Methods and apparatus for additive manufacturing of digitally weaved BIO-textiles through continuous fiber extrusion

Abstract

Methods of making a woven three-dimensional silk material include selecting a digital design parameter set comprising at least one woven three-dimensional silk material property, and weaving, via coextrusion, a silk fibroin material and a continuous fiber according to the digital design parameter set to form a woven three-dimensional silk material possessing the at least one woven three-dimensional silk material property. Silk leathers that are layered structures comprising at least one woven three-dimensional silk material may be made from the disclosed methods.

Description

PATENT Attorney Docket No. T002896 WO -2095.0733 METHODS AND APPARATUS FOR ADDITIVE MANUFACTURING OF DIGITALLY WEAVED BIO-TEXTILES THROUGH CONTINUOUS FIBER EXTRUSION CLAIM TO PRIORITY

[0001] This application relates to, incorporates by reference for all purposes, and claims priority to United States Application Serial Number 63 / 756,507 filed on February 10, 2025, and United States Application Serial Number 63 / 821,840 filed on June 11, 2025.BACKGROUND

[0002] Presently, the fashion and textile industry generated a staggering 1.23 billion tons of waste in 2019 with projections estimating up to 2.23 billion in 2025. This includes silk-based waste, which accounts for 11 million tons, much of which remains unrecycled. Historically, the delicacy of printed silk foam structures has impeded their integration into fabric production despite their numerous advantages. A need exists for new materials overcoming these challenges.SUMMARY

[0003] In some aspects, the techniques described herein relate to the method of the immediately preceding claim, wherein decreasing at least one of the density of lines or the number of 3D printed layers, and / or modifying the weaving pattern, increases the flexibility of the woven three-dimensional silk material, the compressed woven three-dimensional silk material, or the cured compressed woven three-dimensional silk material.

[0004] In some aspects, the techniques described herein relate to a method including: a) pre-treating a thread to enhance mechanical strength; and b) co-extruding the pre-treated thread with a biopolymer to form a woven three-dimensional material.

[0005] These and other systems, methods, objects, features, and advantages of the present disclosure will be apparent to those skilled in the art from the following detailed description of the preferred embodiment and the drawings.

[0006] All documents mentioned herein are hereby incorporated in their entirety by reference. References to items in the singular should be understood to include items in the plural, and vice versa, unless explicitly stated otherwise or clear from the text. Grammatical conjunctions are intended to express any and all disjunctive and conjunctive combinations of conjoined clauses, sentences, words, and the like, unless otherwise stated or clear from the context.BRIEF DESCRIPTION OF THE FIGURES

[0007] The disclosure and the following detailed description of certain embodiments thereof may be understood by reference to the following figures:

[0008] Fig. 1 depicts an exemplary 3D-printed woven-like biocomposite consisting of a triaxial woven-like pattern printed with a coated silk fiber and a silk foam.PATENT Attorney Docket No. T002896 WO -2095.0733

[0009] Fig. 2A depicts a top-down view of an exemplary fiber structure formed of one continuous engineered fiber. The fiber is triaxially interwoven to form a hexagonal rattan pattern. The structure is simultaneously printed with the base material (foam) and then compressed to achieve woven-like mechanical properties.

[0010] Fig. 2B depicts a tilted view of an exemplary fiber structure formed of one continuous engineered fiber. The fiber is triaxially interwoven to form a hexagonal rattan pattern. The structure is simultaneously printed with the base material (foam) and then compressed to achieve woven-like mechanical properties.

[0011] Fig. 3 shows an increasing number of locked intersections from a single yarn (left) to two yarns (middle) to a triaxially-woven pattern (right).

[0012] Fig. 4A depicts a close-up view of a 3D-printed leather-like biocomposite.

[0013] Fig. 4B shows a top-down view of a 3D-printed leather-like biocomposite.

[0014] Fig. 5 depicts two views of a computationally-designed upper shoe textile.

[0015] Fig. 6A shows Scanning Electron Microscopy (SEM) of 15 pure silk fibers held together by a latex coating. Scale bar is 10 pm.

[0016] Fig. 6B shows Scanning Electron Microscopy (SEM) of 15 pure silk fibers held together by a latex coating. Scale bar is 40 pm.

[0017] Fig. 7 shows an exemplary coating device.

[0018] Fig. 8 depicts the additive manufacturing process. The left inset shows an exemplary extruded beam including silk fibers coated with a silk-based foam. The right inset shows the co-extrusion printhead and extruded filament.

[0019] Fig. 9A, Fig. 9B, and Fig. 9C depict a 25 cm × 25 cm soft composite article.

[0020] Fig. 10 shows triaxial (left), biaxial grid 0° (middle), and biaxial grid 45° (right) weavings.

[0021] Fig. 11 shows ultimate tensile strength and fracture strain measurements of 40:20:40 braided cotton and PU synthetic leather.

[0022] Fig. 12A and Fig. 12B show the impact of fiber-to-matrix ratio on fracture strain (Fig. 12A) and ultimate tensile strength (Fig. 12B) as a function of grid resolution.

[0023] Fig. 13A and Fig. 13B show the impact of fiber-to-matrix ratio on stress and fracture strain (Fig. 13A) and stress / strain curves as a function of fiber volume fractions and angle (Fig. 13B).

[0024] Fig. 14A shows the impact of weaving type on ultimate tensile strength and fracture strain.

[0025] Fig. 14B shows the impact of fibers in composites on ultimate tensile strength and fracture strain.

[0026] Fig. 14C shows the impact of material type on ultimate tensile strength and fracture strain.PATENT Attorney Docket No. T002896 WO -2095.0733 DETAILED DESCRIPTION

[0027] Before the present disclosure is described in further detail, it is to be understood that the disclosure is not limited to the particular embodiments described. It is also understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting. The scope of the present disclosure will be limited only by the claims. As used herein, the singular forms "a", "an", and "the" include plural embodiments unless the context clearly dictates otherwise.

[0028] In this application, unless otherwise clear from context, (i) the term “a” may be understood to mean “at least one”; (ii) the term “or” may be understood to mean “and / or”; (iii) the terms “comprising” and “including” may be understood to encompass itemized components or steps whether presented by themselves or together with one or more additional components or steps; and (iv) the terms “about” and “approximately” are used as equivalents and may be understood to permit standard variation as would be understood by those of ordinary skill in the art; and (v) where ranges are provided, endpoints are included.

[0029] The present disclosure provides methods for designing and printing three-dimensional printed mono-material biotextiles as well as the biotextiles themselves (Fig. 1). These biotextiles may be referred to as articles or 3D-printed articles. The methods described herein facilitate the fabrication of continuous fiber-reinforced 3D-printed biocomposites.

[0030] Silk Foam

[0031] Silk foam, as described herein and by U. S. Provisional Patent Application No. 63 / 808,348 and PCT International Application Nos. PCT / US2023 / 083663, PCT / US2025 / 017896, PCT / US2025 / 033223, PCT / US2025 / 026472, PCT / US2025 / 016857, and PCT / US2025 / 033230 which are hereby incorporated by reference in their entireties for all purposes, possesses intriguing structural and physicochemical properties. The compositions described herein are based on the use of an aqueous silk fibroin (SF) solution or SF powder in conjunction with glycerol, xanthan gum from Xanthomonas Campestris, and / or alginic acid sodium salt from brown algae. As an added benefit, silk fibroin is renowned for its capacity to stabilize biological molecules and colorimetric sensors. The potential to 3D print foam structures with high porosity and surface area is likely to augment the sensitivity and versatility of other sensor technologies, particularly in gas sensing applications.

[0032] Weaving and Digital Weaving

[0033] The method and system described herein allow the creation of warp-weft-like or woven-like biofabrics by the simultaneous co-extrusion of a silk-based biopolymer (e.g., a silk foam or a paste) and a continuous fiber or cord. In some embodiments, this is made possible by layer deposition in orthogonal directions until a microarchitecture including the biopolymer and the continuous fiber isPATENT Attorney Docket No. T002896 WO -2095.0733 obtained. The 3D structure is then compressed cross-link through thermal, mechanical and / or chemical bonds. By doing so, this process enables the creation of weaved-like textiles digitally and three-dimensionally-designed. Fig. 2A and Fig. 2B show an exemplary fiber structure formed of one continuous engineered fiber. The fiber is triaxially interwoven to form a pattern (e.g., a hexagonal rattan pattern). The structure is being printed simultaneously with a base material (foam) and then compressed to achieve woven-like mechanical properties.

[0034] This computational workflow allows for function-driven textiles by digitally tuning yarn geometry and density. It can create biaxial (flexible) and triaxial (stiff) weaves with structural integrity through locked intersections. Programmable parameters, including filament type, size, and resolution, allow tailored mechanical performance, with cross-linking achieved via heat, pressure, or chemistry. This approach leads to the fabrication of sustainable, monomaterial composites with inherent strength and tunability. Drawing on natural structural efficiency and circular design principles, it offers a compelling alternative to synthetic composites and animal-derived leathers.

[0035] Impressively, this disclosure makes it possible to create different types of textiles comprising a continuous either biaxially or triaxially interweaved fiber (Fig. 3) over several layers ranging from loop-like textiles to synthetic leathers (Fig. 4A and Fig. 4B). In this way, an infinite number of patterns can be created, including negative Poisson's ratio patterns such as auxetics. This method also makes it possible to computationally generate textiles whose properties gradually vary according to need. These gradients can be enabled within the same sample by modifying the properties of the biopolymer and the continuous fiber. Exemplary, non-limiting properties that may vary with gradients include color, stiffness, porosity, softness, elasticity, tensile strength, and pattern.

[0036] Exemplary, non-limiting weaving patterns that may be selected as part of a digital design parameter set include a rattan pattern, a hexagonal rattan pattern, a Dutch weave, a twill weave, a plain weave, a twill Dutch weave, a lock crimp, an inter-crimp, a twill Dutch double, stranded, a basket wave, a plait, a Poisson’s ratio pattern, a triaxial weave pattern, a biaxial weave pattern, a warp-weft pattern, a satin weave, or combinations thereof. Three exemplary weaving types are shown in Fig. 10 - triaxial (left), biaxial grid 0° (middle), and biaxial grid 45° (right) weavings.

[0037] Methods of Making

[0038] The methods described herein may produce a wide range of articles, including but not limited to, a traditional one-piece upper shoe (Fig. 5) which may be assembled using traditional shoe-making techniques. Additionally or alternatively, the method may allow for direct 3D printing on a shoe using a 6-axis robotic arm, allowing for direct custom assembly of the woven textile into a desired shape.

[0039] In some embodiments, a method of making a woven three-dimensional silk material includes steps a) and b). Step a) includes selecting a digital design parameter set comprising at least one silkPATENT Attorney Docket No. T002896 WO -2095.0733 material property. Step b) includes weaving, via coextrusion, silk fibroin material and a continuous fiber according to the digital design parameter set from step a) to form a woven three-dimensional silk material, which may be referred to as woven material, with the at least one woven three-dimensional silk material property.

[0040] In some embodiments, the method may further include step c) compressing the woven three-dimensional silk material to form a compressed woven three-dimensional silk material, which may be referred to as compressed material. Additionally or alternatively, the method may further include steps d) and e). Step d) includes selecting a curing condition set for the woven three-dimensional silk material or compressed woven three-dimensional silk material. Step e) includes curing the woven three-dimension silk material or compressed woven three-dimensional silk material to form a cured woven three-dimensional silk material or cured compressed woven three-dimensional silk material, which may be referred to as cured material. Additionally or alternatively, the method may further comprise steps f) and g). Step f) includes applying at least one layer of silk foam to at least one woven material, compressed material, or cured material. Step g) includes optionally compressing or thermally compressing the at least one layer of silk foam.

[0041] The methods described herein may result in no chemical waste and / or solvent waste.

[0042] Digital Design Parameters

[0043] The disclosure herein provides for a digital design parameter set used in the weaving of three-dimensional silk materials. Exemplary, non-limiting digital design parameters include a weaving pattern, a number of printed layers, a density of lines, a number of intersections, an angle of vertices at intersections, a Poisson’s ratio, a number of layers of printing, a shape, a selection of the continuous fiber material, a selection of the silk fibroin material, a selection of a silk fibroin material property, a temperature of extrusion, a rate of extrusion, a fan speed, a printing speed, a biaxial interweave, a triaxial interweave, a yarn size, a pitch distance, a desired flexibility, or combinations thereof. A skilled artisan will appreciate that the interplay between digital design parameters may inform selection.

[0044] The digital design parameter set may also include instructions to vary one or more of the digital design parameters throughout the coextrusion. These instructions may result in at least one gradient of color, stiffness, porosity, softness, elasticity, tensile strength, pattern, extensibility, or a combination thereof.

[0045] Silk Fibroin Material

[0046] The silk fibroin material may include at least one of a liquid composition, a whipped silk cream, or a silk meringue as described herein or in United States Patent Application number 18 / 403,477, which is hereby incorporated by reference for all purposes. The silk fibroin may be made from the liquid composition of or made by any one of claims 1 to 3, 24 to 31, 42 to 44, 65 to 72, 83 to 85, 106PATENT Attorney Docket No. T002896 WO -2095.0733 to 113, 124 to 126, 147 to 154, 170 to 176, or 181 to 188 of United States Patent Application 18 / 403,477 as filed. The silk fibroin may be made from the whipped silk cream of or made by the method of any one of claims 4, 8 to 32, 45, 49 to 73, 86, 90 to 114, 127, 131 to 155, or 165 to 188 of United States Patent Application 18 / 403,477 as filed. The silk fibroin may be made from the silk meringue of or made by the method of any one of claims 5, 9 to 32, 46, 50 to 73, 87, 91 to 114, 128, 132 to 155, or 165 to 188 of United States Patent Application 18 / 403,477 as filed.

[0047] In some embodiments, the silk fibroin material may include an additive as described herein. The additive may be a biological additive. The silk fibroin material may be a silk foam or a silk paste.

[0048] The methods and materials described herein provide for selecting a silk fibroin material property. Exemplary, non-limiting examples of silk fibroin material properties include an elastic modulus, an ultimate stress, a tensile toughness, an ultimate strain, a β-sheet content of the silk fibroin, a β-turn content of the silk fibroin, a transmittance, a color, a hardness, a stiffness, a conductivity, a resistance, a thermal conductivity, a permeability, or a permittivity.

[0049] Flexibility of Woven Articles

[0050] The stiffness of triaxial fabrics is primarily due to the locked intersections of the yarns. As seen in Fig. 3, the left panel shows a single red yarn restricted from moving upward by the intersection of the blue and grey yarns. When this intersection is expanded to two (middle panel), the blue yarn is constrained from moving vertically in either direction. Extending this locked intersection configuration to a complete unit (right panel) results in all the yams being restricted from moving in any direction. The performance of triaxial fabric, influenced by these locked intersections, depends on the yarn size and the pitch distance between them. The inventors discovered that by controlling these variables, they could a woven fabric with specific extensibility resulting in a truly engineered fabric structure.

[0051] In some embodiments, modifying the weaving pattern and / or decreasing the density of lines and / or the number of 3D-printed layers may increase the flexibility of the 3D-printed articles. Conversely, increasing the density of lines and / or number of 3D-printed layers may decrease the flexibility of 3D-printed articles.

[0052] In some embodiments, the at least one woven three-dimensional silk material property is a desired flexibility. The digital design parameter set may include the desired flexibility in a plurality of flexibility values. Each flexibility value within the plurality of flexibility values may pair with a corresponding value of line density for the weaving that results in the flexibility value, and without wishing to be bound by any particular theory, it is believed that the woven three-dimensional silk material behaves similarly to leather with respect to this property.

[0053] Compressing ArticlesPATENT Attorney Docket No. T002896 WO -2095.0733

[0054] In some cases, the present disclosure provides a method for making compressed articles. A method of making a compressed article can include compressing the uncompressed article with a force of between 0.25 MPa and 50 MPa for a length of time of between 15 minutes and 6 hours. In some cases, the compressing can be heat-compressing that is performed at an elevated temperature of between 80 °C and 200 °C, including at least 120 °C. In some cases, compressing can be performed without application of heat such as at ambient temperatures (20 - 25 °C). In some cases, the compressing can be performed with a calendar press or a hydraulic press. A method of making a compressed article can include compressing the uncompressed article with a force of between 0.25 MPa and 50 MPa for a length of time of between 5 minutes and 6 hours. A skilled artisan will recognize that the exact parameters of heat-compressing may vary outside of the specified parameters but still provide the desired outcome.

[0055] Compressing the woven material may result in at least one of thermal, mechanical, or chemical crosslinking within the compressed material.

[0056] Curing

[0057] Curing of articles may include curing the woven three-dimension silk material or compressed woven three-dimensional silk material to form a cured woven three-dimensional silk material or cured compressed woven three-dimensional silk material, which may be referred to as cured material. Curing conditions may include applying at least one of heat, a stream of air, light (e.g., ultraviolet light), electron beams, and / or additives. A curing condition set may include the method to be applied (e.g., heating) and a length of time to apply the method. The method may include more than one step (e.g., heating and light) to be carried out sequentially or simultaneously. A method with more than one step may have different curing lengths of time per step or the same curing length of time per step. A skilled artisan will appreciate that the exact curing method and conditions may depend on the composition of the article. Additives may include, without limitation, crosslinking agents, catalysts, photo-initiators, or combinations thereof. The curing conditions may be selected to produce uniform curing throughout the article or spatially graded curing to create regions having different mechanical or functional properties.

[0058] Coextrusion

[0059] The present disclosure provides for a printhead as described herein as well as in PCT / US2025 / 033223 filed on June 11, 2025, which is hereby incorporated by reference for all purposes. The printhead weaves via coextrusion. In some embodiments, the printhead includes a coaxial nozzle and a continuous fiber dispenser. The printhead is configured to co-extrude a silk fibroin material with a continuous fiber. Fig. 8 shows a schematic of one embodiment of a co-flow printhead.PATENT Attorney Docket No. T002896 WO -2095.0733 The coaxial or co-flow nozzle includes two ducts: a first duct for a silk fibroin material (labeled as biofoam filament) extrusion, and a continuous fiber duct for thread extrusion (labeled as silk fiber). The nozzle may be fabricated from biocompatible resins suitable for medical applications (e.g., BioMed Clear V1 resin).

[0060] The first duct has a fitting on a first end. The fitting may be any type of fitting with a protruding component and a receiving component. The fitting may be a screw-type fitting (e.g., a Luer lock fitting). Silk fibroin material flows through as shown by the large arrow. The continuous fiber duct is arranged at an incline with respect to the first duct. The continuous fiber duct is configured to receive and guide the continuous fiber. A second end of the continuous fiber duct is configured to receive the continuous fiber exiting the continuous fiber duct. A co-flow tip is arranged at the second end of the first duct and is configured to receive a co-extruded material for weaving. The silk fibroin material and the continuous fiber may be co-extruded in a single path. A diameter of the silk fibroin material may be greater than a diameter of the continuous fiber in the co-extruded material by between 5 times to 100 times, including at least 10 times, at least 30 times, or at least 50 times.

[0061] In some embodiments of the printhead, the printhead may comprise more than one nozzle for printing of more than one continuous fiber simultaneously. Printheads may also be coupled together to print more than one continuous fiber simultaneously. It should be understood that a person of skill in the art, having the benefit of this disclosure, would know when to couple printheads for certain applications, and further, if the coupled printheads should have the same or different features. It should be understood that coupled printheads may have the same or different features when compared to one another.

[0062] In some embodiments of the printhead, the printhead further includes a thread dispenser, which may also be referred to as a feeder. The thread dispenser transforms the rotational movement of wheels into a linear movement for the continuous fiber. In some cases, the wheels of the thread dispenser may be made of at least one of thermoplastic polyurethane (TPU), silicon rubber, polyurethane, thermoplastic polyethylene, or any suitable material that may be extruded or molded.

[0063] The thread dispenser is a highly precise, custom-made mechanism with several unique features that minimize disruption to the coextrusion, with three features described herein, i) The continuous fiber duct of the thread dispenser ensures deposition of the thread by propulsion, which is uncommon in thread deposition, ii) A precise pipe network prevents buckling effects by ensuring proper alignment of the continuous fiber, iii) The thread dispenser may be augmented with a sensor or sensor system that includes a sensor that detects any unwanted continuous fiber behavior (e.g., poor positioning of the continuous fiber or the continuous fiber deviating from the desired trajectory). If the sensor detects any undesirable behavior of the continuous fiber, the sensor may send a signal to the printer to pausePATENT Attorney Docket No. T002896 WO -2095.0733 printing or initiate a de-bottlenecking maneuver. The vertical alignment of the thread and the length of the continuous fiber duct are also carefully selected to minimize friction and constraints in the thread extrusion method. The sensor may be an infrared (IR) sensor or an optical sensor. For example, an optical sensor may be coupled with a computer executing a machine learning algorithm to detect the presence / absence of the continuous fiber.

[0064] In some embodiments, the continuous fiber may be at least one of a silk yarn, a waste silk product, a filament, an electrode, a counter-electrode, an enzymatically-coated fiber, a non-spun thread, an electrochemical fiber, a conductive fiber, a copper material, a platinum material, a metallic material, a polyester material, a cotton material, a graphite fiber, or a nylon material. A diameter of the continuous fiber may be between 35 pm and 80 pm or between 0.1 mm and 1.5 mm. A diameter of the continuous fiber may be at least 35 pm, at least 60 pm, at least 70 pm, at least 0.2 mm, at least 0.3 mm, at least 0.5 mm, at least 0.7 mm, at least 0.8 mm, or at least 1 mm. A diameter of the continuous fiber may be at most 1.5 mm, at most 1.3 mm, at most 1.1 mm, at most 1 mm, at most 0.6 mm, at most 0.4 mm, at most 80 pm, at most 65 pm, or at most 45 pm.

[0065] In some embodiments, the first duct of the printhead may include a pipe comprising copper and / or aluminum, or other suitable material. The first duct may lead the continuous fiber through the wheels and have a diameter between 1 mm and 2 mm, including at least 1.2 mm, at least 1.4 mm, at least 1.6 mm, or at least 1.8 mm.

[0066] The continuous fiber duct may have a funnel-shaped mouth with a diameter of between 2 mm and 5 mm. The diameter of the funnel-shaped mouth may be at least 2.5 mm, at least 3.5 mm, or at least 4.5 mm. The diameter of the funnel-shaped mouth may be at most 5 mm, at most 4 mm, or at most 3 mm. The continuous fiber duct may have a diameter between 0.1 mm and 1 mm including at least 0.2 mm, at least 0.3 mm, at least 0.5 mm, at least 0.7 mm, or at least 0.8 mm. The continuous fiber duct may be between 5 mm and 100 mm in length, including but not limited to, between 10 mm and 80 mm in length. The continuous fiber duct may be at least 10 mm in length, at least 30 mm in length, at least 50 mm in length, at least 70 mm in length, or at least 90 mm in length. The continuous fiber duct may be at most 95 mm in length, at most 75 mm in length, at most 55 mm in length, at most 35 mm in length, or at most 15 mm in length. The continuous fiber duct may have an angle of incline with respect to the first duct of between 5° and 45°. The angle of incline may be at least 5°, at least 15°, at least 25°, or at least 45°. The angle of incline may be at most 40°, at most 30°, at most 20°, or at most 10°.

[0067] In some embodiments, the printhead described herein may be configured to store the silk fibroin material under a storage pressure, which is optionally greater than atmospheric pressure. The storage pressure may be between 0.25 MPa and 25 MPa. The storage pressure may be at least 0.5 MPa,PATENT Attorney Docket No. T002896 WO -2095.0733 at least 1 MPa, at least 5 MPa, at least 10 MPa, or at least 20 MPa. The storage pressure may be at most 25 MPa, at most 15 MPa, at most 5 MPa, or at most 1 MPa.

[0068] In some embodiments, the printhead described herein may further include at least one vessel for delivering the silk fibroin material to the coextrusion nozzle. The at least one vessel may be configured to connect to the coextrusion nozzle via the fitting. The at least one vessel may be at least one of a cartridge or a syringe that is at least one of removable, refillable, or replaceable. The vessel may be configured to store a volume of the silk fibroin material under the storage pressure. The volume may be between 100 mL and 400 mL. The volume may be at least 100 mL, at least 150 mL, at least 250 mL, or at least 350 mL. The volume may be at most 400 mL, at most 300 mL, at most 200 mL, or at most 100 mL.

[0069] In some embodiments, the printhead described herein may include a mechanical extrusion system. The mechanical extrusion system may include a bipolar stepper motor with a gearbox and a rear axle configured to operate at a torque of between 1 Nm and 4 Nm, including but not limited to, at least 1.5 Nm, at least 2 Nm, or at least 3 Nm.

[0070] In some embodiments, the printhead described herein may include a ventilation system positioned adjacent to the co-extrusion nozzle. Airflow from the ventilation system is directed to the co-extruded material that is being extruded from the co-extrusion nozzle. The ventilation system may include at least one fan, which may be directed towards the co-extruded material being extruded to enable layer-by-layer drying of the co-extruded material. The fan may be directed at an airflow angle with respect to a direction of co-extrusion of between 35° and 65°, including but not limited to, at least 40°, at least 42°, at least 45°, or at least 47°. The fan may have a speed between 3,000 bpm and 4,000 bpm, including but not limited to, at least 3100 bpm, at least 3200 bpm, at least 3500 bpm, or at least 3800 bpm.

[0071] The ventilation system may be configured to deliver airflow at a predetermined fluid velocity optionally between 4 m / s and 10 m / s, including but not limited to, at least 5 m / s, at least 7 m / s, or at least 9 m / s. The predetermined fluid velocity may be variable and may vary with a change in distance from the ventilation system to a surface of the co-extruded material, an extrusion rate from the extrusion nozzle, a layer height, a length of path, an extrusion width, and / or a composition of the silk fibroin material.

[0072] The ventilation system may be between 60 mm and 80 mm from a tip of the coextrusion needle, including but not limited to, at least 65 mm, at least 70 mm, or at least 75 mm. The ventilation system may be between 60 mm and 100 mm from the co-extruded material, including but not limited to, at least 65 mm, at least 80 mm, or at least 95 mm.

[0073] Continuous FibersPATENT Attorney Docket No. T002896 WO -2095.0733

[0074] Exemplary, non-limiting continuous fiber materials include latex, silk cord, silk yarn, a waste silk product, a filament, an electrode, a counter-electrode, an enzymatically-coated fiber, a non-spun thread, an electrochemical fiber, a conductive fiber, a copper material, a platinum material, a metallic material, a polyester material, a cotton material, a graphite fiber, a nylon material, a light-emitting fiber element (e.g., electroluminescent wires, LEDs, or optical fibers), or reeled silk fibers. An exemplary continuous fiber is shown in Fig. 6A and Fig. 6B consisting of 15 pure silk fibers held together by a latex coating. The latex not only binds the fibers together, but also creates a protective layer for the fibers.

[0075] The continuous fiber may be coated with a coating composition to form a coated continuous fiber. Fig. 7 shows an exemplary apparatus that may be used to coat continuous fibers. Exemplary, non-limiting coating compositions include latex, silk fibroin solution, glycerol solution, aqueous gelatin (optionally Type A), alginate, or combinations thereof. Without wishing to be bound by any particular theory, it is believed that adding gelatin permits better printability (as evidenced by extrusion and shape fidelity) by improving the stabilization of the foam material during compression which preserves foam microstructure during extrusion.

[0076] In some embodiments, the coating composition may include a silk fibroin solution containing silk fibroin in a concentration between 0.5% and 40% or between 1% and 25%. The silk fibroin solution may contain silk fibroin in an amount of at least 0.5%, at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, or at least 25%. The silk fibroin solution may contain silk fibroin in an amount of at most 40%, at most 35%, at most 30%, at most 25%, at most 20%, or at most 15%.

[0077] A diameter of the continuous fiber may be between 35 pm and 1.5 mm, including but not limited to, between 35 pm and 80 pm or between 0.1 mm and 1.5 mm. A diameter of the continuous fiber may be at least 35 pm, at least 60 pm, at least 70 pm, at least 0.2 mm, at least 0.3 mm, at least 0.5 mm, at least 0.7 mm, at least 0.8 mm, or at least 1 mm. A diameter of the continuous fiber may be at most 1.5 mm, at most 1.3 mm, at most 1.1 mm, at most 1 mm, at most 0.6 mm, at most 0.4 mm, at most 80 pm, at most 65 pm, or at most 45 pm. The diameter of the continuous fiber may be at least partially a result of the coating composition.

[0078] A coated continuous fiber may possess a resistance to compression that is superior to a comparison resistance to compression of a comparison continuous fiber that lacks a coating but is otherwise identical to the coated continuous fiber. A coated continuous fiber may have a resistance to compression that is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150% greater than the comparison resistance of the comparison continuous fiber that lacks the coating but is otherwise identical to the comparison continuous fiber.PATENT Attorney Docket No. T002896 WO -2095.0733

[0079] Articles Made by Methods Described Herein

[0080] Articles made by any of the methods or processes described herein may be referred to generally as ‘articles’ or ‘3D-printed articles’. Exemplary, non-limiting articles produced herein may include a shoe, a shoe upper, a garment, a textile, an insulating material, an engineered composite, a metamaterial, a biofabric, a 3D structure, a structural material, a building material, an insulating material, a packaging material, a tool, a biomedical implant, a prosthetic, or an orthotic.

[0081] In some embodiments, the article may have a thickness of at least 0.1 mm, at least 0.2 mm, at least 0.3 mm, at least 0.4 mm, or at least 0.5 mm.

[0082] The article may include an article embedded component. The article embedded component may include at least one of a sensor, electronic component, electronic device, haptic switch, electrical wiring, power supply, RFID tag, or combinations thereof.

[0083] In some embodiments, textiles produced herein may include biaxial interweaved fibers and / or triaxial interweaved fibers. Triaxial interweaved fibers, often featuring a 0° / ±60° structure, can be generated with a continuous fiber, creating a stable, equilateral triangular pattern. Triaxial interweaved fibers, while generally being prepared from longer lengths of fibers relative to biaxial weaves, may feature high isotropic strength, superior shear / tear resistance, and minimal bias compared to biaxial weaves. Biaxial woven fibers, typically arranged in a 0° / 90° orientation, can be formed from a continuous fiber. This conventional weave architecture provides high strength and stiffness along the principal fiber directions but exhibits comparatively lower shear stability and greater susceptibility to distortion when subjected to off-axis loading. Biaxial woven fabrics are straightforward to manufacture and offer efficient unidirectional and bidirectional reinforcement, however, their anisotropic mechanical response may lead to increased bias deformation, reduced tear resistance, and less uniform load distribution relative to triaxial architectures. In some embodiments, biaxial or triaxial woven fabrics may be woven from more than one fiber, wherein the properties of the fibers may differ from one another and the properties of the formed fabric may likewise differ from fabrics formed from one continuous fiber.

[0084] In some embodiments, the article may include a triaxial weave pattern and have at least one of an isotropic response to tensile deformation, an isotropic response to shear deformation, an extensibility, or a combination thereof.

[0085] An exemplary article of 25 cm × 25 cm is shown in Fig. 9A, Fig. 9B, and Fig. 9C.

[0086] Mechanical Properties of Articles

[0087] In some embodiments, the articles described herein may have an elastic modulus of between 0.01 GPa and 200 GPa or between 0.1 GPa and 10 GPa. The elastic modulus may be at least 0.01 GPa, at least 0.05 GPa, at least 0.1 GPa, at least 0.5 GPa, at least 1 GPa, at least 3 GPa, at least 5 GPa, atPATENT Attorney Docket No. T002896 WO -2095.0733 least 7.5 GPa, at least 10 GPa, at least 20 GPa, at least 40 GPa, at least 60 GPa, at least 80 GPa, at least 100 GPa, or at least 200 GPa. The elastic modulus may be at most 50 GPa, at most 30 GPa, at most 10 GPa, at most 7 GPa, at most 5 GPa, at most 1 GPa, at most 0.5 GPa, or at most 0.01 GPa.

[0088] In some embodiments, the articles described herein may have an ultimate tensile strength between 0.1 MPa and 5,000 MPa or between 1 MPa and 500 MPa. The ultimate tensile strength may be at least 0.1 MPa, at least 0.3 MPa, at least 0.5 MPa, at least 0.7 MPa, at least 1 MPa, at least 10 MPa, at least 30 MPa, at least 50 MPa, at least 100 MPa, at least 300 MPa, at least 1000 MPa, at least 2,500 MPa, at least 4,000 Mpa, or at least 5000 MPa. The ultimate tensile strength may be at most 4,000 MPa, at most 1,000 MPa, at most 500 MPa, at most 400 MPa, at most 300 MPa, at most 200 MPa, or at most 100 MPa.

[0089] In some embodiments, the articles described herein may have a tensile toughness between 0.1 MJ / m3and 750.0 MJ / m3, or between 0.5 MJ / m3and 5.0 MJ / m3. The tensile toughness may be at least 0.1 MJ / m3, at least 0.3 MJ / m3, at least 0.5 MJ / m3, at least 0.7 MJ / m3, at least 1.0 MJ / m3, at least 1.5 MJ / m3, at least 2.0 MJ / m3, at least 2.5 MJ / m3, at least 20 MJ / m3, at least 100 MJ / m3, at least 250 MJ / m3, at least 400 MJ / m3, at least 500 MJ / m3, at least 600 MJ / m3, or at least 750 MJ / m3The tensile toughness may be at most 550 MJ / m3, at most 450 MJ / m3, at most 350 MJ / m3, at most 250 MJ / m3, at most 150 MJ / m3, at most 50 MJ / m3, at most 10.0 MJ / m3, at most 8 MJ / m3, at most 6 MJ / m3, at most 5 MJ / m3, at most 4 MJ / m3, at most 3 MJ / m3, or at most 2.5 MJ / m3.

[0090] In some embodiments, the articles described herein may have an ultimate strain of between 0.01% and 80% or between 0.1% and 50%. The ultimate strain may be at least 0.01%, at least 0.5%, at least 0.1%, at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, or at least 65%. The ultimate strain may be at most 80%, at most 75%, at most 70%, at most 65%, at most 60%, at most 55%, at most 50%, at most 45%, at most 40%, at most 35%, or at most 30%.

[0091] The articles may contain silk fibroin having a [3-sheet content of less than 25%, less than 20%, less than 15%, less than 10%, less than 8%, less than 6%, or less than 5%.

[0092] The articles may contain P-tum content of between 1% and 50% or between 5% and 35%. The -turn content may be at least 1%, at least 3%, at least 5%, at least 8%, at least 10%, at least 15%, or at least 20%. The P-turn content may be at most 50%, at most 45%, at most 40%, at most 35%, at most 30%, at most 25%, at most 20%, or at most 15%.

[0093] The articles may have a transmittance of 60% or greater or 80% or great within the range of visible light (e.g., 380-780 nm, 380-700 nm, 400-780 nm, 400-700 nm, or other generally accepted definition for visible light). The transmittance may be 50% or greater, 55% or greater, 60% or greater,PATENT Attorney Docket No. T002896 WO -2095.0733 65% or greater, 70% or greater, 75% or greater, 80% or greater, 85% or greater, 90% or greater, or 95% or greater.

[0094] Methods of Pre -Treating Threads

[0095] A method may comprise steps a) and b). Step a) includes pre-treating a thread to enhance mechanical strength and step b) includes co-extruding the pre-treated thread with a biopolymer to form a woven three-dimensional material, which may be referred to as an article herein.

[0096] In some embodiments, the pre-treating includes coating the thread with a solution including glycerol. In other embodiments, the pre-treating includes coating the thread with the coating compositions described herein. In other embodiments, the pre -treating includes coating the thread with at least one of a polymer or a biopolymer. Pre-treating may include one coating or more than one coating. Pre-treating may include coating with multiple coating compositions or solutions applied sequentially or simultaneously. The coating may enhance the mechanical properties of the thread, specifically its longitudinal compressive strength. Threads having the coating can be pushed without folding in on itself during printing. Coatings can be applied to incorporate function (e.g. strain sensor) or a decoration (e.g., a dye).

[0097] The pre-treated thread may be at least one of latex, silk cord, silk yam, a waste silk product, a filament, an electrode, a counter-electrode, an enzymatically-coated fiber, a non-spun thread, an electrochemical fiber, a conductive fiber, a copper material, a platinum material, a metallic material, a polyester material, a cotton material, a graphite fiber, a nylon material, a light-emitting fiber element (e.g., electroluminescent wires, LEDs, or optical fibers), or reeled silk fiber.

[0098] The pre-treated thread may have a resistance to compression that is superior to a comparison resistance to compression of a comparison thread lacking a pre -treatment but otherwise identical to the pre -treated thread. The pre-treated thread may have a resistance to compression that is at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, or at least 150% greater than a comparison resistance to compression of a thread lacking the pre -treatment but is otherwise identical to the pre-treated thread.

[0099] The articles produced by this method may have at least one characteristic that is superior to a corresponding characteristic of a woven three-dimensional material lacking the pre-treated thread. The characteristic may be a resistance to stretching and / or a resistance to sheer stress.

[0100] The article may have a resistance to stretching that is at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, or at least 150% higher than a comparison article lacking the pre-treated thread.PATENT Attorney Docket No. T002896 WO -2095.0733

[0101] The article may have a resistance to shear stress at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, or at least 150% higher than a comparison article lacking the pre-treated thread.

[0102] Silk Leathers

[0103] A silk leather may be a layered structure including at least one woven three-dimensional silk material made by the methods described herein. The silk leather may include a silk foam layer. The layered structure may include at least one of a whipped silk cream layer, a silk meringue layer, a compressed silk meringue layer, a hot-pressed silk meringue layer, or a fabric layer.

[0104] In some aspects, the weight ratio of silk fibroin, polysaccharide, and plasticizer may have an impact on one or more of a mechanical property, a density, or a water content of a resulting material made from the composition. Without wishing to be bound by any particular theory, variations of the weight of the plasticizer may preferentially impact mechanical properties, variations of the weight of the polysaccharide or combination of polysaccharides may preferentially impact density, and variations of the weight of the silk fibroin may preferentially impact water content.

[0105] The mixture of silk fibroin and polysaccharide (e.g., xanthan gum, alginate, etc.) includes a weight ratio of silk fibroin to polysaccharide of between 1:4 and 20:1 or between 1:2 and 10:1. For example, the mixture of silk fibroin and polysaccharide can include a weight ratio of silk fibroin to polysaccharide of at least 1:4, at least 1:3, or at least 1:2. For example, mixture of silk fibroin and polysaccharide can include a weight ratio of silk fibroin to polysaccharide of at most 20:1, at most 19: 1, at most 18: 1, at most 16: 1, at most 15: 1, at most 14: 1, at most 12: 1, at most 11: 1, or at most 10:1.

[0106] The compositions herein may have weight ratios as disclosed in PCT International Application No. PCT / US2025 / 026472, which is incorporated by reference herein in its entirety for all purposes.

[0107] Approximately: as used herein, the term “approximately” or “about,” as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In certain embodiments, the term “approximately” or “about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

[0108] Composition: as used herein, may be used to refer to a discrete physical entity that comprises one or more specified components. In general, unless otherwise specified, a composition may be of any form - e.g., gas, gel, liquid, solid, etc. In some embodiments, “composition” may refer to a combination of two or more entities for use in a single embodiment or as part of the same article. It is not required in all embodiments that the combination of entities result in physical admixture, that is,PATENT Attorney Docket No. T002896 WO -2095.0733 combination as separate co-entities of each of the components of the composition is possible; however many practitioners in the field may find it advantageous to prepare a composition that is an admixture of two or more of the ingredients in a pharmaceutically acceptable carrier, diluent, or excipient, making it possible to administer the component ingredients of the combination at the same time.

[0109] Improve, increase, or reduce: as used herein or grammatical equivalents thereof, indicate values that are relative to a baseline measurement, such as a measurement in a similar composition made according to previously known methods.

[0110] Substantially: as used herein, the term “substantially” refers to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest. One of ordinary skill in the biological arts will understand that biological and chemical phenomena rarely, if ever, go to completion and / or proceed to completeness or achieve or avoid an absolute result. The term “substantially” is therefore used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

[0111] It should be apparent to those skilled in the art that many additional modifications beside those already described are possible without departing from the inventive concepts. In interpreting this disclosure, all terms should be interpreted in the broadest possible manner consistent with the context. Variations of the term "comprising" should be interpreted as referring to elements, components, or steps in a non-exclusive manner, so the referenced elements, components, or steps may be combined with other elements, components, or steps that are not expressly referenced. Embodiments referenced as "comprising" certain elements are also contemplated as "consisting essentially of" and "consisting of" those elements. When two or more ranges for a particular value are recited, this disclosure contemplates all combinations of the upper and lower bounds of those ranges that are not explicitly recited. For example, recitation of a value of between 1 and 10 or between 2 and 9 also contemplates a value of between 1 and 9 or between 2 and 10.

[0112] As used herein, "silk fibroin" refers to silk fibroin protein whether produced by silkworm, spider, or other insect, or otherwise generated (Lucas et al., Adv. Protein Chem., 13: 107-242 (1958)). Any type of silk fibroin can be used in different embodiments described herein. Silk fibroin produced by silkworms, such as Bombyx mori, is the most common and represents an earth-friendly, renewable resource. For instance, silk fibroin used in a silk film may be attained by extracting sericin from the cocoons of B. mori. Organic silkworm cocoons are also commercially available. There are many different silks, however, including spider silk (e.g., obtained from Nephila clavipes), transgenic silks, genetically engineered silks, such as silks from bacteria, yeast, mammalian cells, transgenic animals, or transgenic plants, and variants thereof, that can be used. See, e.g., WO 97 / 08315 and U. S. Pat. No.5,245,012, each of which is incorporated herein by reference in their entireties.PATENT Attorney Docket No. T002896 WO -2095.0733

[0113] According to various embodiments, a variety of functionalizing agents may be used with the silk-containing embodiments described herein (e.g., silk membrane, silk composition, silk articles, silk matrix, silk foam, silk microsphere, liquid composition, whipped silk cream, silk meringue, compressed silk meringue, hot-pressed silk meringue, silk leather, silk powder, silk toner, edible silkbased films, etc.). It should be understood that the examples herein may recite one or a few silkcontaining embodiments but are applicable to any silk-containing embodiment, as applicable. In some embodiments, a functionalizing agent may be any compound or molecule that facilitates the attachment to and / or development (e.g., growth) of one or more endothelial cells on a silk membrane. In some embodiments, a functionalizing agent may be any compound or molecule that facilitates the attachment and / or development (e.g., growth) of one or more megakaryocytes and / or hematopoietic progenitor cells on a silk matrix and / or silk membrane. In some embodiments, a functionalizing agent may be or comprise an agent suitable for facilitating the production of one or more of white blood cells and red blood cells.

[0114] In some embodiments, a functionalizing agent may be or comprise a cell attachment mediator and / or an extracellular matrix protein, for example: collagen (e.g., collagen type I, collagen type III, collagen type IV, or collagen type VI), elastin, fibronectin, vitronectin, laminin, fibrinogen, von Willebrand factor, proteoglycans, decorin, perlecan, nidogen, hyaluronan, and / or peptides containing known integrin binding domains (e.g., “RGD” integrin binding sequence, or variations thereof), that are known to affect cellular attachment.

[0115] In some embodiments, a functionalizing agent may be any soluble molecule produced by endothelial cells. Non-limiting examples include fibroblast growth factor-1 (FGF1) and vascular endothelial growth factors (VEGF).

[0116] According to some embodiments, a plurality of functionalizing agents may be used. For example, in some embodiments wherein production of platelets is desired, provided compositions may comprise the use of laminin, fibronectin and / or fibrinogen, and type IV collagen in order to facilitate the attachment and growth of endothelial cells on a silk membrane (e.g., a porous silk membrane) and / or attachment of megakaryocytes to a silk matrix.

[0117] In some embodiments, a functionalizing agent may be embedded or otherwise associated with a silk membrane and / or silk matrix such that at least a portion of the functionalizing agent is surrounded by a silk membrane and / or silk matrix as contrasted to a functionalizing agent simply being positioned along the surface of a silk membrane and / or silk matrix. In some embodiments, a functionalizing agent is distributed along and / or incorporated in substantially the entire surface area of a silk membrane / silk wall. In some embodiments, a functionalizing agent is distributed and / or incorporated only at one or more discrete portions of a silk membrane / wall and / or silk matrix. In some embodiments, aPATENT Attorney Docket No. T002896 WO -2095.0733 functionalizing agent is distributed in and / or along at least one of the lumen-facing side of a silk wall and the matrix-facing side of a silk wall.

[0118] According to various embodiments, any application-appropriate amount of one or more functionalizing agents may be used. In some embodiments, the amount of an individual functionalizing agent may be between about 1 pg / mL and 1,000 pg / mL (e.g., between about 2 pg / mL and 1,000 pg / mL, 5 pg / mL and 1,000 pg / mL, 10 pg / mL and 1,000 pg / mL, 10 pg / mL and 500 pg / mL, 10 pg / mL and 100 pg / mL). In some embodiments, the amount of an individual functionalizing agent may be at least 1 pg / mL (e.g., at least 5 pg / mL, 10 pg / mL, 15 pg / mL, 20 pg / mL, 25 pg / mL, 50 pg / mL, 100 pg / mL, 200 pg / mL, 300 pg / mL, 400 pg / mL, 500 pg / mL, 600 pg / mL, 700 pg / mL, 800 pg / mL, or 900 pg / mL). In some embodiments, the amount of an individual functionalizing agent is at most 1,000 pg / mL (e.g., 900 pg / mL, 800 pg / mL, 700 pg / mL, 600 pg / mL, 500 pg / mL, 400 pg / mL, 300 pg / mL, 200 pg / mL, 100 pg / mL, 90 pg / mL, 80 pg / mL, 70 pg / mL, 60 pg / mL, 50 pg / mL, 40 pg / mL, 30 pg / mL, 20 pg / mL, 10 pg / mL, or 5 pg / mL).

[0119] In some aspects, the composition comprises one or more sensing agents, such as a sensing dye. The sensing agents / sensing dyes are environmentally sensitive and produce a measurable response to one or more environmental factors. In some aspects, the environmentally- sensitive agent or dye may be present in the composition in an effective amount to alter the composition from a first chemicalphysical state to a second chemical-physical state in response to an environmental parameter (e.g., a change in pH, light intensity or exposure, temperature, pressure or strain, voltage, physiological parameter of a subject, and / or concentration of chemical species in the surrounding environment) or an externally applied stimulus (e.g., optical interrogation, acoustic interrogation, and / or applied heat). In some cases, the sensing dye is present to provide one optical appearance under one given set of environmental conditions and a second, different optical appearance under a different given set of environmental conditions. Suitable concentrations for the sensing agents described herein can be the concentrations for the colorants and additives described elsewhere herein. A person having ordinary skill in the chemical sensing arts can determine a concentration that is appropriate for use in a sensing application of the inks described herein.

[0120] In some aspects, the first and second chemical-physical state may be a physical property of the composition, such as a mechanical property, a chemical property, an acoustical property, an electrical property, a magnetic property, an optical property, a thermal property, a radiological property, or an organoleptic property. Exemplary sensing dyes or agents include, but are not limited to, a pH sensitive agent, a thermal sensitive agent, a pressure or strain sensitive agent, a light sensitive agent, or a potentiometric agent.PATENT Attorney Docket No. T002896 WO -2095.0733

[0121] Exemplary pH sensitive dyes or agents include, but are not limited to, cresol red, methyl violet, crystal violet, ethyl violet, malachite green, methyl green, 2-(p-dimethylaminophenylazo)pyridine, paramethyl red, metanil yellow, 4-phenylazodiphenylamine, thymol blue, metacresol purple, orange IV, 4-o-Tolylazo-o-toluindine, quinaldine red, 2,4-dinitrophenol, erythrosine disodium salt, benzopurpurine 4B, N, N-dimethyl-p-(m-tolylazo) aniline, p-dimethylaminoazobenene, 4,4'-bis(2-amino-l-naphthylazo)-2,2'-stilbenedisulfonic acid, tetrabromophenolphthalein ethyl ester, bromophenol blue, Congo red, methyl orange, ethyl orange, 4-(4-dimethylamino-l-naphylazo)-3-methoxybenzenesulfonic acid, bromocresol green, resazurin, 4- phenylazo- 1-napthylamine, ethyl red 2-( 1 -dimethylaminopheny azo) pyridine, 4-(p-ethoxyphenylazo)-m-pheny lene-diamine monohydrochloride, resorcin blue, alizarin red S, methyl red, propyl red, bromocresol purple, chlorophenol red, p-nitrophenol, alizarin, 2-(2,4-dinitrophenylazo)-l-napthol-3,6-disulfonic acid, bromothymol blue, 6, 8-dinitro-lH-quinazoline-2, 4-dione, brilliant yellow, phenol red, neutral red, m-nitrophenol, cresol red, turmeric, metacresol purple, 4,4'-bis(3-amino-l-naphthylazo)-2,2'-stilbenedisulfonic acid, thymol blue, p-naphtholbenzein, phenolphthalein, o-cresolphthalein, ethyl bis(2,4-dimethylphenyl) ethanoate, thymolphthalein, nitrazine yellow, alizarin yellow R, alizarin, p-(2,4-dihydroxyphenylazo) benzenesulfonic acid, 5,5'-indigodisulfonic acid, 2,4,6-trinitrotoluene, 1,3,5-trinitrobenzene, and clayton yellow.

[0122] Exemplary light responsive dyes or agents include, but are not limited to, photochromic compounds or agents, such as triarylmethanes, stilbenes, azastilbenes, nitrones, fulgides, spiropyrans, napthopyrans, spiro-oxazines, quinones, derivatives, and combinations thereof.

[0123] Exemplary potentiometric dyes include, but are not limited to, substituted amiononaphthylehenylpridinium (ANEP) dyes, such as di-4-ANEPPS, di-8-ANEPPS, and N-(4-Sulfobutyl)-4-(6-(4-(Dibutylamino)phenyl)hexatrienyl)Pyridinium (RH237).

[0124] Exemplary temperature sensitive dyes or agents include, but are not limited to, thermochromic compounds or agents, such as thermochromic liquid crystals, leuco dyes, fluoran dyes, octadecylphosphonic acid.

[0125] Exemplary pressure or strain sensitive dyes or agents include, but are not limited to, spiropyran compounds and agents.

[0126] Exemplary chemi-sensitive dyes or agents include, but are not limited to, antibodies such as immunoglobulin G (IgG) which may change color from blue to red in response to bacterial contamination.

[0127] In some aspects, the compositions comprise one or more additive, dopant, or biologically active agent suitable for a desired intended purpose. In some aspects, the additive or dopant may be present in the composition in an amount effective to impart an optical or organoleptic property to thePATENT Attorney Docket No. T002896 WO -2095.0733 composition. Exemplary additives or dopants that impart optical or organoleptic properties include, but are not limited to, dyes / pigments, flavorants, aroma compounds, granular or fibrous fillers.

[0128] Additionally or alternatively, the additive, dopant, or biologically active agent may be present in the composition in an amount effective to "functionalize" the composition to impart a desired mechanical property or added functionality to the composition. Exemplary additive, dopants, or biologically active agent that impart the desired mechanical property or added functionality include, but are not limited to: environmentally sensitive / sensing dyes; active biomolecules; conductive or metallic particles; micro and nanofibers (e.g., silk nanofibers for reinforcement, carbon nanofibers); nanotubes; inorganic particles (e.g., hydroxyapatite, tricalcium phosphate, bioglasses); drugs (e.g., antibiotics, small molecules, or low molecular weight organic compounds); proteins and fragments or complexes thereof (e.g., enzymes, antigens, antibodies, and antigen-binding fragments thereof); DNA / RNA (e.g., siRNA, miRNA, mRNA); cells and fractions thereof (viruses and viral particles; prokaryotic cells such as bacteria; eukaryotic cells such as mammalian cells and plant cells; fungi).

[0129] In some aspects, the additive or dopant comprises a flavoring agent or flavorant.

[0130] Exemplary flavorants include ester flavorants, amino acid flavorants, nucleic acid flavorants, organic acid flavorants, and inorganic acid flavorants, such as, but not limited to, diacetyl, acetyl propionyl, acetoin, isoamyl acetate, benzaldehyde, cinnamaldehyde, ethyl propionate, methyl anthranilate, limonene, ethyl decadienoate, allyl hexanoate, ethyl maltol, ethylvanillin, methyl salicylate, manzanate, glutamic acid salts, glycine salts, guanylic acids salts, inosinic acid salts, acetic acid, ascorbic acid, citric acid, fumaric acid, lactic acid, malic acid, phosphoric acid, tartaric acid, derivatives, and mixtures thereof.

[0131] In some aspects, the additive or dopant comprises an aroma compound. Exemplary aroma compounds include ester aroma compounds, terpene aroma compounds, cyclic terpenes, and aromatic aroma compounds, such as, but not limited to, geranyl acetate, methyl formate, methyl acetate, methyl propionate, methyl butyrate, ethyl acetate, ethyl butyrate, isoamyl acetate, pentyl butyrate, pentyl pentanoate, octyl acetate, benzyl acetate, methyl anthranilate, myrcene, geraniol, nerol, citral, citronellal, citronellol, linalool, nerolidol, limonene, camphor, menthol, carone, terpineol, alphaionone, thujone, eucalyptol, benzaldehyde, eugenol, cinnamaldehyde, ethyl maltol, vanillin, anisole, anethole, estragole, thymol.

[0132] In some aspects, the additive or dopant comprises a colorant, such as a dye or pigment. In some aspects, the dye or pigment imparts a color or grayscale to the composition. The colorant can be different than the sensing agents and / or sensing dyes below. Any organic and / or inorganic pigments and dyes can be included in the inks. Exemplary pigments suitable for use in the present disclosure include International Color Index or C. I. Pigment Black Numbers 1, 7, 11, and 31, C. I. Pigment BluePATENT Attorney Docket No. T002896 WO -2095.0733 Numbers 15, 15:1, 15:2, 15:3, 15:4, 15:6, 16, 27, 29, 61 and 62, C. I. Pigment Green Numbers 7, 17, 18 and 36, C. I. Pigment Orange Numbers 5, 13, 16, 34 and 36, C. I. Pigment Violet Numbers 3, 19, 23 and 27, C. I. Pigment Red Numbers 3, 17, 22, 23, 48:1, 48:2, 57:1, 81:1, 81:2, 81:3, 81:5, 101, 114, 122, 144, 146, 170, 176, 179, 181, 185, 188, 202, 206, 207, 210 and 249, C. I. Pigment Yellow Numbers 1, 2, 3, 12, 13, 14, 17, 42, 65, 73, 74, 75, 83, 30, 93, 109, 1 10, 128, 138, 139, 147, 142, 151, 154 and 180, D& C Red No. 7, D& C Red No. 6 and D& C Red No. 34, carbon black pigment (such as Regal 330, Cabot Corporation), quinacridone pigments (Quinacridone Magenta (228-0122), available from Sun Chemical Corporation, Fort Lee, N. J.), diarylide yellow pigment (such as AAOT Yellow (274-1788) available from Sun Chemical Corporation); and phthalocyanine blue pigment (such as Blue 15:3 (294-1298) available from Sun Chemical Corporation). The classes of dyes suitable for use in present invention can be selected from acid dyes, natural dyes, direct dyes (either cationic or anionic), basic dyes, and reactive dyes. The acid dyes, also regarded as anionic dyes, are soluble in water and mainly insoluble in organic solvents and are selected, from yellow acid dyes, orange acid dyes, red acid dyes, violet acid dyes, blue acid dyes, green acid dyes, and black acid dyes. European Patent 0745651, incorporated herein by reference, describes a number of acid dyes that are suitable for use in the present disclosure. Exemplary yellow acid dyes include Acid Yellow 1 International Color Index or C. I.10316); Acid Yellow 7 (C. I. 56295); Acid Yellow 17 (C. I. 18965); Acid Yellow 23 (C. I. 19140); Acid Yellow 29 (C. I. 18900); Acid Yellow 36 (C. I. 13065); Acid Yellow 42 (C. I. 22910); Acid Yellow 73 (C. I. 45350); Acid Yellow 99 (C. I. 13908); Acid Yellow 194; and Food Yellow 3 (C. I. 15985). Exemplary orange acid dyes include Acid Orange 1 (C. I. 13090 / 1); Acid Orange 10 (C. I. 16230); Acid Orange 20 (C. I. 14603); Acid Orange 76 (C. I. 18870); Acid Orange 142; Food Orange 2 (C. I. 15980); and Orange B.

[0133] Exemplary red acid dyes include Acid Red 1 (C. I. 18050); Acid Red 4 (C. I. 14710); Acid Red 18 (C. I. 16255); Acid Red 26 (C. I. 16150); Acid Red 27 (C. I. 16185); Acid Red 51 (C. I. 45430, available from BASF Corporation, Mt. Olive, N. J.); Acid Red 52 (C. I. 45100); Acid Red 73 (C. I.27290); Acid Red 87 (C. I. 45380); Acid Red 94 (C. I. 45440) Acid Red 194; and Food Red 1 (C. I.14700). Exemplary violet acid dyes include Acid Violet 7 (C. I. 18055); and Acid Violet 49 (C. I.42640). Exemplary blue acid dyes include Acid Blue 1 (C. I. 42045); Acid Blue 9 (C. I. 42090); Acid Blue 22 (C. I. 42755); Acid Blue 74 (C. I. 73015); Acid Blue 93 (C. I. 42780); and Acid Blue 158A (C. I.15050). Exemplary green acid dyes include Acid Green 1 (C. I. 10028); Acid Green 3 (C. I. 42085); Acid Green 5 (C. I. 42095); Acid Green 26 (C. I. 44025); and Food Green 3 (C. I. 42053). Exemplary black acid dyes include Acid Black 1 (C. I. 20470); Acid Black 194 (Basantol® X80, available from BASF Corporation, an azo / l :2 CR-complex.PATENT Attorney Docket No. T002896 WO -2095.0733

[0134] Exemplary direct dyes for use in the present disclosure include Direct Blue 86 (C. I. 74180); Direct Blue 199; Direct Black 168; Direct Red 253; and Direct Yellow 107 / 132 (C. I. Not Assigned).

[0135] Exemplary natural dyes for use in the present disclosure include Alkanet (C. I. 75520,75530); Annatto (C. I. 75120); Carotene (C. I. 75130); Chestnut; Cochineal (C. I.75470); Cutch (C. I. 75250, 75260); Divi-Divi; Fustic (C. I. 75240); Brazilin (C. I. 75280); Logwood (C. I. 75200); Osage Orange (C. I. 75660); Paprika; Quercitron (C. I. 75720); Saffron (C. I. 75100); Sandal Wood (C. I. 75510, 75540, 75550, 75560); Sumac; and Turmeric (C. I. 75300). Exemplary reactive dyes for use in the present disclosure include Reactive Yellow 37 (monoazo dye); Reactive Black 31 (diazo dye); Reactive Blue 77 (phthalo cyanine dye); and Reactive Red 180 and Reactive Red 108 dyes. Suitable also are the colorants described in The Printing Ink Manual (5th ed., Leach et al. eds. (2007), pages 289-299). Other organic and inorganic pigments and dyes and combinations thereof can be used to achieve the colors desired.

[0136] In addition to or in place of visible colorants, compositions provided herein can contain ETV fluorophores that are excited in the ETV range and emit light at a higher wavelength (typically 400 nm and above). Examples of ETV fluorophores include but are not limited to materials from the coumarin, benzoxazole, rhodamine, napthalimide, perylene, benzanthrones, benzoxanthones or benzothiaxanthones families. The addition of a UV fluorophore (such as an optical brightener for instance) can help maintain maximum visible light transmission. The amount of colorant, when present, generally is between 0.05% to 5% or between 0.1% and 1% based on the weight of the composition.

[0137] For non-white compositions, the amount of pigment / dye generally is present in an amount of from at or about 0.1 wt% to at or about 20 wt% based on the weight of the composition. In some applications, a non-white ink can include 15 wt% or less pigment / dye, or 10 wt% or less pigment / dye or 5 wt% pigment / dye, or 1 wt% pigment / dye based on the weight of the composition. In some applications, a non-white ink can include 1 wt% to 10 wt%, or 5 wt% to 15 wt%, or 10 wt% to 20 wt% pigment / dye based on the weight of the composition. In some applications, a non-white ink can contain an amount of dye / pigment that is 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt%, 16 wt%, 17 wt%, 18 wt%, 19 wt% or 20 wt% based on the weight of the composition.

[0138] For white compositions, the amount of white pigment generally is present in an amount of from at or about 1 wt% to at or about 60 wt% based on the weight of the composition. In some applications, greater than 60 wt% white pigment can be present. Preferred white pigments include titanium dioxide (anatase and rutile), zinc oxide, lithopone (calcined coprecipitate of barium sulfate and zinc sulfide), zinc sulfide, blanc fixe and alumina hydrate and combinations thereof, although any of these can be combined with calcium carbonate. In some applications, a white ink can include 60 wt% or less whitePATENT Attorney Docket No. T002896 WO -2095.0733 pigment, 55 wt% or less white pigment, 50 wt% white pigment, 45 wt% white pigment, 40 wt% white pigment, 35 wt% white pigment, 30 wt% white pigment, 25 wt% white pigment, 20 wt% white pigment, 15 wt% white pigment, or 10 wt% white pigment, based on the weight of the composition. In some applications, a white ink can include 5 wt% to 60 wt%, 5 wt% to 55 wt%, 10 wt% to 50 wt%, 10 wt% to 25 wt%, 25 wt% to 50 wt%, 5 wt% to 15 wt%, or 40 wt% to 60 wt% white pigment based on the weight of the composition. In some applications, a non-white ink can an amount of dye / pigment that is 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15%, 16 wt%, 17 wt%, 18 wt%, 19 wt%, 20 wt%, 21 wt%, 22 wt%, 23 wt%, 24 wt%, 25%, 26 wt%, 27 wt%, 28 wt%, 29 wt%, 30 wt%, 31 wt%, 32 wt%, 33 wt%, 34 wt%, 35%, 36 wt%, 37 wt%, 38 wt%, 39 wt%, 40 wt%, 41 wt%, 42 wt%, 43 wt%, 44 wt%, 45%, 46 wt%, 47 wt%, 48 wt%, 49 wt%, 50 wt%, 51 wt%, 52 wt%, 53 wt%, 54 wt%, 55%, 56 wt%, 57 wt%, 58 wt%, 59 wt% or 60 wt% based on the weight of the composition.

[0139] In some aspects, the additive or dopant comprises a conductive additive. Exemplary conductive additives include, but are not limited to graphite, graphite powder, carbon nanotubes, and metallic particles or nanoparticles, such as gold nanoparticles. In some aspects, the conductive additive is biocompatible and non-toxic.

[0140] In some aspects, the additive is a biologically active agent. The term “biologically active agent” as used herein refers to any molecule which exerts at least one biological effect in vivo. For example, the biologically active agent can be a therapeutic agent to treat or prevent a disease state or condition in a subject. Biologically active agents include, without limitation, organic molecules, inorganic materials, proteins, peptides, nucleic acids (e.g., genes, gene fragments, gene regulatory sequences, and antisense molecules), nucleoproteins, polysaccharides, glycoproteins, and lipoproteins. Classes of biologically active compounds that can be incorporated into the composition provided herein include, without limitation, anticancer agents, antibiotics, analgesics, anti-inflammatory agents, immunosuppressants, enzyme inhibitors, antihistamines, anti-convulsants, hormones, muscle relaxants, antispasmodics, ophthalmic agents, prostaglandins, anti-depressants, anti-psychotic substances, trophic factors, osteoinductive proteins, growth factors, and vaccines.

[0141] The term “active agent” may also be used herein to refer to a biological sample (e.g., a sample of tissue or fluid, such as for instance blood) or a component thereof, and / or to a biologically active entity or compound, and / or to a structural ly or functionally labile entity.

[0142] Exemplary active agents include, but are not limited to, therapeutic agents, diagnostic agents (e.g., contrast agents), and any combinations thereof. In some embodiments, the active agent present in a silk matrix (e.g., a silk microsphere), composition, or the like can include a labile active agent, e.g., an agent that can undergo chemical, physical, or biological change, degradation and / orPATENT Attorney Docket No. T002896 WO -2095.0733 deactivation after exposure to a specified condition, e.g., high temperatures, high humidity, light exposure, and any combinations thereof. In some embodiments, the active agent present in the silk matrix (e.g., a silk microsphere), composition, or the like can include a temperature-sensitive active agent, e.g., an active agent that will lose at least about 30% or more of its original activity or bioactivity, upon exposure to a temperature of at least about 10 °C. or above, including at least about 15 °C. or above, at least about room temperature or above, or at least about body temperature (e.g., about 37 °C.) or above.

[0143] The active agent can be generally present in the silk matrix (e.g., a silk microsphere), composition, or the like in an amount of about 0.01 % (w / w) to about 70% (w / w), about 0.1% (w / w) to about 50% (w / w), or about 1 % (w / w) to about 30% (w / w). The active agent can be present on a surface of the silk matrix (e.g., a silk microsphere), composition, or the like and / or encapsulated and dispersed in the silk matrix (e.g., a silk microsphere), composition, or the like homogeneously, heterogeneously, or in a gradient. In some embodiments, the active agent can be added into the silk solution, which is then subjected to the methods described herein for preparing a silk matrix (e.g., a silk microsphere), composition, or the like. In some embodiments, the active agent can be coated on a surface of the silk matrix (e.g., a silk microsphere), composition, or the like. In some embodiments, the active agent can be loaded in a silk matrix (e.g., a silk microsphere), composition, or the like by incubating the silk microsphere in a solution of the active agent for a period of time, during which an amount of the active agent can diffuse into the silk matrix (e.g., a silk microsphere), composition, or the like, and thus distribute within the silk matrix (e.g., a silk microsphere), composition, or the like.

[0144] In some aspects, the additive is a therapeutic agent. As used herein, the term “therapeutic agent” means a molecule, group of molecules, complex or substance administered to an organism for diagnostic, therapeutic, preventative medical, or veterinary purposes. As used herein, the term “therapeutic agent” includes a “drug” or a “vaccine.” This term includes externally and internally administered topical, localized and systemic human and animal pharmaceuticals, treatments, remedies, nutraceuticals, cosmeceuticals, biologicals, devices, diagnostics and contraceptives, including preparations useful in clinical and veterinary screening, prevention, prophylaxis, healing, wellness, detection, imaging, diagnosis, therapy, surgery, monitoring, cosmetics, prosthetics, forensics and the like. This term can also be used in reference to agriceutical, workplace, military, industrial and environmental therapeutics or remedies comprising selected molecules or selected nucleic acid sequences capable of recognizing cellular receptors, membrane receptors, hormone receptors, therapeutic receptors, microbes, viruses or selected targets comprising or capable of contacting plants, animals and / or humans. This term can also specifically include nucleic acids and compounds comprising nucleic acids that produce a therapeutic effect, for example deoxyribonucleic acid (DNA),PATENT Attorney Docket No. T002896 WO -2095.0733 ribonucleic acid (RNA), nucleic acid analogues (e.g., locked nucleic acid (LNA), peptide nucleic acid (PNA), xeno nucleic acid (XNA)), or mixtures or combinations thereof, including, for example, DNA nanoplexes, siRNA, microRNA, shRNA, aptamers, ribozymes, decoy nucleic acids, antisense nucleic acids, RNA activators, and the like. Generally, any therapeutic agent can be included in the composition provided herein.

[0145] The term “therapeutic agent” also includes an agent that is capable of providing a local or systemic biological, physiological, or therapeutic effect in the biological system to which it is applied. For example, the therapeutic agent can act to control infection or inflammation, enhance cell growth and tissue regeneration, control tumor growth, act as an analgesic, promote anti-cell attachment, and enhance bone growth, among other functions. Other suitable therapeutic agents can include anti-viral agents, hormones, antibodies, or therapeutic proteins. Other therapeutic agents include prodrugs, which are agents that are not biologically active when administered but upon administration to a subject are converted to biologically active agents through metabolism or some other mechanism. Additionally, a silk-based drug delivery composition can contain one therapeutic agent or combinations of two or more therapeutic agents.

[0146] A therapeutic agent can include a wide variety of different compounds, including chemical compounds and mixtures of chemical compounds, e.g., small organic or inorganic molecules; saccharides; oligosaccharides; polysaccharides; biological macromolecules, e.g., peptides, proteins, and peptide analogs and derivatives; peptidomimetics; antibodies and antigen binding fragments thereof; nucleic acids; nucleic acid analogs and derivatives; an extract made from biological materials such as bacteria, plants, fungi, or animal cells; animal tissues; naturally occurring or synthetic compositions; and any combinations thereof. In some aspects, the therapeutic agent is a small molecule.

[0147] The term “bioactivity,” as used herein in reference to an active agent, generally refers to the ability of an active agent to interact with a biological target and / or to produce an effect on a biological target. For example, bioactivity can include, without limitation, elicitation of a stimulatory, inhibitory, regulatory, toxic or lethal response in a biological target. The biological target can be a molecule or a cell. For example, a bioactivity can refer to the ability of an active agent to modulate the effect / activity of an enzyme, block a receptor, stimulate a receptor, modulate the expression level of one or more genes, modulate cell proliferation, modulate cell division, modulate cell morphology, or any combination thereof. In some instances, a bioactivity can refer to the ability of a compound to produce a toxic effect in a cell. Exemplary cellular responses include, but are not limited to, lysis, apoptosis, growth inhibition, and growth promotion; production, secretion, and surface expression of a protein or other molecule of interest by the cell; membrane surface molecule activation including receptorPATENT Attorney Docket No. T002896 WO -2095.0733 activation; transmembrane ion transports; transcriptional regulations; changes in viability of the cell; changes in cell morphology: changes in presence or expression of an intracellular component of the cell; changes in gene expression or transcripts; changes in the activity of an enzyme produced within the cell; and changes in the presence or expression of a ligand and / or receptor (e.g., protein expression and / or binding activity). Methods for assaying different cellular responses are well known to one of skill in the art, e.g., western blot for determining changes in presence or expression of an endogenous protein of the cell, or microscopy for monitoring the cell morphology in response to the active agent, or FISH and / or qPCR for the detection and quantification of changes in nucleic acids. Bioactivity can be determined in some embodiments, for example, by assaying a cellular response.

[0148] In reference to an antibody, the term “bioactivity” includes, but is not limited to, epitope or antigen binding affinity, the in vivo and / or in vitro stability of the antibody, the immunogenic properties of the antibody, e.g., when administered to a human subject, and / or the ability to neutralize or antagonize the bioactivity of a target molecule in vivo or in vitro. The aforementioned properties or characteristics can be observed or measured using art-recognized techniques including, but not limited to, scintillation proximity assays, ELISA, ORIGEN immunoassay (IGEN), fluorescence quenching, fluorescence ELISA, competitive ELISA, SPR analysis including, but not limited to, SPR analysis using a BIAcore biosensor, in vitro and in vivo neutralization assays (see, for example, International Publication No. WO 2006 / 062685), receptor binding, and immunohistochemistry with tissue sections from different sources including human, primate, or any other source as needed. In reference to an immunogen, the “bioactivity” includes immunogenicity, the definition of which is discussed in detail later. In reference to a virus, the “bioactivity” includes infectivity, the definition of which is discussed in detail later. In reference to a contrast agent, e.g., a dye, the “bioactivity” refers to the ability of a contrast agent when administered to a subject to enhance the contrast of structures or fluids within the subject's body. The bioactivity of a contrast agent also includes, but is not limited to, its ability to interact with a biological environment and / or influence the response of another molecule under certain conditions.

[0149] As used herein, the term “small molecule” can refer to compounds that are “natural productlike," however, the term “small molecule" is not limited to “natural product-like" compounds. Rather, a small molecule is typically characterized in that it contains several carbon — carbon bonds and has a molecular weight of less than 5000 Daltons (5 kDa), preferably less than 3 kDa, still more preferably less than 2 kDa, and most preferably less than 1 kDa. In some cases, it is preferred that a small molecule has a molecular weight equal to or less than 700 Daltons.

[0150] Exemplary therapeutic agents include, but are not limited to, those found in Harrison’s Principles of Internal Medicine, 13th Edition, Eds. T. R. Harrison et al. McGraw-Hill N. Y., NY;PATENT Attorney Docket No. T002896 WO -2095.0733 Physicians' Desk Reference, 50th Edition, 1997, Oradell New Jersey, Medical Economics Co.; Pharmacological Basis of Therapeutics, 8th Edition, Goodman and Gilman, 1990; United States Pharmacopeia, The National Formulary, ETSP XII NF XVII, 1990, the complete contents of all of which are incorporated herein by reference.

[0151] Therapeutic agents include the herein disclosed categories and specific examples. It is not intended that the category be limited by the specific examples. Those of ordinary skill in the art will recognize also numerous other compounds that fall within the categories and that are useful according to the present disclosure. Examples include a radiosensitizer, a steroid, a xanthine, a beta-2-agonist bronchodilator, an anti-inflammatory agent, an analgesic agent, a calcium antagonist, an angiotensinconverting enzyme inhibitors, a beta-blocker, a centrally active alpha- agonist, an alpha- 1 -antagonist, an anticholinergic / antispasmodic agent, a vasopressin analogue, an antiarrhythmic agent, an antiparkinsonian agent, an antiangina / antihypertensive agent, an anticoagulant agent, an antiplatelet agent, a sedative, an anxiolytic agent, a peptidic agent, a biopolymeric agent, an antineoplastic agent, a laxative, an antidiarrheal agent, an antimicrobial agent, an antifungal agent, a vaccine, a protein, or a nucleic acid. In a further aspect, the pharmaceutically active agent can be coumarin, albumin, steroids such as betamethasone, dexamethasone, methylprednisolone, prednisolone, prednisone, triamcinolone, budesonide, hydrocortisone, and pharmaceutically acceptable hydrocortisone derivatives; xanthines such as theophylline and doxophylline; beta-2-agonist bronchodilators such as salbutamol, fenterol, clenbuterol, bambuterol, and salmeterol; anti-inflammatory agents, including antiasthmatic anti-inflammatory agents, antiarthritis anti-inflammatory agents, and non-steroidal antiinflammatory agents, examples of which include but are not limited to sulfides, mesalamine, budesonide, salazopyrin, diclofenac, pharmaceutically acceptable diclofenac salts, nimesulide, naproxen, acetaminophen, ibuprofen, ketoprofen and piroxicam; analgesic agents such as salicylates; calcium channel blockers such as nifedipine, amlodipine, and nicardipine: angiotensin converting enzyme inhibitors such as captopril, benazepril hydrochloride, fosinopril sodium, trandolapril, ramipril, lisinopril, enalapril, quinapril hydrochloride, and moexipril hydrochloride; beta-blockers (i.e., beta adrenergic blocking agents) such as sotalol hydrochloride, timolol maleate, esmolol hydrochloride, carteolol, propanolol hydrochloride, betaxolol hydrochloride, penbutolol sulfate, metoprolol tartrate, metoprolol succinate, acebutolol hydrochloride, atenolol, pindolol, and bisoprolol fumarate; centrally active alpha-2-agonists such as clonidine; alpha- 1 -antagonists such as doxazosin and prazosin; anticholinergic / antispasmodic agents such as dicyclomine hydrochloride, scopolamine hydrobromide, glycopyrrolate, clidinium bromide, flavoxate, and oxybutynin; vasopressin analogues such as vasopressin and desmopressin; antiarrhythmic agents such as quinidine, lidocaine, tocainide hydrochloride, mexiletine hydrochloride, digoxin, verapamil hydrochloride, propafenonePATENT Attorney Docket No. T002896 WO -2095.0733 hydrochloride, flecainide acetate, procainamide hydrochloride, moricizine hydrochloride, and disopyramide phosphate; antiparkinsonian agents, such as dopamine, L-Dopa / Carbidopa, selegiline, dihydroergocryptine, pergolide, lisuride, apomorphine, and bromocriptine; antiangina agents and antihypertensive agents such as isosorbide mononitrate, isosorbide dinitrate, propranolol, atenolol and verapamil; anticoagulant and antiplatelet agents such as coumadin, warfarin, acetylsalicylic acid, and ticlopidine; sedatives such as benzodiazepines and barbiturates; anxiolytic agents such as lorazepam, bromazepam, and diazepam; peptidic and biopolymeric agents such as calcitonin, leuprolide and other LHRH agonists, hirudin, cyclosporin, insulin, somatostatin, protirelin, interferon, desmopressin, somatotropin, thymopentin, pidotimod, erythropoietin, interleukins, melatonin, granulocyte / macrophage-CSF, and heparin; antineoplastic agents such as etoposide, etoposide phosphate, cyclophosphamide, methotrexate, 5-fluorouracil, vincristine, doxorubicin, cisplatin, hydroxyurea, leucovorin calcium, tamoxifen, flutamide, asparaginase, altretamine, mitotane, and procarbazine hydrochloride; laxatives such as senna concentrate, casanthranol, bisacodyl, and sodium picosulphate; antidiarrheal agents such as difenoxin hydrochloride, loperamide hydrochloride, furazolidone, diphenoxylate hydrochloride, and microorganisms; vaccines such as bacterial and viral vaccines; antimicrobial agents such as penicillins, cephalosporins, and macrolides, antifungal agents such as imidazolic and triazolic derivatives; and nucleic acids such as DNA sequences encoding for biological proteins, and antisense oligonucleotides.

[0152] Anti-cancer agents include alkylating agents, platinum agents, antimetabolites, topoisomerase inhibitors, antitumor antibiotics, antimitotic agents, aromatase inhibitors, thymidylate synthase inhibitors, DNA antagonists, farnesyltransferase inhibitors, pump inhibitors, histone acetyltransferase inhibitors, metalloproteinase inhibitors, ribonucleoside reductase inhibitors, TNF alpha agonists / antagonists, endothelin A receptor antagonists, retinoic acid receptor agonists, immunomodulators, hormonal and antihormonal agents, photodynamic agents, and tyrosine kinase inhibitors.

[0153] Antibiotics include aminoglycosides (e.g., gentamicin, tobramycin, netilmicin, streptomycin, amikacin, neomycin), bacitracin, carbapenems (e.g., imipenem / cilastatin), cephalosporins, colistin, methenamine, monobactams (e.g., aztreonam), penicillins (e.g., penicillin G, penicillin V, methicillin, nafcillin, oxacillin, cloxacillin, dicloxacillin, ampicillin, amoxicillin, carbenicillin, ticarcillin, piperacillin, mezlocillin, azlocillin), polymyxin B, quinolones, and vancomycin; and bacteriostatic agents such as chloramphenicol, clindamycin, macrolides (e.g., erythromycin, azithromycin, clarithromycin), lincomycin, nitrofurantoin, sulfonamides, tetracyclines (e.g., tetracycline, doxycycline, minocycline, demeclocy cline), and trimethoprim. Also included are metronidazole, fluoroquinolones, and rifampin.PATENT Attorney Docket No. T002896 WO -2095.0733

[0154] Enzyme inhibitors are substances which inhibit an enzymatic reaction. Examples of enzyme inhibitors include edrophonium chloride, N-methylphysostigmine, neostigmine bromide, physostigmine sulfate, tacrine, 1 -hydroxymaleate, iodotubercidin, p-bromotetranisole, 10-(alpha-diethylaminopropionyl)-phenothiazine hydrochloride, calmidazolium chloride, hemicholinium-3,3,5-dinitrocatechol, diacylglycerol kinase inhibitor I, diacylglycerol kinase inhibitor II, 3-phenylpropargylamine, N°-monomethyl-L-arginine acetate, carbidopa, 3 -hydroxy benzylhydrazine, hydralazine, clorgyline, deprenyl, hydroxylamine, iproniazid phosphate, 6-MeO-tetrahydro-9H-pyrido-indole, nialamide, pargyline, quinacrine, semicarbazide, tranylcypromine, N, N-diethylaminoethyl-2,2-diphenylvalerate hydrochloride, 3-isobutyl-l-methylxanthine, papaverine, indomethacin, 2-cyclooctyl-2 -hydroxyethylamine hydrochloride, 2,3-dichloro-α-methylbenzylamine (DCMB), 8,9-dichloro-2,3,4,5-tetrahydro-lH-2-benzazepine hydrochloride, p-aminoglutethimide, p-aminoglutethimide tartrate, 3 -iodotyrosine, alpha-methyltyrosine, acetazolamide, dichlorphenamide, 6-hydroxy-2-benzothiazolesulfonamide, and allopurinol.

[0155] Antihistamines include pyrilamine, chlorpheniramine, and tetrahydrozoline, among others.

[0156] Anti-inflammatory agents include corticosteroids, nonsteroidal anti-inflammatory drugs (e.g., aspirin, phenylbutazone, indomethacin, sulindac, tolmetin, ibuprofen, piroxicam, and fenamates), acetaminophen, phenacetin, gold salts, chloroquine, D-Penicillamine, methotrexate colchicine, allopurinol, probenecid, and sulfinpyrazone.

[0157] Muscle relaxants include mephenesin, methocarbamol, cyclobenzaprine hydrochloride, trihexyphenidyl hydrochloride, levodopa / carbidopa, and biperiden.

[0158] Anti-spasmodics include atropine, scopolamine, oxyphenonium, and papaverine.

[0159] Analgesics include aspirin, phenylbutazone, indomethacin, sulindac, tolmetin, ibuprofen, piroxicam, fenamates, acetaminophen, phenacetin, morphine sulfate, codeine sulfate, meperidine, nalorphine, opioids (e.g., codeine sulfate, fentanyl citrate, hydrocodone bitartrate, loperamide, morphine sulfate, noscapine, norcodeine, normorphine, thebaine, nor-binaltorphimine, buprenorphine, chlornaltrexamine, funaltrexamine, nalbuphine, nalorphine, naloxone, naloxonazine, naltrexone, and naltrindole), procaine, lidocaine, tetracaine and dibucaine. Ophthalmic agents include sodium fluorescein, rose bengal, methacholine, adrenaline, cocaine, atropine, alpha-chymotrypsin, hyaluronidase, betaxolol, pilocarpine, timolol, timolol salts, and combinations thereof.

[0160] Prostaglandins are art recognized and are a class of naturally occurring chemically related long-chain hydroxy fatty acids that have a variety of biological effects.

[0161] Anti -depressants are substances capable of preventing or relieving depression.

[0162] Examples of anti-depressants include imipramine, amitriptyline, nortriptyline, protriptyline, desipramine, amoxapine, doxepin, maprotiline, tranylcypromine, phenelzine, and isocarboxazid.PATENT Attorney Docket No. T002896 WO -2095.0733

[0163] Trophic factors are factors whose continued presence improves the viability or longevity of a cell trophic factors include, without limitation, platelet-derived growth factor (PDGP), neutrophilactivating protein, monocyte chemoattractant protein, macrophage- inflammatory protein, platelet factor, platelet basic protein, and melanoma growth stimulating activity; epidermal growth factor, transforming growth factor (alpha), fibroblast growth factor, platelet- derived endothelial cell growth factor, insulin-like growth factor, glial derived growth neurotrophic factor, ciliary neurotrophic factor, nerve growth factor, bone growth / cartilage-inducing factor (alpha and beta), bone morphogenetic proteins, interleukins (e.g., interleukin inhibitors or interleukin receptors, including interleukin 1 through interleukin 10), interferons (e.g., interferon alpha, beta and gamma), hematopoietic factors, including erythropoietin, granulocyte colony stimulating factor, macrophage colony stimulating factor and granulocyte-macrophage colony stimulating factor; tumor necrosis factors, and transforming growth factors (beta), including beta-1, beta-2, beta-3, inhibin, and activin.

[0164] Hormones include estrogens (e.g., estradiol, estrone, estriol, diethylstilbestrol, quinestrol, chlorotrianisene, ethinyl estradiol, mestranol), anti-estrogens (e.g., clomiphene, tamoxifen), progestins (e.g., medroxyprogesterone, norethindrone, hydroxyprogesterone, norgestrel), antiprogestin (mifepristone), androgens (e.g., testosterone cypionate, fluoxymesterone, danazol, testolactone), antiandrogens (e.g., cyproterone acetate, flutamide), thyroid hormones (e.g., triiodothyronne, thyroxine, propylthiouracil, methimazole, and iodixode), and pituitary hormones (e.g., corticotropin, somatotropin, oxytocin, and vasopressin). Hormones are commonly employed in hormone replacement therapy and / or for purposes of birth control. Steroid hormones, such as prednisone, are also used as immunosuppressants and anti-inflammatories. In some aspects, the additive is an agent that stimulates tissue formation, and / or healing and regrowth of natural tissues, and any combinations thereof. Agents that increase formation of new tissues and / or stimulates healing or regrowth of native tissue at the site of injection can include, but are not limited to, fibroblast growth factor (FGF), transforming growth factor-beta (TGF-beta, platelet-derived growth factor (PDGF), epidermal growth factors (EGFs), connective tissue activated peptides (CTAPs), osteogenic factors including bone morphogenic proteins, heparin, angiotensin II (A-II) and fragments thereof, insulin-like growth factors, tumor necrosis factors, interleukins, colony stimulating factors, erythropoietin, nerve growth factors, interferons, biologically active analogs, fragments, and derivatives of such growth factors, and any combinations thereof.

[0165] In some aspects, the silk composition can further comprise at least one additional material for soft tissue augmentation, e.g., dermal filler materials, including, but not limited to, poly(methyl methacrylate) microspheres, hydroxyapatite, poly(L-lactic acid), collagen, elastin, and glycosaminoglycans, hyaluronic acid, commercial dermal filler products such as BOTOX® (fromPATENT Attorney Docket No. T002896 WO -2095.0733 Allergan), DYSPORT®, COSMODERM®, EVOLENCE®, RADIESSE®, RESTYLANE®, JUVEDERM® (from Allergan), SCULPTRA®, PERLANE®, and CAPTIQEIE®, and any combinations thereof.

[0166] In some aspects, the additive is a wound healing agent. As used herein, a “wound healing agent" is a compound or composition that actively promotes wound healing process.

[0167] Exemplary wound healing agents include, but are not limited to dexpanthenol; growth factors; enzymes; hormones; povidon-iodide; fatty acids; anti-inflammatory agents; antibiotics; antimicrobials; antiseptics; cytokines; thrombin; analgesics; opioids; aminoxyls; furoxans; nitrosothiols; nitrates and anthocyanins; nucleosides, such as adenosine; and nucleotides, such as adenosine diphosphate (ADP) and adenosine triphosphate (ATP); neurotransmitter / neuromodulators, such as acetylcholine and 5-hydroxytryptamine (serotonin / 5-HT); histamine and catecholamines, such as adrenalin and noradrenalin; lipid molecules, such as 5-sphingosine-l -phosphate and lysophosphatidic acid; amino acids, such as arginine and lysine; peptides such as the bradykinins, substance P and calcium gene-related peptide (CGRP); nitric oxide; and any combinations thereof.

[0168] In certain aspects, the active agents provided herein are immunogens. In one aspect, the immunogen is a vaccine. Most vaccines are sensitive to environmental conditions under which they are stored and / or transported. For example, freezing may increase reactogenicity (e.g., capability of causing an immunological reaction) and / or loss of potency for some vaccines (e.g., HepB, and DTaP / IPV / FQB), or cause hairline cracks in the container, leading to contamination. Further, some vaccines (e.g., BCG, Varicella, and MMR) are sensitive to heat. Many vaccines (e.g., BCG, MMR, Varicella, Meningococcal C Conjugate, and most DTaP-containing vaccines) are light sensitive. See, e.g., Galazka et al., Thermostability of vaccines, in Global Programme for Vaccines & Immunization (World Health Organization, Geneva, 1998); Peetermans et al., Stability of freeze-dried rubella virus vaccine (Cendehill strain) at various temperatures, J. Biological Standardization 179 (1973). Thus, the compositions and methods provided herein also provide for stabilization of vaccines regardless of the cold chain and / or other environmental conditions.

[0169] In some aspects, the additive is a cell, e.g., a biological cell. Cells useful for incorporation into the composition can come from any source, e.g., mammalian, insect, plant, etc. In some aspects, the cell can be a human, rat or mouse cell. In general, cells to be used with the compositions provided herein can be any types of cells. In general, the cells should be viable when encapsulated within compositions. In some aspects, cells that can be used with the composition include, but are not limited to, mammalian cells (e.g. human cells, primate cells, mammalian cells, rodent cells, etc.), avian cells, fish cells, insect cells, plant cells, fungal cells, spore cells, bacterial cells, and hybrid cells. In some aspects, exemplary cells that can be used with the compositions include platelets, activated platelets,PATENT Attorney Docket No. T002896 WO -2095.0733 stem cells, totipotent cells, pluripotent cells, and / or embryonic stem cells. In some aspects, exemplary cells that can be encapsulated within compositions include, but are not limited to, primary cells and / or cell lines from any tissue. For example, cardiomyocytes, myocytes, hepatocytes, keratinocytes, melanocytes, neurons, astrocytes, embryonic stem cells, adult stem cells, hematopoietic stem cells, hematopoietic cells (e.g. monocytes, neutrophils, macrophages, etc.), ameloblasts, fibroblasts, chondrocytes, osteoblasts, osteoclasts, neurons, sperm cells, egg cells, liver cells, epithelial cells from lung, epithelial cells from gut, epithelial cells from intestine, liver, epithelial cells from skin, etc., and / or hybrids thereof, can be included in the silk / platelet compositions disclosed herein. Those skilled in the art will recognize that the cells listed herein represent an exemplary, not comprehensive, list of cells. Cells can be obtained from donors (allogenic) or from recipients (autologous). Cells can be obtained, as a non-limiting example, by biopsy or other surgical means known to those skilled in the art.

[0170] In some aspects, the cell can be a genetically modified cell. A cell can be genetically modified to express and secrete a desired compound, e.g. a bioactive agent, a growth factor, differentiation factor, cytokines, and the like. Methods of genetically modifying cells for expressing and secreting compounds of interest are known in the art and easily adaptable by one of skill in the art.

[0171] Differentiated cells that have been reprogrammed into stem cells can also be used.

[0172] For example, human skin cells reprogrammed into embryonic stem cells by the transduction of Oct3 / 4, Sox2, c-Myc and Klf4 (Junying Yu, et. ah, Science, 2007, 318, 1917-1920 and Takahashi K. et. al, Cell, 2007, 131, 1-12).

[0173] The methods and systems described herein may be deployed in part or in whole through a machine having a computer, computing device, processor, circuit, and / or server that executes computer readable instructions, program codes, instructions, and / or includes hardware configured to functionally execute one or more operations of the methods and systems disclosed herein. The terms computer, computing device, processor, circuit, and / or server, as utilized herein, should be understood broadly.

[0174] Any one or more of the terms computer, computing device, processor, circuit, and / or server include a computer of any type, capable to access instructions stored in communication thereto such as upon a non-transient computer readable medium, whereupon the computer performs operations of systems or methods described herein upon executing the instructions. In certain embodiments, such instructions themselves comprise a computer, computing device, processor, circuit, and / or server. Additionally or alternatively, a computer, computing device, processor, circuit, and / or server may be a separate hardware device, one or more computing resources distributed across hardware devices, and / or may include such aspects as logical circuits, embedded circuits, sensors, actuators, input and / or output devices, network and / or communication resources, memory resources of any type, processingPATENT Attorney Docket No. T002896 WO -2095.0733 resources of any type, and / or hardware devices configured to be responsive to determined conditions to functionally execute one or more operations of systems and methods herein.

[0175] Network and / or communication resources include, without limitation, local area network, wide area network, wireless, internet, or any other known communication resources and protocols. Example and non-limiting hardware, computers, computing devices, processors, circuits, and / or servers include, without limitation, a general purpose computer, a server, an embedded computer, a mobile device, a virtual machine, and / or an emulated version of one or more of these. Example and non-limiting hardware, computers, computing devices, processors, circuits, and / or servers may be physical, logical, or virtual. A computer, computing device, processor, circuit, and / or server may be: a distributed resource included as an aspect of several devices; and / or included as an interoperable set of resources to perform described functions of the computer, computing device, processor, circuit, and / or server, such that the distributed resources function together to perform the operations of the computer, computing device, processor, circuit, and / or server. In certain embodiments, each computer, computing device, processor, circuit, and / or server may be on separate hardware, and / or one or more hardware devices may include aspects of more than one computer, computing device, processor, circuit, and / or server, for example as separately executable instructions stored on the hardware device, and / or as logically partitioned aspects of a set of executable instructions, with some aspects of the hardware device comprising a part of a first computer, computing device, processor, circuit, and / or server, and some aspects of the hardware device comprising a part of a second computer, computing device, processor, circuit, and / or server.

[0176] A computer, computing device, processor, circuit, and / or server may be part of a server, client, network infrastructure, mobile computing platform, stationary computing platform, or other computing platform. A processor may be any kind of computational or processing device capable of executing program instructions, codes, binary instructions and the like. The processor may be or include a signal processor, digital processor, embedded processor, microprocessor or any variant such as a co-processor (math co-processor, graphic co-processor, communication co-processor and the like) and the like that may directly or indirectly facilitate execution of program code or program instructions stored thereon. In addition, the processor may enable execution of multiple programs, threads, and codes. The threads may be executed simultaneously to enhance the performance of the processor and to facilitate simultaneous operations of the application. By way of implementation, methods, program codes, program instructions and the like described herein may be implemented in one or more threads. The thread may spawn other threads that may have assigned priorities associated with them; the processor may execute these threads based on priority or any other order based on instructions provided in the program code. The processor may include memory that stores methods, codes, instructions andPATENT Attorney Docket No. T002896 WO -2095.0733 programs as described herein and elsewhere. The processor may access a storage medium through an interface that may store methods, codes, and instructions as described herein and elsewhere. The storage medium associated with the processor for storing methods, programs, codes, program instructions or other type of instructions capable of being executed by the computing or processing device may include but may not be limited to one or more of a CD-ROM, DVD, memory, hard disk, flash drive, RAM, ROM, cache and the like.

[0177] A processor may include one or more cores that may enhance speed and performance of a multiprocessor. In embodiments, the process may be a dual core processor, quad core processors, other chip-level multiprocessor and the like that combine two or more independent cores (called a die).

[0178] The methods and systems described herein may be deployed in part or in whole through a machine that executes computer readable instructions on a server, client, firewall, gateway, hub, router, or other such computer and / or networking hardware. The computer readable instructions may be associated with a server that may include a file server, print server, domain server, internet server, intranet server and other variants such as secondary server, host server, distributed server and the like. The server may include one or more of memories, processors, computer readable transitory and / or non-transitory media, storage media, ports (physical and virtual), communication devices, and interfaces capable of accessing other servers, clients, machines, and devices through a wired or a wireless medium, and the like. The methods, programs, or codes as described herein and elsewhere may be executed by the server. In addition, other devices required for execution of methods as described in this application may be considered as a part of the infrastructure associated with the server.

[0179] The server may provide an interface to other devices including, without limitation, clients, other servers, printers, database servers, print servers, file servers, communication servers, distributed servers, and the like. Additionally, this coupling and / or connection may facilitate remote execution of instructions across the network. The networking of some or all of these devices may facilitate parallel processing of program code, instructions, and / or programs at one or more locations without deviating from the scope of the disclosure. In addition, all the devices attached to the server through an interface may include at least one storage medium capable of storing methods, program code, instructions, and / or programs. A central repository may provide program instructions to be executed on different devices. In this implementation, the remote repository may act as a storage medium for methods, program code, instructions, and / or programs.

[0180] The methods, program code, instructions, and / or programs may be associated with a client that may include a file client, print client, domain client, internet client, intranet client and other variants such as secondary client, host client, distributed client and the like. The client may include one or more of memories, processors, computer readable transitory and / or non-transitory media, storage media,PATENT Attorney Docket No. T002896 WO -2095.0733 ports (physical and virtual), communication devices, and interfaces capable of accessing other clients, servers, machines, and devices through a wired or a wireless medium, and the like. The methods, program code, instructions, and / or programs as described herein and elsewhere may be executed by the client. In addition, other devices utilized for execution of methods as described in this application may be considered as a part of the infrastructure associated with the client.

[0181] The client may provide an interface to other devices including, without limitation, servers, other clients, printers, database servers, print servers, file servers, communication servers, distributed servers, and the like. Additionally, this coupling and / or connection may facilitate remote execution of methods, program code, instructions, and / or programs across the network. The networking of some or all of these devices may facilitate parallel processing of methods, program code, instructions, and / or programs at one or more locations without deviating from the scope of the disclosure. In addition, all the devices attached to the client through an interface may include at least one storage medium capable of storing methods, program code, instructions, and / or programs. A central repository may provide program instructions to be executed on different devices. In this implementation, the remote repository may act as a storage medium for methods, program code, instructions, and / or programs.

[0182] The methods and systems described herein may be deployed in part or in whole through network infrastructures. The network infrastructure may include elements such as computing devices, servers, routers, hubs, firewalls, clients, personal computers, communication devices, routing devices and other active and passive devices, modules, and / or components as known in the art. The computing and / or non-computing device(s) associated with the network infrastructure may include, apart from other components, a storage medium such as flash memory, buffer, stack, RAM, ROM and the like. The methods, program code, instructions, and / or programs described herein and elsewhere may be executed by one or more of the network infrastructural elements.

[0183] The methods, program code, instructions, and / or programs described herein and elsewhere may be implemented on a cellular network having multiple cells. The cellular network may either be frequency division multiple access (FDMA) network or code division multiple access (CDMA) network. The cellular network may include mobile devices, cell sites, base stations, repeaters, antennas, towers, and the like.

[0184] The methods, program code, instructions, and / or programs described herein and elsewhere may be implemented on or through mobile devices. The mobile devices may include navigation devices, cell phones, mobile phones, mobile personal digital assistants, laptops, palmtops, netbooks, pagers, electronic books readers, music players, and the like. These mobile devices may include, apart from other components, a storage medium such as a flash memory, buffer, RAM, ROM and one or more computing devices. The computing devices associated with mobile devices may be enabled to executePATENT Attorney Docket No. T002896 WO -2095.0733 methods, program code, instructions, and / or programs stored thereon. Alternatively, the mobile devices may be configured to execute instructions in collaboration with other devices. The mobile devices may communicate with base stations interfaced with servers and configured to execute methods, program code, instructions, and / or programs. The mobile devices may communicate on a peer to peer network, mesh network, or other communications network. The methods, program code, instructions, and / or programs may be stored on the storage medium associated with the server and executed by a computing device embedded within the server. The base station may include a computing device and a storage medium. The storage device may store methods, program code, instructions, and / or programs executed by the computing devices associated with the base station.

[0185] The methods, program code, instructions, and / or programs may be stored and / or accessed on machine readable transitory and / or non-transitory media that may include: computer components, devices, and recording media that retain digital data used for computing for some interval of time; semiconductor storage known as random access memory (RAM); mass storage typically for more permanent storage, such as optical discs, forms of magnetic storage like hard disks, tapes, drums, cards and other types; processor registers, cache memory, volatile memory, non-volatile memory; optical storage such as CD, DVD; removable media such as flash memory (e.g., USB sticks or keys), floppy disks, magnetic tape, paper tape, punch cards, standalone RAM disks, Zip drives, removable mass storage, off-line, and the like; other computer memory such as dynamic memory, static memory, read / write storage, mutable storage, read only, random access, sequential access, location addressable, file addressable, content addressable, network attached storage, storage area network, bar codes, magnetic ink, and the like.

[0186] Certain operations described herein include interpreting, receiving, and / or determining one or more values, parameters, inputs, data, or other information. Operations including interpreting, receiving, and / or determining any value parameter, input, data, and / or other information include, without limitation: receiving data via a user input; receiving data over a network of any type; reading a data value from a memory location in communication with the receiving device; utilizing a default value as a received data value; estimating, calculating, or deriving a data value based on other information available to the receiving device; and / or updating any of these in response to a later received data value. In certain embodiments, a data value may be received by a first operation, and later updated by a second operation, as part of the receiving a data value. For example, when communications are down, intermittent, or interrupted, a first operation to inteipret, receive, and / or determine a data value may be performed, and when communications are restored an updated operation to interpret, receive, and / or determine the data value may be performed.PATENT Attorney Docket No. T002896 WO -2095.0733

[0187] Certain logical groupings of operations herein, for example methods or procedures of the current disclosure, are provided to illustrate aspects of the present disclosure. Operations described herein are schematically described and / or depicted, and operations may be combined, divided, reordered, added, or removed in a manner consistent with the disclosure herein. It is understood that the context of an operational description may require an ordering for one or more operations, and / or an order for one or more operations may be explicitly disclosed, but the order of operations should be understood broadly, where any equivalent grouping of operations to provide an equivalent outcome of operations is specifically contemplated herein. For example, if a value is used in one operational step, the determining of the value may be required before that operational step in certain contexts (e.g. where the time delay of data for an operation to achieve a certain effect is important), but may not be required before that operation step in other contexts (e.g. where usage of the value from a previous execution cycle of the operations would be sufficient for those purposes). Accordingly, in certain embodiments an order of operations and grouping of operations as described is explicitly contemplated herein, and in certain embodiments re-ordering, subdivision, and / or different grouping of operations is explicitly contemplated herein.

[0188] The methods and systems described herein may transform physical and / or or intangible items from one state to another. The methods and systems described herein may also transform data representing physical and / or intangible items from one state to another.

[0189] The elements described and depicted herein, including in flow charts, block diagrams, and / or operational descriptions, depict and / or describe specific example arrangements of elements for purposes of illustration. However, the depicted and / or described elements, the functions thereof, and / or arrangements of these, may be implemented on machines, such as through computer executable transitory and / or non-transitory media having a processor capable of executing program instructions stored thereon, and / or as logical circuits or hardware arrangements. Example arrangements of programming instructions include at least: monolithic structure of instructions; standalone modules of instructions for elements or portions thereof; and / or as modules of instructions that employ external routines, code, services, and so forth; and / or any combination of these, and all such implementations are contemplated to be within the scope of embodiments of the present disclosure Examples of such machines include, without limitation, personal digital assistants, laptops, personal computers, mobile phones, other handheld computing devices, medical equipment, wired or wireless communication devices, transducers, chips, calculators, satellites, tablet PCs, electronic books, gadgets, electronic devices, devices having artificial intelligence, computing devices, networking equipment, servers, routers and the like. Furthermore, the elements described and / or depicted herein, and / or any other logical components, may be implemented on a machine capable of executing program instructions.PATENT Attorney Docket No. T002896 WO -2095.0733 Thus, while the foregoing flow charts, block diagrams, and / or operational descriptions set forth functional aspects of the disclosed systems, any arrangement of program instructions implementing these functional aspects are contemplated herein. Similarly, it will be appreciated that the various steps identified and described above may be varied, and that the order of steps may be adapted to particular applications of the techniques disclosed herein. Additionally, any steps or operations may be divided and / or combined in any manner providing similar functionality to the described operations. All such variations and modifications are contemplated in the present disclosure. The methods and / or processes described above, and steps thereof, may be implemented in hardware, program code, instructions, and / or programs or any combination of hardware and methods, program code, instructions, and / or programs suitable for a particular application. Example hardware includes a dedicated computing device or specific computing device, a particular aspect or component of a specific computing device, and / or an arrangement of hardware components and / or logical circuits to perform one or more of the operations of a method and / or system. The processes may be implemented in one or more microprocessors, microcontrollers, embedded microcontrollers, programmable digital signal processors or other programmable device, along with internal and / or external memory. The processes may also, or instead, be embodied in an application specific integrated circuit, a programmable gate array, programmable array logic, or any other device or combination of devices that may be configured to process electronic signals. It will further be appreciated that one or more of the processes may be realized as a computer executable code capable of being executed on a machine readable medium.

[0190] The computer executable code may be created using a structured programming language such as C, an object oriented programming language such as C++, or any other high-level or low-level programming language (including assembly languages, hardware description languages, and database programming languages and technologies) that may be stored, compiled or interpreted to run on one of the above devices, as well as heterogeneous combinations of processors, processor architectures, or combinations of different hardware and computer readable instructions, or any other machine capable of executing program instructions.

[0191] Thus, in one aspect, each method described above and combinations thereof may be embodied in computer executable code that, when executing on one or more computing devices, performs the steps thereof. In another aspect, the methods may be embodied in systems that perform the steps thereof, and may be distributed across devices in a number of ways, or all of the functionality may be integrated into a dedicated, standalone device or other hardware. In another aspect, the means for performing the steps associated with the processes described above may include any of the hardware and / or computer-readable instructions described above. All such permutations and combinations are contemplated in embodiments of the present disclosure.PATENT Attorney Docket No. T002896 WO -2095.0733

[0192] Unless otherwise specified or indicated by context, the terms “a”, “an”, and “the” mean “one or more.” For example, “a molecule” should be interpreted to mean “one or more molecules”.

[0193] As used herein, “about”, “approximately”, “substantially”, and “significantly” will be understood by persons of ordinary skill in the art and will vary to some extent on the context in which they are used. If there are uses of the term which are not clear to persons of ordinary skill in the art given the context in which it is used, “about” and “approximately” will mean plus or minus ≤10% of the particular term and “substantially” and “significantly” will mean plus or minus >10% of the particular term.

[0194] As used herein, the terms “include” and “including” have the same meaning as the terms “comprise” and “comprising.” The terms “comprise” and “comprising” should be interpreted as being “open” transitional terms that permit the inclusion of additional components further to those components recited in the claims. The terms “consist” and “consisting of’ should be interpreted as being “closed” transitional terms that do not permit the inclusion of additional components other than the components recited in the claims. The term “consisting essentially of’ should be interpreted to be partially closed and allowing the inclusion only of additional components that do not fundamentally alter the nature of the claimed subject matter.

[0195] All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0196] All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein.

[0197] Preferred aspects of this invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations of those preferred aspects may become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventors expect a person having ordinary skill in the art to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than as specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.PATENT Attorney Docket No. T002896 WO -2095.0733

[0198] While the invention has been illustrated and described in detail in the foregoing drawings and description, the same is to be considered as illustrative and not restrictive in character, it being understood that only illustrative embodiments thereof have been shown and described and that all changes and modifications that come within the spirit of the invention are desired to be protected. For example, any of the features or functions of any of the embodiments disclosed herein may be incorporated into any of the other embodiments disclosed herein.

[0199] The following examples illustrate some embodiments and aspects of the invention. It will be apparent to those skilled in the relevant art that various modifications, additions, substitutions, and the like can be performed without altering the spirit or scope of the invention, and such modifications and variations are encompassed within the scope of the invention as defined in the claims which follow. The following examples do not in any way limit the invention.EXAMPLES

[0200] Example 1: Mechanical Testing

[0201] The process of continuous fiber extrusion (CFE) of silk-based biopolymer leads to the manufacturing of multidimensional laminate composites. These composites have been mechanically tested following ASTM D3039 under tension to characterize their ultimate tensile strength and fracture strain as well as their yielding and rupture behaviors.

[0202] Among the parameters assessed, ultimate tensile strength and fracture strain gave a good indication of the composite’s behaviors. These properties were mainly influenced by the fiber type, the matrix composition, the fiber volume fraction and the weaving pattern.

[0203] Testing has shown that a 3D printed silk-based composite can be compared to similarly tested polyurethane (PU) leather from the market with some compositions having higher ultimate tensile strength.

[0204] A 40:20:40 (silk:alginate:glycerol) braided cotton biaxial weave woven three-dimensional silk material was compared to a polyurethane synthetic leather. Overall, the biaxial weave performed better in mechanical testing compared to the PU synthetic leather (Fig. 11). The biaxial weave had an ultimate tensile strength of around 20 MPa while the polyurethane synthetic leather had an ultimate tensile strength of around 13 MPa.

[0205] Fig. 12A and 12B depict data for composite test specimens that were fabricated with two different grid resolutions corresponding to fiber orientations of 0° and 45°. Specimens were produced across multiple fiber-volume -fraction ranges, including approximately 7.6-8.2% and 3.6-3.7%.

[0206] Tensile testing was performed to determine the fracture strain of each configuration. The results are shown in Fig. 12A. For each fiber-volume-fraction range, the fracture strain values for the 0° orientation and the 45° orientation were recorded.PATENT Attorney Docket No. T002896 WO -2095.0733

[0207] Ultimate tensile strength (UTS) was also measured for the same sets of specimens. The UTS results are shown in Fig. 12B. The data demonstrate the influence of fiber- volume fraction and fiber orientation on the measured tensile properties of the composite materials. The fiber volume fraction increases because the weave is tighter.

[0208] Figs. 13A and 13B depict data for composite structures having layered fiber arrangements that were evaluated to characterize the effects of fiber-volume fraction and fiber orientation on material performance. Specimens were prepared with three representative fiber-volume-fraction ranges of approximately 4.7-4.9%, 7.8-8.2%, and 11.7-13.1%.

[0209] Mechanical testing was conducted to measure both stress at failure and fracture strain. Fig. 13A shows, for a weave with 0° fiber orientation, stress values reported relative to the left vertical axis and fracture strain values reported relative to the right vertical axis. Each fiber-volume-fraction range exhibits distinct performance characteristics.

[0210] Stress-strain behavior for multiple layered configurations is shown in Fig. 13B. Curves correspond to different fiber orientations, including 0° (along the fibers) and 45° (diagonally), as well as different fiber-volume-fraction ranges. Insets illustrate the associated fiber-layering patterns, and arrows denote the applied loading direction. These results demonstrate the influence of layering configuration and fiber orientation on overall material stiffness, yield behavior, and fracture response. The increase in fiber volume fraction results in the superimposition of several printed layers.

[0211] Figs. 14A, 14B, and 14C depict composite specimens that were fabricated using two different weaving patterns: a triaxial weave, a grid pattern with fibers oriented at 0°, and a grid pattern with fibers oriented at 45°. Mechanical testing was conducted to determine ultimate tensile strength (UTS) and fracture strain for each configuration.

[0212] In Fig. 14A, stress is presented on the left vertical axis and fracture strain on the right vertical axis. The triaxial weave and grid pattern tested at 0° exhibited comparatively higher stress relative to the grid configuration tested at 45°, while the grid pattern tested at 45° exhibited higher fracture strain than the grid pattern tested at 0° and the triaxial weave. These results demonstrate the influence of fiber-interlacing patterns on mechanical performance.

[0213] In Fig. 14B, the contribution of different fiber materials to ultimate tensile strength and fracture strain was assessed. Specimens were produced using the following matrix-fiber formulations: cotton fibers in a 20:20:60 ratio matrix, silk fibers in a 20:20:60 ratio matrix, and silk cord in a 20:20:60 matrix.

[0214] UTS and fracture strain measurements were obtained, and the results are plotted in Fig. 14B. For all formulations, UTS values are shown using the left vertical axis and fracture strain using the right vertical axis. Silk-containing composites demonstrated increased UTS relative to cotton-PATENT Attorney Docket No. T002896 WO -2095.0733 containing composites, with the silk cord formulation exhibiting higher fracture strain than the silk formulation.

[0215] In Fig. 14C, multiple fiber materials were incorporated into triaxial weave structures to assess their mechanical behavior. Tested materials included silk cord in three ratios (20:20:60, 40:20:40, and 33:33:33) as well as a polyurethane (PU) synthetic leather variant.

[0216] Results are presented in Fig. 14C, which displays UTS values on the left vertical axis and fracture-strain values on the right vertical axis. The PU synthetic leather formulation exhibited the highest UTS and one of the highest fracture-strain values among the tested materials.

[0217] Example 2: Composition and Fiber Coating

[0218] Aqueous gelatin (Type A) solution showed good results. Solutions of concentration in a range of 1 to 10% were tested and were very effective in stiffening the fiber before printing. Gelatin was also included in foam compositions. Foam materials (10% total solid content) of composition 10:10:20:60 (Silk fibroin: Gelatin: Alginate: Glycerol) and 20:20:40:20 have shown good results. Adding gelatin permits better printability via improved extrusion and shape fidelity, which increases stabilization of the foam material during compression. This preserves the foam microstructure during extrusion. EQUIVALENTS AND SCOPE

[0219] The recitation of a listing of elements in any definition of a variable herein includes definitions of that variable as any single element or combinations (or subcombinations) of listed elements. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof. Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein.

[0220] In addition to the features described above and elsewhere herein, the present disclosure also includes the following clauses:Clause 1. A method of making a woven three-dimensional silk material, the method comprising: (a) selecting a digital design parameter set comprising at least one woven three-dimensional silk material property; and (b) weaving, via coextrusion, a silk fibroin material and a continuous fiber according to the digital design parameter set to form a woven three-dimensional silk material possessing the at least one woven three-dimensional silk material property.Clause 2. The method of clause 1, the method further comprising: c) compressing the woven three-dimensional silk material, thereby forming a compressed woven three-dimensional silk material. Clause 3. The method of any one of the preceding clauses, the method further comprising: d) selecting a curing condition set for at least one of the woven three-dimensional silk material or the compressed woven three-dimensional silk material; and e) curing the woven three-dimensional silkPATENT Attorney Docket No. T002896 WO -2095.0733 material or compressed woven three-dimensional silk material according to the curing condition set to form a cured woven three-dimensional silk material or a cured compressed woven three-dimensional silk material.Clause 4. The method of any one of the preceding clauses, the method further comprising: f) applying at least one layer of a silk foam to at least one woven three-dimensional silk material, the compressed woven three-dimensional silk material, or the cured compressed woven three-dimensional silk material; and g) optionally compressing or thermally compressing the at least one layer of the silk foam.Clause 5. The method of any one of clause 2 to the immediately preceding clause, wherein the compressing of step c) results in at least one of thermal, mechanical, or chemical crosslinking.Clause 6. The method of any one of the preceding clauses, wherein coextrusion comprises coextruding the silk fibroin material with the continuous fiber through a co-extrusion nozzle comprising: a first duct having a first diameter, the first duct comprising a fitting on a first end; a continuous thread duct having a continuous thread duct diameter, the continuous thread duct arranged at an incline with respect to the first duct, the continuous thread duct configured to receive and guide the continuous fiber; and wherein a second end of the first duct is configured to receive the continuous fiber exiting the continuous thread duct; and a co-flow tip arranged at the second end of the first duct and configured to receive a co-extruded material for weaving, the co-extruded material comprising the silk fibroin material with the continuous fiber.Clause 7. The method of any one of the preceding clauses, wherein the digital design parameter set comprises at least one of a weaving pattern, a number of printed layers, a density of lines, a number of intersections, an angle of vertices at intersections, a Poisson's ratio, a number of layers of printing, a shape, a selection of the continuous fiber material, a selection of the silk fibroin material, a selection of a silk fibroin material property, a temperature of extrusion, a rate of extrusion, a fan speed, a printing speed, a biaxial interweave, a triaxial interweave, a yarn size, a pitch distance, or combinations thereof.Clause 8. The method of the immediately preceding clause, wherein decreasing at least one of the density of lines or the number of 3D printed layers, and / or modifying the weaving pattern, increases the flexibility of the woven three-dimensional silk material, the compressed woven three-dimensional silk material, or the cured compressed woven three-dimensional silk material.Clause 9. The method of the clause 7, wherein the selection of a silk fibroin material property comprises at least one of an elastic modulus, an ultimate stress, a tensile toughness, an ultimate strain, a β-sheet content of the silk fibroin, a β-turn content of the silk fibroin, a transmittance, aPATENT Attorney Docket No. T002896 WO -2095.0733 color, a hardness, a stiffness, a conductivity, a resistance, a thermal conductivity, a permeability, or a permittivity.Clause 10. The method of the clause 7, wherein the at least one woven three-dimensional silk material property is a desired flexibility, and the digital design parameter set includes the desired flexibility in a plurality of flexibility values, each flexibility value of the plurality of flexibility values paired with a corresponding value of density of lines for the weaving that results in the flexibility value.Clause 11. The method of the clause 7, wherein the weaving pattern is a rattan pattern, a hexagonal rattan pattern, a dutch weave, a twill weave, a plain weave, a twill dutch weave, a lock crimp, an inter-crimp, a twill dutch double, stranded, a basket wave, a plait, a Poisson's ratio pattern, a triaxial weave pattern, a biaxial weave pattern, a warp-weft pattern, or a satin weave.Clause 12. The method of any one of the preceding clauses, wherein the continuous printing deposits layers in orthogonal directions, thereby forming a micro-architecture.Clause 13. The method of any one of the preceding clauses, wherein the continuous fiber is selected from the group consisting of latex, silk cord, silk yarn, a waste silk product, a filament, an electrode, a counter-electrode, an enzymatically-coated fiber, a non-spun thread, an electrochemical fiber, a conductive fiber, a copper material, a platinum material, a metallic material, a polyester material, a cotton material, a graphite fiber, a nylon material, a light-emitting fiber element (e.g., electroluminescent wires, LEDs, or optical fibers), or reeled silk fibers.Clause 14. The method of any one of the preceding clauses, wherein the continuous fiber has a diameter of between 0.1 mm and 1.5 mm.Clause 15. The method of any one of the preceding clauses, wherein the continuous fiber is coated with a coating composition to form a coated continuous fiber, the coating composition comprising at least one of latex, a silk fibroin solution, or a glycerol solution.Clause 16. The method of the immediately preceding clause, wherein the diameter of the coated continuous fiber is a result of the coating composition.Clause 17. The method of clause 15, wherein the silk fibroin solution comprises silk fibroin in a concentration of between 1% and 25%.Clause 18. The method of clause 15, wherein the coated continuous fiber possesses a resistance to compression that is superior to a comparison resistance to compression of a comparison continuous fiber lacking a coating but otherwise identical to the coated continuous fiber.Clause 19. The method of clause 15, wherein the coated continuous fiber has a resistance to compression that is at least 30%, at least 50%, at least 75%, at least 100%, or at least 150% greaterPATENT Attorney Docket No. T002896 WO -2095.0733 than a comparison resistance to compression of a comparison continuous fiber lacking a coating but otherwise identical to the coated continuous fiber.Clause 20. The method of any one of the preceding clauses, wherein the digital design parameter set includes instructions to vary one or more of the digital design parameters throughout the coextrusion. Clause 21. The method of the immediately preceding clause, wherein the variation of the digital design parameter set results in a gradient of at least one of color, stiffness, porosity, softness, elasticity, tensile strength, pattern, extensibility, or a combination thereof.Clause 22. The method of any one of the preceding clauses, wherein the method does not produce chemical waste or solvent waste.Clause 23. The method of any one of the preceding clauses, wherein the silk fibroin material comprises an additive.Clause 24. The method of the immediately preceding clause, wherein the additive is an active agent. Clause 25. The method of any one of the preceding clauses, wherein the silk fibroin material is a silk foam or a silk paste.Clause 26. The method of clause 23, wherein the additive is a biological additive.Clause 27. An article made by the method of any one of the preceding clauses.Clause 28. The article of the immediately preceding clause, wherein the article is at least one of a shoe, a shoe upper, a garment, a textile, an insulating material, an engineered composite, a metamaterial, a biofabric, a 3D structure, a structural material, a building material, an insulating material, a packaging material, a tool, a biomedical implant, a prosthetic, or an orthotic.Clause 29. The article of the immediately preceding clause, wherein the textile comprises a biaxial interweaved fiber or a triaxial interweaved fiber.Clause 30. The article of any one of clauses 27 to the immediately preceding clause, wherein the article has a thickness of at least 0.5 mm.Clause 31. The article of any one of clauses 27 to the immediately preceding clause, wherein the article comprises an embedded component.Clause 32. The article of any one of clauses 27 to the immediately preceding clause, wherein the article further comprises at least one of a sensor, electronic component, electronic device, haptic switch, electrical wiring, power supply, RFID tag, or combinations thereof.Clause 33. The article of any one of clauses 27 to the immediately preceding clause, wherein the article comprises a triaxial weave pattern, wherein the article has at least one of an isotropic response to tensile deformation, an isotropic response to shear deformation, an extensibility, or a combination thereof.PATENT Attorney Docket No. T002896 WO -2095.0733 Clause 34. The article of any one of clauses 27 to the immediately preceding clause, wherein the article has one or more of the following properties: an elastic modulus between 0.1-10 GPa; an ultimate stress between 1-500 MPa; a tensile toughness of between 0.5-5.0 MJ / m3; an ultimate strain of between 0.1-50%; silk fibroin having a β-sheet content of greater than 5%; silk fibroin having a β-turn content of less than 15%; a transmittance of 80% or greater over a visual range; and combinations thereof.Clause 35. The article of any one of clauses 27 to claim 33, wherein prior to drying or in an absence of drying the article is wet and has one or more of the following properties: an elastic modulus between 10-1000 MPa; an ultimate stress between 0.1-100 MPa; a tensile toughness of between 10.0-100.0 MJ / m3; an ultimate strain of between 35-500%; silk fibroin having a β-sheet content of less than 10%; silk fibroin having a β-turn content of between 5-35%; a transmittance of 80% or greater over a visual range; and combinations thereof.Clause 36. A method comprising: a) pre-treating a thread to enhance mechanical strength; and b) coextruding the pre -treated thread with a biopolymer to form a woven three-dimensional material. Clause 37. The method of clause 36, wherein pre-treating comprises coating the thread with a solution comprising latex, gelatin, alginate, and / or silk fibroin.Clause 38. The method of the immediately preceding clause, wherein the solution further comprises glycerol.Clause 39. The method of clause 36, wherein pre-treating comprises coating the thread with a solution comprising glycerol.Clause 40. The method of clause 36, wherein pre-treating comprises coating the thread with at least one of a polymer or a biopolymer.Clause 41. The method of clause 36, wherein the woven three-dimensional material possesses at least one characteristic that is superior to a corresponding characteristic of a woven three-dimensional material lacking the pre-treated thread.Clause 42. The method of clause 41, wherein the characteristic is a resistance to stretching, and the woven three-dimensional material has a resistance to stretching that is at least 30%, at least 50%, at least 75%, at least 100%, or at least 150% higher than a woven three-dimensional material lacking the pre-treated thread.Clause 43. The method of clause 41, wherein the characteristic is a resistance to shear stress, and the woven three-dimensional material has a resistance to shear stress that is at least 30%, at least 50%, at least 75%, at least 100%, or at least 150% higher than a woven three-dimensional material lacking the pre-treated thread.PATENT Attorney Docket No. T002896 WO -2095.0733 Clause 44. The method of clause 36, wherein the pre-treated thread is at least one of latex, silk cord, silk yarn, a waste silk product, a filament, an electrode, a counter-electrode, an enzymatically-coated fiber, a non-spun thread, an electrochemical fiber, a conductive fiber, a copper material, a platinum material, a metallic material, a polyester material, a cotton material, a graphite fiber, a nylon material, a light-emitting fiber element (e.g., electroluminescent wires, LEDs, or optical fibers), or reeled silk fiber.Clause 45. The method of clause 36, wherein the pre-treated thread possesses a resistance to compression that is superior to a comparison resistance to compression of a comparison thread lacking a pre-treatment but otherwise identical to the pre-treated thread.Clause 46. The method of clause 36, wherein the pre-treated thread has a resistance to compression that is at least 30%, at least 50%, at least 75%, at least 100%, or at least 150% greater than a comparison resistance to compression of a thread lacking the pre-treatment but otherwise identical to the pre -treated thread.Clause 47. The method of clause 1, wherein the silk fibroin material comprises at least one of a liquid composition, a whipped silk cream, or a silk meringue.Clause 48. The method of clause 1, wherein the silk fibroin material is made from the liquid composition of any one of claims 1 to 3, 24 to 31, 42 to 44, 65 to 72, 83 to 85, 106 to 113, 124 to 126, 147 to 154, 170 to 176, or 181 to 188 of United States Patent Application 18 / 403,477.Clause 49. The method of clause 1, wherein the silk fibroin material is made from the whipped silk cream of or made by the method of any one of claims 4, 8 to 32, 45, 49 to 73, 86, 90 to 114, 127, 131 to 155, or 165 to 188 of United States Patent Application 18 / 403,477.Clause 50. The method of clause 1, wherein the silk fibroin material is made from the silk meringue of or made by the method of any one of claims 5, 9 to 32, 46, 50 to 73, 87, 91 to 114, 128, 132 to 155, or 165 to 188 of United States Patent Application 18 / 403,477.Clause 51. A silk leather that is a layered structure comprising at least one woven three-dimensional silk material made by the method of any one of clauses 1 - 26 and 36 - 50.Clause 52. The silk leather of clause 51, wherein the layered structure comprises a silk foam layer. Clause 53. The silk leather of clause 51 or 52, wherein the layered structure comprises at least one of a whipped silk cream layer, a silk meringue layer, a compressed silk meringue layer, a hot-pressed silk meringue layer, or a fabric layer.Clause 54. The method of clause 37, wherein the gelatin comprises Type A gelatin.

[0221] The scope of the present invention is not intended to be limited to the above Description, but rather is as set forth in the following claims:

Claims

1. PATENT Attorney Docket No. T002896 WO -2095.0733 CLAIMSWhat is claimed is:

1. A method of making a woven three-dimensional silk material, the method comprising:a) selecting a digital design parameter set comprising at least one woven three- dimensional silk material property; andb) weaving, via coextrusion, a silk fibroin material and a continuous fiber according to the digital design parameter set to form a woven three-dimensional silk material possessing the at least one woven three-dimensional silk material property, optionally wherein the silk fibroin material comprises at least one of a liquid composition, a whipped silk cream, or a silk meringue2. The method of claim 1, the method further comprising:c) compressing the woven three-dimensional silk material, thereby forming a compressed woven three-dimensional silk material.

3. The method of any one of the preceding claims, the method further comprising:d) selecting a curing condition set for at least one of the woven three-dimensional silk material or the compressed woven three-dimensional silk material; ande) curing the woven three-dimensional silk material or compressed woven three- dimensional silk material according to the curing condition set to form a cured woven three- dimensional silk material or a cured compressed woven three-dimensional silk material.

4. The method of any one of the preceding claims, the method further comprising:f) applying at least one layer of a silk foam to at least one woven three-dimensional silk material, the compressed woven three-dimensional silk material, or the cured compressed woven three-dimensional silk material; andg) optionally compressing or thermally compressing the at least one layer of the silk foam.

5. The method of any one of claim 2 to the immediately preceding claim, wherein the compressing of step c) results in at least one of thermal, mechanical, or chemical crosslinking.

6. The method of any one of the preceding claims, wherein coextrusion comprises co-extruding the silk fibroin material with the continuous fiber through a co-extrusion nozzle comprising:a first duct having a first diameter, the first duct comprising a fitting on a first end;a continuous thread duct having a continuous thread duct diameter, the continuous thread duct arranged at an incline with respect to the first duct, the continuous thread duct configured toPATENT Attorney Docket No. T002896 WO -2095.0733 receive and guide the continuous fiber, wherein a second end of the first duct is configured to receive the continuous fiber exiting the continuous thread duct; anda co-flow tip arranged at the second end of the first duct and configured to receive a coextruded material for weaving, the co-extruded material comprising the silk fibroin material with the continuous fiber.

7. The method of any one of the preceding claims, wherein the digital design parameter set comprises at least one of a weaving pattern, a number of printed layers, a density of lines, a number of intersections, an angle of vertices at intersections, a Poisson’s ratio, a number of layers of printing, a shape, a selection of the continuous fiber material, a selection of the silk fibroin material, a selection of a silk fibroin material property, a temperature of extrusion, a rate of extrusion, a fan speed, a printing speed, a biaxial interweave, a triaxial interweave, a yarn size, a pitch distance, or combinations thereof,optionally wherein the selection of a silk fibroin material property comprises at least one of an elastic modulus, an ultimate stress, a tensile toughness, an ultimate strain, a β-sheet content of the silk fibroin, a β-turn content of the silk fibroin, a transmittance, a color, a hardness, a stiffness, a conductivity, a resistance, a thermal conductivity, a permeability, or a permittivity,optionally wherein the at least one woven three-dimensional silk material property is a desired flexibility, and the digital design parameter set includes the desired flexibility in a plurality of flexibility values, each flexibility value of the plurality of flexibility values paired with a corresponding value of density of lines for the weaving that results in the flexibility value, and optionally wherein the weaving pattern is a rattan pattern, a hexagonal rattan pattern, a dutch weave, a twill weave, a plain weave, a twill dutch weave, a lock crimp, an inter-crimp, a twill dutch double, stranded, a basket wave, a plait, a Poisson’s ratio pattern, a triaxial weave pattern, a biaxial weave pattern, a warp-weft pattern, or a satin weave, andoptionally wherein decreasing at least one of the density of lines or the number of 3D printed layers, and / or modifying the weaving pattern, increases the flexibility of the woven three-dimensional silk material, the compressed woven three-dimensional silk material, or the cured compressed woven three-dimensional silk material.

8. The method of any one of the preceding claims, wherein the continuous printing deposits layers in orthogonal directions, thereby forming a micro-architecture.

9. The method of any one of the preceding claims, wherein the continuous fiber is selected from the group consisting of latex, silk cord, silk yarn, a waste silk product, a filament, an electrode, a counter-electrode, an enzymatically-coated fiber, a non-spun thread, an electrochemical fiber, a conductive fiber, a copper material, a platinum material, a metallic material, a polyester material, aPATENT Attorney Docket No. T002896 WO -2095.0733 cotton material, a graphite fiber, a nylon material, a light-emitting fiber element (e.g., electroluminescent wires, LEDs, or optical fibers), or reeled silk fibers, and / orhas a diameter of between 0.1 mm and 1.5 mm, and / oris coated with a coating composition to form a coated continuous fiber, the coating composition comprising at least one of latex, a silk fibroin solution, or a glycerol solution, optionally wherein the diameter of the coated continuous fiber is a result of the coating composition, and / or wherein the silk fibroin solution comprises silk fibroin in a concentration of between 1% and 25%, and / orwherein the coated continuous fiber possesses a resistance to compression that is superior to a comparison resistance to compression of a comparison continuous fiber lacking a coating but otherwise identical to the coated continuous fiber, and / orwherein the coated continuous fiber has a resistance to compression that is at least 30%, at least 50%, at least 75%, at least 100%, or at least 150% greater than a comparison resistance to compression of a comparison continuous fiber lacking a coating but otherwise identical to the coated continuous fiber.

10. The method of any one of the preceding claims, wherein the digital design parameter set includes instructions to vary one or more of the digital design parameters throughout the coextrusion.

11. The method of the immediately preceding claim, wherein the variation of the digital design parameter set results in a gradient of at least one of color, stiffness, porosity, softness, elasticity, tensile strength, pattern, extensibility, or a combination thereof.

12. The method of any one of the preceding claims, wherein the method does not produce chemical waste or solvent waste.

13. The method of any one of the preceding claims, wherein the silk fibroin material comprises an additive, optionally wherein the additive is an active agent or a biological additive.

14. The method of any one of the preceding claims, wherein the silk fibroin material is a silk foam or a silk paste.

15. An article made by the method of any one of the preceding claims, optionallywherein the article is at least one of a shoe, a shoe upper, a garment, a textile, an insulating material, an engineered composite, a metamaterial, a biofabric, a 3D structure, a structural material, a building material, an insulating material, a packaging material, a tool, a biomedical implant, a prosthetic, or an orthotic, optionally wherein the textile comprises a biaxial interweaved fiber or a triaxial interweaved fiber.

16. The article of claim 14, wherein the article comprises a thickness of at least 0.5 mm, and / or an embedded component, and / orPATENT Attorney Docket No. T002896 WO -2095.0733 at least one of a sensor, electronic component, electronic device, haptic switch, electrical wiring, power supply, RFID tag, or combinations thereof, and / ora triaxial weave pattern, wherein the article has at least one of an isotropic response to tensile deformation, an isotropic response to shear deformation, an extensibility, or a combination thereof.

17. The article of any one of claims 14 to the immediately preceding claim,wherein the article has one or more of the following properties:an elastic modulus between 0.1-10 GPa:an ultimate stress between 1-500 MPa;a tensile toughness of between 0.5-5.0 MJ / m3;an ultimate strain of between 0.1-50%;silk fibroin having a [l-sheet content of greater than 5%;silk fibroin having a [3-turn content of less than 15%;a transmittance of 80% or greater over a visual range; andcombinations thereof.

18. The article of any one of claims 14 to claim 15,wherein prior to drying or in an absence of drying the article is wet and has one or more of the following properties:an elastic modulus between 10-1000 MPa;an ultimate stress between 0.1-100 MPa;a tensile toughness of between 10.0-100.0 MJ / m3;an ultimate strain of between 35-500%;silk fibroin having a P-sheet content of less than 10%;silk fibroin having a P-turn content of between 5-35%;a transmittance of 80% or greater over a visual range; andcombinations thereof.

19. A method comprising:a) pre-treating a thread to enhance mechanical strength; andb) co-extruding the pre-treated thread with a biopolymer to form a woven three-dimensional material, optionally wherein pre-treating comprises coating the thread with a solution comprising latex, gelatin, alginate, and / or silk fibroin,optionally wherein the gelatin comprises Type A gelatin, optionallywherein the solution further comprises glycerol,optionally wherein pre-treating comprises coating the thread with a solution comprising glycerol,PATENT Attorney Docket No. T002896 WO -2095.0733 optionally wherein pre-treating comprises coating the thread with at least one of a polymer or a biopolymer, optionally wherein the woven three-dimensional material possesses at least one characteristic that is superior to a corresponding characteristic of a woven three-dimensional material lacking the pre -treated thread, optionally wherein the characteristic is a resistance to stretching, and the woven three-dimensional material has a resistance to stretching that is at least 30%, at least 50%, at least 75%, at least 100%, or at least 150% higher than a woven three-dimensional material lacking the pre-treated thread, and / or wherein the characteristic is a resistance to shear stress, and the woven three-dimensional material has a resistance to shear stress that is at least 30%, at least 50%, at least 75%, at least 100%, or at least 150% higher than a woven three-dimensional material lacking the pretreated thread, andoptionally wherein the pre-treated thread is at least one of latex, silk cord, silk yarn, a waste silk product, a filament, an electrode, a counter-electrode, an enzymatically-coated fiber, a non-spun thread, an electrochemical fiber, a conductive fiber, a copper material, a platinum material, a metallic material, a polyester material, a cotton material, a graphite fiber, a nylon material, a light-emitting fiber element (e.g., electroluminescent wires, LEDs, or optical fibers), or reeled silk fiber, and / or wherein the pre-treated thread possesses a resistance to compression that is superior to a comparison resistance to compression of a comparison thread lacking a pre-treatment but otherwise identical to the pre-treated thread, and / orwherein the pre-treated thread has a resistance to compression that is at least 30%, at least 50%, at least 75%, at least 100%, or at least 150% greater than a comparison resistance to compression of a thread lacking the pre-treatment but otherwise identical to the pre-treated thread.

20. A silk leather that is a layered structure comprising at least one woven three-dimensional silk material made by the method of any one of claims 1 - 13 and 18 - 19, optionally wherein the layered structure comprises a silk foam layer, optionally wherein the layered structure comprises at least one of a whipped silk cream layer, a silk meringue layer, a compressed silk meringue layer, a hot-pressed silk meringue layer, or a fabric layer.

Images