Compositions of cold slurries with hyaluronic acid and methods therefor

Abstract

Disclosed herein is a composition comprising an amount of water; hyaluronic acid; and a first excipient, wherein the composition is configured to form into a flowable cold slum when the composition is exposed to a temperature below 0° C.

Description

[Technical field]

[0001] Technical Field This application claims priority under 35 USC § 119(c) to U.S. Ser. No. 63 / 309291, filed February 11, 2022, the contents of which are incorporated by reference in their entirety.

[0002] The present disclosure relates generally to compositions and methods for producing flowable and / or injectable cold slurry-forming biomaterials. More specifically, the present invention relates to the formulation of a biocompatible solution preferably comprising a liquid (e.g., saline, water, or phosphate buffered saline), glycerol, hyaluronic acid, a poloxamer, and optionally a lipid. [Background technology]

[0003] background Cold slurries (e.g., ice slurries) are known in the art as compositions made of sterile ice particles of water, varying amounts of excipients or additives, such as freezing point depressants, hydrotropic molecules, and optionally one or more active pharmaceutical ingredients, as described in U.S. Application No. 15 / 505,042 ("'042 Application"; Publication No. US2017 / 0274011), the disclosure of which relating to the formulation of cold slurry compositions is incorporated herein by reference in its entirety. Prior art cold slurries can be delivered, preferably by injection, to tissues of a subject, preferably a human patient, to cause selective or non-selective cryotherapy and / or cryolipolysis for prophylactic, therapeutic, or cosmetic surgery purposes. Injectable cold slurries can be used to treat a variety of disorders that require inhibition of nerve conduction. For example, U.S. Application No. 15 / 505,039 ("the '039 Application"; Publication No. US2017 / 0274078), which is incorporated by reference in its entirety for its disclosure of reversible inhibition of nerve conduction, discloses the use of the slurry to induce reversible degeneration of nerves (e.g., via Wallerian degeneration) by causing lipid crystallization in the myelin sheath of the nerve. The '039 Application also discloses the use of the injectable cold slurry to treat a variety of other disorders requiring somatic or autonomic nerve inhibition, such as motor spasms, hypertension, hyperhidrosis, and urinary incontinence.

[0004] A method for preparing the cold slurry is shown in U.S. Application No. 16 / 080,092 ("the '092 application"; Publication No. US2019 / 0053939), which is incorporated herein by reference in its entirety. However, the method disclosed in the '092 application requires the point of care to manufacture the cold slurry by installing a medical ice slurry manufacturing system. This technology also requires the point of care to employ steps to maintain the sterility of the cold slurry during manufacture and prior to administration. Alternative methods for preparing the cold slurry are disclosed in U.S. Patent No. 11,241,330 and International Publication No. WO 2022 / 261494 A1 ("'494 PCT"). The disclosures in WO 2022 / 211904 A1 ("'904 PCT") regarding manufacturing methods are incorporated herein by reference. The disclosures in the '494 PCT relating to cold slurry compositions are incorporated herein by reference. The disclosures in this application are compatible with the methods and systems disclosed in International Publication No. WO 2017 / 147367 A1 ("'367 PCT") and the '904 PCT.

[0005] The '904 PCT discloses a method for easily transporting sterile biomaterial to the point of care using standard transport techniques, where the biomaterial can be transformed into a flowable, injectable cold slurry at the point of care without compromising the sterility of the biomaterial at the point of care, without requiring manufacturing facilities to be available at the point of care. The disclosure of the present application is compatible with the methods and systems disclosed in the '904 PCT.

[0006] There is a need for compositions and methods that allow for simple transport, storage and preparation of flowable and / or injectable cold slurries at clinical points of care, without compromising the sterility of the biomaterial (e.g., the solution that is transformed into the cold slurry) during preparation, without requiring specialized manufacturing equipment to be available at the point of care, and without compromising the sterility of the biomaterial at the point of care. The present disclosure addresses this need by providing improved cold slurry compositions and preparation methods that allow the biocompatible solution to be received at the point of care in a container that is easily transported and stored, and that can be located in a standard freezer, and can optionally perform further physical agitation of the contents of the container to convert the biocompatible solution into a therapeutic substance, e.g., a flowable and / or injectable cold slurry. The present disclosure describes compositions and methods that can provide an adequate and consistent amount of ice particles after exposure to freezing temperatures, allowing for high injection reliability of the cold slurry through a syringe needle. Summary of the Invention

[0007] overview In one aspect, the present disclosure provides a composition comprising an amount of water, hyaluronic acid, and a first excipient, the composition being configured to form into a flowable cold slurry comprising a plurality of ice crystals when exposed to a temperature below 0° C.

[0008] In some embodiments, the composition further comprises a water-soluble surfactant. In further embodiments, the water-soluble surfactant is a poloxamer molecule. In some embodiments, the composition comprises a plurality of poloxamer molecules. In some embodiments, the composition comprises a poloxamer particle, the poloxamer particle comprising a plurality of poloxamer molecules. In further embodiments, the poloxamer particle is a micelle. In certain embodiments, the poloxamer molecule is selected from the group consisting of poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 183, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer 235, poloxamer 237, poloxamer 238, poloxamer 282, poloxamer 284, poloxamer 288, poloxamer 331, poloxamer 333, poloxamer 334, poloxamer 335, poloxamer 338, poloxamer 401, poloxamer 402, poloxamer 403, poloxamer 407, poloxamer 105 benzoate, poloxamer 182 dibenzoate, and combinations thereof. In some embodiments, the poloxamer is poloxamer 407 and the concentration of poloxamer 407 is about 0.1% (w / w) to 10% (w / w). In some embodiments, the concentration of poloxamer 407 in the composition is about 0.5% (w / w). As used herein, (w / w) and (w / v) are interchangeable.

[0009] In certain embodiments, the composition further comprises a first excipient selected from the group consisting of a salt, an ion, lactated Ringer's solution, a sugar, a biocompatible surfactant, a polyol, and combinations thereof.

[0010] In some embodiments, the first excipient is glycerol. In some embodiments, the concentration of glycerol in the composition is about 12% to 25% (w / w). In some embodiments, the concentration of glycerol in the composition is about 19% (w / w).

[0011] In some embodiments, the composition further comprises a second excipient, hi some embodiments, the second excipient is sodium chloride or sodium phosphate to form saline or phosphate buffered saline.

[0012] In some embodiments, the composition further comprises a third excipient. In some embodiments, the third excipient is a water-insoluble substance. In some embodiments, the water-insoluble substance is a lipid. In some embodiments, the lipid is selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), egg sphingomyelin (DPSM), dipalmitoyl phosphatidylcholine (DPPC), dicetyl phosphate (DCP), La-phosphatidylcholine (soybean PC), phosphatidylethanolamine, (PE), phosphatidylserine (PS), and phosphatidylglycerol (PG).

[0013] In certain aspects, the compositions disclosed herein are configured to form a plurality of ice crystals when exposed to a temperature of about -25°C to about -5°C.

[0014] In certain aspects, the compositions disclosed herein are configured to have an injection force of less than about 30 lbf when injected through a 16G needle, a 17G needle, an 18G needle, a 19G needle, a 20G needle, a 22G needle, a 23G needle, or a 24G needle.

[0015] In certain aspects, the compositions disclosed herein are configured to have an injection force of less than about 30 lbf when injected through a 17G or 18G needle.

[0016] In one aspect, the method for preparing cold slurry for administration to patient at clinical point of care is provided.In one embodiment, the method comprises: preparing a composition comprising hyaluronic acid and an amount of water; adding a first excipient to the composition, the excipient comprises water-soluble surfactant; the composition is configured to form the cold slurry comprising a plurality of ice particles when the composition is cooled to a temperature below about 0 ℃.

[0017] In some aspects, the water-soluble surfactant is a hydrotropic molecule. In some embodiments, the hydrotropic molecule is a poloxamer molecule. In certain embodiments, the poloxamer molecule is selected from the group consisting of poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 183, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer 235, poloxamer 237, poloxamer 238, poloxamer 282, poloxamer 284, poloxamer 288, poloxamer 331, poloxamer 333, poloxamer 334, poloxamer 335, poloxamer 338, poloxamer 401, poloxamer 402, poloxamer 403, poloxamer 407, poloxamer 105 benzoate, poloxamer 182 dibenzoate, and combinations thereof. In some embodiments, the poloxamer is poloxamer 407 and the concentration of poloxamer 407 is about 0.1% (w / w) to 10% (w / w). In some embodiments, the concentration of poloxamer 407 in the composition is about 0.5% (w / w).

[0018] In certain aspects, the methods provided herein further comprise adding a second excipient to the composition, wherein the composition including the second excipient is configured to form a cold slurry when the composition is cooled to a temperature below about 0° C. In certain embodiments, the second excipient is selected from the group consisting of a salt, an ion, lactated Ringer's solution, a sugar, a biocompatible surfactant, a polyol, and combinations thereof.

[0019] In some embodiments, the second excipient is glycerol. In some embodiments, the concentration of glycerol in the composition is about 12% to 25% (w / w). In some embodiments, the concentration of glycerol in the composition is about 19% (w / w).

[0020] In certain aspects, the methods provided herein further comprise adding a third excipient to the composition, wherein the composition comprising the second excipient and the third excipient is configured to form a cold slurry when the composition is cooled to a temperature below about 0° C. In certain embodiments, the third excipient is sodium chloride or sodium phosphate to form saline or phosphate buffered saline.

[0021] In certain aspects, the methods provided herein further comprise adding a fourth excipient to the composition, wherein the composition comprising the second excipient, the third excipient, and the fourth excipient is configured to form a cold slurry when the composition is cooled to a temperature below about 0° C.

[0022] In some embodiments, the fourth excipient is a water-insoluble substance. In some embodiments, the water-insoluble substance is a lipid. In some embodiments, the lipid is selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), egg sphingomyelin (DPSM), dipalmitoyl phosphatidylcholine (DPPC), dicetyl phosphate (DCP), La-phosphatidylcholine (soybean PC), phosphatidylethanolamine, (PE), phosphatidylserine (PS), and phosphatidylglycerol (PG).

[0023] In some aspects, the methods provided herein further include a composition configured to form a plurality of ice crystals when the composition is exposed to a temperature of about -25°C to about -5°C.

[0024] In some aspects, the methods provided herein include a composition configured to have an injection force of less than about 30 lbf when injected through a 16G, 17G, 18G, 19G, 20G, 22G, 23G or 24G needle. In some embodiments, the composition is configured to have an injection force of less than about 30 lbf when injected through a 17G or 18G needle.

[0025] In one aspect, a method for preparing cold slurry for administration to patient at clinical point of care is provided.In one embodiment, the method comprises: receiving a composition comprising a freezing point depressant and hyaluronic acid; and cooling the composition to a temperature below about 0°C to form cold slurry, and the cold slurry comprises a plurality of ice particles.In one embodiment, the freezing point depressant is glycerol.

[0026] In some aspects, the composition further comprises an amount of a poloxamer molecule. In certain embodiments, the poloxamer molecule is selected from the group consisting of poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 183, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer 235, poloxamer 237, poloxamer 238, poloxamer 282, poloxamer 284, poloxamer 288, poloxamer 331, poloxamer 333, poloxamer 334, poloxamer 335, poloxamer 338, poloxamer 401, poloxamer 402, poloxamer 403, poloxamer 407, poloxamer 105 benzoate, poloxamer 182 dibenzoate, and combinations thereof. In some embodiments, the poloxamer is poloxamer 407 and the concentration of poloxamer 407 is about 0.1% (w / w) to 10% (w / w). In some embodiments, the concentration of poloxamer 407 in the composition is about 0.5% (w / w).

[0027] In some aspects, the step of receiving the composition includes receiving the composition in a container.

[0028] In some embodiments, the container is a first syringe. In some embodiments, the method further comprises connecting the first syringe to a second syringe and processing the cold slurry via a back-and-forth cycle, the back-and-forth cycle comprising extruding the cold slurry from the first syringe to the second syringe and extruding the cold slurry from the second syringe to the first syringe.

[0029] In one aspect, the method further includes treating the cold slurry through a second, third, or fourth back-and-forth cycle.

[0030] In some aspects, the container is a container configured for topical application. In some embodiments, the container configured for topical application is a first tube.

[0031] In one aspect, the method further includes connecting the first tube to a second tube and processing the slurry through a reciprocating cycle, the reciprocating cycle including extruding the cold slurry from the first tube to the second tube and extruding the cold slurry from the second tube to the first tube.

[0032] In certain aspects, the methods provided herein further include monitoring the temperature of the cold slurry.

[0033] In one aspect, the monitoring step includes viewing a temperature sensitive indicator on a syringe or container holding the cold slurry, the temperature sensitive indicator configured to indicate the temperature of the cold slurry.

[0034] In some embodiments, the temperature sensitive indicator is a temperature sensitive sticker. In some embodiments, the temperature sensitive indicator provides a visual indication when the cold slurry reaches a predetermined temperature.

[0035] In some embodiments, the predetermined temperature is about −15° C. In some embodiments, the predetermined temperature is about −19° C. to −11° C., about −18° C. to −12° C., about −17° C. to −13° C., or about −16° C. to −14° C.

[0036] In one aspect, the temperature sensitive indicator is configured to provide a visual indication when the cold slurry is at a temperature cooler than a predetermined temperature. In one aspect, the temperature sensitive indicator is configured to provide a visual indication when the cold slurry is at a temperature warmer than a predetermined temperature.

[0037] In one aspect, the monitoring step includes viewing a thermometer.

[0038] In one aspect, the monitoring step includes viewing a temperature monitoring component embedded within a vessel holding the cold slurry, hi one embodiment, the temperature component is provided within the vessel or along the flow path.

[0039] In some aspects, the monitoring step further includes listening for an audio indicator configured to indicate when the composition has reached a predetermined temperature.

[0040] In some aspects, the composition is terminally sterilized. In some embodiments, the composition is terminally sterilized by autoclaving or steam sterilization. In some embodiments, autoclaving or steam sterilization comprises subjecting the composition to a temperature of about 118°C to 121°C. In some embodiments, the temperature is about 118°C.

[0041] In one aspect, a cold slurry delivery system is provided. In one embodiment, the cold slurry delivery system includes a container for holding a slurry composition, the container including a sterile barrier and a temperature indicator, and the container is configured to allow manual stirring of the slurry composition without breaking the sterile barrier.

[0042] In some embodiments, the container is a syringe or a tube.

[0043] In one aspect, the container is configured to be coupled to a second container.

[0044] In one aspect, the container and the second container are configured to be coupled using a connector.

[0045] In some aspects, the container and the second container include a first syringe and a second syringe, and the first syringe and the second syringe each include a male luer component. In some embodiments, the connector includes a female luer component.

[0046] In certain embodiments, the first syringe and the second syringe are connected using a female luer component, and the slurry composition can be transferred from the first syringe to the second syringe to manually stir the slurry composition.

[0047] In one aspect, a temperature sensitive indicator is provided on the container.

[0048] In one aspect, the temperature sensitive indicator includes a temperature sensitive sticker.

[0049] In some aspects, the temperature sensitive indicator provides a visual indication when the slurry composition reaches a predetermined temperature. In some embodiments, the predetermined temperature is about -15°C. In some embodiments, the predetermined temperature is about -19°C to -11°C, about -18°C to -12°C, about -17°C to -13°C, or about -16°C to -14°C.

[0050] In one aspect, the temperature sensitive indicator provides a visual indication when the slurry composition is at a temperature cooler than a predetermined temperature.

[0051] In one aspect, the temperature sensitive indicator provides a visual indication when the slurry composition is warmer than a predetermined temperature. [Brief description of the drawings]

[0052] BRIEF DESCRIPTION OF THE DRAWINGS The following figures illustrate exemplary embodiments of the present invention. [Figure 1] FIG. 1 is a table showing exemplary formulations of the cold slurry compositions described herein. [Diagram 2] FIG. 2 is a graph showing the ice content characterization of cold slurry compositions described herein and cold slurry compositions containing glycerol and PBS, prepared, for example, by the methods disclosed in the '367 PCT. [Diagram 3] FIG. 3 shows an embodiment of a composition comprising a plurality of poloxamer micelles, glycerol and a hyaluronic acid gel. [Figure 4A] FIG. 4A-B. FIG. 4A is an image showing an exemplary arrangement of two syringes for processing a slurry composition in a back and forth process between the two syringes, according to certain embodiments described herein. [Figure 4B] FIG. 4B shows a process flow diagram for preparing a pourable cold slurry using a back-and-forth ("BAF") process by subjecting the contents to three BAF cycles, according to certain embodiments described herein. [Diagram 5]5A-D. FIG. 5A shows an exemplary syringe arrangement including a temperature sensitive indicator for measuring the temperature of the contents of the syringe, where the indicator displays a range of temperatures, according to embodiments described herein. FIG. 5B shows an exemplary syringe arrangement including a temperature sensitive indicator that displays a single specific temperature, according to embodiments described herein. FIG. 5C shows an exemplary syringe arrangement including an external temperature sensitive indicator that displays "Process Now" if the contents of the syringe reach a predetermined temperature. FIG. 5D shows an exemplary syringe arrangement including multiple temperature sensitive indicators that display messages reading "Wait," "Processing," and "Discard" if the contents of the syringe reach a series of predetermined temperatures. [Figure 6] FIG. 6 is a graph showing the injection force (lbf) for injecting a slurry as a function of freezer temperature, where the syringe contains a composition prepared according to the present disclosure through a 17G needle. [Figure 7A] 7A-B. FIG. 7A is a graph showing the relationship between equilibrium temperature (as a proxy for ice content) and pour temperature for compositions prepared according to the present disclosure. [Figure 7B] FIG. 7B is a graph showing the relationship between injection force (lbf) and injection temperature for compositions prepared according to the present disclosure when injected through a 17G needle. [Figure 8] FIG. 8 is a graph showing the relationship between injection force (lbf) and post-treatment temperature when injected through an 18G needle in compositions prepared according to the present disclosure and subjected to 1, 2, 3 or 4 BAF cycles. [Figure 9] FIG. 9 is a plot showing injection force (lbf) versus post-treatment temperature in compositions prepared according to the present disclosure subjected to two BAF cycles and injected through either a 17G or 18G needle. [Figure 10] FIG. 10 is a graph showing the ice content characterization of the compositions described herein. [Figure 11]FIG. 11 is a table summarizing the pour force (lbf) and pour certainty for compositions described herein when prepared according to different processing methods. [Figure 12] FIG. 12 is a plot showing the injection force required for cold slurry compositions described herein that are terminally sterilized using gamma irradiation versus non-gamma irradiated compositions. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0053] Detailed Description The present disclosure relates generally to compositions and methods for producing flowable and / or injectable cold slurry-forming biomaterials. More specifically, disclosed herein are compositions comprising water, hyaluronic acid, and at least one excipient or additive. In some embodiments, the at least one excipient or additive is Pluronic acid. TM (also known as "poloxamers"). As used herein, the term "excipient" refers to any substance that is not itself a therapeutic agent, but is used as a diluent, adjuvant and / or vehicle for delivery of a therapeutic agent (in this case the therapeutic agent is ice) to a subject or patient, and / or a substance added to a composition to improve its handling, stability or storage characteristics. The terms "excipient" and "additive" are used interchangeably herein. In some embodiments, the solution may also include liposomes, lipids or other lipid structures (e.g., lamellar or non-lamellar structures, bilayer and non-bilayer structures, e.g., lipid nanoparticles, micelles, etc.), water-insoluble substances (i.e. substances that do not dissolve in water) or water-soluble surfactants, e.g., hydrotropic molecules (e.g., polysorbates).

[0054] In some embodiments, the flowable and / or pourable or topically applied composition contains a significant amount of ice to provide therapeutic benefits for various applications.For example, therapeutic applications of cold slurries are disclosed in U.S. Application Nos. 16 / 288,073 and 16 / 327,266, the disclosures of which relating to various therapeutic applications are incorporated herein by reference.

[0055] In some embodiments, the final product administered to a human patient or subject (e.g., a non-patient human or non-human animal) via injection is a cold slurry composed of sterile ice particles in water and varying amounts of excipients / additives, such as hyaluronic acid, poloxamers, and / or freezing point depressants. For example, the percentage of ice particles in the cold slurry may constitute less than about 10% by weight of the slurry, about 10% to about 20% by weight, about 20% to about 30% by weight, about 30% to about 40% by weight, about 40% to about 60% by weight, greater than about 60% by weight, etc. The size of the ice particles can optionally be determined by the composition of the components, such as water-soluble surfactants (e.g., hydrotropic molecules), poloxamers (e.g., Pluronic TM The ice particles are controlled by adding lipids such as F127 or P407 and / or lipids to allow flowability through various sized containers (e.g., needle gauge sizes of about 7 to about 43). Various sized containers are described in U.S. Application No. 15 / 505,042 (Publication No. US2017 / 027401l), the disclosure of which relating to containers for infusion is incorporated herein by reference. Additionally, other methods can be used to condition the size of the ice particles to allow flowability and / or infusion through various sized containers (e.g., using filters or transferring the composition back and forth between two syringes). In some embodiments, the majority of the ice particles have a diameter that is less than about half the inner diameter of the lumen or container used for infusion. For example, the ice particles can be about 1.5 mm or less in diameter for use with a 3 mm catheter.

[0056] In some embodiments, the cold slurry may include one or more excipients. The excipients may comprise less than about 10% volume per volume (v / v) of the cold slurry, between about 10% v / v and about 20% v / v, between about 20% v / v and about 30% v / v, between about 30% v / v and 40% v / v, and greater than about 40% v / v. Various added excipients may be used to change the phase change temperature of the cold slurry (e.g., reduce the freezing point), change the ice percentage of the cold slurry, change the viscosity of the cold slurry, prevent ice particle agglomeration, prevent dendritic ice formation (i.e., crystals with multiple branching "tree-like" formations, such as those found in snowflakes), maintain ice particles separated, increase the thermal conductivity of the fluid phase, or improve the overall prophylactic, therapeutic, or cosmetic efficacy of the flowable and / or injectable cold slurry. In the compositions described herein, such excipients may include hyaluronic acid, poloxamers, polysorbates (or other water soluble surfactants, e.g., hydrotropes), water insoluble materials, lipids (e.g., lipid particles), all of which prevent ice particles from agglomerating, prevent dendritic ice formation (i.e., crystals having multiple branching "tree-like" formations, such as those found in snowflakes), or keep the ice particles separate so that the cold slurry is flowable and / or pourable when removed from the freezer.

[0057] One or more freezing point depressants may be added to sterile water as an excipient to form a cold slurry with a freezing point below 0° C. (e.g., about −10° C.). Lowering the freezing point of the cold slurry allows it to remain fluid and pourable while still containing an effective proportion of ice particles. Suitable freezing point depressants include salts (e.g., sodium chloride, betadex sulfobutylether sodium), ions, lactated Ringer's solution, sugars (e.g., glucose, sorbitol, mannitol, hetastarch, sucrose, (2-hydroxypropyl)-β-cyclodextrin or combinations thereof), biocompatible surfactants such as glycerol (also known as glycerin or glycerine), other polyols (e.g., polyvinyl alcohol, polyethylene glycol 300, polyethylene glycol 400, propylene glycol), other sugar alcohols, or urea, and the like. Other exemplary freezing point depressants are disclosed in U.S. Application No. 15 / 505,042 (Publication No. US2017 / 027401l), the disclosure of which relating to slurry composition components is hereby incorporated by reference in its entirety.

[0058] The present disclosure describes compositions that, when frozen, result in a flowable and / or pourable cold slurry. In some embodiments, the compositions of the present disclosure do not require processing or manipulation to be flowable and / or pourable. However, manipulation may be used in other embodiments to further improve flowability and pourability or promote consistency. In some embodiments, the composition includes a suspension of a fluid with a high water content (e.g., about 70%-80%, about 80%-90% or higher than about 90%), a solute (e.g., glycerol) used to depress the freezing point, and hyaluronic acid. In some embodiments, to further improve the flowability and pourability of the cold slurry, the solution may include one or more of a lipid, a water-insoluble compound, or a water-soluble surfactant, such as a hydrotropic compound (e.g., polysorbate) or a poloxamer (e.g., P407). In some embodiments, the solution further includes additional excipients, such as sodium chloride or sodium phosphate, for example to form saline or phosphate buffered saline.

[0059] The present disclosure provides various compositions. In some embodiments, the composition comprises an effective amount of hyaluronic acid to generate a flowable and / or pourable cold slurry. Without intending to be bound by any particular theory, it is believed that the hyaluronic acid in the composition facilitates the formation of small ice crystals when the solution is exposed to freezing temperatures (e.g., about -20°C to -15°C, about -15°C to -10°C, about -10°C to -5°C, or about -10°C in some embodiments). In some embodiments, the composition comprising hyaluronic acid is transformed into a flowable and / or pourable cold slurry with ice particles when placed in a standard freezer, without the need to apply any mechanical agitation or further processing to the cold slurry.

[0060] In some embodiments, the composition further comprises an effective amount of a water-soluble surfactant, such as a hydrotropic compound (e.g., polysorbate), to produce a flowable and / or pourable ice slurry. In some embodiments, the water-soluble surfactant is a poloxamer (or Pluronic TM ) molecule. In some embodiments, any surfactant with a hydrophilic-lipophilic balance (HLB) value greater than 10 is considered water soluble. In some embodiments, the water soluble surfactant in the composition is at a concentration of about 0.01% (w / w) to 0.5% (w / w), about 0.5% (w / w) to 1% (w / w), about 1% (w / w) to 2% (w / w), about 2% (w / w) to 5% (w / w), about 5% (w / w) to 10% (w / w), or greater than about 10% (w / w). Without intending to be bound by any particular theory, it is believed that the water soluble surfactant further acts to prevent ice particles from growing too large when the composition is exposed to freezing temperatures (e.g., below about -5°C); large ice particles can prevent the composition from being flowable or pourable.

[0061] In some embodiments, the composition further comprises an effective amount of one or more of hyaluronic acid, any hydrotropic molecule, and lipid or water-insoluble particles to generate a flowable and / or injectable cold slurry in the form of an emulsion. In some embodiments, the emulsion is any composition described herein that includes lipids. In some embodiments, the lipids in the composition are assembled into lipid particles having one or more morphologies known in the art (e.g., lamellar or non-lamellar structures, bilayer and non-bilayer structures, such as liposomes, lipid nanoparticles, micelles, etc.). The lipid particle morphology of the present disclosure may be determined by any method known in the art, such as by CryoTEM. In some embodiments, the lipid particles in the composition are about 5 μm to about 300 μm in diameter. In some embodiments, the lipid particles are about 250 μm in diameter. In some embodiments, the lipid particles in the composition are about 5 μm to 20 μm in diameter or about 8 μm to 14 μm in diameter. Without intending to be bound by any particular theory, it is believed that the lipid or water-insoluble particles prevent the ice particles from growing so large that the composition is no longer flowable or pourable when the composition is exposed to freezing temperatures.

[0062] In some embodiments, the hyaluronic acid is of natural or synthetic origin. In some embodiments, the hyaluronic acid has a molecular weight of about 250 kDa to 5,000 kDa. In some embodiments, the hyaluronic acid has a molecular weight of about 1,000 kDa. In some embodiments, the concentration of the hyaluronic acid in the composition is about 0.01% (w / w) to 2% (w / w). In some embodiments, the concentration of the hyaluronic acid in the composition is about 0.1% (w / w) to 1% (w / w). In some embodiments, the concentration of the hyaluronic acid in the composition is about 0.5% (w / w) to 1.5% (w / w). In some embodiments, the concentration of the hyaluronic acid in the composition is about 0.05% (w / w) to 0.75% (w / w). In some embodiments, the concentration of the hyaluronic acid in the composition is about 0.5% (w / w). In some embodiments, the hyaluronic acid in the composition is in the form of a hyaluronic acid gel (i.e., having a high viscosity).

[0063] In some embodiments, the excipient is selected from the group consisting of salts, ions, lactated Ringer's solution, sugars, biocompatible surfactants, polyols, and combinations thereof. In some embodiments, the excipient is a polyol. In some embodiments, the polyol is glycerol. In some embodiments, the glycerol concentration of the composition is about 12%-25% (w / w). In some embodiments, the glycerol concentration of the composition is about 20% (w / w).

[0064] In some embodiments, the composition includes a second excipient, such as sodium chloride or sodium phosphate to form saline or phosphate buffered saline.

[0065] In some embodiments, the composition comprises a third excipient. In some embodiments, the third excipient is a water-soluble surfactant. In some embodiments, the third excipient is a hydrotropic compound. In some embodiments, the third excipient is a polysorbate.

[0066] In some embodiments, the composition comprises a Pluronic TM(also referred to as "poloxamers"). In some embodiments, the poloxamers form macromolecular assemblies, such as micelles. In some embodiments, the macromolecular assemblies are micelles (or "poloxamer micelles"). In some embodiments, the Pluronics or poloxamers include Pluronic L31, Pluronic L35, Pluronic F38, Pluronic L43, Pluronic L44, Pluronic L61, Pluronic F68, Pluronic F77, Pluronic L81, Pluronic P84, Pluronic P85, Pluronic F77, Pluronic F87, Pluronic L92, Pluronic F98, Pluronic L101, Pluronic P103, Pluronic P104, Pluronic P105, Pluronic F108, Pluronic L121, Pluronic P123, Pluronic F127, poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 183, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer 235, poloxamer 237, poloxamer 238, poloxamer 282, poloxamer 284, poloxamer 288, poloxamer 331, poloxamer 333, poloxamer 334, poloxamer 335, poloxamer 338, poloxamer 401, poloxamer 402, poloxamer 403, poloxamer 407, poloxamer 105 benzoate, poloxamer 182 dibenzoate, and combinations thereof. In some embodiments, the Pluronic or poloxamer is Pluronic F127. In some embodiments, the poloxamer is poloxamer 407.

[0067] In some embodiments, the concentration of the poloxamer is about 0% to 10% (w / w). In some embodiments, the concentration of the poloxamer is about 0.1% to 10% (w / w). In some embodiments, the concentration of the poloxamer is about 5% (w / w). In some embodiments, the Pluronic TM Pluronic TM F127, Pluronic TM The concentration of F127 is about 5% (w / w). In some embodiments, the poloxamer is poloxamer 407, and the concentration of poloxamer 407 is about 5% (w / w).

[0068] In some embodiments, the composition comprises a lipid. In some embodiments, the composition comprises a plurality of lipids in the form of liposomes formed with phospholipids (e.g., soybean PC). The lipids can be of any type (e.g., phospholipids, cholesterol, complex lipids, or combinations thereof) or the composition can comprise any other water-insoluble substance instead of lipids. The lipids (or lipid particles) or water-insoluble substances are present in a relatively high concentration, preferably about 6% (w / w) to 28% (w / w) of the composition. Without intending to be bound by any particular theory, it is believed that the lipids (or lipid particles, e.g., liposomes) or water-insoluble substances prevent large ice crystal formation and therefore generate an emulsion when the composition is exposed to freezing temperatures (about -25°C to -15°C, about -15°C to -10°C, about -15°C to -5°C, about -10°C to -5°C, or about -10°C in some embodiments). This allows the composition to have ice particles while also being flowable and / or pourable. In some embodiments, the composition further comprises lipid particles. In some embodiments, the lipid particles are liposomes. In some embodiments, the lipid particles are micelles. In some embodiments, the lipid particles are composed of phospholipids. In some embodiments, the phospholipids are selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), egg sphingomyelin (DPSM), dipalmitoylphosphatidylcholine (DPPC), dicetylphosphate (DCP), La-phosphatidylcholine (PC), phosphatidylethanolamine, (PE), phosphatidylserine (PS), phosphatidylglycerol (PG), La-phosphatidylcholine (soybean PC) and combinations thereof. In some embodiments, the lipid is La-phosphatidylcholine (soybean PC). In some embodiments, the lipid concentration in the composition is about 0% to 30% (w / w).

[0069] In some embodiments, the composition further comprises ethanol. In some embodiments, the concentration of ethanol in the composition is about 0.01% to 0.1%. In some embodiments, the concentration of ethanol in the composition is about 0.07% or less.

[0070] In some embodiments, the composition comprises glycerol, hyaluronic acid and Pluronic acid in water, saline or phosphate buffered saline. TM In some embodiments, the composition comprises glycerol, hyaluronic acid and Pluronic F127 in saline. TM In some embodiments, the composition comprises about 20% (w / w) glycerol, about 0.5% (w / w) hyaluronic acid, and about 5% (w / w) Pluronic F127 in water, saline, or phosphate buffered saline. TM In some embodiments, the composition comprises about 20% (w / w) glycerol, about 0.5% (w / w) hyaluronic acid, and about 5% (w / w) Pluronic Acid in water, saline, or phosphate buffered saline. TM Hyaluronic acid, including F127, has a molecular weight of approximately 1000 kDa.

[0071] In some embodiments, the composition comprises glycerol, PBS, hyaluronic acid and poloxamer 407 (Pluronic TM In one embodiment, the composition comprises glycerol having a concentration of about 18.9% (w / w), 1x PBS having a concentration of about 75.6% (w / w), hyaluronic acid having a molecular weight of 1000 kDa and a concentration of about 5% (w / w), and poloxamer 407 (Pluronic F127) having a concentration of about 0.5% (w / w). TMF127). See, e.g., FIG. 1. In some embodiments, the composition comprises a temperature set point of about -14°C. See supra. In some embodiments, the composition has an ice content, the ice content being about 50%. In some embodiments, the composition is the composition shown in FIG. 1. In some embodiments, the composition comprises a relatively small percentage of hyaluronic acid, about 0.5% (w / w) to 1.5% (w / w), the molecular weight of the hyaluronic acid being about 1,000 kDa. In such embodiments, the composition comprises a concentration of glycerol of about 15% (w / w) to 25% (w / w) and a small percentage of polysorbate of about 0.25% (w / w) to 1.5% (w / w). This allows the composition to have ice particles while remaining flowable and / or pourable once frozen.

[0072] In some embodiments, the composition is filled into a container having a volume of less than 10 mL and a shape that creates the maximum surface area of ​​the container wall. Without intending to be bound by any particular theory, it is believed that the large surface area to volume ratio facilitates an increased freezing rate and further prevents the formation of large ice crystals, thus improving flowability and injectability. In some embodiments, the total injection volume of the cold slurry into the patient, optionally via multiple containers and multiple injections, is about 5 mL to 10 mL, about 10 mL to 20 mL, about 20 mL to 30 mL, about 30 mL to 40 mL, about 40 mL to 50 mL, about 50 mL to 60 mL, about 60 mL to 70 mL, or greater than about 70 mL. In some embodiments, the total injection volume is about 60 mL.

[0073] Methods of producing cold slurries by formulating solutions that prevent the formation of large ice crystals are described in PCT Application Nos. PCT / US20 / 43280 and '494 PCT, which are incorporated herein by reference in their entirety. Described herein is an unexpected method of forming a cold slurry by producing a composition having hyaluronic acid, water and at least one excipient, and the production of the cold slurry does not require any mechanical manipulation or stirring of the composition. In some embodiments, the composition further comprises a poloxamer. In some embodiments, the composition comprises a plurality of lipids. In some embodiments, the injectability of the composition is improved by utilizing small mechanical stirring.

[0074] In some embodiments, the compositions described herein are homogenous mixtures, the composition medium throughout the container is uniform, and the components are evenly distributed. In some embodiments, the addition of poloxamer, hyaluronic acid, and glycerol prevents the formation of large ice crystals, and the flowable cold slurry can be injected into a subject immediately after removal of the cold slurry from a freezer or another cooling environment.

[0075] The compositions provided herein may be provided in a syringe or other container. The syringe or other container may also include a visible temperature indicator that may allow visual monitoring of the temperature of the slurry or the approximate temperature of the slurry. The temperature indicator may be a temperature-sensitive display, sticker, marker, crayon, lacquer, pellet, etc., such as a reversible temperature indicator that may dynamically track temperature changes. The temperature indicator may be located inside the syringe or other container (e.g., a pellet placed directly in the internal solution), on the internal wall of the syringe or other container, on the external wall of the syringe or other container, or in any location that allows visual tracking of the temperature of the contents inside the syringe or other container. In some embodiments, the composition is provided in a syringe, and the syringe is placed in a freezer. The syringe is removed from the freezer after a sufficient time (e.g., 24 hours) for ice crystals to form in the syringe. After removing the syringe from the freezer, the syringe contents are monitored, for example, using a temperature indicator (e.g., an infrared sensor or an external temperature-sensitive indicator). Once the syringe contents reach a predetermined temperature, the syringe is connected to a second syringe and subjected to BAF treatment (e.g., three BAF cycles). The slurry is then injected into a patient or subject. In some embodiments, alternative methods of processing the slurry can be used, such as forcing the slurry through a filter; including a wire between two syringe openings around which the slurry needs to be injected; providing an internal component in the syringe that provides such treatment, such as a magnet, an internal blade, or an internal forming wire (e.g., a spring). In some embodiments, the BAF treatment or other treatment is done after the contents of the container are removed from the freezer and reach a predetermined temperature. In some embodiments, the BAF treatment or other treatment is done immediately after the container is removed from the freezer and before it reaches a predetermined temperature. Without intending to be bound by any particular theory, it is believed that the additional mechanical treatment reduces the ice crystal size in the cold slurry, making it easier for the ice crystals to be injected through the needle.

[0076] Referring to FIG. 2, two different cold slurry compositions (batches) are characterized with respect to their temperature profile over time and ice content. The different cold slurry batches were heated to 40° C. and placed in a copper plate with thermocouple wires that measured the change in temperature of the cold slurry over time. The data plotted are: 1) with 12.6% (w / w) glycerol and PBS (trace A); C and A M ) and 2) containing 0.5% (w / w) soy-PC, 0.08% (w / w) EtOH, 0.75% (w / w) hyaluronic acid (1,000 kDa), 16% (w / w) glycerol and PBS (trace B C and B. M Figure 1 shows the temperature change over time in seconds for two different cold slurry compositions (represented by 100°C). Temperature was measured at two different locations for each cold slurry: one thermocouple embedded inside the copper plate (trace A) and one thermocouple embedded inside the copper plate (trace B). C , B C ), and the other thermocouple is placed in the middle of the copper plate exposed on the outside of the plate (trace A M , B M When the batch of cold slurry is first introduced into the copper plate, the thermocouple wire (trace A) embedded inside the plate is C , B C ) first measures the warm temperature of the heated plate (e.g., 31° C. for trace Ac at time 0) and then a lower temperature (e.g., 31° C. for trace A at about 2 min) due to the cooling effect of the introduced cold slurry. C The plate is allowed to reach equilibrium at 20°C for 100°C. Meanwhile, the thermocouple wire (trace A) is placed in the middle of the plate. M , B M ), when the cold slurry is first introduced into the copper plate, the wire is exposed so that the cold slurry immediately comes into contact with the thermocouple wire. This results in an initial negative temperature reading at the intermediate position (e.g., trace A at time 0) due to the wire coming into contact with the crystallized cold slurry. Mabout -4 °C), and then the cold slurry is placed on a heated plate (e.g., trace A at about 6 min). M Allow equilibration at warmer temperatures (approximately 14 °C) to initiate dissolution. The thermocouple wire exposed to the outside of the plate (trace A) M , B M ) can be used to detect phase transitions during which the crystallized cold slurry begins to dissolve. The graph shows the phase transitions for the slurry composition with hyaluronic acid (trace B). C and B. M ) has a progressive phase transition. The graph also shows that the cold slurry batches (Trace B) having hyaluronic acid compositions C and B. M ) reach equilibrium in a similar time frame (as measured by the two thermocouple wire positions) and at similar temperatures of about 10 °C to 14 °C depending on the thermocouple position (inner / middle). On the other hand, cold slurries with different compositions (lacking hyaluronic acid; trace A C and A M ) has a different temperature profile than the composition containing hyaluronic acid, reaching equilibrium more quickly at a temperature of about 15° C.-17° C. depending on the location of the thermocouple (inside / middle). Thus, FIG. 2 shows that cold slurries can have different compositions that can be designed to have different temperature profiles or can be designed to perform equally well.

[0077] FIG. 3 shows an embodiment of a composition comprising poloxamer micelles, glycerol and a hyaluronic acid gel. In this embodiment, the composition comprises a Pluronic acid having an aqueous core (e.g., containing water). TM (e.g. Pluronic TMF127). In some embodiments, the external medium of the micelles comprises a hyaluronic acid gel in a water-glycerol and optionally ethanol solution (not shown). In some embodiments, water droplets are trapped within the external medium (i.e., outside the micelles in the hyaluronic acid gel in a water-glycerol and optionally ethanol solution (not shown)). The embodiment of FIG. 3 is referred to herein as a "micellar dispersion" or "micellar gel". The micellar dispersion or micellar gel comprises surfactant micelles that trap water, where the micelles are suspended in the hyaluronic acid gel. In some embodiments, the form of the micelles in the composition is a normal micelle (as shown in FIG. 3) or a reverse micelle. In some embodiments, a lipid micelle composition comprising hyaluronic acid is provided.

[0078] In some embodiments, the at least partially crystallized composition comprising hyaluronic acid, glycerol, water and saline (or PBS) comprises sufficient ice particles to be flowable and / or pourable without the addition of other excipients when exposed to freezing temperatures (i.e., placed in a freezer). In some embodiments, the at least partially crystallized composition comprising hyaluronic acid, glycerol and water comprises sufficient ice particles to be flowable and / or pourable without the addition of other excipients when exposed to freezing temperatures (i.e., placed in a freezer). In alternative embodiments, the addition of poloxamer, polysorbate or lipid further improves the flowability and pourability of the partially crystallized composition. It has also been found that increasing the rate of freezing of the material to a faster rate further improves the flowability and pourability of the composition. Optimizing the freezing rate includes selecting the material of the container (e.g., syringe) in which the composition is placed, the outer shape of the container, and the selection of the cooling environment or freezer (e.g., the humidity of the freezer can be adjusted to improve the flowability and / or pourability of the resulting cold slurry). It has also been found that the pourability or flowability of the partially crystallized composition can be improved by spacing the containers apart when placed in the freezer.

[0079] In some embodiments, the composition (e.g., in the form of a liquid solution) can be packaged and sealed in a container such as a syringe. The syringe can be filled aseptically (e.g., using aseptic procedures) or the syringe can be prefilled, sealed, and then terminally sterilized (e.g., using autoclave or steam sterilization). The composition can also be provided in any other sealed container that can be terminally sterilized, such as a tube used for topical ointments or a larger container that is then used to fill multiple syringes. In some embodiments, the prefilled syringe or other container can be terminally sterilized using gamma irradiation or autoclave or steam sterilization at a temperature of about 118°C. In some embodiments, the prefilled syringe or other container can be terminally sterilized using gamma irradiation or autoclave or steam sterilization at a temperature of about 118°C to 121°C. In a preferred embodiment, when using hyaluronic acid with a molecular weight of 1,000 kDa, the composition is terminally sterilized using steam, since sterilization using radiation affects the molecular weight of hyaluronic acid. In some embodiments, the composition is terminally sterilized using hyaluronic acid having a molecular weight of 1,000 kDa, the composition is terminally sterilized using a low dose of radiation or using radiation to irradiate a still frozen cold slurry.

[0080] In some embodiments, the composition inside the syringe (or other container) is transformed into a flowable and / or injectable cold slurry by placing the pre-filled syringe (or other container) in a standard freezer or other refrigerated environment. In some embodiments, a structure is provided for storing one or more pre-filled syringes (or other containers) in a freezer or other refrigerated environment, the structure being configured to store a first pre-filled syringe (or container) of the one or more pre-filled syringes (or other containers) at a predetermined distance from a second pre-filled syringe (or container) of the one or more pre-filled syringes (or other containers). In some embodiments, a structure is provided for storing multiple pre-filled syringes (or containers) at a predetermined distance from each other to facilitate achieving uniform and rapid cooling of the cold slurry. See, e.g., Example 3. In some embodiments, the predetermined distance is at least 0.5 inches. In some embodiments, the predetermined distance is about 0.5 inches. In some embodiments, the predetermined spacing is about 0.5 inches, about 0.75 inches, about 1 inch, about 1.25 inches, about 1.5 inches, about 1.75 inches, about 2 inches, about 2.25 inches, about 2.5 inches, about 2.75 inches, about 3 inches, about 3.25 inches, about 3.5 inches, about 3.75 inches, about 4 inches, or greater than about 4 inches. In some embodiments, the structure is a rack, tray, or box. In some embodiments, the composition can be flash frozen using liquid nitrogen or other liquid cooling methods to accelerate the process.

[0081] After freezing, in some embodiments, the syringe or container can be removed from the freezer, cooling environment or other freezing method, and the cold slurry can be immediately injected or applied, optionally by topical application, for therapeutic benefit. In some embodiments, the slurry can be directly applied to tissue after an invasive surgical procedure. In some embodiments, the cold slurry can be injected directly from the syringe using a needle. In some embodiments, the needle is a 16G needle, a 17G needle, an 18G needle, a 19G needle, a 20G needle, a 21G needle, a 22G needle, a 23G needle or a 24G needle. The cold slurry can also be removed from the container for topical application, such as by squeezing the container to disperse the cold slurry at the target treatment site when the container is removed from the freezer. In some embodiments, the cold slurry is in a flowable or injectable form immediately after removal from the freezer without any further mechanical manipulation.

[0082] In some embodiments, after removal from the freezer, the cold slurry is subjected to mechanical manipulation to improve flowability or injectability, and then optionally injected or applied by topical application for therapeutic benefit. In some embodiments, as shown in FIG. 4A, the cold slurry can be provided in a first syringe or container that is configured to be connected to a second syringe or container via a female luer adapter to facilitate "round trip" (BAF) processing of the cold slurry. See, e.g., Example 5. In some embodiments, as shown in FIG. 4A, a system is provided that includes a first sterile syringe or other container that includes a male luer component and contains the cold slurry, and a second sterile syringe or container that includes a male luer component and does not contain the cold slurry, and the first syringe or container and the second syringe or container are connected by a sterile female luer connector. In some embodiments, the BAF process comprises: (1) pushing the cold slurry from a first syringe into a second syringe, and (2) pushing the cold slurry from the second syringe back into the first syringe to complete one "back and forth" cycle. In some embodiments, the cold slurry is subjected to one, two, three, four or more BAF cycles. See Example 5; FIG. 8. In some embodiments, the cold slurry is subjected to two BAF cycles. In some embodiments, the cold slurry is subjected to three BAF cycles. An exemplary method including three back and forth cycles is shown in FIG. 4B. The exemplary method shown in FIG. 4B includes freezing the first syringe for 24 hours, where the syringe holds the slurry composition, and then removing the syringe from the freezer. The temperature of the current cold slurry is then observed using temperature monitoring (e.g., using a temperature sensitive sticker as shown in FIGS. 5A-5D) until the cold slurry reaches a predetermined temperature. Once the predetermined temperature is reached, the slurry is processed using a second syringe connected to the first syringe using a Luer connector and subjected to three BAF cycles. After the three BAF cycles, the slurry is injected. This process can be repeated as necessary to prepare the number of syringes required for processing.Forcing the syringe contents from one syringe to another is a well-known technique that does not compromise the sterility of the syringe contents when using sterilized components. Optionally, additional elements can be added between the syringes to further break apart the ice crystals. For example, one of the syringe openings can be covered with a filter or with a wire to break apart the ice crystals. It is understood that the method shown in FIG. 4B can be used with the method and system described in International Publication No. WO 2022 / 055934, in which a syringe containing a biocompatible composition described herein is transported to a point-of-care at ambient temperature and then placed in a freezer at the point-of-care to transform the composition into a cold slurry containing a plurality of ice crystals. The disclosure in the '934 PCT regarding methods of transporting and transforming a biocompatible composition is incorporated herein by reference.

[0083] In some embodiments, after being removed from the freezer, the syringe is set aside and warmed to a predetermined temperature for injection or topical application. In some embodiments, the predetermined temperature is reached after the syringe contents are subjected to mechanical agitation. In some embodiments, the syringe or other container has an external temperature indicator configured to indicate when the predetermined temperature is reached, for example, by a color change or by displaying a message. In some embodiments, the temperature indicator is a temperature-sensitive sticker or the like. The temperature indicator may allow for visual monitoring of the temperature of the contents of the syringe or the approximate temperature of the contents of the syringe. The temperature indicator, such as a temperature-sensitive sticker or the like, may display a range of temperatures, and a color change in an area corresponding to the predetermined temperature range indicates that the predetermined temperature has been reached. See, for example, FIG. 5A. The temperature indicator may be a temperature-sensitive indicator, such as a reversible temperature indicator that may dynamically track temperature changes, a sticker, a marker, a crayon, a lacquer, a pellet, or the like. The temperature indicator may be located inside the container (e.g., pellets placed directly into the internal solution), on the inside wall of the container, on the outside wall of the container, or any location that allows visual tracking of the temperature of the contents inside the container. In some embodiments, the predetermined temperature is about -15°C. In some embodiments, the predetermined temperature is about -19°C to -11°C, about -18°C to -12°C, about -17°C to -13°C, or about -16°C to -14°C. In some embodiments, a color change in the area corresponding to a temperature cooler than the predetermined temperature indicates that the cold slurry is colder than the predetermined temperature and indicates that the cold slurry should be warmed to the predetermined temperature before injection or application. In some embodiments, the cooler temperature is about -20°C. In some embodiments, the cooler temperature is about -20°C to -25°C, about -25°C to -30°C, or less than about -30°C. In some embodiments, a color change in an area corresponding to a temperature warmer than a predetermined temperature indicates that the cold slurry is warmer than desired for injection or application to produce a therapeutic effect, hi some embodiments, the syringe may be re-frozen if the warmer temperature is reached.For example, in some embodiments, the warmer temperature is about -10°C to -8°C, about -10°C to -9°C, about -9°C to -8°C, about -10°C to -7°C, about -10°C to -6°C, about -10°C to -5°C, about -10°C to -4°C, about -10°C to -3°C, about -10°C to -2°C, about -10°C to -1°C, about -10°C to 0°C, -9°C to 8°C, about -9°C to -7°C, The temperature may be about -9°C to -6°C, about -9°C to -5°C, about -9°C to -4°C, about -9°C to -3°C, about -9°C to -2°C, about -9°C to -1°C, about -9°C to 0°C, about -8°C to -7°C, about -8°C to -6°C, about -8°C to -5°C, about -8°C to -4°C, about -8°C to -3°C, about -8°C to -2°C, about -8°C to -1°C, about -8°C to 0°C, or warmer than 0°C. In some embodiments, the warmer temperature is at least -10°C to -8°C or warmer. In some embodiments, the warmer temperature is at least -9°C to -8°C or warmer. In some embodiments, the warmer temperature is at least -10°C to -9°C or warmer. In some embodiments, the indicator, such as a temperature sensitive sticker, displays a temperature or a range of temperatures indicating the time when the cold slurry reaches a predetermined temperature. See, e.g., FIG. 5B. In some embodiments, the indicator, such as a temperature sensitive sticker, includes an area that displays a message indicating that a predetermined temperature has been reached. See, e.g., FIG. 5C. For example, in some embodiments, the indicator may display a message such as "Process Now", "Process", "Agitate", "Go" or any other suitable message. In some embodiments, the indicator, such as a temperature sensitive sticker, includes an area that displays a message indicating that the cold slurry is cooler than a predetermined temperature or warmer than a predetermined temperature. See, e.g., FIG. 5D. For example, in some embodiments, the indicator may display a message such as "Wait", "Hold", "Not Yet", "Too Cold" or any other suitable message indicating that the cold slurry is cooler than a predetermined temperature.In some embodiments, the indicator may display a message such as "Discard", "Too Warm", "Throw Away", "Refreeze" or any other suitable message indicating that the cold slurry is warmer than a predetermined temperature. In some embodiments, a method is provided that includes temperature monitoring and reciprocating mechanical stirring as shown in FIG. 4B.

[0084] In one aspect, a composition is provided, the composition requires an injection force of less than about 30 lbs of plunger force to inject the composition through a syringe of a 16G needle, a 17G needle, an 18G needle, a 19G needle, a 20G needle, a 21G needle, a 22G needle, a 23G needle, or a 24G needle. In some embodiments, the composition provided herein requires an injection force of less than about 30 lbs of plunger force to inject the composition through a 17G or 18G needle. The compositions disclosed herein were prepared according to FIG. 2 and tested to see if there is a relationship between freezer temperature and injection force. As shown in FIG. 6, the composition in the syringe had more ice particles at warmer temperatures, so a colder freezer temperature requires a higher injection force to inject the cold slurry from the syringe. To determine the ice content of the cold slurry, a method and apparatus were designed that allows the ice content to be calculated based on the final equilibrium temperature of the apparatus after a predetermined amount of cold slurry is dispensed into the apparatus as described in Example 3. With reference to FIG. 7A, as the equilibrium temperature increases, the injection temperature increases. As further shown in FIG. 7B, as the injection temperature decreases, the injection force increases due to the increase in ice content. As the temperature of the cold slurry increases, the injection force decreases. This result was observed for all syringe sizes (e.g., 3cc, 6cc) and needle gauges (e.g., 17G, 18G) tested. See, e.g., FIGS. 6, 7B, 8-9. Furthermore, as the temperature increases, it is possible to increase the overall reliability of the injection, i.e., to reduce the number of failed injections (e.g., injections where a spike in injection force is observed and the slurry cannot be expelled from the syringe). By monitoring the temperature of the cold slurry, it is possible to optimize the injectability (reducing the injection force) without reducing the ice content below the therapeutic threshold for a given application.It has been found that the reliability of injection is greatly increased by following the steps of (1) placing the syringe in a freezer to form a cold slurry containing a plurality of ice crystals; (2) waiting for the cold slurry to reach a predetermined temperature after removal from the freezer; and (3) subjecting the cold slurry to three BAF cycles between two syringes. These results were consistently observed for injections of ≧5 mL (e.g., injections of 6 cc). In another aspect, the injection force is reduced by mechanical agitation of the cold slurry according to the methods described herein, e.g., subjecting the cold slurry to one or more back and forth cycles. See, e.g., Figures 8, 10-11; Example 5.

[0085] With reference to Figure 1, 6 cc cold slurry compositions (batches) prepared according to the following description are characterized with respect to their temperature profile and ice content. Different cold slurry batches were placed on a copper plate heated to 40°C with thermocouple wires measuring the change in temperature of the cold slurry over time. The plotted data represents the average temperature profile and ice content of 18.9% (w / w) glycerol, 75.6% (w / w) 1xPBS, 5.0% (w / w) 1000 kDa hyaluronic acid and 0.5% (w / w) poloxamer 407 (Pluronic TM 1 shows the temperature change over time for three batches of cold slurry compositions containing 1,2-dichlorophenyl ether (FeOH) and 1,2-dichlorophenyl ether (F127). The three batches of cold slurry compositions were then subjected to two BAF treatment cycles as described herein. Two different locations for each cold slurry: embedded inside the copper plate (trace A); C , B C , C C ) and the middle of the copper plate exposed on the outside of the plate (trace A M , B M , C M When the batch of cold slurry was first introduced into the copper plate, the temperature was measured by a thermocouple wire (trace A) embedded inside the plate. C , B C , C C) is initially measured at the warm temperature of the heated plate (e.g., trace A at time 0). C , B C and C C 35-40°C for 1 min), and then a lower temperature (e.g., trace A at about 2 min) was measured due to the cooling effect of the introduced cold slurry. C , B C and C C Meanwhile, the thermocouple wire (trace A) located in the middle of the plate reaches equilibrium at 20°C. M , B M , C M ), when the cold slurry is first introduced to the copper plate, it immediately contacts the thermocouple wire since the thermocouple wire is exposed. This initially results in a negative temperature reading at the middle location (e.g., trace A at time 0) due to the crystallized cold slurry contacting the wire. M for −4° C.), then at a warmer temperature when the cold slurry begins to dissolve on the heated plate (e.g., trace A at about 6 min). M , B M and C M Induce equilibrium at approximately 12 °C to 14 °C for 10 min. The thermocouple wires exposed on the outside of the plate (trace A) M , B M , C M ) can be used to detect a phase transition as the crystallized cold slurry begins to dissolve. The graph shows that all cold slurry compositions tested (each prepared as described in FIG. 1 and subjected to two BAF cycles) have a progressive phase transition. The graph also shows that cold slurry batches prepared in this manner consistently reach equilibrium in a similar time frame and at similar temperatures (trace A as measured by two thermocouple wire locations). M , B M and C M 10 thus shows that cold slurries designed in accordance with the present disclosure produce consistent temperature profiles and ice content when subjected to mechanical agitation.

[0086] With reference to FIG. 11, characterization of the slurry compositions prepared in FIG. 1, subjected to different processing techniques, and injected using 18G or 17G needles is provided. For all tests, the cold slurries were held in 5cc syringes. The robustness is characterized by quantifying the percentage of injections that did not exhibit an injection force spike beyond a predefined limit. For the results shown in FIG. 11, the predefined limit is 40 lbf. First, the injection force was measured for cold slurries frozen at -20°C and not subjected to further processing. The injection robustness was less than 20% using the 18G needle and 90% using the 17G needle, meaning that the cold slurry could be injected through the 18G needle less than 20% of the time and through the 17G needle 90% of the time. Second, the injection force was measured for cold slurries frozen at -20°C and subjected to three BAF cycles after removal from the freezer without waiting and temperature monitoring. The injection certainty was 78% using the 18G needle and higher than 95% using the 17G needle. Finally, the injection force was measured for cold slurries frozen at -20°C, warmed to -15°C as determined by temperature monitoring, and subjected to three BAF cycles. The injection certainty was higher than 95% using the 18G needle and the 17G needle. Thus, FIG. 11 shows that injection certainty of higher than 95% can be achieved when the slurry is subjected to three BAF cycles after being frozen at -20°C and injected using a 17G needle. FIG. 11 further shows that certainty of higher than 95% can be achieved using both the 18G needle and the 17G needle when the slurry is subjected to three BAF cycles after being frozen at -20°C and warmed to -15°C as determined by temperature monitoring.

[0087] Referring to FIG. 12, results from an experiment testing the effect of sterilizing a syringe containing a slurry composition using radiation (gamma / e-beam sterilization) are shown. The results showed that radiation sterilization using gamma / e-beam sterilization resulted in an increase in the injection force required to expel the cold slurry from the syringe. See FIG. 12. Without wishing to be bound by a particular theory, this increase in the required injection force may be due to the effect of the molecular weight of the hyaluronic acid in the composition. In some experiments, syringes containing the slurry composition were sterilized using autoclave sterilization (steam sterilization) at various temperatures (e.g., about 118°C or about 121°C). Unlike radiation sterilization, it was observed that autoclave or steam sterilization did not increase the injection force required to expel the cold slurry from the syringe.

[0088] The compositions described herein can be used for various applications. After the composition according to some embodiments of the present disclosure is exposed to freezing temperatures and forms a flowable cold slurry, the cold slurry can be administered locally to the area for therapeutic treatment. A method of local administration of cold slurry to the ocular surface is described in International Patent Application PCT / US21 / 24514, the disclosure of which is incorporated herein by reference in its entirety for therapeutic use of the slurry. The compositions described herein can also be used to form flowable and / or injectable cold slurries that can be injected into targeted treatment areas for therapeutic effects. The injection method for cold slurries described in International Patent Application US2017 / 0274078, the disclosure of which is incorporated herein by reference in its entirety for therapeutic use of the injected slurry.

[0089] The devices, systems, compositions and methods disclosed herein are not limited in scope to the specific embodiments described herein. Indeed, various modifications of the devices, systems and methods in addition to those described will become apparent to those skilled in the art from the foregoing description. EXAMPLES

[0090] Working Example Example 1 - Method for preparing a cold slurry composition containing hyaluronic acid and poloxamer A cold slurry composition was prepared containing the following components: glycerol, hyaluronic acid, lipids and poloxamer (e.g., Pluronic TM F127) were tested. The compositions were prepared with a glycerol content ranging from about 12-25% w / w. The compositions further included a hyaluronic acid content ranging from about 0.1-1% w / w of hyaluronic acid having a molecular weight ranging from 250-5000 kDa. The lipid content in the compositions ranged from 0-30% w / w. The poloxamer content (e.g. Pluronic TM F127) ranged from 0 to 30% w / w.

[0091] The formulations for the cold slurry compositions were evaluated using two main test methods. First, the compositions were tested to measure the injection force through various needle gauges (e.g., 17G or 18G). A force test stand was used to apply force to the syringe plunger at a given rate and peak force required to squeeze all the material through the needle (e.g., 17G or 18G needle). Second, an apparatus for testing the heat capacity of cold slurry compositions was used to characterize the ice content for the cold slurry compositions. After evaluation, the compositions were tested for the following properties: 20% w / w glycerol, 0.5% w / w 1000kDa hyaluronic acid and 5% w / w Pluronic acid. TM Formulations containing F127 were prepared in saline, see Figure 1.

[0092] Example 2 - Evaluation of cold slurry cooling techniques and freezer spacing A cold slurry composition was prepared as described in Example 2. In order to optimize performance, different preparation methods were tested to determine the effect of different preparation methods on injection force described below. In this example, an experiment was conducted to determine whether the freezing temperature affected the injection force required to expel the cold slurry composition from the syringe. The results showed that a lower freezing temperature required a higher injection force. See FIG. 6. Furthermore, flash freezing the cold slurry composition at a temperature of about -60°C required an increase in injection force compared to the control. In a particular experiment, the cold slurry in multiple syringes was placed in a freezer by spacing the syringes apart to achieve uniform and rapid cooling of the cold slurry. When the syringes were placed very close together, freezing was observed to be adversely affected and larger ice crystals were observed. In contrast, a reduced injection force was observed when injecting the cold slurry from syringes spaced farther apart in the freezer.

[0093] Example 3 - Injectable frozen slurry temperature monitoring for performance optimization at the point of care Although lower injection forces are generally preferred for injecting cold slurries according to the present disclosure, it is desirable that the cold slurry be within a precise temperature range and not too warm to produce a therapeutic effect. It is desirable for the cold slurry to retain sufficient ice content, as this allows the cold slurry to extract energy from the target tissue to provide a therapeutic effect. If the ice content of the cold slurry is too low, the therapeutic effect may be reduced. Conversely, if the ice content of the cold slurry is too high, the cold slurry may not be injectable. In this example, an experiment was conducted to determine the ice content of the slurry using a copper plate. The ice content was calculated based on the final equilibrium temperature of the device after a given volume of slurry was dispensed into the device. As shown in FIG. 7A, the relationship between the injection temperature and the final equilibrium temperature is inversely related, such that as the injection temperature increases, the ice content of the cold slurry decreases. See FIG. 7A.

[0094] Further experiments showed that the injection force decreased as the temperature of the cold slurry increased. See FIG. 7B. Lower ice content also produced more reliable injections, i.e., the number of failed injections was reduced when the cold slurry had a lower ice content. However, as noted above, it is desirable for the cold slurry to retain sufficient ice content and temperature to retain therapeutic efficacy. Results showed that it is possible to optimize injection performance by controlling the ice content by targeting a predetermined injection temperature. Thus, a temperature monitoring device and system was designed that includes an indicator that provides an indication when the cold slurry has reached a predetermined temperature, where the predetermined temperature is selected to reduce the injection force required to expel the cold slurry from the syringe without reducing the ice content of the cold slurry below the therapeutic temperature. See, e.g., FIG. 5A-D; also see FIG. 4B. Additionally, additional temperature monitoring devices and systems have been designed that include multiple indicators, where the multiple indicators provide an indication when the cold slurry is at a temperature that is too cold for injection; when the cold slurry reaches a predetermined temperature, where the predetermined temperature is selected to reduce the injection force required to expel the cold slurry from the syringe without reducing the ice content of the cold slurry below the therapeutic temperature; or when the cold slurry reaches a temperature that is too warm to achieve a desired therapeutic effect. See, e.g., Figures 5A, 5B-C; see also Figure 4B.

[0095] Example 4 - Method for handling cold slurries between multiple syringes to improve injectability In certain experiments, different mechanical processing methods were tested. In some experiments, the cold slurry composition was prepared by connecting a first syringe and a second syringe using a connector, e.g., a Luer connector, and extruding the contents of the first syringe "shut-shut" into the second syringe. See Figs. 4A-B, 8. For example, in an experiment, a first syringe containing a male Luer component and containing the cold slurry composition was connected to a second syringe containing a male Luer component and not containing the cold slurry composition using a female-to-female Luer connector to connect the first and second syringes. See Figs. 4A-B. The slurry was shuttled back and forth between the first and second syringes, where one shuttle cycle included transporting the cold slurry from the first syringe to the second syringe and then transporting it back from the second syringe to the first syringe. It was observed that treating the slurry by extruding it back and forth significantly improved performance in injection tests and reduced the injection force required to expel the cold slurry from the syringe. See Figures 8-11. Without wishing to be bound by a particular theory, it is believed that performing one or more cycles of reciprocation reduces the injection force by reducing the size of the ice crystals in the cold slurry, distributing the ice crystals more uniformly within the cold slurry, and / or imparting shear forces to the cold slurry composition to increase the temperature of the cold slurry. An experiment was conducted to compare the injection force required to expel the slurry from the syringe after one, two, three, or four cycles of reciprocation. Figure 8. The experiment showed a significant reduction in the injection force required to inject the cold slurry through a 6 mL syringe fitted with a 17 G needle. Ibid. A control cold slurry not subjected to reciprocating cycles required an injection force of about 50-95 lbf. See, e.g., Figure 11. Cold slurries subjected to a single back and forth cycle required reduced injection forces ranging from about 6 to 38 lbf. Id.; see also Figures 8 and 9.Experiments showed that the injection temperature of the cold slurry was inversely related to the injection force (see FIG. 7B), but performing one or more double cycles further reduced the injection force required when the data was normalized to temperature. See FIG. 8. Results showed that increasing the number of consecutive double cycles to two or three cycles further reduced the injection force. Ibid. It was observed that after three double cycles, no further reduction in injection force was observed in slurries subjected to a fourth double cycle. Ibid. Further experiments showed that cold slurries subjected to two double cycles were consistently injectable using injection pressures of less than about 30 lbf over a range of post-treatment temperatures using 17G and 18G needles. See FIG. 9.

[0096] Further experiments included a processing element in a connector, e.g., a Luer connector, between the first and second syringes to push the contents of the first syringe back and forth into the second syringe as described above. It was observed that including a processing element in the connector produced equivocal results on the required injection force.

[0097] Example 5 - Sterilization Method for Cold Slurry Compositions Containing Hyaluronic Acid In certain experiments, various sterilization methods were investigated. In some experiments, the syringes containing the slurry composition were sterilized using radiation (gamma / e-beam sterilization). The results showed that radiation sterilization using gamma / e-beam sterilization resulted in an increase in the injection force required to expel the slurry from the syringe. See FIG. 12. Without wishing to be bound by a particular theory, this increase in the injection force required may be due to an effect on the molecular weight of hyaluronic acid in the composition. In some experiments, the syringes containing the slurry composition were sterilized using autoclave sterilization (steam sterilization) at various temperatures (e.g., about 118°C or about 121°C). Unlike radiation sterilization, it was observed that autoclave or steam sterilization did not increase the injection force required to expel the slurry from the syringe.

[0098] Example 6 - Method of making a cold slurry composition containing hyaluronic acid and soy PC Procedure for making 0.5% soy PC / HA / PBS / glycerol or 0.5% Tween 80 / HA / PBS / glycerol for shedding tests 1. Preparation of PBS / glycerol solution Weigh 20 g glycerol into a 125 mL narrow-mouth Erlenmeyer flask with a magnetic stir bar, add 80 mL 1x PBS, and seal the flask neck with parafilm to prevent evaporation of the solvent. Stir the liquid at 500 rpm for 4 h to obtain PBS / glycerol (41 wt / wt or 5 / 1 vol / vol). 2. Preparation of 0.75% HA / PBS / glycerol solution Weigh out a specific amount of HA 1000 kD (0.15 g–0.75 g to make 20 g–100 g solutions) into a 125 mL narrow-mouth Erlenmeyer flask with a magnetic stir bar (8 mm x 35 mm) and add a specific amount of PBS / glycerol solution (19.85 g–99.25 g to make 20 g–100 g solutions). Seal the mouth of the flask with parafilm to prevent evaporation of the solvent. Stir the liquid at 1200 rpm for 12–24 h to obtain 0.75% HA / PBS / glycerol. 3. Dissolution of soy PC in ethanol Weigh 1 g of soy PC into a 5 mL glass vial and add 0.2 mL ethanol with a pipette, vortex it, and then incubate it at 45 °C for 12-24 h to result in a light yellow, clear solution. 4. Preparation of 0.5% soy PC / HA / PBS / glycerol Example for preparing a 20g sample Weigh out 0.116 g of the soy PC in ethanol prepared above into a 125 mL narrow-mouth Erlenmeyer flask using a disposable transfer pipette. Add 19.884 g of 0.75% HA / PBS / glycerol. Add a magnetic stir bar (8 mm x 35 mm) and seal the mouth of the flask with parafilm to prevent evaporation of the solvent. Stir the liquid at 1200 rpm for 4-24 h to obtain 0.5% soy PC / HA / PBS / glycerol. 5. Preparation of 0.5% Tween 80 / HA / PBS / Glycerol Example for preparing a 20g sample Weigh 0.1 g Tween 80 into a 125 mL narrow-mouth Erlenmeyer flask using a disposable transfer pipette. Add 19.9 g of 0.75% HA / PBS / glycerol. Add a magnetic stir bar (8 mm x 35 mm) and seal the mouth of the flask with parafilm to prevent evaporation of the solvent. Stir the liquid at 1200 rpm for 4-24 h to obtain 0.5% Tween 80 / HA / PBS / glycerol. 6. The 0.5% Soy PC / HA / PBS / Glycerol or 0.5% Tween 80 / HA / PBS / Glycerol prepared above is transferred to a 20mL glass vial and sonicated for 10m. The resulting liquid is then loaded into 3x3mL syringes with 21G needles. The syringes are placed in a freezer at -20°C. After 12-24 h at -20 °C, remove the syringe from the freezer and couple it with an 18 G needle for the expulsion test. The test should be performed within 1 min of removal from the freezer.

Claims

1. A certain amount of water; hyaluronic acid; and 1. A composition comprising a first excipient, the composition being configured to form into a flowable cold slurry comprising a plurality of ice crystals when exposed to a temperature of 0° C. or below.

2. The composition further comprises a water-soluble surfactant; wherein Optionally, the water-soluble surfactant is a poloxamer molecule; Optionally, the composition comprises a plurality of poloxamer molecules; or Optionally, the composition comprises a poloxamer particle, the poloxamer particle comprising a plurality of poloxamer molecules; wherein:

2. The composition of claim 1, wherein the poloxamer particles are micelles.

3. 3. The composition of claim 2, wherein the poloxamer molecule is selected from the group consisting of poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 183, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer 235, poloxamer 237, poloxamer 238, poloxamer 282, poloxamer 284, poloxamer 288, poloxamer 331, poloxamer 333, poloxamer 334, poloxamer 335, poloxamer 338, poloxamer 401, poloxamer 402, poloxamer 403, poloxamer 407, poloxamer 105 benzoate, poloxamer 182 dibenzoate, and combinations thereof.

4. The composition of claim 3, wherein the poloxamer is poloxamer 407 and the concentration of poloxamer 407 is about 0.1% (w / w) to 10% (w / w), and optionally the concentration of poloxamer 407 in the composition is about 0.5% (w / w).

5. The method of claim 1, wherein the first excipient is selected from the group consisting of salts, ions, lactated Ringer's solution, sugars, biocompatible surfactants, polyols, and combinations thereof; optionally, the first excipient is glycerol; optionally, the concentration of glycerol in the composition is about 12% to 25% (w / w); 2. The composition of claim 1, optionally wherein the concentration of glycerol in the composition is about 19% (w / w).

6. The composition further comprises a second excipient; 10. The composition of claim 1, wherein the optional second excipient is sodium chloride or sodium phosphate to form saline or phosphate buffered saline.

7. The composition further comprising a third excipient; wherein: optionally, the third excipient is a water-insoluble material; Optionally, the water-insoluble substance is a lipid; 7. The composition of claim 6, wherein the lipid is optionally selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), egg sphingomyelin (DPSM), dipalmitoylphosphatidylcholine (DPPC), dicetylphosphate (DCP), La-phosphatidylcholine (soybean PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylglycerol (PG).

8. The composition of claim 1, wherein the first excipient is glycerol and the composition further comprises a water-soluble surfactant that is poloxamer 407 and a second excipient that is phosphate-buffered saline.

9. Configured to form a plurality of ice crystals when exposed to a temperature of about -25°C to about -5°C; or configured to have an injection force of less than about 30 lbf when injected through a 16G needle, a 17G needle, an 18G needle, a 19G needle, a 20G needle, a 22G needle, a 23G needle, or a 24G needle; or 10. The composition of claim 1, configured to have an injection force of less than about 30 lbf when injected through a 17G or 18G needle.

10. A method for preparing a cold slurry comprising the composition of claim 1 for administration to a patient at the clinical point of care, the method comprising: preparing a composition comprising hyaluronic acid and an amount of water; adding a first excipient to the composition, wherein the excipient comprises a water-soluble surfactant; and placing the composition in a freezer Including; The method, wherein the composition is configured to form a cold slurry comprising a plurality of ice particles when the composition is cooled to a temperature below about 0°C.

11. The method further comprising adding a second excipient to the composition, wherein the composition including the second excipient forms a cold slurry when the composition is cooled to a temperature below about 0°C; Optionally, the second excipient is selected from the group consisting of salts, ions, lactated Ringer's solution, sugars, biocompatible surfactants, polyols, and combinations thereof; Optionally, the second excipient is glycerol; Optionally, the method further comprises adding a third excipient to the composition, wherein the composition comprising the second excipient and the third excipient forms a cold slurry when the composition is cooled to a temperature below about 0°C; 11. The method of claim 10, optionally further comprising adding a fourth excipient to the composition, wherein the composition comprising the second excipient, the third excipient, and the fourth excipient is configured to form a cold slurry when the composition is cooled to a temperature below about 0°C.

12. 11. The method of claim 10, wherein the step of placing the composition in a freezer comprises exposing the composition to a temperature of about -25°C to about -5°C, and wherein the composition is configured to form a plurality of ice crystals when exposed to a temperature of about -25°C to about -5°C.

13. 13. The method of claim 12, wherein the step of placing the composition in a freezer comprises freezing at -20°C.

14. The method of claim 13, further comprising a step of warming the composition to a temperature of approximately -15°C and / or a step of subjecting the composition to three back-and-forth BAF cycles, wherein one BAF cycle comprises (1) extruding the cold slurry from a first syringe into a second syringe and (2) extruding the cold slurry from the second syringe back into the first syringe.

15. A cold slurry delivery system comprising a container holding the slurry composition of claim 1, the container comprises a sterile barrier and a temperature indicator, the container being configured to allow manual stirring of the slurry composition without disrupting the sterile barrier; Optionally, the container is a syringe or a tube; Optionally, the container is configured to be coupled to a second container; Optionally, the container and the second container are configured to be coupled using a connector; Optionally, the container and the second container comprise a first syringe and a second syringe, the first syringe and the second syringe each comprising a male luer component; Optionally, the connector comprises a female luer component; Optionally, the first syringe and the second syringe are connected using a female luer component, and the slurry composition can be transferred from the first syringe to the second syringe to manually stir the slurry composition.

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