PRESSURE-SENSITIVE HYDROGEL AND METHOD OF USE

Abstract

Embodiments of the disclosure may include a pressure sensitive hydrogel composition. The composition may include a liquid solvent, a polymer, and an acid gas. The composition may be capable of having a fluid phase in which the acid gas is dissolved in the solvent and the polymer is dissolved in the solvent, and the composition may be capable of having a gel phase in which the acid gas is not dissolved in the liquid solvent and the polymer is precipitated out of the solvent. The composition may also include a chemical compound or a pharmaceutical agent that can be released after the composition is delivered to a target tissue region.

Claims

1. A method of making a pressure-sensitive hydrogel composition, comprising: exposing a polymer and a solvent to an acid gas; and subjecting the polymer, the solvent, and the acid gas to an increase in pressure to dissolve the acid gas in the solvent to form an acidic solution, causing the polymer to dissolve into the solution to form a fluid under a first pH, subjecting the fluid to a decrease in pressure causes the acid gas to come out of solution to form a gel under a second pH higher than the first pH, wherein the fluid and the gel are reversibly interchangeable.

2. The method of claim 1, wherein the polymer is selected from the group consisting of collagen, gelatin, cellulose, hyaluronic acid, casein, alginate, fibrinogen, thrombin, and any combination thereof.

3. A method of repairing tissue, comprising: delivering and pressurizing the pressure-sensitive hydrogel composition of claim 1 to a target tissue region; and depressurizing the hydrogel composition, wherein the hydrogel composition comprises the polymer, the solvent and the acid gas, wherein the pressurized hydrogel composition is in a liquid state, and wherein the depressurized hydrogel composition is in a gel state.

4. The method of claim 3, wherein the pressurized hydrogel composition has a lower pH than the depressurized hydrogel composition.

5. The method of claim 3, wherein depressurizing the hydrogel composition includes exposing the hydrogel composition to ambient conditions.

6. The method of claim 3, wherein the polymer and the acid gas are dissolved in the solvent when pressurized, and the acid gas comes out of the solvent when the hydrogel composition is depressurized.

7. The method of claim 3, wherein the hydrogel composition further includes a pharmaceutical agent, wherein the pharmaceutical agent is dispersed in the gel when the hydrogel composition is depressurized, and the pharmaceutical agent is released after the hydrogel composition is delivered to the target tissue region.

8. The method of claim 1, wherein the acid gas is selected from the group consisting of hydrogen chloride, chlorine, sulfur dioxide, thiosulfate, nitrogen dioxide, hydrogen sulfide, hydrogen fluoride, carbon dioxide, and any combination thereof.

9. The method of claim 1, wherein the solvent is selected from the group consisting of water, saline solutions, buffer solutions, alcohol, esters, ketones, hydrocarbons, ether, and any combination thereof.

10. The method of claim 3, wherein the polymer is selected from the group consisting of collagen, gelatin, cellulose, hyaluronic acid, casein, alginate, fibrinogen, thrombin, and any combination thereof.

11. The method of claim 3, wherein the acid gas is selected from the group consisting of hydrogen chloride, chlorine, sulfur dioxide, thiosulfate, nitrogen dioxide, hydrogen sulfide, hydrogen fluoride, carbon dioxide, and any combination thereof.

12. The method of claim 3, wherein the solvent is selected from the group consisting of water, saline solutions, buffer solutions, alcohol, esters, ketones, hydrocarbons, ether, and any combination thereof.

Description

IV. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate exemplary embodiments of the present disclosure, and together with the description, serve to explain the principles of the disclosure.

[0020] FIG. 1 depicts the appearance of six exemplary hydrogels in a gel state, according to embodiments of the present disclosure;

[0021] FIG. 2 depicts the exemplary hydrogels of FIG. 1 approximately 30 minutes after being released from a syringe;

[0022] FIG. 3 depicts the appearance of five exemplary hydrogels in a gel state, according to embodiments of the present disclosure;

[0023] FIG. 4 depicts the exemplary hydrogels of FIG. 3 approximately 30 minutes after being released from a syringe;

[0024] FIG. 5 depicts the appearance of seven exemplary hydrogels in a gel state, according to embodiments of the present disclosure;

[0025] FIG. 6 depicts the exemplary hydrogels of FIG. 5 approximately 30 minutes after being released from a syringe;

[0026] FIG. 7 depicts the appearance of four exemplary hydrogels in a gel state, according to embodiments of the present disclosure;

[0027] FIG. 8 depicts the exemplary hydrogels of FIG. 7 approximately 30 minutes after being released from a syringe;

[0028] FIG. 9 depicts the appearance of four exemplary hydrogels in a gel state, according to embodiments of the present disclosure;

[0029] FIG. 10 depicts the exemplary hydrogels of FIG. 9 approximately 30 minutes after being released from a syringe;

[0030] FIG. 11a depicts the appearance of an exemplary hydrogel in a fluid state, according to embodiments of the present disclosure;

[0031] FIG. 11b depicts the appearance of the exemplary hydrogel of FIG. 11a in a gel state;

[0032] FIG. 12a depicts the appearance of the exemplary hydrogel of FIG. 11a in a fluid state;

[0033] FIG. 12b depicts the appearance of the exemplary hydrogel of FIG. 11a in a gel state; and

[0034] FIG. 13 depicts an exemplary drug delivery hydrogel, according to an exemplary embodiment of the present disclosure.

V. DETAILED DESCRIPTION

[0035] Reference will now be made in detail to the exemplary embodiments of the present disclosure described below and illustrated in the accompanying drawings. Wherever possible, the same reference numbers will be used throughout the drawings to refer to same or like parts.

[0036] While the present disclosure is described herein with reference to illustrative embodiments of pressure-sensitive hydrogels for medical applications, it is understood that the devices and methods of the present disclosure may be employed for any suitable application, including, but not limited to, agricultural, industrial, environmental, or commercial applications. Further, while collagen hydrogels using polymer networks are described, any suitable network component, including colloidal networks, e.g., may be utilized, and the hydrogels may include any suitable natural or synthetic polymers. Likewise, gels according to the present disclosure are not limited to hydrogels, and may include any other suitable liquid other than, or in addition to, water. Those having ordinary skill in the art and access to the teachings provided herein will recognize additional modifications, applications, embodiments, and substitution of equivalents that all fall within the scope of the disclosure. Accordingly, the disclosure is not to be considered as limited by the foregoing or following descriptions.

[0037] As used herein, the term fluid may refer to a substance whose molecules move freely past one another and has no fixed shape, e.g., a gas or a liquid. The term dissolve may refer to either partial or complete dissolving of a solute into a solvent, and solution may refer to a mixture created when a solute dissolves (either partially or fully) into a solvent. As used herein, the term hydrogel will be understood to mean both a gel in which water is a solvent, as well as a gel in which one or more solvents other than water are used.

[0038] The solubility of an acid gas may increase as the pressure of the surrounding environment increases. When not dissolved into solution, acid gases may not substantially affect the pH of surrounding liquids. When dissolved within a liquid, acid gases may increase the acidity of the resulting solution, thus, decreasing the solution's pH value. Accordingly, under higher pressures, an acid gas exposed to a liquid may become more soluble, and as a result, may dissolve into solution. In turn, the dissolving of the acid gas into solution may decrease the pH of the solution. If the pressure is then decreased, the acid gas may come back out of solution, reducing the solution's acidity and thus raising the solution's pH value.

[0039] Embodiments of the present disclosure utilize the solubility traits of synthetic and/or natural polymers to create new pressure-sensitive hydrogels. In some embodiments, hydrogels may include substantially neutral, aqueous polymer solutions. The polymers may be soluble in acidic conditions and may be insoluble or less soluble as the pH becomes less acidic. The aqueous polymer solutions may be exposed to acid gas. When exposed to higher pressures, the acid gas may be forced to dissolve into the aqueous polymer solution. As the acid gas dissolves into the solution, it may lower the pH value of the resulting solution, which may in turn cause the polymer to substantially dissolve into the acidic, aqueous solution. This may create a fluid solution of dissolved acid gas and polymer. A subsequent decrease in pressure may decrease the solubility of the acid gas, causing the acid gas to come out of solution. The precipitation of acid gas out of solution may in turn increase the pH value of the aqueous solution, e.g., in some examples, restoring it to a substantially neutral pH, or to a less-acidic or basic pH. The decrease in the solution's acidity may in turn cause the polymer to precipitate out of solution, causing the mixture to transition from a fluid to a gel. In this way, synthetic or natural polymers may be used to form hydrogels capable of transitioning from a fluid phase to a gel phase in response to a pressure stimulus using novel mechanisms different than those previously utilized.

[0040] Embodiments of the present disclosure may take advantage of the different solubility traits of different synthetic polymers and/or natural polymers, such as collagen, to create pressure-sensitive hydrogels. As discussed previously, collagen may be soluble under acidic conditions with pH values of less than neutral. In some embodiments, a collagen hydrogel may include collagen and a solvent (e.g., water) exposed to an acid gas under ambient pressure. Under ambient pressure, the acid gas may not dissolve, or may only partially dissolve, into solution, and thus exposure to the acid gas under ambient pressure may not significantly decrease the pH value of the solution. Due to the higher (e.g., substantially neutral) pH, the collagen may not dissolve into solution at ambient pressure, or may only partially dissolve, or may form an inhomogeneous state. At this point, the collagen hydrogel may exist in a gel phase.

[0041] Exposing the mixture to an increase in pressure may increase the solubility of the acid gas, creating an acid-gas solution and causing the pH of the resulting solution to drop. The decrease in pH may cause the collagen to dissolve, e.g., into a substantially homogenous solution. Causing the collagen to dissolve into solution may transition the hydrogel to a fluid state. Thus, under relatively lower-pressure conditions, the hydrogel may transition to a gel phase, and under relatively higher-pressure conditions, the hydrogel may transition to a fluid phase.

[0042] A subsequent decrease in pressure may once again decrease the solubility of the acid gas, causing it to come out of solution, and causing the pH value of the solution to rise. In some embodiments, the pH value of the solution may be neutralized, or may become less acidic or basic, as the acid gas comes out of solution. This may in turn cause the collagen to precipitate, at least in part, and may cause the mixture to transition back from a fluid to a gel. Accordingly, a pressure-sensitive hydrogel may be prepared according to the mechanisms described above.

[0043] As described above, the pressure-sensitive hydrogels exist in a liquid state when acid gas is forced into solution under increased pressures, causing the pH value of the solution to drop. The hydrogels transition to a gel state when the acid gas comes out of solution, and the polymer precipitates out due to the increase in pH value. Because the hydrogel sets to form a gel as the acid gas comes out of solution, gas may be trapped as it comes out of solution, forming bubbles within the set hydrogel. The formation of bubbles may cause the hydrogel to be porous when in the gel phase. In some embodiments, the bubbles may not connect with one another, and while the hydrogel may be porous, it may simultaneously be able to act as a sealant. In some embodiments, the porous nature of the gel may make the resulting hydrogel breathable, i.e., allowing air and/or gases to pass through, while in some embodiments the resulting hydrogel may be impermeable to air and/or gases.

[0044] Embodiments of the present disclosure may include any suitable natural or synthetic polymer or polymers that are more soluble under a first range of pH values, e.g., more acidic conditions having a relatively lower pH, and are less soluble under a second, different range of pH values, e.g., less acidic conditions, neutral conditions, or basic conditions having a relatively higher pH compared to the first pH range. Under the first range of pH values, a polymer hydrogel may exist in a fluid state, and under the second range, the polymer hydrogel may transition to a gel state. A hydrogel made from such a polymer may include or may be exposed to an acid gas, and as the solubility of the acid gas increases with an increase in pressure, the increased acidity of the resulting acid-gas solution may increase the solubility of the polymer, causing the hydrogel to transition to a fluid phase. The pH of the solution may thus be controlled via pressure, and accordingly, pressure may be manipulated to trigger transitioning of the hydrogel from a gel state to a fluid state, and/or vice versa. Accordingly, pressure-sensitive hydrogels may be created by manipulating the ability to dissolve or precipitate certain polymers based on changes in pH and the ability to manipulate pH by forcing acid gases into and out of solution in response to changes in pressure.

[0045] As discussed above, any suitable polymers or combination of polymers may be used to form a pressure-sensitive hydrogel. For example, polymers such as gelatin, cellulose, hyaluronic acid, chitosan, casein, alginate, fibrinogen, or thrombin may be used. The type or types of polymers used to form a pressure-sensitive hydrogel may be determined, at least in part, on the application of the hydrogel, the type of acid gas used, or the type of liquid solvent used. In turn, the type of polymer selected may affect the pH ranges needed to dissolve or precipitate the polymer into and out of solution. For example, an acidic pH of less than 6 may be used to dissolve the polymer, and an approximately neutral pH level of 6 or greater may be used to precipitate the polymer out of solution. In some embodiments, a more acidic pH and a less-acidic pH may be used to dissolve and precipitate the polymer, respectively, or an acidic pH and a basic pH may be used to dissolve and precipitate the polymer, respectively. In some embodiments, the difference between the dissolving pH and the precipitating pH may be small, while in other embodiments, the difference between the dissolving pH and the precipitating pH may be larger. The type of polymer used and the ranges of pH values required to dissolve or precipitate that polymer may affect the liquid solvent or the acid gas used in conjunction with that polymer to form a suitable pressure-sensitive hydrogel.

[0046] In some embodiments, multiple polymers may be used to form a single hydrogel. For example, in some embodiments, polymers with different solubility characteristics at different pH ranges may be included in the same hydrogel. Thus, exposure to different pH ranges may cause the hydrogel to transition to a gel state at different pressures as different amounts of acid gas are forced to dissolve into solution because of the change in pressure. In some embodiments, the polymers used may form gels of different firmness. For example, exposing a multi-polymer hydrogel to a first set of pressures may cause a first amount of acid gas to dissolve, creating a first pH range that causes a first polymer, but not a second polymer, to dissolve into solution. The second, non-dissolved polymer may cause a first gel state with a first viscosity. Exposing the hydrogel to a second set of pressures may cause a second, lesser amount of acid gas to dissolve, creating a second pH range and causing the first polymer to also precipitate out of solution, in addition to, or instead of, the second polymer. The first polymer may precipitate to form a second gel state having a second, different viscosity from the first gel state. Thus, a pressure-sensitive hydrogel may be created that is capable of transitioning to gel states with different viscosities, depending on the amount of pressure that the hydrogel is subjected to.

[0047] In some embodiments, hydrogels incorporating more than one polymer may provide redundancy. For example, some polymers may denature under different conditions, for example, different temperatures or pH ranges, or may have different shelf-lives. The inclusion of multiple types of polymers in a single hydrogel may increase the overall stability of the hydrogel or increase the likelihood that the hydrogel will transition from a fluid to a gel state as intended when subjected to a change in pressure. Such hydrogels may be useful, for example, when delivering the hydrogel to different environments (e.g., different areas in the body, when used in conjunction with different treatments or drugs), or when distribution or storage conditions to an end user or by a user of the hydrogel may be unpredictable or uncontrollable.

[0048] Further, any suitable type or types of acid gas may be used with embodiments of the present disclosure. For example, hydrogen chloride, chlorine, sulfur dioxide, thiosulfate, nitrogen dioxide, hydrogen sulfide, hydrogen fluoride, carbon dioxide, or any suitable combination thereof, may be used as a suitable acid gas. In some embodiments, weaker gases may be utilized because they may be easier to force into and precipitate out of solution. Further, in some embodiments, the acid gas may be in a solid or a liquid state when introduced to the mixture. For example, if carbon dioxide is used as the acid gas, it may be introduced to the fluid hydrogel mixture either in a gaseous, a liquid state, or solid state (i.e., dry ice). If introduced in a liquid or solid state, carbon dioxide may either completely or partially (so that some solid or liquid still remains) transition to an acid gas and then dissolve into solution. The pH values achieved by forcing different acid gases into solution may vary, for example, the pH values may range from approximately 7 to approximately 2, depending on the type of acid gas used. Thus, the type of acid gas used may affect the type of polymer used. For example, collagen may dissolve at a pH of approximately 3 and may precipitate at a pH of approximately 5. The type or types of acid gas used may be determined, at least in part, by the hydrogel application, the type of solvent used, or the type of polymer used.

[0049] In some embodiments, the type of acid gas used may affect the amount of pressure required to dissolve the acid gas into and out of solution, and thus the pressure at which the hydrogel will transition to a gel state or a liquid state. In some embodiments, the type of acid gas used may affect the pH change induced when the acid gas is dissolved. In some embodiments, multiple acid gases may be used in a single hydrogel. For example, a first acid gas may dissolve into solution under a first range of pressures and/or may create a first pH range when dissolved. A second acid gas may dissolve into solution under a second range of pressures and/or may cause a second pH range when dissolved. Accordingly, the different acid gases may cause the hydrogel to transition to fluid or a gel at different pressure ranges. The different acid gases may also cause more or less dissolving of the polymer, or more or less homogeneity of the polymer solution. This may in turn affect the viscosity of the hydrogel when transitioning to the gel state. In some embodiments, using multiple acid gases may also facilitate the use of multiple polymers within a hydrogel.

[0050] In addition, any suitable type of liquid solvent may be used to form a hydrogel according to the present disclosure. While the present disclosure refers primarily to hydrogels, and thus brings to mind the use of water as the solvent, any suitable liquid solvent may be used, for example, saline solutions, buffer solutions, alcohol (e.g., methanol or ethanol), esters, ketones, hydrocarbons, and ether, or any suitable combination thereof. In some embodiments, a buffer solution (e.g., Hank's Balanced Salt Solution, Dulbecco's Phosphate-Buffered Saline, Ringer Solution, Tri-HCl, etc.) may be used alone or in combination with another solvent, e.g., water. The type or types of solvents used may be determined, at least in part, by the hydrogel application, the type of acid gas used, or the type of polymer used. In turn, the type of acid gas used, the type of polymer used, and the ranges of pH needed to dissolve or precipitate the polymer may affect the type of solvent chosen.

[0051] The amount of pressure necessary to transition a hydrogel from a fluid state to a gel state and/or back to a fluid state may depend, at least in part, on the type of solvent, polymer, or acid gas used in a given hydrogel embodiment. In some embodiments, the transition point from a fluid to a gel may be defined as a predetermined amount above ambient pressure, and the hydrogel may exist as a gel at a pressure substantially equal to ambient pressure. In other embodiments, the transition point may be above ambient pressure, while the hydrogel may also exist as a gel above ambient pressure, or the transition point may be below ambient pressure, or the transition point may be at substantially ambient pressure and the hydrogel may exist as a gel below ambient pressure. Further, as will be recognized, ambient pressure may be relative depending on geographic location, and the exact transition point may vary slightly depending on ambient conditions, for example, geographic location or temperature.

[0052] The amount of pressure necessary to transition between the fluid and gel states may be chosen, at least in part, according to the intended application of the hydrogel. In some embodiments, the transition from a fluid to a gel or from a gel to a fluid may be gradual and may occur over a range of transition pressures. In other embodiments, the hydrogel phase transition may occur more abruptly as a given pressure threshold is crossed. In some embodiments, the transition point from a fluid to a gel phase may be different than the transition point of a gel to a fluid phase or vice versa, for example, in abruptness or in the exact pressure required. The gradual or abrupt nature of the transition may be affected, at least in part, by the polymer or solvent used, or by environmental conditions, such as the surrounding temperature or humidity, for example. In some embodiments, the transition from a fluid to a gel may take less than a minute, while in some embodiments, the transition may take several minutes. For example, the transition may take approximately 30 seconds, 1 to 2 minutes, 10 minutes, or up to two hours or longer.

[0053] Additionally, according to the gas laws, there is a known relationship between temperature and pressure. Thus, both of these variables may affect the gelling properties of a pressure-sensitive hydrogel. Under different temperatures and different pressures, hydrogels may have different gelling rates or different gelling properties (e.g., viscosity, handling, etc.). For example, exposing a hydrogel to a higher temperature may cause the hydrogel to transition to a gel more quickly than the same hydrogel when exposed to a lower temperature. The effects of temperature and pressure may also depend at least in part on the type of polymer included in a given hydrogel. For example, a collagen-based hydrogel may take approximately 1 to 2 minutes to gel at approximately body temperature (roughly 37.5 C.) and a pressure of 1 atm. By changing the temperature, different gels may have different optimal transition pressures. Accordingly, by controlling the temperature in addition to the pressure, the gelling rate and/or the gelling characteristics may be more finely controlled. The exact optimum pressure ranges for transitioning a hydrogel and for maintaining either a gel or liquid state may depend on the polymer used and the acid gas used and may be unpredictable.

[0054] The transition of an exemplary, pressure-sensitive hydrogel from a fluid to a gel may be reversible in some embodiments. For example, exposing the hydrogel to a decrease in pressure may cause the hydrogel to transition from a fluid to a gel, and subsequently increasing the pressure may cause the hydrogel to transition from a gel back to a fluid. In some embodiments, the transition may be irreversible. For example, exposing the hydrogel to a decrease in pressure may cause the hydrogel to transition from a fluid to a gel, and subsequently increasing the pressure may not cause the hydrogel to transition back to a fluid. In such embodiments, once transitioned, the hydrogel may maintain its gel state independent of the surrounding pressure.

EXAMPLES OF PRESSURE-SENSITIVE HYDROGELS

Examples 1-6

[0055] A series of six hydrogels was formed by mixing 0.2 g of collagen and 4 g of solid carbon dioxide (dry ice) with one of the following buffer solutions in a 20 ml stainless-steel bottle: (i) 10 ml of Hank's Balanced Salt Solution (HBSS), (ii) Dulbecco's Phosphate-Buffered Saline (DPBS), (iii) normal saline, (iv) Ringer Solution, (v) Tri-HCl buffer, and (vi) water. The bottle was sealed with a screwed valve in order to prevent leakage of the carbon dioxide. At equilibrium, under room temperature (approximately 30 C.), the dry ice inside of the bottle was transformed into either a homogeneous gas state or a gas-liquid coexistent state. The bottle was allowed to remain at room temperature for approximately 18-20 hours.

[0056] The valve was then unscrewed, and the collagen was released from the stainless-steel bottle, allowing the hydrogel to return to atmospheric pressure. The gelling ability and handling properties of each of the released gels was analyzed, and the visual appearance of each gel was observed and photographed.

[0057] Gelling Ability

[0058] Gelling ability, or gelation property, is generally defined as the ability of a hydrogel to form a gel and retain its shape over a period of time. Gelling ability may be assessed by visual observation of the shape and ability of the gel to maintain a given shape once set. This visual observation can then be scored and/or ranked. Gelling ability was assessed according to the following procedure. For each of the six hydrogels, 0.39 ml of the gel was put into a 96-well plate. Triplicates were made for all groups. The plate was put into an ELISA reader after approximately 10 minutes of incubation at room temperature, and results were read with a measurement wavelength of 450 nm.

[0059] Table 1 (below) shows the semi-qualitative evaluation of gelling ability. Gelation property measures the ability of a gel to maintain a given shape after gelling. For example, a 0 rating may indicate that while a hydrogel gelled, it failed to keep its shape.

TABLE-US-00001 TABLE 1 Semi-Qualitative Evaluation of Gelling Ability OD 450 nm Gelation Property 4.00 or more +++ 3.00-3.99 ++ 2.50-2.99 + 2.49 or less 0

[0060] Handling Property

[0061] Handling property is generally defined as the ability of a hydrogel to gel at the desired time. For example, if a delivery device is used to store and then release a hydrogel, a hydrogel that gels while still in the delivery device would not have good handling properties, because it would have setand therefore its shape would have been determinedin the device rather than when released from the device for use in a given application. Handling property is a measure of the manipulability and operation of a given hydrogel. Handling property was assessed according to the following procedure. For each of the six hydrogels, 1.5 ml of the gel was put into a syringe of which the tip had been removed. The gel-filled syringe was incubated at room temperature for approximately 10 minutes. The gel was pushed out from the syringe, and the integrity of the gel structure was evaluated and observed.

[0062] If the gel could remain intact as a cylinder shape after being pushed out of the syringe, this indicated that the gel was not set until it was released from the stainless-steel bottle. If the gel broke into small pieces after being pushed out of the syringe, this indicated that the gel set before it was released from the bottle and could not be modified into a desired shape. Photographs were taken after pushing the gels out of the syringe for observation of visual appearance. Photographs were again taken of the gels after approximately 30 minutes of incubation at room temperature.

[0063] Table 2 (below) shows the results of gelling ability, handling property, and visual appearance of the gels released from the stainless-steel bottle containing solid carbon dioxide with the various buffer solutions. The appearances of the resulting gels are depicted in FIGS. 1 and 2. FIG. 1 shows the appearance of collagen mixed with solid carbon dioxide and each of the six buffer solutions after approximately 10 minutes of gelling in the syringe. FIG. 2 depicts the appearance of collagen mixed with solid carbon dioxide and each of the six buffer solutions approximately 30 minutes after being pushed out of the syringe.

TABLE-US-00002 TABLE 2 Gelling Ability and Handling of Buffer Solution Gels Example 1 2 3 4 5 6 Collagen(g) 0.2 0.2 0.2 0.2 0.2 0.2 CO2 (g) 4 4 4 4 4 4 HBSS(ml) 10 Potassium Chloride (5.33 mM) Potassium Phosphate monobasic (0.441 mM) Sodium Bicarbonate (4.17 mM) Sodium Chloride (137.83 mM) Sodium Phosphate dibasic anhydrous(0.338 mM) D-Glucose (5.56 mM) DPBS(ml) 10 Potassium Chloride (2.67 mM) Potassium Phosphate monobasic (1.47 mM) Sodium Chloride (137.93 mM) Sodium Phosphate dibasic (8.06 mM) Saline(ml) 10 Sodium Chloride (153.84 mM) Ringer solution(ml) 10 Sodium Chloride (147 mM) Potassium Chloride (4.02 mM) Calcium Chloride Dihydrate (2.24 mM) Tris-HCl buffer(ml) 10 Tris base(50 mM) Purified water(ml) 10 Operation Yes Yes Yes Yes Yes OD450 nm 2.762 3.851 3.088 3.096 2.706 2.038 Gelation property + ++ ++ ++ + 0

Examples 7-11

[0064] A series of five hydrogels was formed by mixing 10 ml of HBSS and 4 g of solid carbon dioxide (dry ice) with one of the following amounts of collagen in a 20 ml stainless-steel bottle: (i) 0.05 g of collagen, (ii) 0.1 g of collagen, (iii) 0.2 g of collagen, (iv) 0.3 g of collagen, and (v) 0.4 g of collagen. The bottle was sealed with a screwed valve in order to prevent leakage of the carbon dioxide. At equilibrium, under room temperature (approximately 30 C.), the dry ice inside of the bottle was transformed into either a homogeneous gas state or a gas-liquid coexistent state. The bottle was allowed to remain at room temperature for approximately 18-20 hours.

[0065] The valve was then unscrewed, and the collagen was released from the stainless-steel bottle, allowing the hydrogel to return to atmospheric pressure. The gelling ability and handling properties of each of the released gels was analyzed, and the visual appearance of each gel was observed and photographed.

[0066] Table 3 (below) shows the results of gelling ability, handling property, and visual appearance of the gels released from the stainless-steel bottle containing solid carbon dioxide and HBSS with the different amounts of collagen. The appearances of the resulting gels are depicted in FIGS. 3 and 4. FIG. 3 shows the appearance of solid carbon dioxide and HBSS mixed with the different amounts of collagen after approximately 10 minutes of gelling in the syringe. FIG. 4 depicts the appearance of solid carbon dioxide and HBSS with the different amounts of collagen approximately 30 minutes after being pushed out of the syringe.

TABLE-US-00003 TABLE 3 Gelling Ability and Handling of Collagen Gels Example 7 8 9 10 11 Collagen(g) 0.05 0.1 0.2 0.3 0.4 CO2 (g) 4 4 4 4 4 HBSS(ml) 10 10 10 10 10 Potassium Chloride (5.33 mM) Potassium Phosphate monobasic (0.441 mM) Sodium Bicarbonate (4.17 mM) Sodium Chloride (137.93 mM) Sodium Phosphate dibasic anhydrous(0.338 mM) D-Glucose (5.56 mM) Operation Yes Yes Yes Yes Yes OD450 nm 1.602 2.989 2.995 3.697 4 Gelation property 0 + + ++ +++

Examples 12-18

[0067] A series of seven hydrogels was formed by mixing 10 ml of HBSS and 2 g of collagen with one of the following amounts of solid carbon dioxide (dry ice) in a 20 ml stainless-steel bottle: (i) 0.5 g of solid carbon dioxide, (ii) 1 g of solid carbon dioxide, (iii) 2 g of solid carbon dioxide, (iv) 3 g of solid carbon dioxide, (v) 4 g of solid carbon dioxide, (vi) 5 g of solid carbon dioxide, and (vii) 6 g of solid carbon dioxide. The bottle was sealed with a screwed valve in order to prevent leakage of the carbon dioxide. At equilibrium, under room temperature (approximately 30 C.), the dry ice inside of the bottle was transformed into either a homogeneous gas state or a gas-liquid coexistent state. The bottle was allowed to remain at room temperature for approximately 18-20 hours.

[0068] The valve was then unscrewed, and the collagen was released from the stainless-steel bottle, allowing the hydrogel to return to atmospheric pressure. The gelling ability and handling properties of each of the released gels was analyzed, and the visual appearance of each gel was observed and photographed.

[0069] Table 4 (below) shows the results of gelling ability, handling property, and visual appearance of the gels released from the stainless-steel bottle containing collagen and HBSS with the different amounts of solid carbon dioxide. The appearances of the resulting gels are depicted in FIGS. 5 and 6. FIG. 5 shows the appearance of collagen and HBSS mixed with the different amounts of solid carbon dioxide after approximately 10 minutes of gelling in the syringe. FIG. 6 depicts the appearance of collagen and HBSS with the different amounts of solid carbon dioxide approximately 30 minutes after being pushed out of the syringe.

[0070] Using different amounts of carbon dioxide to evaluate pH-sensitivity may create different pressures if the remaining variables, e.g., bottle size, are kept the same across experiment groups. Accordingly, Table 4 also includes the theoretical pressure calculation for each experiment group. For example, while not measured during the experimental trials, and while not bound to a particular range, the pressure may be approximately 55.5 atm when 1 g of carbon dioxide is mixed with 10 ml of HBSS, and the carbon dioxide may begin to dissolve into solution at approximately 56 atm and 20 C.

TABLE-US-00004 TABLE 4 Gelling Ability and Handling of CO.sub.2 Gels Example 12 13 14 15 16 17 18 Collagen(g) 0.2 0.2 0.2 0.2 0.2 0.2 0.2 CO2 (g) 0.5 1 2 3 4 5 6 Pressure@20 C.(atm) 27.77 55.5 56 56 56 56 56 HBSS(ml) 10 10 10 10 10 10 10 Potassium Chloride (5.33 mM) Potassium Phosphate monobasic (0.441 mM) Sodium Bicarbonate (4.17 mM) Sodium Chloride (137.93 mM) Sodium Phosphate dibasic anhydrous(0.338 mM) D-Glucose (5.56 mM) Operation Yes Yes Yes Yes Yes Yes Yes OD450 nm 2.836 2.831 2.888 2.865 2.882 2.879 2.946 Gelation property + + + + + + +

Examples 19-22

[0071] A series of four hydrogels was formed by mixing 10 ml of water, 0.2 g of collagen, and 4 g of solid carbon dioxide with one of the following amounts of sodium bicarbonate (NaHCO.sub.3) in a 20 ml stainless-steel bottle: (i) 3 mg of NaHCO.sub.3, (ii) 7.5 mg of NaHCO.sub.3, (iii) 15 mg of NaHCO.sub.3, and (iv) 30 mg of NaHCO.sub.3. The bottle was sealed with a screwed valve in order to prevent leakage of the carbon dioxide. At equilibrium, under room temperature (approximately 30 C. or less), the dry ice inside of the bottle was transformed into either a homogeneous gas state or a gas-liquid coexistent state. The bottle was allowed to remain at room temperature for approximately 18-20 hours.

[0072] The valve was then unscrewed, and the collagen was released from the stainless-steel bottle, allowing the hydrogel to return to atmospheric pressure. The gelling ability and handling properties of each of the released gels was analyzed, and the visual appearance of each gel was observed and photographed.

[0073] Table 5 (below) shows the results of gelling ability, handling property, and visual appearance of the gels released from the stainless-steel bottle containing collagen and solid carbon dioxide with the different concentrations of NaHCO.sub.3. The appearances of the resulting gels are depicted in FIGS. 7 and 8. FIG. 7 shows the appearance of collagen and solid carbon dioxide mixed with the different concentrations of NaHCO.sub.3 after approximately 10 minutes of gelling in the syringe. FIG. 8 depicts the appearance of collagen and solid carbon dioxide with the different concentrations of NaHCO.sub.3 approximately 30 minutes after being pushed out of the syringe.

TABLE-US-00005 TABLE 5 Gelling Ability and Handling of NaHCO.sub.3 Gels Example 19 20 21 22 Collagen(g) 0.2 0.2 0.2 0.2 CO2 (g) 4 4 4 4 Purified water(ml) 10 10 10 10 NaHCO3(mg) 3 7.5 15 30 Operation No No No No OD450 nm 2.302 2.535 2.593 2.621 Gelation property 0 + + +

Examples 23-26

[0074] A series of four hydrogels was formed by mixing 10 ml of water, 0.2 g of collagen, and 4 g of solid carbon dioxide with one of the following amounts of sodium hydroxide (NaOH) in a 20 ml stainless-steel bottle: (i) 0.4 mg of NaOH, (ii) 4 mg of NaOH, (iii) 20 mg of NaOH, and (iv) 40 mg of NaOH. The bottle was sealed with a screwed valve in order to prevent leakage of the carbon dioxide. At equilibrium, under room temperature (approximately 30 C.), the dry ice inside of the bottle was transformed into either a homogeneous gas state or a gas-liquid coexistent state. The bottle was allowed to remain at room temperature for approximately 18-20 hours.

[0075] The valve was then unscrewed, and the collagen was released from the stainless-steel bottle, allowing the hydrogel to return to atmospheric pressure. The gelling ability and handling properties of each of the released gels was analyzed, and the visual appearance of each gel was observed and photographed.

[0076] Table 6 (below) shows the results of gelling ability, handling property, and visual appearance of the gels released from the stainless-steel bottle containing collagen and solid carbon dioxide with the different concentrations of NaOH. The appearances of the resulting gels are depicted in FIGS. 9 and 10. FIG. 9 shows the appearance of collagen and solid carbon dioxide mixed with the different concentrations of NaOH after approximately 10 minutes of gelling in the syringe. FIG. 10 depicts the appearance of collagen and solid carbon dioxide with the different concentrations of NaOH approximately 30 minutes after being pushed out of the syringe.

TABLE-US-00006 TABLE 6 Gelling Ability and Handling of NaOH Gels Example 23 24 25 26 Collagen(g) 0.2 0.2 0.2 0.2 CO2 (g) 4 4 4 4 Purified water(ml) 10 10 10 10 NaOH(mg) 0.4 4 20 40 Operation Yes Yes Yes Yes OD450 nm 2.4 2.587 2.657 2.701 Gelation property 0 + + +

Example 27

[0077] As discussed above, other polymers may be used instead of, or in addition to, collagen. For example, chitosan may be used in place of collagen. Chitosan powder may be dissolved at an acidic pH below 7 and may then begin precipitating out of solution to form a gel as the pH becomes less acidic and approaches neutral. In Example 27, 0.06 g of chitosan was dissolved into 3 ml of 0.1 M HCl to prepare a 2% chitosan solution, as shown in FIG. 11a. The solution was incubated at a temperature of approximately 37 C. to allow the chitosan to dissolve into the 0.1 M HCl. After 1 hour, 3 ml of 0.1 M NaHCO.sub.3 was added to the chitosan solution to neutralize the solution. The mixture was then incubated at 37 C. for two hours, and during that time, the chitosan precipitated out of solution and formed a chitosan gel, as is shown in FIG. 11b.

[0078] After the two-hour incubation period, the gel was mixed with 4 g of solid carbon dioxide in a 20 ml stainless steel bottle. The bottle was sealed with a screwed valve to inhibit leakage of carbon dioxide. The chitosan gel was incubated at room temperature for approximately 18 to 20 hours. During this time, the increased pressure caused the carbon dioxide to dissolve into solution, decreasing the pH value and causing the chitosan gel to dissolve back into solution. The chitosan hydrogel was then released from the stainless steel bottle, as shown in FIG. 12a, and the visual appearance was observed. The chitosan hydrogel was incubated at 37 C. following release. During that time, the chitosan solution transformed into a gel as the pH neutralized and the chitosan precipitated out of solution. FIG. 12b depicts the visual appearance of the chitosan hydrogel in a gel after the two hours of incubation.

Example 28

[0079] As discussed above, in some embodiments, the hydrogel may also serve as a delivery mechanism for a chemical compound. Such an exemplary hydrogel may deliver one or more pharmaceutical agents locally in the area to which the hydrogel is applied, e.g., to a target tissue region. This may allow a user to deliver a targeted dose of drug to a region of the body instead of, or in addition to, using the hydrogel to manipulate and/or repair tissue. In such embodiments, a pressurized hydrogel may be delivered to a region of the body, may be allowed to depressurize and gel in that body region, and may release a pharmaceutical agent contained in the hydrogel over a period of time as the hydrogel transitions from one state to another or as it degrades in the body. The pharmaceutical agent may be dispersed in the hydrogel when the hydrogel is in a depressurized gel state and may be released after the hydrogel is delivered to the target tissue region.

[0080] In one exemplary embodiment, caffeine was used as a model chemical compound to demonstrate the ability of hydrogels in accordance with this disclosure to act as a delivery mechanism for chemical compounds. In example 28, the hydrogel was formed by mixing 0.2 g of collagen, 4 g of solid carbon dioxide, and 10 ml of HBSS buffer with 0.0015 g of caffeine in a 20 ml stainless bottle. The bottle was sealed with a screwed valve in order to prevent leakage of the carbon dioxide. At equilibrium, under room temperature (approximately 30 C. or less), the dry ice inside of the bottle was transformed into either a homogeneous gas state or a gas-liquid coexistent state. The bottle was allowed to remain at room temperature for approximately 18-20 hours.

[0081] The valve was then unscrewed, the collagen was released from the stainless-steel bottle into a petri dish, and the resulting foamy material was allowed to cure at 37 C. for 3 minutes. The cured hydrogel was then cut into 1 cm2 cm4 mm pieces. The pieces were then placed into a 50 ml tube together with 10 ml of ammonium acetate buffer and 20 l of collagenase solution (10 mg/ml). The tube was then incubated in a water bath maintained at 37 C. Samples of supernatant were collected from the tube at a series of time points, and the concentration of caffeine in the supernatant sample at each time point was measured. The results of these measurements are shown in FIG. 13.

[0082] FIG. 13 demonstrates that the caffeine was released into the supernatant from the hydrogel over time. This time release of a chemical compound may mimic how the hydrogel would be degraded by collagenase inside of the human body. In this embodiment, approximately 80% of the caffeine in the hydrogel was released within the first 3 hours, while remaining caffeine was gradually released as the hydrogel degraded. The concentration of caffeine in the supernatant plateaued after about 24-48 hours of degradation in the collagenase solution, which may generally correspond with the time it takes for this specific hydrogel to degrade. This example demonstrates the ability of an exemplary hydrogel embodiment to deliver and release a drug in a predetermined amount. For example, in some embodiments, a hydrogel may deliver an initial burst release of a drug, e.g., due to the presence of drug absorbed on the surface of the set hydrogel, as well as a more gradual release of drug over time once applied to a target tissue region. It is believed that the drug is released as the hydrogel degrades, for example, by collagenase, within the body. While the caffeine example described above demonstrates the capacity for the disclosed hydrogel to act as a delivery mechanism, it is expected that the ability of the hydrogel to deliver a specific chemical compound may depend on, among other things, the properties of the specific hydrogel selected, the environment in which it is used, the nature and properties of the specific drug or combination of drugs, the dosing necessary for therapeutic benefits, and/or the intended therapeutic treatment, for example.

[0083] Embodiments of the present disclosure may provide several benefits over currently available hydrogels. For example, embodiments of the disclosure may provide a hydrogel capable of gelling in situ that is easy to manufacture and easy to transition between fluid and gel states using conventional processes. Many in-situ gelling hydrogels are controlled by temperature, which can require that the products be stored under regulated, low-temperature conditions prior to use. Moreover, the temperature may be difficult to control during use, and the hydrogel operation temperature window may be narrow. By using pressure to manipulate pH, embodiments of the present disclosure may offer hydrogel products that can be stored under room temperature conditions or under a wider range of temperatures. Further, in-situ gelling may be easily achieved once the pressurized condition is released and the hydrogel is re-exposed to ambient pressure.

[0084] The flowable and injectable nature of hydrogels according to the present disclosure make this type of hydrogel easy to transport, easy to deliver, and conformable for a wide variety of uses. These pressure-sensitive hydrogels are well-suited for applications that require the filling of three-dimensional areas. While other available hydrogels may be limited for use as a sealant and may require use of an accompanying scaffold, these pressure-sensitive hydrogels may be capable acting as both a sealant and a scaffold. Exemplary, three-dimensional uses may include medical applications, such as repairing wounds (e.g., after surgery or following resections, or after traumatic injuries such as gun shots) or filling defects, e.g., for bone or cartilage repair. In some embodiments, hydrogels may be used for dural repair or as dural substitutes, e.g., following excision of a brain tumor, craniotomy, or spinal surgery. They may also be used as three-dimensional scaffolds for biomedical and tissue engineering applications.

[0085] Further, because hydrogels of the present disclosure are porous, they may offer additional advantages over other hydrogel sealants. For example, many sealants adhere to the surrounding tissues, applying pressure to these tissues. The hydrogel may swell and expand in volume, causing pressure when in biological conditions. This pressure may build up and cause damage to the surrounding tissue or may cause unwanted pressure differentials on either side of the sealant. The pressure-sensitive hydrogels disclosed herein form tiny bubbles within the gel as it sets. The bubbles may not connect, and so while the gel may provide an adequate sealant, the porous nature of the hydrogel in the gel state may reduce or eliminate the pressure otherwise caused by conventional sealants. This may be particularly useful in medical applications, for example, when used in dural repair, so as to not apply pressure to portions of the central nervous system.

[0086] In addition to being biocompatible, hydrogels of the present disclosure also have tissue adhesion properties. Tissue may not be able to re-grow into most of the currently available hydrogels, so even when used with scaffolding, traditional hydrogels may stay separate within the body of a patient. Alternatively, tissue may be able to grow into embodiments of these novel pressure-sensitive hydrogels. This may allow for more natural, and/or stronger healing and regrowth over time, again providing advances in medical and three-dimensional applications.

[0087] Hydrogels of the present disclosure may also be used to manipulate, move, or stabilize tissue, to lift and/or separate tissue or layers of tissue (e.g., to aid in resection), or for tissue augmentation. Hydrogels may be used to coat, encapsulate, absorb, or dissolve substances, or to act as a barrier to moisture, for example. Hydrogels may also be used for treatment, for example, to aid in drug delivery, including, e.g., localized and/or slow-release delivery. Use for treatment may be alone or in combination with one of the above uses (e.g., to fill a wound and deliver an antiseptic).

[0088] In medical applications, a pressurized, fluid hydrogel may be delivered to an area of the body, and upon delivery, it may be exposed to a lower pressure, e.g., ambient pressure. Exposure to ambient pressure may cause the acid gas to come out of solution, causing the hydrogel to gel within the body. The acid gas that comes out of solution as the hydrogel sets may then be metabolized by the body, may be either passively or actively removed from the body, or may be left within the body. This may depend on the type of acid gas used. For example, if carbon dioxide is chosen as the acid gas, it may be metabolized by the body, because carbon dioxide is a common metabolite. In some embodiments, the resulting hydrogel may not be directly ejected into the body. Accordingly, after ejection, much of the acid gas may dissipate into atmosphere, with only a portion of the acid gas being trapped inside the hydrogel. Additionally, the remaining acid gas may be released over a period of time as the hydrogel degrades. In such embodiments, there may not be a burst of acid gas at the initial stage of hydrogel delivery and implantation, since most of the acid gas will dissipate into the atmosphere or will only slowly release over time. The smaller quantity of remaining acid gas may have no significant effect on the body when it is slowly released as the hydrogel degrades. In some embodiments, though, this remaining quantity may be actively removed from the body, or, in the case of gases such as carbon dioxide, may be metabolized by the body.

[0089] In some embodiments, other materials may be included in the hydrogel. For example, dyes may be added to aid in user visualization. Pharmaceutical agents may be included in the hydrogel to achieve localized and/or time-released delivery of treatment to the surrounding area as the hydrogel degrades within the body. Examples of such agents may include anesthetics, anti-inflammatories, antiseptics, or medications that facilitate tissue regeneration, prevent infection (e.g., antibiotics), or treat diseases (e.g., cancer). Other additive materials may include chemicals, ceramics, and biomaterials, such as growth factor, fibrinogen, platelet-rich plasma, or other suitable materials or combination of materials. In some embodiments, chemicals capable of controlling or adjusting pH and osmolality, such as, e.g., sodium bicarbonate, sodium chloride, calcium chloride, or potassium chloride, may be included. Such additives may act as a buffer, e.g., as the hydrogel degrades and the acid gas is released.

[0090] The many features and advantages of the present disclosure are apparent from the detailed specification, and thus, it is intended by the appended claims to cover all such features and advantages of the present disclosure that fall within the true spirit and scope of the present disclosure. Further, since numerous modifications and variations will readily occur to those skilled in the art, it is not desired to limit the present disclosure to the exact construction and operation illustrated and described, and accordingly, all suitable modifications and equivalents may be resorted to, falling within the scope of the present disclosure.

[0091] Moreover, those skilled in the art will appreciate that the conception upon which this disclosure is based may readily be used as a basis for designing other structures, methods, and systems for carrying out the several purposes of the present disclosure. Accordingly, the claims are not to be considered as limited by the foregoing description.