidic device. The introduced cells may include healthy cells, diseased cells, or both. The cells may be from human cell lines, non-human cell lines, or both. The cells may be from primary cell cultures, or organ explants, or the like. Components added to the microfluidic device may include, for example, anti-inflammatory drugs, anti-fibrotic drugs, steroids, or immune-related drugs, or any combination thereof.

[0027] The first biological composition may include serum, ascorbic acid, retinoic acid, proline, bleomycin, cyclophosphamide, amiodarone, procainamide, penicillamine, gold, or nitrofurantoin, or any combination thereof. The first biological composition may include a compound, such as an agent associated with causing fibrosis, such as pulmonary fibrosis, or liver fibrosis, or both. The first biological composition may include an environmental substance, such as asbestos, silica, titania, zirconia, graphite, coal dust, mineral dust, or any combination thereof. The first biological composition may include bacteria, fungi, components thereof, or combinations thereof. The first biological composition may include fibrosis-promoting biomolecules, complement proteins, blood coagulation-related proteins, bacterial cell wall proteins, or lipopolysaccharides (LPS), or any combination thereof. The second biological composition may include at least one type of biological cell capable of promoting fibrosis. The second biological composition may include a factor that promotes differentiation of a biological cell into a biological cell type that can promote fibrosis, a factor that promotes expression of a biomolecule that promotes fibrosis, or a factor that promotes proliferation of a biological cell type that can promote fibrosis, or any combination thereof. The second biological composition may include serum, platelets, leukocytes, macrophages, monocytes, peripheral blood mononuclear cells (PBMCs), stem cells, mast cells, dendritic cells, neutrophils, granulocytes, T cells, or B cells, or any combination thereof. The third biological composition may include a population of molecules that can form a gradient that radiates from the third channel into the first channel, or into both the first and second channels. The third biological composition may include a cytokine, a chemokine, a cell nutrient, a transcription factor, or a growth factor, or any combination thereof.The third biological composition may be, for example, CCL-2 / MCP-1, CCL3, CCL11, fibroblast growth factor (FGF), interferon, IFN-γ, interleukin (IL), IL-4, IL-1, IL-2, IL-5, IL-6, IL-7, IL-9, IL-13, IL-17A, IL-17F, IL-21, IL-22, IL-25, platelet-derived growth factor (PDGF), transforming growth factor (TGF)-alpha, transforming growth factor (TGF)-beta, granulocyte colony-stimulating factor (G-CSF), macrophage colony-stimulating growth factor (M-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), prothrombin, or thrombin, or any combination thereof. At least one type of molecule of the gradient may include a cytokine, a chemokine, a cell nutrient, a transcription factor, or a growth factor, or any combination thereof. At least one type of molecule of the gradient may include transforming growth factor beta (TGF-β). TGF-β may include TGF-β1, TGF-β2, or TGF-β3, or any combination thereof. The first biological composition, the second biological composition, or the third biological composition, or any combination thereof, may include labeled cells, or labeled biomolecules, or both. The first biological composition, the second biological composition, or the third biological composition, or any combination thereof, may include a compound with known or potential anti-fibrotic efficacy.

[0028] The fibrosis diagnostic system can be used to model one or more types of fibrosis. Fibrosis can be associated with any tissue or organ, such as, for example, lung, heart, liver, pancreas, kidney, bladder, skin, skeletal muscle, or smooth muscle, or any combination thereof. Examples of fibrosis include endomyocardial fibrosis, cystic pancreatic fibrosis, diatomaceous fibrosis, diffuse interstitial fibrosis, idiopathic fibrosis, endomyocardial fibrosis, graphitic fibrosis, idiopathic pulmonary fibrosis, idiopathic retroperitoneal fibrosis, mediastinal fibrosis, neoplastic fibrosis, nodular subepidermal fibrosis, pancreatic vesical fibrosis, periureteral fibrosis, postfibrotic fibrosis, proliferative fibrosis, pulmonary fibrosis, replacement fibrosis, retroperitoneal fibrosis, root sleeve fibrosis, or any combination thereof. Fibrosis can be the result of tissue autografts, tissue allografts, or tissue xenografts, or any combination thereof. Fibrosis may be the result of inorganic materials, such as metals, ceramics, minerals, or any combination thereof. Fibrosis may be characterized by the deposition of collagen on the target object. One or more types of collagen may characterize fibrosis. Collagen may include, for example, collagen I, collagen II, collagen III, collagen IV, collagen V, collagen VI, collagen VII, collagen VIII, collagen IX, collagen X, collagen XI, collagen XII, collagen XIII, collagen XIV, collagen XV, collagen XVI, collagen XVII, collagen XVIII, collagen XIX, collagen XX, collagen XXI, collagen XXII, collagen XXIII, collagen XXIV, collagen XXV, collagen XXVI, collagen XXVII, or collagen XXVIII, or any combination thereof. One or more types of procollagen, or collagen metabolites, or both, may characterize fibrosis.

[0029] The fibrosis diagnostic system can be preloaded with one or more components, such as a reagent, or a target object, or both. For example, the fibrosis diagnostic system can include a substrate, a first biological composition, a second biological composition, or a third biological composition, or any combination thereof. The fibrosis diagnostic system can include a target object. The fibrosis diagnostic system can include a gradient of at least one molecule across the first channel, the second channel, or the third channel, or any combination thereof. One or more components of the substrate, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, can be immobilized on the channel surface.

[0030] The fibrosis diagnostic system may include a detector. The detector may include a sensor. The sensor may be configured to detect a characteristic indicative of fibrosis or a characteristic of the target object. The detector may be configured to receive the microfluidic device, for example, in a receptacle or on a microscope stage. The detector may include a detector housing comprising a detector interior. The detector may include a receptacle configured to receive the microfluidic device. For example, the receptacle may comprise an opening in the detector housing. Any number or type of receptacles, or both, may be used. The receptacle may be configured to receive one or more microfluidic devices. A plurality of receptacles may be provided to receive a corresponding plurality of microfluidic devices.

[0031] The fibrosis diagnostic system may include a thermal subsystem. The thermal subsystem may include a heater, or a cooler, or both. The thermal subsystem may be configured to regulate a temperature of the microfluidic device. The detector may include a thermal subsystem. The thermal subsystem may be configured to regulate a temperature of the microfluidic device, for example, when in the receptacle.

[0032] The fibrosis diagnostic system may include a fluidic subsystem configured for fluid communication with the microfluidic device. The fluidic subsystem may be configured to load one or more components into the microfluidic device. The fluidic subsystem may be configured to supply a solid, liquid, or gas, or any combination thereof, to the microfluidic device. The fluidic subsystem may be configured to load a target object, a substrate, a first biological composition, a second biological composition, or a third biological composition, or any combination thereof, to the microfluidic device. The fluidic subsystem may be configured to supply one or more nutrients to the microfluidic device. The fluidic subsystem may be configured to regulate a pressure inside the microfluidic device, for example, in one or more of the channels. The pressure inside the microfluidic device, for example, the channels, may be above or below atmospheric pressure. A pressure differential may be applied between channels, within the channels, or both. A pressure in, above, or below the osmotic pressure may be applied. The fluidic subsystem may be configured to regulate a gradient across the first channel, the second channel, or the third channel, or any combination thereof. The gradient can be controlled, for example, by adding gradient solutes / molecules or by applying pressure, or both. Pressure can be applied, for example, at an osmotic pressure associated with the gradient molecules, above an osmotic pressure associated with the gradient molecules, or below an osmotic pressure associated with the gradient molecules. The fluidic subsystem can be configured to remove one or more components from the microfluidic device. The detector can, for example, comprise the fluidic subsystem. The fluidic subsystem can be configured to be in fluid communication with the microfluidic device, for example, when the microfluidic device is in the receptacle. The fluidic subsystem can include channels, valves, manifolds, or pumps, or any combination thereof. The fluidic subsystem can comprise a handheld pipe. The fluidic subsystem can be configured to provide a desired flow rate of solids, liquids, or gases, or any combination thereof, through one or more of the multiple channels.The flow rate can be, for example, less than about 0.001 μL / s, between about 0.001 μL / s and about 100 mL / s, between about 0.01 μL / s and about 10 mL / s, between about 0.1 μL / s and about 1.0 mL / s, between about 1.0 μL / s and about 100 μL / s, between about 10 μL / s and about 50 μL / s, or greater than 100 mL / s, or any value therebetween, or any intermediate range. The fluidic subsystem can include or be configured to fluidly connect a supply of one or more reagents, for example, multiple reservoirs or a single reservoir.

[0033] The sensor may be configured to detect particles, waves, or both emitted from the target object, or the microfluidic device, or both, indicative of fibrosis. The sensor may be configured to detect electromagnetic radiation. The electromagnetic radiation may include, for example, ultraviolet, visible, or infrared light, or any combination thereof. The electromagnetic radiation may include fluorescence, or phosphorescence, or both. The detector may include a particle source, or a wave source, or both, configured to transmit particles, waves, or both to the target object, or the microfluidic device, or both. The fibrosis diagnostic system may include an electromagnetic radiation source. For example, the detector may comprise an electromagnetic radiation source. The electromagnetic radiation source may include, for example, a light emitting diode, a laser, an incandescent light source, an arc light source, a halogen light source, a chemical vapor light source, or a fluorescent light source, or any combination thereof. The detector may comprise an optical subsystem configured to pass radiation, affect radiation, or both. The optical subsystem may include one or more lenses, one or more filters, one or more mirrors, or one or more diffraction gratings, or any combination thereof. The optical subsystem, or the sensor, or both, may include a microscope or one or more components thereof. The optical subsystem may include a sensor, or an electromagnetic radiation source, or both. The sensor may include a microphone, and the fibrosis diagnostic system may include an acoustic wave source. The acoustic waves may include ultrasound waves, and the microphone may detect the ultrasound waves. The sensor may include a force sensor, an actuator, or both. Fibrosis of the target object may result in a fibrous structure having a tensile strength, or a compressive strength, or both, that may be detected by the sensor.

[0034] The sensor may be configured to generate an output signal based on an input signal detected by the sensor. The detector may include a transmitter, receiver, transceiver, or any combination thereof configured to receive information or transmit information, or both. The information may include an output signal. The information may include an operational signal for operation of the sensor or other components of the system. The information may travel over a network. The network may be a local network or a long distance network. The network may be wired, or wireless, or both. The detector or other components of the system may include a data port. Any desired number of data ports may be provided. The system may further include a processor configured to receive and process the information. The processor may be configured to control one or more subsystems or components of the fibrosis diagnostic system. For example, the processor may be configured to control the sensor, the electromagnetic radiation source, the optical subsystem, the thermal subsystem, or the fluid subsystem, or any combination thereof. The processor may be configured to receive the output of the signal from the sensor, receive the operational signal from a user input, or both. The detector may include, for example, a processor. The processor may include a local processor, or a remote processor, or both. The processor may be a microprocessor. The processor may be a central processing unit (CPU). The processor may include one or more processing chips. The system may include a memory configured for communication with the processor. The memory may be local or remote. The memory may be temporary or non-temporary or both. The memory may include random access memory (RAM) or read only memory (ROM) or both. The memory may be configured to store a program that controls the fibrosis diagnostic system or components thereof. The processor may include, for example, a memory. The detector may include, for example, a memory.

[0035] The fibrosis diagnostic system may comprise a user interface. The system may include any number of user interfaces, or any type of user interface, or both. The user interface may be interactive, non-interactive, or both, e.g., the user interface may include one or more components that are interactive and one or more components that are not interactive. The user interface may be standalone or may be integrated into one or more system subsystems or components. The user interface may include a monitor, buttons, a keyboard, a mouse, an optical signal, a camera, an eyepiece, a speaker, or a microphone, or any combination thereof. The monitor may include, for example, a touch screen. The camera may be a still photo camera, or a video camera, or both. The eyepiece may be part of a microscope system or subsystem. For example, two eyepiece lenses may be provided to allow binocular viewing. The detector may comprise, for example, a user interface.

[0036] The fibrosis diagnostic system may include a power source, or a power interface, or both. The power source may comprise a battery, e.g., a rechargeable battery. The power interface may comprise, e.g., an inverter for receiving an AC wall current and outputting a DC current. The detector may include, e.g., a power source, or a power interface, or both.

[0037] The fibrosis diagnostic system may include a kit or may be provided as a kit. A kit may include two or more components of the system. A kit may include multiple supplies of a particular system component, e.g., multiple multi-fluidic devices. A kit may include, for example, a microfluidic device, and one or more reagents for use with the microfluidic device. A kit may include, for example, a microfluidic device, and a substrate, a first biological composition, a second biological composition, or a third biological composition, or any combination thereof. Whether or not provided as a kit, one or more components or subsystems of the fibrosis diagnostic system may include an identifier. The identifier may include a serial number, a product number, a bar code, an RFID tag, a color code, a name, a number, or an alphanumeric code, or any combination thereof.

[0038] The present disclosure provides methods of using a fibrosis diagnostic system, its subsystems, and its components. For example, a method of analyzing a target object for susceptibility to fibrosis is provided. The method may include one or more of the following steps: The target object, a substrate, and a first biological composition may be disposed in a first channel of a microfluidic device. The second biological composition may be disposed in a second channel of the microfluidic device. The third biological composition may be disposed in a third channel of the microfluidic device. The target object, or the first channel, or both, may be analyzed to determine the presence or absence of fibrosis associated with the target object.

[0039] The analysis may be performed after a predetermined period of time following placement of the target object, substrate, first biological composition, second biological composition, or third biological composition, or any combination thereof, into the microfluidic device. The predetermined period of time may be sufficient to allow for the formation of a gradient of at least one type of molecule. The analysis may be performed more than once. Any two consecutive analyses may be separated by a predetermined period of time. The period of time may be counted as seconds, minutes, hours, days, or weeks. The period of time may be, for example, less than about 1 minute, about 1 minute to about 1 month, about 1 hour to about 2 weeks, about 3 hours to about 1 week, about 6 hours to about 3 days, about 12 hours to about 1 day, more than about 1 month, or any period of time therebetween, or a range of periods therebetween. The analysis may include removing one or more components from the microfluidic device.

[0040] The analysis may include an immunoassay. The analysis may include the use of an antibody. The antibody may be labeled with a detectable label. The analysis may include application of a primary antibody and a secondary antibody that includes a detectable label and can bind to the primary antibody. The primary antibody can bind to collagen, alpha smooth muscle actin, or both. The analysis may include a histological assay. The cells and / or tissues can be fixed for the assay, for example, prior to staining. Any suitable fixative can be used, for example, formaldehyde fixative, neutral buffered formalin (NBF), paraformaldehyde (PFA), or methanol. The fixative can be an aldehyde fixative, mercury fixative, alcohol-based fixative, oxidizing agent fixative, or picrate fixative, or any combination thereof; further examples of fixatives include acetone, phosphate-buffered formalin, zinc formalin, calcium formaldehyde, saline formaldehyde, alcohol formalin, alcoholic formaldehyde acetate, zinc formaldehyde, Alcian blue, Bouin's fixative, B-5 ​​fixative, Carnoy's fixative, Clark's fixative, Gendre's fixative, Helly's fixative, Holland's fixative, Methacan, and Zenker's fixative. The cells and / or tissues can be partially or completely dehydrated for the assay. The analysis can include embedding, or antigen retrieval, or both. Any suitable stain or combination of stains can be used. The stain can include a dye. Picric acid can be used. The analysis can include, for example, applying Picrosirius Red dye alone or in combination with one or more additional stains. Examples of stains include carmine, hematin, hematoxylin, fluorescein, eosin, silver nitrate, Giemsa stain, Golgi stain, Gram stain, Kluva-Barrera stain, Calorie's CT stain, Masson's stain, Periodic Acid Schiff (PAS) stain, Kalogeno's stain, Trichrome stain, AZAN trichrome stain, toluidine blue, and Behoev's van Gyson stain, and any combination thereof.

[0041] Analyzing may include visually inspecting the target object, or the first channel, or both, for one or more indications of fibrosis associated with the target object. Analyzing may include using a microscope. Analyzing may include using one or more spectroscopic techniques. For example, analyzing may include nonlinear optical (NLO) multiphoton microscopy, second harmonic generation (SHG) imaging, third harmonic generation (THG) imaging, two-photon excited fluorescence (TPEF), two-photon excited fluorescence (TPEF), cell tracker dye imaging, visible light imaging, (ELISA), Griess assay, histopathology, immunostaining, clear tissue microscopy, or proteomic profiling of the homogenized hydrogel, or any combination thereof. Analyzing may include detecting one or more indications of fibrosis associated with the target object using a detector. Analyzing may include inserting the microfluidic device into a receptacle.

[0042] The method may include preparing a target object, a substrate, a first biological composition, a second biological composition, or a third biological composition, or any combination thereof, from a patient sample. The method may include implanting the target object into or on a patient. For example, the method may include implanting a medical device, or tissue, or both, into or on a patient, and the target object is a representative model of the medical device, or tissue, or both. The method may include removing the medical device, or tissue, or both, from the patient, and the target object is a representative model of the medical device, or tissue, or both.

[0043] The method may include administering an antifibrotic agent to the patient. Analyzing may include determining a quantitative measure of fibrosis. The dose of antifibrotic agent administered to the patient may be based on the quantitative measure of fibrosis. The antifibrotic agent may be administered to the patient before, simultaneously with, or after implanting the target object, tissue associated with the target object, or medical device associated with the target object, or any combination thereof. One or more therapeutic agents may be administered to the patient in addition to, or as an alternative to, the antifibrotic agent. The patient may be, for example, a mammalian patient, or a human patient, or both.

[0044] A method of screening a therapy for effectiveness in treating fibrosis is provided. The method may include one or more of the following steps: The target object, the substrate, and the first biological composition may be disposed in a first channel of a microfluidic device. The second biological composition may be disposed in a second channel of the microfluidic device. The third biological composition may be disposed in a third channel of the microfluidic device. The therapy may be implemented in the microfluidic device. The target object, or the first channel, or both, may be analyzed to determine the presence, absence, or extent of fibrosis associated with the target object. The effectiveness of the therapy may be determined based on the presence, absence, or difference in extent of fibrosis without implementing the therapy. The therapy may include a therapeutic compound, and implementing may include disposing the therapeutic compound in the first channel, the second channel, or the third channel, or any combination thereof. The therapeutic compound may include a pharmaceutical compound, or a biologic, or both. The therapy may include modification of the target object. The therapy may include disposing a medical device in the first channel, the second channel, or the third channel, or any combination thereof. The therapy can include irradiating the microfluidic device. The method can include treating the patient with the screened therapy. The screening methods provided herein include high-throughput screening methods.

[0045] A method of screening a therapy for effectiveness in treating cancer is provided by the present disclosure. The method may include one or more of the following steps: A target object, a substrate, and a first biological composition may be disposed in a first channel of a microfluidic device, and the target object may include neoplastic cells. A second biological composition may be disposed in a second channel of the microfluidic device. A third biological composition may be disposed in a third channel of the microfluidic device. The therapy may be implemented on the microfluidic device. The target object, or the first channel, or both, may be analyzed to determine the presence, absence, or extent of fibrosis associated with the target object. The efficacy of the therapy may be determined based on the presence of cancer, the absence of cancer, or a difference in the extent of cancer without implementing the therapy. Other screening methods using the fibrosis diagnostic system are also provided. For example, the screening method may be used to monitor immunological phenomena generally associated with foreign bodies, as well as more specific phenomena such as graft-versus-host disease (GVHD) or autoimmune syndromes.

[0046] A non-transitory computer readable medium is provided by the present disclosure. For example, a non-transitory computer readable medium is provided that stores a program including instructions for performing the methods of the present disclosure. A non-transitory computer readable medium is provided that stores a program including instructions for controlling a fibrosis diagnostic system or subsystem, or a component thereof, or any combination thereof.

[0047] FIG. 1A is a perspective view of a microfluidic device 110 according to the present disclosure. The microfluidic device 110 can be used in the fibrosis diagnostic system, or method, or both of the present disclosure. The microfluidic device 110 can include a device housing 112, an exterior surface 114 of the device housing 112. The device housing 112 can define a device interior 116 that can include a plurality of channels 118. The plurality of channels 118 can include a first channel 120 between a second channel 140 and a third channel 160. A first septum 130 can separate or otherwise separate the first channel 120 from the second channel 140. A second septum 150 can separate or otherwise separate the first channel 120 from a third channel 150. The first septum 130 and the second septum 150 can have any design, such as a porous membrane or an array of pillars (micropillars) as described herein. First channel 120 may include an interior 122, a first end 123, a first end 123 proximal to first channel port 124, a second end 125, and a second end 125 proximal to second channel port 126. First channel 120 may include a surface 127, which may be modified, for example, with a surface modification 129 configured to receive and retain one or more target objects. First channel 120 may include a third channel port 118 between first channel port 124 and second channel port 126 to facilitate insertion and removal of one or more target objects from interior 122. Second channel 140 may include an interior 142, a first end 143, a first end 143 proximal to first channel port 144, a second end 145, and a second end 145 proximal to second channel port 146. Third channel 160 can include an interior 162, a first end 163, the first end 163 proximal to a first channel port 164, a second end 165, and a second end 165 proximal to a second channel port 166. The channel port can provide a passage between the channel interior and the device surface 116 through the device housing 112 and an otherwise external region of the microfluidic device 110.FIG. 1B is a top view of the microfluidic device shown in FIG. 1A.

[0048] FIG. 2A is a perspective view of a fibrosis diagnostic system 200 showing a microfluidic device 210 disengaged from a detector 270 according to the present disclosure. The microfluidic device 210 can comprise features described for, for example, the microfluidic device 110 or any other microfluidic device described herein. The detector 270 can comprise a detector housing 272, a detector surface 274, a detector interior 276, and a receptacle 278 that allows full or partial access of the microfluidic device 210 to the detector interior 276. The depiction of the detector 270 is for discussion purposes only and other designs suitable for a given embodiment can be employed. For example, the detector 270 can employ a more open design, such as an open microscope design that is open receptive in the form of a stage for placement of the microfluidic device 210. The detector 270 can include a user interface 280, and the illustrations of FIGS. 2A, 2B are illustrative and non-limiting. The user interface of the system 200 can be incorporated into the detector 270 or provided as a stand-alone component, or both. The user interface 280 is shown to include elements such as a monitor 282, a button 284, signals 286, 287, and data ports 288, 289. The screen 282 can include, for example, a screen. The screen can be provided as a touch screen. The screen can display a graphic user interface (GUI). The button 284 can, for example, allow control of one or more functions of the detector 270, for example, to turn the detector 270 on / off or to start imaging the microfluidic device 210, or both. Although a single button is shown, any number of buttons can be provided or omitted, for example, a button for each function, or multiple buttons in the form of a keyboard or keypad. The button or multiple buttons can be replaced with other types of actuators, such as toggle switches. The signals 286, 287 can include, for example, lights, such as lights of respective colors.Each signal may correspond, for example, to a particular operational state of the detector 270, such as a power state, engagement of the microfluidic device 210, or imaging of the microfluidic device 210, or any combination thereof. Alternatively or additionally, the signal may change color to correspond to a particular operational state. Although two signals are shown, any number of buttons may be provided or omitted. One or more of the signals may take the form of a speaker, or a microphone, or both. Data ports 288, 289 may be provided for any number of different functions, such as data transfer, power transmission, and connection to peripheral user interface components. Although two data ports are shown, any number of data ports may be provided or omitted. In an embodiment including a microscope, the user interface 280 may include one or more eyepieces configured for viewing of the microfluidic device 210 by a user.

[0049] FIG. 2A includes several exemplary breakaways within the detector housing 272 to illustrate various components that may be located within the detector interior 276. A fluidic subsystem 279 is shown that may be provided to supply various components to the microfluidic device 210 when held within the receptacle 278. A thermal subsystem 299, e.g., a heater, is shown that may be provided to regulate the temperature of the microfluidic device 210 when held within the receptacle 278. A sensor 290 is shown that may detect one or more signals from the microfluidic device 210, e.g., radiation resulting from excitation radiation emitted from the radiation source 291. The number and arrangement of the sensors and light sources may vary. An optical subsystem 292 is also shown and may include one or more optical elements, such as lenses, mirrors, diffraction gratings, and filters. The optical subsystem 292 may include one or both of the sensor 290 and the radiation source 291. A processor 294 is shown that may process information related to the operation of the detector 270. For example, a transceiver 296 is shown that can receive and transmit signals to and from a user interface 280, a sensor 290, a processor 294, a network via data ports 288, 289, or a network wireless, or any combination thereof. A power source 298 is shown that can take the form of a standalone power source, or a power interface for transmitting / transmitting power from an external power source, or both. Figure 2B is a perspective view of the fibrosis diagnostic system 200 of Figure 2A, showing a microfluidic device 210 engaged with a detector 270 through a receptacle 278 in accordance with the present disclosure.

[0050] FIG. 3 is a plan view of a microfluidic device 310 according to the present disclosure. The microfluidic device 310 can be used in the fibrosis diagnostic system, or method, or both of the present disclosure. The microfluidic device 310 can include a device housing 312, an exterior surface 314 of the device housing 112. The device housing 312 can define a device interior 316 that can include a plurality of channels. The plurality of channels can include a first channel 320 between a second channel 340 and a third channel 360. A first septum 330 can separate or otherwise separate the first channel 320 from the second channel 340. The first septum 330 is shown to have a micropillar design including a plurality of pillars (micropillars) 332 that define a plurality of openings 334 therebetween. A second septum 350 can separate or otherwise separate the first channel 320 from the third channel 350. The second septum 350 is shown to have a micropillar design including a plurality of pillars (micropillars) 352 defining a plurality of openings 354 therebetween. The plurality of channels may include features similar or identical to those shown for the plurality of channels 118 of FIG. 1A. For example, the first channel 320 may include a first channel port 324 proximal to the first end, a second channel port 326 proximal to the second end, and a third channel port 318 between the first channel port 324 and the second channel port 326 to facilitate insertion and removal of one or more target objects from the container 329 of the first channel 320. The second channel 340 may include a first channel port 344 proximal to the first end and a second channel port 346 proximal to the second end. The third channel 360 may include a first channel port 364 proximal to the first end and a second channel port 366 proximal to the second end. The channel ports can provide passage between channel interiors through the device housing 312 and the device surface 316 and areas otherwise external to the microfluidic device 310 .

[0051] FIG. 4 is a plan view of a microfluidic device 410 according to the present disclosure. The microfluidic device 410 can be used in the fibrosis diagnostic system, or method, or both of the present disclosure. The microfluidic device 410 can include a device housing 412, an exterior surface 414 of the device housing 412. The device housing 412 can define a device interior 416 that can include a plurality of channels. The plurality of channels can include a first channel 420 between a second channel 440 and a third channel 460. A first septum 430 can separate or otherwise separate the first channel 420 from the second channel 440. A second septum 450 can separate or otherwise separate the first channel 420 from the third channel 450. The plurality of channels can include features similar or identical to those shown for the plurality of channels 118 of FIG. 1A. Additionally, one or more of the channels can be divided into channel segments with respective segments differing from one another in one or more parameters. For example, the first channel 420 can include a first channel port 424 proximal to the first end, a second channel port 326 proximal to the second end, and a third channel port 418 between the first channel port 424 and the second channel port 426 to facilitate insertion and removal of one or more target objects from a container 429 of the first channel 420. The first channel 420 can have three segments 420a, 420b, and 420c. The first channel port 424 can be located in the second segment 420b, the second channel port 426 can be located in the third segment 420c, and the third channel port can be located in the first segment 420a. The first segment 420a can be adjacent to both ceptors 430, 450 along its length. The second channel 440 can include a first channel port 444 proximal to the first end and a second channel port 446 proximal to the second end.The second channel 440 can have three segments 440a, 440b, and 440c, with the first channel port 444 located in the second segment 440b and the second channel port 446 located in the third segment 440c. The second segment 440b can have an angle α relative to the first segment 440a. The third segment 440c can have an angle β relative to the first segment 440a. The angles alpha and β can be the same or different. They can be acute, right, or obtuse angles. The second segment 440b can have an angle ψ relative to the second segment 420b. The third segment 440c can have an angle χ relative to the third segment 420c. The angles ψ and χ can be the same or different. They can be acute, right, or obtuse angles. The third channel 460 may include a first channel port 464 proximal to the first end and a second channel port 466 proximal to the second end. The third channel 440 may have three segments 460a, 460b, and 460c, with the first channel port 464 located in the second segment 460b and the second channel port 466 located in the third segment 460c. The second segment 460b may have an angle γ relative to the first segment 460a. The third segment 460c may have an angle δ relative to the first segment 460a. The angles γ and δ may be the same or different. They may be the same or different from the angles α and β. They may be acute, right, or obtuse. The second segment 460b may have an angle φ relative to the second segment 420b. The third segment 460c may have an angle ω relative to the third segment 420c. The angles φ and ω can be the same or different. They can be the same as or different from the angles ψ and χ. They can be acute, right, or obtuse. The channel ports can provide passage between the channel interior through the device housing 412 and the device surface 416 and an otherwise external region of the microfluidic device 410.

[0052] FIG. 5 is a plan view of a microfluidic device 510 according to the present disclosure. The microfluidic device 510 can be used in the fibrosis diagnostic system, or method, or both of the present disclosure. The microfluidic device 510 can include a device housing 512, an exterior surface 514 of the device housing 512. The device housing 512 can define a device interior 516 that can include a plurality of channels. The plurality of channels can include a first channel 520 between a second channel 540 and a third channel 560. A first septum 530 can separate or otherwise separate the first channel 520 from the second channel 540. A second septum 550 can separate or otherwise separate the first channel 520 from the third channel 550. The plurality of channels can include features similar or identical to those shown for the plurality of channels 118 of FIG. 1A. Additionally, one or more of the channels can be divided into channel segments with respective segments differing from one another in one or more parameters. For example, the first channel 520 can include a first channel port 424 proximal to the first end, a second channel port 526 proximal to the second end, and a third channel port 518 between the first channel port 524 and the second channel port 526 to facilitate insertion and removal of one or more target objects from a container 529 in the first channel 520. The first channel 520 can have three segments 520a, 520b, and 520c. The first segment 520a can include three regions 520a1, 520a2, and 520a3. The first channel port 524 can be located in the second segment 520b, the second channel port 526 can be located in the third segment 520c, and the third channel port can be located in the first segment 520a. The first region 520a1 can be adjacent to both septa 530, 550 along its length. Region 520a2 may be between region 520a1 and second segment 520b. Region 520a3 may be between region 520a1 and third segment 520c.Regions 520a2 and 520a3 may vary in cross-sectional area, width, or diameter, or any combination thereof, along their respective lengths. For example, regions 520a2 and 520a3 may be defined by angles θ and ρ, respectively. Angles θ and ρ may be the same or different. They may be acute, right, or obtuse. Second channel 540 may include a first channel port 544 proximal to the first end and a second channel port 546 proximal to the second end. Second channel 540 may have three segments 540a, 540b, and 540c, with first channel port 544 located in second segment 440b and second channel port 546 located in third segment 540c. Third channel 560 may include a first channel port 564 proximal to the first end and a second channel port 566 proximal to the second end. The third channel 540 can have three segments 560a, 560b, and 560c, with a first channel port 564 located in the second segment 560b and a second channel port 566 located in the third segment 560c. The channel ports can provide a passage between a channel interior through the device housing 512 and the device surface 516 or an otherwise external region of the microfluidic device 510.

[0053] FIG. 6A is a top perspective view of a microfluidic device 610 according to the present disclosure. FIG. 6B is a front perspective view of the microfluidic device 610 shown in FIG. 6A. FIG. 6C is an enlarged perspective view of a portion of the microfluidic device 610 shown in FIG. 6A. FIG. 6D is an enlarged plan view of a portion of the microfluidic device 610 shown in FIG. 6A. FIG. 6E is a further enlarged plan view of a portion of the enlarged view shown in FIG. 6D. FIGS. 6A-6E are discussed in detail below in Example 1. FIG. 7A is a schematic cross-sectional view of a loaded microfluidic device 710 at a first time point according to the present disclosure. FIG. 7B is a schematic cross-sectional view of the loaded microfluidic device 710 shown in FIG. 7A at a second time point. FIGS. 7A and 7B are discussed in detail below in Example 1.

[0054] FIG. 8 is a schematic diagram of a fibrosis diagnostic system 800 according to the present disclosure. The fibrosis diagnostic system 800 can include various components and subsystems, such as a microfluidic device 810, a reagent 820, a target object 830, a fluidic subsystem 840, a thermal subsystem 850, a sensor 860, a processor 870, a memory 880, and a user interface 890. These various components and subsystems can be arranged, configured, combined, multiplied, or omitted, or any combination thereof, without departing from the present disclosure. One or more components or subsystems of the system can be provided within one or more sterile enclosures or other enclosures that separate the contents thereof from the surrounding environment. FIG. 9 and FIG. 10 show two examples of such a system.

[0055] FIG. 9 is a schematic diagram of a fibrosis diagnostic system 900 according to the present disclosure. In the system 900, a kit 905 is provided that includes a microfluidic device 910 and a reagent 920. A target object preparation subsystem 915 is provided that is configured to form a target object 930 for insertion into the microfluidic device 910 with the reagent 920. A microfluidic device loader 925 is provided that may include a microfluidic subsystem for loading the reagent 920, or the target object 930, or both, into the microfluidic device 910. An incubator 935 is provided that may include a thermal subsystem 950 configured to regulate the temperature of the microfluidic device 910 and its contents during a fibrosis diagnostic assay. A detector 945 that includes a sensor 960 is provided for imaging the microfluidic device 910 or its contents, or both, during a fibrosis diagnostic assay. A computing device 945 is provided that may include a processor 970, a memory 980, and a user interface 990 that may be used to control the system 900 and process information associated therewith. One or more system components or subsystems may be interconnected via a network 955 .

[0056] 10 is a schematic diagram of a fibrosis diagnostic system 1000 according to the present disclosure. The system can include a loaded microfluidic device 1065 comprising a microfluidic device 1010 containing a reagent 1020 and a target object 1030. The system can include a fluidic subsystem 1040, a thermal subsystem 1050, a sensor 1060, a processor 1070, a memory 1080, and a detector 1075 comprising a user interface 1075. After passing from loading the microfluidic device 1010, the loaded microfluidic device 1065 or components thereof can be inserted into or onto the detector 1075 to analyze the target object 1030 for possible fibrosis. EXAMPLES

[0057] A three-channel microfluidic device 610, 710 (figures 6A-6E, 7A, 7A, and 7B) is provided for use in co-culture of fibroblasts and macrophages with a target object (721), e.g., three-dimensional cancer spheroids or biomaterial, embedded in a substrate (722a). The microfluidic mold / housing 612, 712 was prepared using standard photolithography techniques, and the device was fabricated in polydimethylsiloxane (PDMS). The central (first) channel 620 contains a fibroblast-embedded three-dimensional matrigel, into which the biomaterial implant or three-dimensional spheroid is placed through holes (ports) 628, 728. The outer (second and third) channels 640, 660 (740, 760) serve to introduce macrophages and chemokines, respectively. Each channel is separated from adjacent channels by respective septa 630, 650 (730, 750) and contains an array of micropillars 652 (shown in FIG. 6B, with corresponding micropillars 632 shown in FIG. 6D and FIG. 6E), 752 forming multiple openings 654 (634) between them. A recess 629, 729 below the central channel 620, 720 allows for reliable placement of biomaterials or spheroids within the device. The versatile device design allows for the introduction of multiple cell types, combinations of three-dimensional, two-dimensional, and fluid cultures, as well as the introduction of various biomaterials, nanomaterials, cytokines / chemokines, therapeutics, and other stimuli that may affect fibrosis formation. The device design can also be modified by adjusting the number of channels, channel height, channel length, channel width, recess height, recess shape, type of micropillars, size of micropillars, spacing between adjacent micropillars, and other aspects of the device feature dimensions. Using fluorescently labeled molecules of different sizes, cytokine gradients can be generated across device 610, allowing for potential stimulation of cells by cytokines. Device 610 and similar devices can be used to study fibrosis in various target objects, for example, melanoma spheroids and alginate-based biomaterials. EXAMPLES

[0058] L929 mouse fibroblasts and RAW264.7 mouse macrophages were fluorescently labeled with red and green CELLTRACKER® (Thermo Fisher Scientific) dyes, respectively. Extensive testing was performed to optimize cell seeding density, culture time, and biomaterial implantation. Cells were co-cultured two-dimensionally or in a microfluidic device as described in Example 1, and then cancer spheroids or alginate-based biomaterials were implanted in the two-dimensional cultures and in the microfluidic device. Immunostaining, fluorescence microscopy, Griess assay, and histological analysis were used to evaluate immune cell activation and fibrosis formation in response to spheroids and implants. Griess assays were performed on medium harvested from cultured cells. Co-culture of L929 fibroblasts and RAW264.7 macrophages at various ratios resulted in similar levels of nitrite production as cultures of RAW264.7 macrophages alone, indicating that L929 cells do not activate RAW264.7 cells. In contrast, stimulating macrophages with lipopolysaccharide (LPS) resulted in macrophage activation, as indicated by nitrite production. Co-culture of RAW264.7 and L929 cells was visualized with CELLTRACKER® dye. No morphological changes were observed as a result of co-culture. F-actin visualization of RAW264.7 cells treated with LPS was performed. Morphological changes were observed with increasing LPS concentrations. Co-culture of L929 and RAW264.7 cells was performed in a microfluidic device. L929 cells were cultured in 3D within a Matrigel channel, while macrophages were introduced in 2D from an adjacent channel. After 24 hours of co-culture, the cells were primarily retained within their respective channels.

[0059] Preliminary 2D experiments found that only L929 fibroblasts did not activate RAW264.7 macrophages and confirmed the effect of lipopolysaccharide (LPS) stimulation on macrophage morphology and activation. These measurements were then projected onto a three-dimensional microfluidic system. The effect of cell number and incubation time on B16-F10 melanoma spheroid formation was also determined. That information was used to select optimal parameters to form spheroids for use in the microfluidic device. F-actin staining of fibroblasts was also performed.

[0060] Device characterization results and data from microfluidic co-culture of fibroblasts and macrophages with melanoma spheroids and alginate-based biomaterials are projected. EXAMPLES

[0061] During the experiments, diffusion of fluorescently labeled dextran molecules of different sizes from the side (lateral) channels into the central channel of the microfluidic device was observed. Experiments were performed with 10k molecular weight (MW) dextran. Experiments were also performed with 10k, 70k MW dextran. Dextran diffusion was measured at 0 h, 3 h, and 24 h. Dextran molecules with 10k and 70k molecular weights successfully diffused from the side channels (from the second or third channel towards the first (central) channel) across the Matrigel-filled channels of the device over a 24-h period. A linear gradient of dextran concentration (measured by fluorescence intensity) was observed across the entire device. These experiments confirm the possibility of the generation of a cytokine gradient within the device. EXAMPLES

[0062] Macrophages and fibroblasts were co-cultured in the side and central channels of the microfluidic device, respectively, and a suture was provided in the central channel during co-culture to monitor any development of fibrosis on the suture. The suture can optionally be placed in the microfluidic device during assembly of the device. EXAMPLES

[0063] Development of melanoma spheroids for placement in microfluidic devices was performed. Melanoma spheroid diameter was measured as a function of cell density and incubation time. Live / dead staining of melanoma spheroids formed at 8 cells / μL (i), 27 cells / μL (ii), and 54 cells / μL (iii) was performed 72 hours after incubation. Live and dead cells were labeled with distinct colors. EXAMPLES

[0064] Biomaterial placement in a microfluidic device was investigated. Alginate beads were implanted into the microfluidic device implant wells (recesses / containers). Alginate was chosen as a model biomaterial since it is known to lead to fibrosis. Suture placement in the fibrosis chip. Matrigel successfully gelled around the sutures. These results confirm the feasibility of biomaterial placement in the fibrosis chip. Suture placement is possible in the device without incorporating an implant well. EXAMPLES

[0065] Induction of collagen deposition by fibroblasts in a two-dimensional model was studied. To determine whether L929 fibroblasts deposit collagen in response to stimuli, well plate experiments were performed according to the protocol described in Good et al., “A High Content, Phenotypic ‘Scar-In-A-Jar’ Assay for Rapid Quantification of Collagen Fibrillogenesis Using Disease-Derived Pulmonary Fibroblasts,” BMC Biomedical Engineering, 1:14, 1-10 (2019), which is incorporated herein by reference in its entirety. Immunostaining experiments were performed on L929 cells and primary mouse dermal fibroblasts exposed to medium containing L-ascorbic acid, FICOLL®, and TGF-β1. Some increase in collagen I deposition and α-SMA expression by primary cells was observed in response to TGF-β1 treatment. In contrast, L929 cells, which have no α-SMA expression, did not increase collagen I deposition in response to TGF-β1 treatment. Furthermore, the type of L-ascorbic acid used had some effect on collagen I deposition. These findings indicate the relevance and effect of cell type and L-ascorbic acid type on extracellular matrix deposition in vitro. EXAMPLES

[0066] One method that can be used to visualize and quantify collagen deposition in microfluidic devices is picrosirius red staining. Picrosirius red staining was performed on paraformaldehyde-fixed decellularized extracellular matrix deposited by WI-38 human lung cells on 2D surfaces to reveal collagen deposition (Figure 11). Part A of Figure 11 shows the results 1 and 9 days after plating the cells on Permanox plastic 4-chamber slides (250,000 cells / chamber). Cells were grown in chambers with or without fibronectin coating and with or without L-ascorbic acid. Cells grown on Permanox plastic 4-chamber slides (250,000 cells / chamber) show increased collagen deposition with L-ascorbic acid treatment, as well as longer culture times (9 days vs. 1 day). In the absence of fibronectin coating, no visible collagen deposition remained after 14-15 minutes of decellularization. Part B of FIG. 11 shows the results of plating WI-38 cells grown in 96-well plates (44,444 cells / well) 9 days after plating with a decellularization time of 5 minutes. The cells appear to deposit collagen fibers that are retained at the short decellularization time of 5 minutes, even in the absence of fibronectin coating. Part C of FIG. 11 shows the results of plating WI-38 cells grown in 8-chamber glass slides (97,222 cells / chamber) 9 days after plating with a decellularization time of 5 minutes. The cells appear to deposit collagen fibers that are retained at the short decellularization time of 5 minutes, even in the absence of fibronectin coating. EXAMPLES

[0067] The use of different materials within a microfluidic device is of interest as it has been found that cells can behave differently on plastic surfaces compared to glass surfaces (Figures 12-17). The material used in the fabrication of the microfluidic device can play a role in how the cells behave, in addition to any material that is placed within the device. The processing of the cells after culture (e.g., fixation methods and decellularization) can also affect the visualization of proteins of interest such as collagen I and alpha smooth muscle cell actin (α-SMA) (Figures 12-17). A variety of different processing and staining methods can be used with the microfluidic device.

[0068] Collagen I and α-SMA deposition by WI-38 cells in 96-well plates was studied. Cells were seeded at 44,444 cells / well and grown on medium containing L-ascorbic acid. Cells were grown in wells with either no coating or fibronectin coating. Cells were grown in the presence or absence of 5 ng / mL TGF-β1. Cells were incubated in 4% paraformaldehyde (PFA) (Figure 12) or methanol (Figure 13). Fluorescent images of "fused" collagen I, α-SMA, and nuclei are shown in consecutive columns. Images shown are at 20X magnification. Figures 14 and 15 show the quantified fluorescence intensity of collagen I and α-SMA channels. Image quantification was performed with ImageJ and the mean image intensity was calculated for each channel. Statistical analysis was performed with GraphPad Prism using two-way ANOVA with multiple comparisons. A significant difference between collagen I intensity in PFA-fixed and methanol-fixed wells is evident at low magnification but not at high magnification. In contrast, a significant difference in α-SMA intensity in PFA-fixed and methanol-fixed wells is evident at all magnifications tested. Figures 16 and 17 show the results of a cell study similar to those shown in Figures 12 and 13, but the cells were grown in eight-chamber glass slides. Cells were seeded at a concentration of 97,222 cells / chamber for regular media treatment and 18,421 cells / chamber for Ficoll Media treatment. Again, improved staining is observed in methanol-fixed cells compared to PFA-fixed cells. Furthermore, collagen I appears to be fibrillar in methanol-fixed cells but localized around the nucleus in PFA-fixed cells.

[0069] The present disclosure may include any combination of these various features or embodiments, as described in sentences and / or paragraphs, above and / or below. Any combination of features disclosed herein is considered to be part of the present disclosure.

[0070] 1. A fibrosis diagnostic system comprising: 1. A microfluidic device comprising: a device housing having an apparatus interior; A plurality of channels within the device, a first channel configured to receive a target object, a substrate, and a first biological composition configured to promote fibrosis of the target object; a second channel configured to receive a second biological composition configured to promote fibrosis of the target object; a first septum separating the first channel and the second channel and including a first plurality of openings providing fluid communication between the first channel and the second channel; a third channel configured to receive a third biological composition configured to promote fibrosis of the target object; a second septum separating the first channel and a third channel and including a second plurality of openings providing fluid communication between the first channel and the third channel; and A fibrosis diagnostic system comprising: a plurality of channel ports within the device housing in fluid communication with a plurality of channels, the plurality of channel ports comprising a first set of channel ports in fluid communication with a first channel, a second set of channel ports in fluid communication with a second channel, and a third set of channel ports in fluid communication with a third channel, each set of channel ports comprising one or more channel ports.

[0071] 2. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the system is configured to form a gradient of at least one type of molecule across the first channel, the second channel, or the third channel, or any combination thereof.

[0072] 3. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the at least one type of molecule comprises a chemotactic molecule.

[0073] 4. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the chemotactic molecule promotes movement of at least one cell type present in the first biological composition, the second biological composition, or the third biological composition, or any combination thereof.

[0074] 5. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, comprises at least one type of molecule.

[0075] 6. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the second and third sets of channel ports each comprise two or more channel ports.

[0076] 7. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein each set of channel ports comprises two or more channel ports.

[0077] 8. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the second and third sets of channel ports each comprise a first channel port and a second channel port, the first channel port being proximal to a first end of its respective channel and the second channel port being proximal to a second end of its respective channel.

[0078] 9. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein each of the set of channel ports comprises a first channel port and a second channel port, the first channel port being proximal to a first end of its respective channel and the second channel port being proximal to a second end of its respective channel.

[0079] 10. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first channel comprises a central channel port configured to receive the target object within the first channel.

[0080] 11. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the central channel port is between the first end of the first channel and the second end of the second channel.

[0081] 12. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein one or more of the plurality of channel ports comprises a seal, valve, or closure, or any combination thereof, configured for movement between an open state and a closed state.

[0082] 13. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, comprising a retaining element comprising one or more of the first, second, and third channels, the retaining element configured to retain at least one component insertable into one or more of the first, second, and third channels.

[0083] 14. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the retaining element comprises a modification of the channel surface.

[0084] 15. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the modification comprises a biomolecule.

[0085] 16. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the biomolecule comprises a polypeptide, a polysaccharide, a lipid, or a polynucleotide, or any combination thereof.

[0086] 17. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the modification comprises a polymer, metal, or ceramic, or any combination thereof.

[0087] 18. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the device housing comprises a polymer, metal, or ceramic, or any combination thereof.

[0088] 19. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the device housing comprises a first polymer and the modification comprises a second polymer, and wherein the first and second polymers differ from one another in chemical composition.

[0089] 20. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the modification comprises a physical structure.

[0090] 21. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the physical structure and the device housing comprise a common material.

[0091] 22. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the modification comprises a container for receiving the target object.

[0092] 23. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the vessel comprises a channel surface recess.

[0093] 24. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the physical structure comprises a plurality of physical structures.

[0094] 25. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the plurality of physical structures comprises a plurality of recesses in the channel surface, or a plurality of protrusions from the channel surface, or both.

[0095] 26. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first channel, the second channel, or the third channel, or any combination thereof, comprises a curved cross-section, or a straight cross-section, or both.

[0096] 27. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first channel, the second channel, or the third channel, or any combination thereof, comprises a constant width, or a constant diameter, or both.

[0097] 28. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first channel, the second channel, or the third channel, or any combination thereof, comprises a variable width, or a fixed diameter, or both.

[0098] 29. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein two or more channels of the plurality of channels have a common length.

[0099] 30. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein two or more of the plurality of channels are parallel to one another.

[0100] 31. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first channel is between the second channel and the third channel.

[0101] 32. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein two or more of the first, second, and third channels differ from one another in terms of volume, surface area, cross-sectional area, length, width, or diameter, or any combination thereof.

[0102] 33. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the width, diameter, or both of the first channel is greater than the width, diameter, or both of the second channel, or the third channel, or both.

[0103] 34. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein a cross-sectional area, width, diameter, or any combination thereof of one or more of the first, second, and third channels varies along its length.

[0104] 35. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein one or more of the first, second, and third channels comprises a first peripheral segment proximate a first end of the channel, a second peripheral segment proximate a second end of the channel, and a central segment between the first and second peripheral segments.

[0105] 36. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the central segment comprises a cross-sectional area, width, or diameter, or any combination thereof, greater than a cross-sectional area, width, or diameter, or any combination thereof, of one or both of the first and second peripheral segments.

[0106] 37. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the central segment of the first channel comprises a cross-sectional area, width, or diameter, or any combination thereof, that is greater than a cross-sectional area, width, or diameter, or any combination thereof, of one or both of the first and second peripheral segments of the first channel.

[0107] 38. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein one or both of the first and second peripheral segments are collinear with the central segment relative to their respective central longitudinal axes.

[0108] 39. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein one or both of the first and second peripheral segments form an angle other than 180° with the central segment relative to their respective central longitudinal axes.

[0109] 40. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the angle comprises a right angle, an acute angle, or an obtuse angle, or any combination thereof.

[0110] 41. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein two or more of the plurality of channels are in a common plane.

[0111] 42. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first, second, and third channels of the plurality of channels are in a common plane.

[0112] 43. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the length of the first septum is less than the length of the first channel, or the length of the second channel, or both.

[0113] 44. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the length of the second septum is less than the length of the first channel, or the length of the third channel, or both.

[0114] 45. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first septum, the second septum, or both, comprise a membrane.

[0115] 46. ​​The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the membrane comprises a porous membrane.

[0116] 47. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the membrane comprises a semi-permeable membrane.

[0117] 48. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the membrane comprises a biological membrane, or a synthetic membrane, or both.

[0118] 49. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first septum includes a first plurality of rows separated by a first plurality of openings, the second septum includes a second plurality of rows separated by a second plurality of second openings, or both.

[0119] 50. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first plurality of rows, or the second plurality of rows, or both, have a straight cross-section, a curved cross-section, or both.

[0120] 51. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first plurality of rows, or the second plurality of rows, or both, are trapezoidal.

[0121] 52. A fibrosis diagnostic system according to any preceding or subsequent embodiment / feature / aspect, comprising: the openings of the first plurality of openings vary in number, size, shape, or spacing, or any combination thereof; or the openings of the second plurality of openings differ in number, size, shape, or spacing, or any combination thereof; or The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect,

[0122] 53. A fibrosis diagnostic system according to any preceding or subsequent embodiment / feature / aspect, comprising: the openings of the first plurality of openings vary in number, size, shape, or spacing, or any combination thereof, along one or more dimensions of the first septum; or the openings of the second plurality of openings vary in number, size, shape, or spacing, or any combination thereof, along one or more dimensions of the second septum; or The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect,

[0123] 54. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first plurality of openings and the second plurality of openings are different from one another.

[0124] 55. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the device housing comprises a substrate, the plurality of channels and the plurality of channel ports being formed in the substrate.

[0125] 56. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the device housing comprises silicone.

[0126] 57. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the device housing comprises polydimethylsiloxane (PDMS).

[0127] 58. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the device housing comprises a flexible material, or a rigid material, or both.

[0128] 59. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the device housing comprises a conductor, or an insulator, or both.

[0129] 60. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the substrate comprises a first material and one or more channel surfaces comprise a second material different from the first material.

[0130] 61. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the second material is distinct from the contents of the one or more channels in which it is present.

[0131] 62. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the second material is a coating on the first material.

[0132] 63. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein one or more channel surfaces are coated with a plurality of distinct materials.

[0133] 64. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein one or more channel surfaces are coated with a plurality of coatings.

[0134] 65. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein at least two of the plurality of coatings differ in material composition.

[0135] 66. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the substrate comprises a first material, and the plurality of coatings comprises a first coating on one or more channel surfaces comprising a second material adjacent to the substrate, and a second coating comprising a third material overlying the first coating.

[0136] 67. The fibrosis diagnostic system of any one of the preceding or subsequent embodiments / features / aspects, wherein the first and second materials are different from each other and the second and third materials are different from each other.

[0137] 68. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first, second, and third materials are different from one another.

[0138] 69. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the plurality of coatings comprises at least two layers having different average thicknesses from each other.

[0139] 70. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the material composition, thickness, or patterning, or a combination thereof, of the coating on one or more channel surfaces varies along the length of one or more channels.

[0140] 71. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein two or more of the channels differ in the material composition of their surface coating.

[0141] 72. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first channel comprises a first coating, the second channel comprises a second coating, and the third channel comprises a third coating, and the first, second, and third coatings differ in material composition, thickness, or patterning, or a combination thereof.

[0142] 73. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first, second, and third channels are coated with the same material.

[0143] 74. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the one or more coatings comprise an organic material, or an inorganic material, or both.

[0144] 75. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the one or more coatings comprise a polymer.

[0145] 76. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the one or more coatings comprise a biomolecule.

[0146] 77. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the one or more coatings comprise a protein, or a nucleic acid, a lipid, or a carbohydrate, or any combination thereof.

[0147] 78. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the one or more coatings comprise fibronectin, an extracellular matrix-derived composition, a basement membrane extract, a decellularized extracellular matrix, a solubilized basement membrane preparation, a mixture of extracellular matrix proteins, a tumor-derived composition, Matrigel, or hyaluronic acid, or any combination thereof.

[0148] 79. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the substrate comprises silicon and one or more channel surfaces are coated with a coating comprising a silane.

[0149] 80. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein one or more channel surfaces are subjected to silanization.

[0150] 81. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein one or more channel surfaces are treated with plasma.

[0151] 82. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the plasma comprises oxygen.

[0152] 83. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the one or more coatings are applied by emulsion, spraying, brushing, injection, precipitation, polymerization, or deposition, or any combination thereof.

[0153] 84. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein one or more channel surfaces comprises a laminate.

[0154] 85. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the device housing comprises a transparent material, a translucent material, a pigmented material, a colored material, a polarized material, a material that selectively transmits wavelengths of electromagnetic radiation, a material that selectively refracts wavelengths of electromagnetic radiation, a material that selectively reflects wavelengths of electromagnetic radiation, or an opaque material, or any combination thereof.

[0155] 86. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the device housing includes a window permitting visibility into the interior of the device.

[0156] 87. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the field of view is of one or more channels of the plurality of channels.

[0157] 88. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the field of view includes a view of the first channel, or the target object, or both.

[0158] 89. A fibrosis diagnostic system according to any preceding or subsequent embodiment / feature / aspect, wherein the device housing comprises multiple windows allowing multiple views into the interior of the device.

[0159] 90. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the target object comprises an organic material, or an inorganic material, or both.

[0160] 91. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the target object comprises tissue, or a medical device, or both.

[0161] 92. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the target object comprises a spheroid, an organoid, an alginate-based biomaterial, a scaffold-supported cell structure, or a bio-printed structure, or any combination thereof.

[0162] 93. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the target object comprises neoplastic cells, malignant cells, or benign cells, or any combination thereof.

[0163] 94. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the target comprises one or more coatings.

[0164] 95. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein one or more coatings of the target are distinct from the substrate.

[0165] 96. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the one or more coatings on the target differ from the one or more coatings on the one or more channel surfaces in terms of material composition, thickness, or patterning, or a combination thereof.

[0166] 97. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the one or more coatings on the target are the same as the one or more coatings on the one or more channel surfaces with respect to material composition, thickness, or patterning, or a combination thereof.

[0167] 98. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the target is coated with a plurality of different materials.

[0168] 99. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the target is coated with multiple coatings.

[0169] 100. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein at least two of the plurality of coatings differ in material composition, thickness, or patterning, or a combination thereof.

[0170] 101. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the plurality of coatings comprises a first coating comprising a first material adjacent to the target, and a second coating comprising a second material overlying the first coating.

[0171] 102. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the target is coated prior to placement within the microfluidic device.

[0172] 103. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the substrate comprises a hydrogel.

[0173] 104. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the matrix comprises fibronectin, an extracellular matrix-derived composition, a basement membrane extract, a decellularized extracellular matrix, a solubilized basement membrane preparation, a mixture of extracellular matrix proteins, a tumor-derived composition, Matrigel, or hyaluronic acid, or any combination thereof.

[0174] 105. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the substrate comprises a natural polymer, an artificial polymer, or a crosslinked polymer, or any combination thereof.

[0175] 106. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the substrate comprises a polysaccharide, a polypeptide, a lipid, a polynucleotide, hyaluronic acid, hyaluronic acid methacrylate, hyaluronic acid diacrylate, fibronectin, fibrinogen, collagen, or collagen methacrylate, or any combination thereof.

[0176] 107. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first biological composition comprises fibroblasts, or endothelial cells, or both.

[0177] 108. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first biological composition comprises serum, ascorbic acid, retinoic acid, proline, or bleomycin, or any combination thereof.

[0178] 109. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first biological composition comprises a fibrosis-promoting biomolecule, a complement protein, a blood coagulation-related protein, a bacterial cell wall protein, or lipopolysaccharide (LPS), or any combination thereof.

[0179] 110. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the second biological composition comprises at least one type of biological cell capable of promoting fibrosis.

[0180] 111. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the second biological composition comprises a factor that promotes differentiation of a biological cell into a type of biological cell that can promote fibrosis, a factor that promotes expression of a biomolecule that promotes fibrosis, or a factor that promotes proliferation of a type of biological cell that can promote fibrosis, or any combination thereof.

[0181] 112. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the second biological composition comprises serum, platelets, leukocytes, macrophages, monocytes, peripheral blood mononuclear cells (PBMCs), stem cells, mast cells, dendritic cells, neutrophils, granulocytes, T cells, or B cells, or any combination thereof.

[0182] 113. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the third biological composition comprises a population of molecules capable of forming a gradient emitted from the third channel into the first channel, or into both the first and second channels.

[0183] 114. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the third biological composition comprises a cytokine, a chemokine, a cell nutrient, a transcription factor, or a growth factor, or any combination thereof.

[0184] 115. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the third biological composition comprises CCL-2 / MCP-1, CCL3, CCL11, fibroblast growth factor (FGF), interferon, IFN-gamma, interleukin (IL), IL-4, IL-1α, IL-1β, IL-2, IL-5, IL-6, IL-7, IL-9, IL-13, IL-17A, IL-17F, IL-21, IL-22, IL-25, platelet-derived growth factor (PDGF), TGF-alpha, TGF-β, granulocyte colony-stimulating factor (G-CSF), macrophage colony-stimulating factor (M-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), prothrombin, or thrombin, or any combination thereof.

[0185] 116. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein at least one type of molecule in the gradient comprises a cytokine, a chemokine, a cell nutrient, a transcription factor, or a growth factor, or any combination thereof.

[0186] 117. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein at least one type of molecule in the gradient comprises transforming growth factor β (TGF-β).

[0187] 118. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein TGF-β comprises TGF-β1.

[0188] 119. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, comprises labeled cells, or labeled biomolecules, or both.

[0189] 120. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, comprises a compound with known or potential anti-fibrotic efficacy.

[0190] 121. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, further comprising a substrate, a first biological composition, a second biological composition, or a third biological composition, or any combination thereof.

[0191] 122. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, further comprising a target object.

[0192] 123. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, further comprising a gradient of at least one molecule across the first channel, the second channel, or the third channel, or any combination thereof.

[0193] 124. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein one or more components of the substrate, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, is anchored to the channel surface.

[0194] 125. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, further comprising a detector, the detector comprising a sensor configured to detect fibrosis or a characteristic indicative thereof of the target object.

[0195] 126. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the detector is configured to receive a microfluidic device.

[0196] 127. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the detector comprises a detector housing comprising a detector interior.

[0197] 128. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the detector further comprises a receptacle configured to receive a microfluidic device, the receptacle comprising an opening within the detector housing.

[0198] 129. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, further comprising a thermal subsystem.

[0199] 130. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the thermal subsystem comprises a heater, or a cooler, or both.

[0200] 131. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the thermal subsystem is configured to regulate the temperature of the microfluidic device.

[0201] 132. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the detector comprises a thermal subsystem.

[0202] 133. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the thermal subsystem is configured to regulate a temperature of the microfluidic device when in the receptacle.

[0203] 134. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, further comprising a fluidic subsystem configured for fluid communication with the microfluidic device.

[0204] 135. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the fluidic subsystem is configured to load one or more components into the microfluidic device.

[0205] 136. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the fluidic subsystem is configured to deliver a solid, liquid, or gas, or any combination thereof, to the microfluidic device.

[0206] 137. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the fluidic subsystem is configured to load a target object, a substrate, a first biological composition, a second biological composition, or a third biological composition, or any combination thereof, into the microfluidic device.

[0207] 138. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the fluidic subsystem is configured to supply one or more nutrients to the microfluidic device.

[0208] 139. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the fluidic subsystem is configured to regulate pressure within the microfluidic device.

[0209] 140. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the fluidic subsystem is configured to regulate a gradient across the first channel, the second channel, or the third channel, or any combination thereof.

[0210] 141. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the fluidic subsystem is configured to remove one or more components from the microfluidic device.

[0211] 142. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the detector comprises a fluidic subsystem.

[0212] 143. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the fluidic subsystem is configured to be in fluid communication with the microfluidic device when the microfluidic device is within the receptacle.

[0213] 144. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the sensor is configured to detect particles, or waves, or both, emitted from the target object, or the microfluidic device, or both.

[0214] 145. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the sensor is configured to detect electromagnetic radiation.

[0215] 146. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the electromagnetic radiation comprises ultraviolet light, visible light, or infrared light, or any combination thereof.

[0216] 147. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the electromagnetic radiation comprises fluorescence, or phosphorescence, or both.

[0217] 148. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the detector further comprises a particle source, or a wave source, or both, and is configured to transmit particles, waves, or both to the target object, or the microfluidic device, or both.

[0218] 149. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, further comprising an electromagnetic radiation source.

[0219] 150. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the detector comprises an electromagnetic radiation source.

[0220] 151. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the electromagnetic radiation source comprises a light emitting diode, a laser, an incandescent light source, an arc light source, a halogen light source, a chemical vapor light source, or a fluorescent light source, or any combination thereof.

[0221] 152. A fibrosis diagnostic system according to any of embodiments / features / aspects 1-10, wherein the detector further comprises an optical subsystem configured to pass radiation or affect radiation, or both.

[0222] 153. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the optical subsystem includes one or more lenses, one or more filters, one or more mirrors, or one or more diffraction gratings, or any combination thereof.

[0223] 154. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the optical subsystem, or the sensor, or both, includes a microscope or one or more components thereof.

[0224] 155. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the optical subsystem comprises a sensor, or an electromagnetic radiation source, or both.

[0225] 156. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the sensor is configured to generate an output signal based on an input signal detected by the sensor.

[0226] 157. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the detector further includes a transmitter, receiver, transceiver, or any combination thereof, or both, configured to receive information or transmit information.

[0227] 158. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the information comprises an output signal.

[0228] 159. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the information includes an operational signal for operation of the sensor.

[0229] 160. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the detector further comprises a data port.

[0230] 161. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, further comprising a processor configured to receive and process information.

[0231] 162. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the processor is configured to control one or more subsystems or components of the fibrosis diagnostic system.

[0232] 163. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the processor is configured to control a sensor, an electromagnetic radiation source, an optical subsystem, a thermal subsystem, or a fluidic subsystem, or any combination thereof.

[0233] 164. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the processor is configured to receive a signal output from the sensor, receive an operational signal from a user input, or both.

[0234] 165. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the detector comprises a processor.

[0235] 166. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, further comprising a memory configured for communication with the processor.

[0236] 167. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the memory is configured to store a program that controls the fibrosis diagnostic system or a component thereof.

[0237] 168. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the processor comprises a memory.

[0238] 169. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the detector comprises a memory.

[0239] 170. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, further comprising a user interface.

[0240] 171. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the user interface includes a monitor, a button, a keyboard, a mouse, an optical signal, a camera, an eyepiece, a speaker, or a microphone, or any combination thereof.

[0241] 172. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the detector comprises a user interface.

[0242] 173. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, further comprising a power source, or a power interface, or both.

[0243] 174. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, wherein the detector includes a power source, or a power interface, or both.

[0244] 175. The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, further comprising a kit, the kit comprising a microfluidic device and one or more reagents for use with the microfluidic device.

[0245] 176. The method further comprises the steps of: A microfluidic device; The fibrosis diagnostic system of any preceding or subsequent embodiment / feature / aspect, comprising a substrate, a first biological composition, a second biological composition, or a third biological composition, or any combination thereof.

[0246] 177. A method for analyzing a target object for susceptibility to fibrosis, comprising: disposing a target object, a substrate, and a first biological composition in a first channel of a microfluidic device according to any preceding or subsequent embodiment / feature / aspect; disposing a second biological composition in a second channel of the microfluidic device; disposing a third biological composition in a third channel of the microfluidic device; analyzing the target object, or the first channel, or both, to determine the presence or absence of fibrosis associated with the target object.

[0247] 178. The method of any preceding or subsequent embodiment / feature / aspect, wherein the analysis is performed after a predetermined period of time after placing the target object, substrate, first biological composition, second biological composition, or third biological composition, or any combination thereof, into the microfluidic device.

[0248] 179. The method of any preceding or subsequent embodiment / feature / aspect, wherein the predetermined period of time is sufficient to allow for the formation of a gradient of at least one type of molecule.

[0249] 180. The method of any preceding or subsequent embodiment / feature / aspect, wherein the analysis is performed more than once.

[0250] 181. A method according to any preceding or subsequent embodiment / feature / aspect, wherein any two successive analyses are separated by a predetermined period of time.

[0251] 182. The method of any preceding or subsequent embodiment / feature / aspect, wherein analyzing includes removing one or more components from the microfluidic device.

[0252] 183. The method of any preceding or subsequent embodiment / feature / aspect, wherein the analysis comprises an immunoassay.

[0253] 184. The method of any preceding or subsequent embodiment / feature / aspect, wherein analyzing comprises use of an antibody.

[0254] 185. The method of any preceding or subsequent embodiment / feature / aspect, wherein the antibody is labeled.

[0255] 186. The method of any preceding or subsequent embodiment / feature / aspect, wherein the analysis comprises application of a primary antibody and a secondary antibody that comprises a detectable label and that binds to the primary antibody.

[0256] 187. The method of any preceding or subsequent embodiment / feature / aspect, wherein the primary antibody binds to collagen, alpha smooth muscle actin, or both.

[0257] 188. The method of any preceding or subsequent embodiment / feature / aspect, wherein the analysis comprises a histological assay.

[0258] 189. The method of any preceding or subsequent embodiment / feature / aspect, wherein the analysis comprises application of picrosirius red dye.

[0259] 190. The method of any preceding or subsequent embodiment / feature / aspect, wherein analyzing includes visually inspecting the target object, or the first channel, or both, for one or more indications of fibrosis associated with the target object.

[0260] 191. The method of any preceding or subsequent embodiment / feature / aspect, wherein analyzing includes using a microscope.

[0261] 192. The method of any preceding or subsequent embodiment / feature / aspect, wherein analyzing includes using one or more spectroscopic techniques.

[0262] 193. The method of any preceding or subsequent embodiment / feature / aspect, wherein the analysis includes nonlinear optical (NLO) multiphoton microscopy, second harmonic generation (SHG) imaging, third harmonic generation (THG) imaging, two-photon excited fluorescence (TPEF), cell tracker dye imaging, visible light imaging, (ELISA), Griess assay, histopathology immunostaining, cleared tissue microscopy, or proteomic profiling of homogenized hydrogels, or any combination thereof.

[0263] 194. The method of any preceding or subsequent embodiment / feature / aspect, wherein analyzing includes detecting one or more indications of fibrosis associated with the target object using a detector.

[0264] 195. The method of any preceding or subsequent embodiment / feature / aspect, wherein analyzing includes inserting a microfluidic device into a receptacle.

[0265] 196. The method of any preceding or subsequent embodiment / feature / aspect, further comprising preparing a target object, a substrate, a first biological composition, a second biological composition, or a third biological composition, or any combination thereof, from a patient sample.

[0266] 197. The method of any preceding or subsequent embodiment / feature / aspect, further comprising implanting the target object in or on the patient.

[0267] 198. The method of any preceding or subsequent embodiment / feature / aspect, further comprising implanting a medical device, or tissue, or both, into or onto a patient, and wherein the target object is a representative model of the medical device, or tissue, or both.

[0268] 199. The method of any preceding or subsequent embodiment / feature / aspect, further comprising removing the medical device, or tissue, or both, from the patient, and wherein the target object is a representative model of the medical device, or tissue, or both.

[0269] 200. The method of any preceding or subsequent embodiment / feature / aspect, further comprising administering to the patient an anti-fibrotic agent.

[0270] 201. The method of any preceding or subsequent embodiment / feature / aspect, wherein analyzing comprises determining a quantitative measure of fibrosis.

[0271] 202. The method of any preceding or subsequent embodiment / feature / aspect, wherein the dosage of anti-fibrotic agent administered to the patient is based on a quantitative measure of fibrosis.

[0272] 203. The method of any preceding or subsequent embodiment / feature / aspect, wherein an anti-fibrotic agent is administered to the patient prior to, simultaneously with, or after implantation of the target object, tissue associated with the target object, or a medical device associated with the target object, or any combination thereof.

[0273] 204. The method of any preceding or subsequent embodiment / feature / aspect, further comprising administering to the patient one or more therapeutic agents in addition to, or as an alternative to, the antifibrotic agent.

[0274] 205. The method of any preceding or subsequent embodiment / feature / aspect, wherein the patient is a mammalian patient, or a human patient, or both.

[0275] 206. A method for screening a therapy for efficacy in treating fibrosis, comprising: disposing a target object, a substrate, and a first biological composition in a first channel of a microfluidic device according to any preceding or subsequent embodiment / feature / aspect; disposing a second biological composition in a second channel of the microfluidic device; disposing a third biological composition in a third channel of the microfluidic device; Administering a therapy on the microfluidic device; analyzing the target object, or the first channel, or both, to determine the presence, absence, or extent of fibrosis associated with the target object; and determining the effectiveness of the therapy based on the presence, absence, or extent of fibrosis without administering the therapy.

[0276] 207. The method of any preceding or subsequent embodiment / feature / aspect, wherein the therapy includes a therapeutic compound and the performing includes placing the therapeutic compound in the first channel, the second channel, or the third channel, or any combination thereof.

[0277] 208. The method of any preceding or subsequent embodiment / feature / aspect, wherein the therapeutic compound comprises a pharmaceutical compound, or a biological compound, or both.

[0278] 209. The method of any preceding or subsequent embodiment / feature / aspect, wherein therapy includes modification of a target object.

[0279] 210. The method of any preceding or subsequent embodiment / feature / aspect, wherein therapy includes placing a medical device in the first channel, the second channel, or the third channel, or any combination thereof.

[0280] 211. The method of any preceding or subsequent embodiment / feature / aspect, wherein the therapy comprises irradiating the microfluidic device.

[0281] 212. The method of any preceding or subsequent embodiment / feature / aspect, wherein the therapy further comprises treating the patient with a screened therapy.

[0282] 213. A method for screening a therapy for effectiveness in treating cancer, comprising: placing a target object, a substrate, and a first biological composition into a first channel of a microfluidic device of any preceding or subsequent embodiment / feature / aspect, wherein the target object comprises neoplastic cells; disposing a second biological composition in a second channel of the microfluidic device; disposing a third biological composition in a third channel of the microfluidic device; Administering a therapy on the microfluidic device; analyzing the target object, or the first channel, or both, to determine the presence, absence, or extent of fibrosis associated with the target object; and determining the effectiveness of the therapy based on the presence, absence, or extent of cancer without administering the therapy.

[0283] 214. A non-transitory computer-readable medium storing a program including instructions for performing a method according to any preceding or subsequent embodiment / feature / aspect.

[0284] 215. A non-transitory computer readable medium storing a program including instructions for controlling a fibrosis diagnostic system, or a subsystem thereof, or a component thereof, or any combination thereof, of a fibrosis diagnostic system according to any preceding or subsequent embodiment / feature / aspect.

[0285] Applicant specifically incorporates the entire contents of all references cited in this disclosure.Furthermore, when an amount, concentration, or other value or parameter is given as either a range, a preferred range, or a list of a preferred upper value and a preferred lower value, this should be understood as specifically disclosing all ranges formed from any pair of any upper value or preferred value and any lower value or preferred value, regardless of whether the range is separately disclosed.When a range of numerical values ​​is recited herein, unless otherwise stated, the range is intended to include its endpoints, and all integers and fractions within the range.The scope of the present disclosure is not limited to the specific values ​​recited when defining the range.

[0286] All references cited herein are incorporated by reference as if each reference was specifically and individually indicated to be incorporated by reference. The citation of any reference is for its disclosure prior to the filing date and should not be construed as an admission that the present disclosure is not entitled to any particular subject matter disclosed herein.

[0287] It will be appreciated that each of the above elements, or two or more elements taken together, may find useful application in other types of methods different from those described above. Without further analysis, the foregoing will fully reveal the gist of the present disclosure, so that others may readily adapt it for various uses while still falling within the scope of the claimed subject matter. The above-described embodiments are presented by way of example only.

Claims

1. A fibrosis diagnostic system comprising:

1. A microfluidic device comprising: a device housing having an interior; a plurality of channels within the device, a first channel configured to receive a target object, a substrate, and a first biological composition configured to promote fibrosis of the target object; a second channel configured to receive a second biological composition configured to promote fibrosis of the target object; a first septum separating the first channel and the second channel and including a first plurality of openings providing fluid communication between the first channel and the second channel; a third channel configured to receive a third biological composition configured to promote fibrosis of the target object; a second septum separating the first channel and a third channel and including a second plurality of openings providing fluid communication between the first channel and the third channel; A fibrosis diagnostic system comprising: a plurality of channel ports within the device housing in fluid communication with the plurality of channels, the plurality of channel ports comprising a first set of channel ports in fluid communication with the first channel, a second set of channel ports in fluid communication with the second channel, and a third set of channel ports in fluid communication with the third channel, each set of channel ports comprising one or more channel ports.

2. 2. The fibrosis diagnostic system of claim 1, wherein the system is configured to form a gradient of at least one type of molecule across the first channel, the second channel, or the third channel, or any combination thereof.

3. The fibrosis diagnostic system of claim 1 or 2, wherein the at least one type of molecule comprises a chemotactic molecule.

4. The fibrosis diagnostic system according to any one of claims 1 to 3, wherein the chemotactic molecule promotes the migration of at least one cell type present in the first biological composition, the second biological composition, or the third biological composition, or any combination thereof.

5. The fibrosis diagnostic system of any one of claims 1 to 4, wherein the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, comprises the at least one type of molecule.

6. The fibrosis diagnostic system of any one of claims 1 to 5, wherein the second and third sets of channel ports each comprise two or more channel ports.

7. The fibrosis diagnostic system of any one of claims 1 to 6, wherein each set of channel ports comprises two or more channel ports.

8. The fibrosis diagnostic system of any one of claims 1 to 7, wherein the second and third sets of channel ports each comprise a first channel port and a second channel port, the first channel port being proximal to a first end of its respective channel, and the second channel port being proximal to a second end of its respective channel.

9. A fibrosis diagnostic system according to any one of claims 1 to 8, wherein each of the sets of channel ports comprises a first channel port and a second channel port, the first channel port being proximal to a first end of its respective channel, and the second channel port being proximal to a second end of its respective channel.

10. The fibrosis diagnostic system of any one of claims 1 to 9, wherein the first channel comprises a central channel port configured to receive the target object within the first channel.

11. The fibrosis diagnostic system of any one of claims 1 to 10, wherein the central channel port is between a first end of the first channel and a second end of the second channel.

12. The fibrosis diagnostic system of any of claims 1 to 11, wherein one or more of the plurality of channel ports comprises a seal, valve, or closure, or any combination thereof, configured for movement between an open state and a closed state.

13. A fibrosis diagnostic system according to any one of claims 1 to 12, comprising a holding element comprising one or more of the first, second, and third channels, the holding element configured to hold at least one component insertable into one or more of the first, second, and third channels.

14. The fibrosis diagnostic system of any of claims 1 to 13, wherein the retaining element comprises a modification of the channel surface.

15. The fibrosis diagnostic system according to any one of claims 1 to 14, wherein the modification comprises a biomolecule.

16. The fibrosis diagnostic system according to any one of claims 1 to 15, wherein the biomolecule comprises a polypeptide, a polysaccharide, a lipid, or a polynucleotide, or any combination thereof.

17. The fibrosis diagnostic system according to any one of claims 1 to 16, wherein the modification comprises a polymer, a metal, or a ceramic, or any combination thereof.

18. The fibrosis diagnostic system of any of claims 1 to 17, wherein the device housing comprises a polymer, a metal, or a ceramic, or any combination thereof.

19. 19. The fibrosis diagnostic system of any of claims 1 to 18, wherein the device housing comprises a first polymer and the modification comprises a second polymer, and the first and second polymers differ from each other in chemical composition.

20. The fibrosis diagnostic system according to any one of claims 1 to 19, wherein the modification comprises a physical structure.

21. The fibrosis diagnostic system of any of claims 1 to 20, wherein the physical structure and the device housing comprise a common material.

22. The fibrosis diagnostic system according to any one of claims 1 to 21, wherein the modification comprises a container for receiving the target substance.

23. The fibrosis diagnostic system according to any one of claims 1 to 22, wherein the container has a recess on the channel surface.

24. The fibrosis diagnostic system of any one of claims 1 to 23, wherein the physical structure comprises a plurality of physical structures.

25. The fibrosis diagnostic system of any one of claims 1 to 24, wherein the plurality of physical structures comprises a plurality of recesses in the channel surface, or a plurality of protrusions from the channel surface, or both.

26. 26. The fibrosis diagnostic system of any of claims 1 to 25, wherein the first channel, the second channel, or the third channel, or any combination thereof, comprises a curved cross-section, or a straight cross-section, or both.

27. The fibrosis diagnostic system of any of claims 1 to 26, wherein the first channel, the second channel, or the third channel, or any combination thereof, comprises a constant width, or a constant diameter, or both.

28. 28. The fibrosis diagnostic system of any of claims 1 to 27, wherein the first channel, the second channel, or the third channel, or any combination thereof, comprises a variable width, or a fixed diameter, or both.

29. The fibrosis diagnostic system of any one of claims 1 to 28, wherein two or more of the plurality of channels have a common length.

30. The fibrosis diagnostic system according to any one of claims 1 to 29, wherein two or more of the plurality of channels are parallel to each other.

31. The fibrosis diagnostic system according to any one of claims 1 to 30, wherein the first channel is located between the second channel and the third channel.

32. The fibrosis diagnostic system of any one of claims 1 to 31, wherein two or more of the first, second, and third channels differ from each other in terms of volume, surface area, cross-sectional area, length, width, or diameter, or any combination thereof.

33. A fibrosis diagnostic system according to any preceding claim, wherein the width, diameter, or both of the first channel is larger than the width, diameter, or both of the second channel, or the third channel, or both.

34. 34. A fibrosis diagnostic system according to any preceding claim, wherein the cross-sectional area, width, diameter, or any combination thereof of one or more of the first, second, and third channels varies along its length.

35. The fibrosis diagnostic system of any one of claims 1 to 34, wherein one or more of the first, second, and third channels comprises a first peripheral segment proximate a first end of the channel, a second peripheral segment proximate a second end of the channel, and a central segment between the first and second peripheral segments.

36. The fibrosis diagnostic system of any of claims 1 to 35, wherein the central segment comprises a cross-sectional area, width, or diameter, or any combination thereof, that is greater than a cross-sectional area, width, or diameter, or any combination thereof, of one or both of the first peripheral segment and the second peripheral segment.

37. 37. The fibrosis diagnostic system of any of claims 1 to 36, wherein the central segment of the first channel comprises a cross-sectional area, width, or diameter, or any combination thereof, that is larger than a cross-sectional area, width, or diameter, or any combination thereof, of one or both of the first and second peripheral segments of the first channel.

38. 38. A fibrosis diagnostic system according to any preceding claim, wherein one or both of the first and second peripheral segments are collinear with the central segment relative to their respective central longitudinal axes.

39. 39. A fibrosis diagnostic system according to any preceding claim, wherein one or both of the first and second peripheral segments form an angle other than 180° with the central segment relative to their respective central longitudinal axes.

40. The fibrosis diagnostic system of any of claims 1 to 39, wherein the angle comprises a right angle, an acute angle, or an obtuse angle, or any combination thereof.

41. The fibrosis diagnostic system of any one of claims 1 to 40, wherein two or more of the plurality of channels are in a common plane.

42. The fibrosis diagnostic system of any one of claims 1 to 41, wherein the first, second, and third channels of the plurality of channels are in a common plane.

43. A fibrosis diagnostic system according to any one of claims 1 to 42, wherein the length of the first septum is shorter than the length of the first channel, or the length of the second channel, or both.

44. A fibrosis diagnostic system according to any one of claims 1 to 43, wherein the length of the second septum is shorter than the length of the first channel, or the length of the third channel, or both.

45. The fibrosis diagnostic system of any of claims 1 to 44, wherein the first septum, the second septum, or both, comprise a membrane.

46. The fibrosis diagnostic system of any one of claims 1 to 45, wherein the membrane comprises a porous membrane.

47. The fibrosis diagnostic system of any one of claims 1 to 46, wherein the membrane comprises a semi-permeable membrane.

48. The fibrosis diagnostic system of any one of claims 1 to 47, wherein the membrane comprises a biological membrane, a synthetic membrane, or both.

49. 49. The fibrosis diagnostic system of any one of claims 1 to 48, wherein the first septum comprises a first plurality of rows separated by the first plurality of openings, the second septum comprises a second plurality of rows separated by the second plurality of second openings, or both.

50. 50. The fibrosis diagnostic system of any of claims 1 to 49, wherein the first plurality of rows, or the second plurality of rows, or both, have a straight cross-section, a curved cross-section, or both.

51. 51. The fibrosis diagnostic system of any one of claims 1 to 50, wherein the first plurality of rows, or the second plurality of rows, or both, are trapezoidal.

52. The fibrosis diagnostic system according to any one of claims 1 to 51, the openings of the first plurality of openings vary in number, size, shape, or spacing, or any combination thereof; or the openings of the second plurality of openings differ in number, size, shape, or spacing, or any combination thereof; or The fibrosis diagnostic system according to any one of claims 1 to 51, wherein the fibrosis diagnostic system is both.

53. A fibrosis diagnostic system according to any one of claims 1 to 52, comprising: the openings of the first plurality of openings vary in number, size, shape, or spacing, or any combination thereof, along one or more dimensions of the first septum; or the openings of the second plurality of openings vary in number, size, shape, or spacing, or any combination thereof, along one or more dimensions of the second septum; or The fibrosis diagnostic system according to any one of claims 1 to 52, wherein the fibrosis diagnostic system is both.

54. The fibrosis diagnostic system of any one of claims 1 to 53, wherein the first plurality of openings and the second plurality of openings are different from each other.

55. The fibrosis diagnostic system of any preceding claim, wherein the device housing comprises a substrate, and the plurality of channels and the plurality of channel ports are formed within the substrate.

56. The fibrosis diagnostic system of any one of claims 1 to 55, wherein the device housing comprises silicone.

57. The fibrosis diagnostic system of any one of claims 1 to 56, wherein the device housing comprises polydimethylsiloxane (PDMS).

58. The fibrosis diagnostic system of any of claims 1 to 57, wherein the device housing comprises a flexible material, a rigid material, or both.

59. The fibrosis diagnostic system of any one of claims 1 to 58, wherein the device housing comprises a conductor, an insulator, or both.

60. 60. The fibrosis diagnostic system of any one of claims 1 to 59, wherein the substrate comprises a first material and one or more channel surfaces comprise a second material different from the first material.

61. 61. A fibrosis diagnostic system according to any preceding claim, wherein the second material is different from the contents of the one or more channels in which it resides.

62. The fibrosis diagnostic system of any one of claims 1 to 61, wherein the second material is a coating on the first material.

63. The fibrosis diagnostic system of any one of claims 1 to 62, wherein one or more channel surfaces are coated with a plurality of different materials.

64. 64. The fibrosis diagnostic system of any one of claims 1 to 63, wherein one or more channel surfaces are coated with multiple coatings.

65. A fibrosis diagnostic system according to any preceding claim, wherein at least two of the plurality of coatings differ in material composition.

66. 66. A fibrosis diagnostic system as described in any one of claims 1 to 65, wherein the substrate comprises a first material and the plurality of coatings comprises a first coating comprising a second material adjacent to the substrate on one or more channel surfaces, and a second coating comprising a third material overlying the first coating.

67. 67. The fibrosis diagnostic system of any preceding claim, wherein the first and second materials are different from each other, and the second and third materials are different from each other.

68. The fibrosis diagnostic system of any one of claims 1 to 67, wherein the first, second, and third materials are different from each other.

69. The fibrosis diagnostic system of any one of claims 1 to 68, wherein the plurality of coatings comprises at least two layers having different average thicknesses.

70. A fibrosis diagnostic system as described in any one of claims 1 to 69, wherein the material composition, thickness, or patterning, or a combination thereof, of the coating on one or more channel surfaces varies along the length of one or more channels.

71. A fibrosis diagnostic system according to any preceding claim, wherein two or more of the channels differ in the material composition of their surface coating.

72. 72. The fibrosis diagnostic system of any of claims 1 to 71, wherein the first channel comprises a first coating, the second channel comprises a second coating, and the third channel comprises a third coating, and the first, second, and third coatings differ in material composition, thickness, or patterning, or a combination thereof.

73. The fibrosis diagnostic system of any one of claims 1 to 72, wherein the first, second, and third channels are coated with the same material.

74. 74. The fibrosis diagnostic system of any one of claims 1 to 73, wherein the one or more coatings comprise an organic material, or an inorganic material, or both.

75. The fibrosis diagnostic system of any one of claims 1 to 74, wherein one or more coatings comprise a polymer.

76. The fibrosis diagnostic system of any one of claims 1 to 75, wherein one or more coatings comprise a biomolecule.

77. 77. The fibrosis diagnostic system of any of claims 1 to 76, wherein the one or more coatings comprise a protein, or a nucleic acid, a lipid, or a carbohydrate, or any combination thereof.

78. 78. The fibrosis diagnostic system of any of claims 1 to 77, wherein the one or more coatings comprise fibronectin, an extracellular matrix-derived composition, a basement membrane extract, a decellularized extracellular matrix, a solubilized basement membrane preparation, a mixture of extracellular matrix proteins, a tumor-derived composition, Matrigel, or hyaluronic acid, or any combination thereof.

79. 79. The fibrosis diagnostic system of any one of claims 1 to 78, wherein the substrate comprises silicon and one or more channel surfaces are coated with a coating comprising silane.

80. 80. A fibrosis diagnostic system according to any one of claims 1 to 79, wherein one or more channel surfaces are subjected to silanization.

81. The fibrosis diagnostic system of any one of claims 1 to 80, wherein one or more channel surfaces are treated with plasma.

82. The fibrosis diagnostic system of any one of claims 1 to 81, wherein the plasma contains oxygen.

83. 83. The fibrosis diagnostic system of any of claims 1 to 82, wherein the one or more coatings are applied by emulsion, spraying, brushing, injection, precipitation, polymerization, or vapor deposition, or any combination thereof.

84. A fibrosis diagnostic system according to any preceding claim, wherein one or more channel surfaces comprise a laminate.

85. 85. The fibrosis diagnostic system of any of claims 1 to 84, wherein the device housing comprises a transparent material, a translucent material, a tinted material, a colored material, a polarized material, a material that selectively transmits wavelengths of electromagnetic radiation, a material that selectively refracts wavelengths of electromagnetic radiation, a material that selectively reflects wavelengths of electromagnetic radiation, or an opaque material, or any combination thereof.

86. A fibrosis diagnostic system according to any preceding claim, wherein the device housing comprises a window allowing visibility into the interior of the device.

87. A fibrosis diagnostic system according to any preceding claim, wherein the field of view is of one or more of the plurality of channels.

88. 88. A fibrosis diagnostic system according to any preceding claim, wherein the field of view includes a field of view of the first channel, or the target object, or both.

89. A fibrosis diagnostic system according to any preceding claim, wherein the device housing comprises multiple windows allowing multiple views of the interior of the device.

90. The fibrosis diagnostic system of any one of claims 1 to 89, wherein the target object comprises an organic material, an inorganic material, or both.

91. The fibrosis diagnostic system of any one of claims 1 to 90, wherein the target object comprises tissue, or a medical device, or both.

92. The fibrosis diagnostic system of any of claims 1 to 91, wherein the target object comprises a spheroid, an organoid, an alginate-based biomaterial, a scaffold-supported cellular structure, or a bioprinted structure, or any combination thereof.

93. 93. The fibrosis diagnostic system of any of claims 1 to 92, wherein the target objects comprise neoplastic cells, malignant cells, or benign cells, or any combination thereof.

94. The fibrosis diagnostic system of any one of claims 1 to 93, wherein the target comprises one or more coatings.

95. A fibrosis diagnostic system according to any preceding claim, wherein the one or more coatings of the target are different from the substrate.

96. A fibrosis diagnostic system as described in any one of claims 1 to 95, wherein the one or more coatings on the target differ from one or more coatings on one or more channel surfaces in terms of material composition, thickness, or patterning, or a combination thereof.

97. A fibrosis diagnostic system as described in any one of claims 1 to 96, wherein the one or more coatings of the target are the same as one or more coatings of one or more channel surfaces in terms of material composition, thickness, or patterning, or a combination thereof.

98. 98. A fibrosis diagnostic system according to any preceding claim, wherein the target is coated with a plurality of different materials.

99. 99. The fibrosis diagnostic system of any of claims 1 to 98, wherein the target is coated with multiple coatings.

100. 100. A fibrosis diagnostic system according to any preceding claim, wherein at least two of said plurality of coatings differ in material composition, thickness, or patterning, or a combination thereof.

101. A fibrosis diagnostic system as described in any one of claims 1 to 100, wherein the multiple coatings comprise a first coating comprising a first material adjacent to the target and a second coating comprising a second material overlying the first coating.

102. 102. The fibrosis diagnostic system of any of claims 1 to 101, wherein the target is coated prior to placement in the microfluidic device.

103. The fibrosis diagnostic system of any one of claims 1 to 102, wherein the substrate comprises a hydrogel.

104. The fibrosis diagnostic system of any of claims 1 to 103, wherein the matrix comprises fibronectin, an extracellular matrix-derived composition, a basement membrane extract, an extracellular matrix, a decellularized extracellular matrix, a solubilized basement membrane preparation, a mixture of extracellular matrix proteins, a tumor-derived composition, Matrigel, or hyaluronic acid, or any combination thereof.

105. The fibrosis diagnostic system of any of claims 1 to 104, wherein the substrate comprises a natural polymer, an artificial polymer, or a cross-linked polymer, or any combination thereof.

106. The fibrosis diagnostic system of any one of claims 1 to 105, wherein the substrate comprises a polysaccharide, a polypeptide, a lipid, a polynucleotide, hyaluronic acid, hyaluronic acid methacrylate, hyaluronic acid diacrylate, fibronectin, fibrinogen, collagen, or methacrylated collagen, or any combination thereof.

107. The fibrosis diagnostic system of any one of claims 1 to 106, wherein the first biological composition comprises fibroblasts, or endothelial cells, or both.

108. The fibrosis diagnostic system of any one of claims 1 to 107, wherein the first biological composition comprises serum, ascorbic acid, retinoic acid, proline, or bleomycin, or any combination thereof.

109. The fibrosis diagnostic system of any of claims 1 to 108, wherein the first biological composition comprises a fibrosis-promoting biomolecule, a complement protein, a blood coagulation-related protein, a bacterial cell wall protein, or lipopolysaccharide (LPS), or any combination thereof.

110. The fibrosis diagnostic system of any one of claims 1 to 109, wherein the second biological composition comprises at least one type of biological cell capable of promoting fibrosis.

111. The fibrosis diagnostic system of any of claims 1 to 110, wherein the second biological composition comprises a factor that promotes differentiation of a biological cell into a biological cell type that can promote fibrosis, a factor that promotes expression of a biological molecule that promotes fibrosis, or a factor that promotes proliferation of a biological cell type that can promote fibrosis, or any combination thereof.

112. The fibrosis diagnostic system of any of claims 1 to 111, wherein the second biological composition comprises serum, platelets, leukocytes, macrophages, monocytes, peripheral blood mononuclear cells (PBMCs), stem cells, mast cells, dendritic cells, neutrophils, granulocytes, T cells, or B cells, or any combination thereof.

113. A fibrosis diagnostic system according to any one of claims 1 to 112, wherein the third biological composition comprises a population of molecules capable of forming a gradient radiating from the third channel into the first channel or into both the first channel and the second channel.

114. The fibrosis diagnostic system of any of claims 1 to 113, wherein the third biological composition comprises a cytokine, a chemokine, a cell nutrient, a transcription factor, or a growth factor, or any combination thereof.

115. The fibrosis diagnostic system of any one of claims 1 to 114, wherein the third biological composition comprises CCL-2 / MCP-1, CCL3, CCL11, fibroblast growth factor (FGF), interferon, IFN-gamma, interleukin (IL), IL-4, IL-1α, IL-1β, IL-2, IL-5, IL-6, IL-7, IL-9, IL-13, IL-17A, IL-17F, IL-21, IL-22, IL-25, platelet-derived growth factor (PDGF), TGF-alpha, TGF-β, granulocyte colony-stimulating factor (G-CSF), macrophage colony-stimulating factor (M-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), prothrombin, or thrombin, or any combination thereof.

116. 116. The fibrosis diagnostic system of any of claims 1 to 115, wherein the at least one type of molecule of the gradient comprises a cytokine, a chemokine, a cell nutrient, a transcription factor, or a growth factor, or any combination thereof.

117. 117. A fibrosis diagnostic system according to any preceding claim, wherein said at least one type of molecule of said gradient comprises transforming growth factor beta (TGF-β).

118. The fibrosis diagnostic system of any one of claims 1 to 117, wherein the TGF-β comprises TGF-β1.

119. The fibrosis diagnostic system of any of claims 1 to 118, wherein the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, comprises labeled cells, or labeled biomolecules, or both.

120. The fibrosis diagnostic system of any of claims 1 to 119, wherein the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, comprises a compound with known or potential anti-fibrotic efficacy.

121. The fibrosis diagnostic system of any of claims 1 to 120, further comprising the substrate, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof.

122. A fibrosis diagnostic system according to any preceding claim, further comprising the target object.

123. The fibrosis diagnostic system of any of claims 1 to 122, further comprising a gradient of at least one molecule across the first channel, the second channel, or the third channel, or any combination thereof.

124. A fibrosis diagnostic system as described in any one of claims 1 to 123, wherein one or more components of the substrate, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, are attached to a channel surface.

125. A fibrosis diagnostic system according to any preceding claim, further comprising a detector, said detector comprising a sensor configured to detect fibrosis or characteristics indicative thereof in said target object.

126. A fibrosis diagnostic system according to any preceding claim, wherein the detector is configured to receive the microfluidic device.

127. A fibrosis diagnostic system according to any preceding claim, wherein the detector comprises a detector housing comprising a detector interior.

128. A fibrosis diagnostic system according to any preceding claim, wherein the detector further comprises a receptacle configured to receive the microfluidic device, the receptacle comprising an opening within the detector housing.

129. 129. The fibrosis diagnostic system of any of claims 1 to 128, further comprising a thermal subsystem.

130. 130. The fibrosis diagnostic system of any of claims 1 to 129, wherein the thermal subsystem comprises a heater, or a cooler, or both.

131. 131. The fibrosis diagnostic system of any of claims 1 to 130, wherein the thermal subsystem is configured to regulate the temperature of the microfluidic device.

132. 132. The fibrosis diagnostic system of any preceding claim, wherein the detector comprises the thermal subsystem.

133. 133. The fibrosis diagnostic system of any of claims 1-132, wherein the thermal subsystem is configured to regulate the temperature of the microfluidic device when in the receptacle.

134. 134. A fibrosis diagnostic system according to any preceding claim, further comprising a fluidic subsystem configured to be in fluid communication with the microfluidic device.

135. 135. The fibrosis diagnostic system of any of claims 1 to 134, wherein the fluidic subsystem is configured to load one or more components into the microfluidic device.

136. 136. A fibrosis diagnostic system according to any preceding claim, wherein the fluidic subsystem is configured to deliver a solid, liquid, or gas, or any combination thereof, to the microfluidic device.

137. 137. The fibrosis diagnostic system of any of claims 1 to 136, wherein the fluidic subsystem is configured to load the target object, the substrate, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, into the microfluidic device.

138. 138. A fibrosis diagnostic system according to any preceding claim, wherein the fluidic subsystem is configured to supply one or more nutrients to the microfluidic device.

139. A fibrosis diagnostic system according to any preceding claim, wherein the fluidic subsystem is configured to regulate pressure within the microfluidic device.

140. 140. A fibrosis diagnostic system according to any preceding claim, wherein the fluidic subsystem is configured to regulate a gradient across the first channel, the second channel, or the third channel, or any combination thereof.

141. 141. A fibrosis diagnostic system according to any preceding claim, wherein the fluidic subsystem is configured to remove one or more components from the microfluidic device.

142. 142. A fibrosis diagnostic system according to any preceding claim, wherein the detector comprises the fluidic subsystem.

143. 143. A fibrosis diagnostic system according to any preceding claim, wherein the fluidic subsystem is configured to be in fluid communication with the microfluidic device when the microfluidic device is within the receptacle.

144. A fibrosis diagnostic system according to any preceding claim, wherein the sensor is configured to detect particles, or waves, or both, emitted from the target object, or the microfluidic device, or both, that are indicative of fibrosis.

145. A fibrosis diagnostic system according to any preceding claim, wherein the sensor is configured to detect electromagnetic radiation.

146. 146. A fibrosis diagnostic system according to any preceding claim, wherein the electromagnetic radiation comprises ultraviolet light, visible light, or infrared light, or any combination thereof.

147. A fibrosis diagnostic system according to any preceding claim, wherein the electromagnetic radiation comprises fluorescence, or phosphorescence, or both.

148. A fibrosis diagnostic system according to any preceding claim, wherein the detector further comprises a particle source, or a wave source, or both, and is configured to transmit particles, waves, or both to the target object, or the microfluidic device, or both.

149. A fibrosis diagnostic system according to any preceding claim, further comprising an electromagnetic radiation source.

150. A fibrosis diagnostic system according to any preceding claim, wherein the detector comprises an electromagnetic radiation source.

151. 151. A fibrosis diagnostic system according to any preceding claim, wherein the electromagnetic radiation source comprises a light emitting diode, a laser, an incandescent light source, an arc light source, a halogen light source, a chemical vapor light source, or a fluorescent light source, or any combination thereof.

152. 152. A fibrosis diagnostic system according to any preceding claim, wherein the detector further comprises an optical subsystem configured to pass radiation, affect radiation, or both.

153. A fibrosis diagnostic system according to any preceding claim, wherein the optical subsystem comprises one or more lenses, one or more filters, one or more mirrors, or one or more diffraction gratings, or any combination thereof.

154. A fibrosis diagnostic system according to any preceding claim, wherein the optical subsystem, or the sensor, or both, comprises a microscope or one or more components thereof.

155. A fibrosis diagnostic system according to any preceding claim, wherein the optical subsystem comprises the sensor, or the electromagnetic radiation source, or both.

156. A fibrosis diagnostic system according to any preceding claim, wherein the sensor is configured to generate an output signal based on an input signal detected by the sensor.

157. A fibrosis diagnostic system according to any preceding claim, wherein the detector further comprises a transmitter, receiver, transceiver, or any combination thereof, configured to receive information, transmit information, or both.

158. A fibrosis diagnostic system according to any preceding claim, wherein the information comprises the output signal.

159. A fibrosis diagnostic system according to any preceding claim, wherein the information comprises an operational signal for operation of the sensor.

160. A fibrosis diagnostic system according to any preceding claim, wherein the detector further comprises a data port.

161. 161. A fibrosis diagnostic system according to any preceding claim, further comprising a processor configured to receive and process information.

162. A fibrosis diagnostic system according to any preceding claim, wherein the processor is configured to control one or more subsystems or components of the fibrosis diagnostic system.

163. A fibrosis diagnostic system according to any preceding claim, wherein the processor is configured to control the sensor, the electromagnetic radiation source, the optical subsystem, the thermal subsystem, or the fluidic subsystem, or any combination thereof.

164. A fibrosis diagnostic system according to any preceding claim, wherein the processor is configured to receive the signal output from the sensor, receive an operating signal from a user input, or both.

165. A fibrosis diagnostic system according to any preceding claim, wherein the detector comprises the processor.

166. A fibrosis diagnostic system according to any preceding claim, further comprising a memory configured for communication with the processor.

167. A fibrosis diagnostic system according to any preceding claim, wherein the memory is configured to store a program that controls the fibrosis diagnostic system or a component thereof.

168. A fibrosis diagnostic system according to any preceding claim, wherein the processor comprises the memory.

169. A fibrosis diagnostic system according to any preceding claim, wherein the detector comprises the memory.

170. A fibrosis diagnostic system according to any preceding claim, further comprising a user interface.

171. The fibrosis diagnostic system of any of claims 1 to 170, wherein the user interface comprises a monitor, buttons, a keyboard, a mouse, an optical signal, a camera, an eyepiece, a speaker, or a microphone, or any combination thereof.

172. A fibrosis diagnostic system according to any preceding claim, wherein the detector comprises the user interface.

173. A fibrosis diagnostic system according to any preceding claim, further comprising a power source, or a power interface, or both.

174. A fibrosis diagnostic system according to any preceding claim, wherein the detector comprises the power source, or the power interface, or both.

175. A fibrosis diagnostic system according to any preceding claim, further comprising a kit, said kit comprising said microfluidic device and one or more reagents for use with said microfluidic device.

176. Further comprising a kit, said kit comprising: the microfluidic device; The fibrosis diagnostic system of any of claims 1 to 175, comprising the substrate, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof.

177. 1. A method of analyzing a target object for susceptibility to fibrosis, comprising: disposing the target object, the substrate, and the first biological composition in the first channel of a microfluidic device according to any one of claims 1 to 176; disposing the second biological composition in the second channel of the microfluidic device; disposing the third biological composition in the third channel of the microfluidic device; analyzing the target object, or the first channel, or both, to determine the presence or absence of fibrosis associated with the target object.

178. 178. The method of any of claims 1-177, wherein the analysis is performed a predetermined period of time after placing the target object, the substrate, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, into the microfluidic device.

179. A method according to any preceding claim, wherein the predetermined period of time is sufficient to allow formation of the gradient of at least one type of molecule.

180. 180. The method of any preceding claim, wherein the analysis is performed more than once.

181. A method according to any preceding claim, wherein any two consecutive analyses are separated by a predetermined period of time.

182. The method of any preceding claim, wherein said analyzing comprises removing one or more components from said microfluidic device.

183. The method of any preceding claim, wherein the assay comprises an immunoassay.

184. The method of any of claims 1 to 183, wherein said analyzing comprises the use of an antibody.

185. The method of any one of claims 1 to 184, wherein the antibody is labeled.

186. 186. The method of any preceding claim, wherein said analyzing comprises application of a primary antibody and a secondary antibody that comprises a detectable label and that binds to said primary antibody.

187. The method of any of claims 1-186, wherein the primary antibody binds to collagen, alpha smooth muscle actin, or both.

188. The method of any preceding claim, wherein the analysis comprises a histological assay.

189. The method of any preceding claim, wherein the analysis comprises application of Picrosirius Red dye.

190. A method according to any preceding claim, wherein the analyzing step comprises visually inspecting the target object, or the first channel, or both, for one or more indications of fibrosis associated with the target object.

191. The method of any preceding claim, wherein said analyzing comprises using a microscope.

192. The method of any preceding claim, wherein said analyzing comprises using one or more spectroscopic techniques.

193. 193. The method of any of claims 1-192, wherein the analysis comprises nonlinear optical (NLO) multiphoton microscopy, second harmonic generation (SHG) imaging, third harmonic generation (THG) imaging, two-photon excited fluorescence (TPEF), two-photon excited fluorescence (TPEF), cell tracker dye imaging, visible light imaging, (ELISA), Griess assay, histopathology, immunostaining, cleared tissue microscopy, or proteomic profiling of homogenized hydrogels, or any combination thereof.

194. 194. The method of any preceding claim, wherein said analyzing comprises using said detector to detect one or more indications of fibrosis associated with said target object.

195. The method of any preceding claim, wherein said analyzing comprises inserting said microfluidic device into said receptacle.

196. 200. The method of any of claims 1-195, further comprising preparing the target object, the substrate, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, from a patient sample.

197. The method of any preceding claim, further comprising implanting the target object in or on a patient.

198. The method of any preceding claim, further comprising implanting a medical device, or tissue, or both, into or onto a patient, wherein the target object is a representative model of the medical device, or tissue, or both.

199. The method of any preceding claim, further comprising removing a medical device, or tissue, or both, from the patient, wherein the target object is a representative model of the medical device, or tissue, or both.

200. 200. The method of any preceding claim, further comprising administering to the patient an anti-fibrotic agent.

201. The method of any preceding claim, wherein said analyzing comprises determining a quantitative measure of fibrosis.

202. 202. The method of any preceding claim, wherein the dosage of anti-fibrotic agent administered to the patient is based on the quantitative measure of fibrosis.

203. The method of any of claims 1 to 202, wherein the anti-fibrotic agent is administered to the patient before, simultaneously with, or after implantation of the target object, tissue associated with the target object, or a medical device associated with the target object, or any combination thereof.

204. The method of any preceding claim, further comprising administering to the patient one or more therapeutic agents in addition to or in the alternative to the anti-fibrotic agent.

205. 205. The method of any of claims 1 to 204, wherein the patient is a mammalian patient, or a human patient, or both.

206. 1. A method of screening a therapy for effectiveness in treating fibrosis, said method comprising: placing the target object, the substrate, and the first biological composition in the first channel of a microfluidic device according to any one of claims 1 to 205; disposing the second biological composition in the second channel of the microfluidic device; disposing the third biological composition in the third channel of the microfluidic device; administering the therapy on the microfluidic device; and analyzing the target object, or the first channel, or both, to determine the presence, absence, or extent of fibrosis associated with the target object; determining the effectiveness of said therapy based on the presence, absence, or degree of said fibrosis without administering said therapy.

207. The method of any of claims 1-206, wherein the therapy comprises a therapeutic compound, and the performing comprises placing the therapeutic compound in the first channel, the second channel, or the third channel, or any combination thereof.

208. The method of any of claims 1-207, wherein the therapeutic compound comprises a pharmaceutical compound, or a biologic, or both.

209. The method of any preceding claim, wherein the therapy comprises modification of the target object.

210. The method of any of claims 1-209, wherein the therapy comprises placing a medical device in the first channel, the second channel, or the third channel, or any combination thereof.

211. The method of any preceding claim, wherein the therapy comprises irradiating the microfluidic device.

212. The method of any of claims 1 to 211, wherein said therapy further comprises treating a patient with said screened therapy.

213. 1. A method of screening a therapy for effectiveness in treating cancer, said method comprising: disposing the target object, the substrate, and the first biological composition into the first channel of the microfluidic device of any one of claims 1 to 212, wherein the target object comprises a neoplastic cell; disposing the second biological composition in the second channel of the microfluidic device; disposing the third biological composition in the third channel of the microfluidic device; administering the therapy on the microfluidic device; and analyzing the target object, or the first channel, or both, to determine the presence, absence, or extent of fibrosis associated with the target object; determining the effectiveness of said therapy based on the presence, absence, or severity of cancer without administering said therapy.

214. A non-transitory computer readable medium storing a program including instructions for performing the method of any of claims 1 to 213.

215. A non-transitory computer readable medium storing a program including instructions for controlling a fibrosis diagnostic system according to any one of claims 1 to 214, or a subsystem thereof, or a component thereof, or any combination thereof.

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