Patent Description:
The past half-century has witnessed development in the use of minimally invasive, endovascular techniques in medicine. Interventions predicated upon real-time image guidance to direct flexible catheters from an easily accessible, superficial blood vessel to a remote blood vessel deep within the body have revolutionized the clinical management of diseases involving many organs. A common paradigm in endovascular procedures is the performance of vascular embolization, a technique in which an occlusive agent is delivered through a catheter to obstruct flow within a target blood vessel.

<CIT> discloses hemostatic compositions comprising gelatin or a derivative thereof and silicate nanoparticles, for example injectable and self-healing hemostats useful for clotting blood.

<CIT> discloses a shear thinning injectable composition for treating tumours comprising: an agent, water, a contrast agent and fumed silica.

The present invention, according to the appended set of claims, encompasses shear-thinning compositions for use in treating a hemorrhage, where the hemorrhage is an internal hemorrhage associated with a medical disorder selected from the group comprising benign cancer, malignant cancer, metastatic cancer, or any combination thereof. In some embodiments, the cancer is selected from the group consisting of breast cancer, prostate cancer, benign prostatic hyperplasia, esophageal cancer, liver cancer, colon cancer, endometrial cancer, brain cancer, bladder cancer, cancer of the uterus, fibroid (leiomyoma) uterus, cancer of the ovary, lung cancer, sarcoma, bone cancer, pancreatic cancer, renal cancer, and stomach cancer.

According to the appended set of claims, the invention relates to a shear-thinning composition and a therapeutic agent or a diagnostic agent for use in treating cancer in a subject in need thereof. In some embodiments, the cancer is selected from the group consisting of breast cancer, prostate cancer, benign prostatic hyperplasia, esophageal cancer, liver cancer, colon cancer, endometrial cancer, brain cancer, bladder cancer, cancer of the uterus, fibroid (leiomyoma) uterus, cancer of the ovary, lung cancer, sarcoma, bone cancer, pancreatic cancer, renal cancer, and stomach cancer. In some embodiments, the cancer comprises a solid tumor.

According to the invention, the therapeutic agent is selected from the group consisting of a chemotherapeutic agent and a sensitizing drug to an adjuvant therapy. In some embodiments, the therapeutic agent or diagnostic agent is preloaded onto at least one surface of the shear-thinning composition prior to the treating. In some embodiments, the therapeutic agent or diagnostic agent preloaded into the shear-thinning composition prior to the treating. In some embodiments, the therapeutic agent or diagnostic agent is released from the shear-thinning composition upon contacting the cancer with the shear-thinning composition.

In some embodiments, the shear-thinning composition can be administered by injection. In some embodiments, the administering is performed using a catheter or a syringe. In some embodiments, the administration comprises an image guided endovascular procedure or an image guided percutaneous procedure.

In some embodiments, the shear-thinning composition is preloaded onto at least one surface of a medical device prior to the treating. In some embodiments, the shear-thinning composition is preloaded into a medical device prior to the treating. In some embodiments, the shear-thinning composition is preloaded into a catheter prior to the treating. In some embodiments, the shear-thinning composition is preloaded into a syringe prior to the treating.

According to the invention, the shear-thinning composition comprises gelatin or a derivative thereof, and silicate nanoparticles. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of gelatin or a derivative thereof, and silicate nanoparticles together. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of gelatin or a derivative thereof, and silicate nanoparticles together. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of gelatin or a derivative thereof, and silicate nanoparticles together. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of gelatin or a derivative thereof, and silicate nanoparticles together. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of gelatin or a derivative thereof, and silicate nanoparticles together.

In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the silicate nanoparticles. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the silicate nanoparticles. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the silicate nanoparticles. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the silicate nanoparticles. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the silicate nanoparticles. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the silicate nanoparticles.

In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the gelatin or a derivative thereof. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the gelatin or a derivative thereof. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the gelatin or a derivative thereof. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the gelatin or a derivative thereof. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the gelatin or a derivative thereof. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the gelatin or a derivative thereof. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the gelatin or a derivative thereof. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the gelatin or a derivative thereof.

In some embodiments, the ratio of silicate nanoparticles to gelatin or a derivative thereof, is from about <NUM> to about <NUM>.

In some embodiments, the silicate nanoparticles comprise silicate nanoplatelets. In some embodiments, the silicate nanoplatelets comprise a positively charged edge and a negatively charged surface. In some embodiments, the overall charge of the silicate nanoparticles is negative.

In some embodiments, the silicate nanoparticles are from about <NUM> to about <NUM> in diameter. In some embodiments, the silicate nanoparticles are from about <NUM> to about <NUM> in diameter. In some embodiments, the silicate nanoparticles are about <NUM> to <NUM> in diameter. In some embodiments, the silicate nanoparticles are from about <NUM> to about <NUM> in thickness. In some embodiments, the silicate nanoparticles are about <NUM> in thickness.

According to the invention, the gelatin derivative is methacrylated gelatin (GelMA), acrylated gelatin, or thiolated gelatin. In some embodiments, the gelatin derivative is methacrylated gelatin (GelMA).

In some embodiments, the shear-thinning composition comprises:.

According to the invention, the shear-thinning composition further comprises water. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the water. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the water. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the water. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the water. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the water. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the water. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the water. In some embodiments, the shear-thinning composition comprises about <NUM> percent to about <NUM> percent by weight of the water.

In some embodiments, the shear-thinning composition is a gel. In some embodiments, the shear-thinning composition is a hydrogel.

In some embodiments, the gelatin is derived from a mammalian source. In some embodiments, the gelatin is type-A porcine gelatin.

In some embodiments, the yield stress of the shear-thinning composition is from about <NUM> Pa to about <NUM> Pa. In some embodiments, the yield stress of the shear-thinning composition is from about <NUM> Pa to about <NUM> Pa. In some embodiments, the yield stress of the shear-thinning composition is from about <NUM> Pa to about <NUM> Pa. In some embodiments, the yield stress of the shear-thinning composition is from about <NUM> Pa to about <NUM> Pa. In some embodiments, the yield stress of the shear-thinning composition is from about <NUM> Pa to about <NUM> Pa. In some embodiments, the yield stress of the shear-thinning composition is from about <NUM> Pa to about <NUM> Pa. In some embodiments, the shear-thinning composition flows upon application of a pressure greater than the yield stress.

case of conflict, the present specification, including definitions, will control.

Shear-thinning compositions are able to flow when adequate force is applied to them, similar to commercial products like toothpaste or ketchup. Once the force is removed, the composition is able to recover its properties to remain stable in its new environment, resistant to physiological degradation or mechanical forces that could disrupt the composition.

Technologies used to-date to address hemorrhaging blood vessels include, for example, the use of metallic coils. These metallic coils are costly, generally require the availability of appropriate sizes at the time of procedure, require appropriate catheters to deliver the coils and may require additional devices to deploy the coils, which may be electrical, magnetic or mechanical in nature. Additionally, metallic coils require technical expertise in their use, and may lead to the sacrifice of a vascular bed. Accordingly, there are numerous clinical conditions where shear-thinning compositions could be applicable, which include, but are not limited to:.

Additional uses of shear-thinning compositions include, but are not limited to the occlusion of abnormal veins, for example, congested gastric and esophageal veins resulting from portal hypertension or from pelvic congestion syndrome and treatment of varicose veins. Shear-thinning compositions may be delivered via percutaneous approach such as a needle or maybe navigated to region of interest within the vasculature via endovascular approach.

In a previous study, shear-thinning compositions were assessed for rheological behavior, physiological stability, and activity (see e.g., <CIT>). Although the application of the shear-thinning composition on rat liver surface was hemostatic in nature, the study did not require injectable shear-thinning behavior and therefore a concentrated formulation was tested. In contrast, endovascular embolization and other small bore catheter based applications require easy injectability through long catheters (up to <NUM>), rapid gelation to create an occlusive seal of the arterial lumen without fragmentation, a formulation that does not cause CT, MRI or US artifact and a gamma irradiated material that is sterile and easy to use. These conditions were not previously assessed or characterized, supporting further investigations into the suitability of the new shear-thinning composition formulations with enhanced properties as injectable hydrogel embolic agents. Indeed, the shear-thinning compositions form a complete, impenetrable cast of the vessel after injection, which is sufficient to occlude the vessel or the aneurysm without relying on thrombosis.

Accordingly, the present invention, according to the appended set of claims, provides shear-thinning compositions for use in treating vascular pathologies selected from hemorrhages associated with cancer, cancer (e.g., arterial embolization of solid tumors). According to the invention, the compositions provided herein may be administered in combination with an additional therapeutic agent for use in the treatment of a cancer. In some embodiments the compositions provided herein are injectable and can be administered via a catheter. In some embodiments, the compositions provided herein form a physiologically stable artificial matrix and can promote the natural clotting cascade. For example, the compositions provided herein flow with minimal applied pressure during injection, providing a method of application that avoids additional patient trauma. Moreover, in some embodiments, the compositions provided herein, once applied to a treatment site, solidify to prevent biomaterial loss (e.g., blood loss) to unaffected areas.

For the terms "for example" and "such as" and grammatical equivalences thereof, the phrase "and without limitation" is understood to follow unless explicitly stated otherwise. As used herein, the term "about" is meant to account for variations due to experimental error. All measurements reported herein are understood to be modified by the term "about", whether or not the term is explicitly used, unless explicitly stated otherwise. As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

A "therapeutically effective amount" of a composition with respect to the subject method of treatment, refers to an amount of the composition(s) in a preparation which, when administered as part of a desired dosage regimen (to a patient or a subject, e.g., a human) alleviates a symptom, ameliorates a condition, or slows the onset of disease conditions according to clinically acceptable standards for the disorder or condition to be treated or the cosmetic purpose, e.g., at a reasonable benefit/risk ratio applicable to any medical treatment.

As used herein, the term "treating" or "treatment" includes reversing, reducing, or arresting one or more of the symptoms, clinical signs, and underlying pathology of a condition in a manner to improve or stabilize a patient's condition.

As used herein, the term "medical condition" refers to a condition or disorder in a subject who is experiencing or displaying the pathology or symptomatology of a disease, condition, or disorder (i.e., symptoms), for example, pain, dysfunction, or distress. It is understood that one of ordinary skill (e.g., a clinician, physician, veterinarian, and the like), is able to diagnose a medical condition.

As used herein, the term "vascular condition" refers to a condition or disorder in a subject related to the vascular system of the subject (i.e., the circulatory system). Example vascular conditions may include, but are not limited to, an aneurysm, a hemorrhage, a venous congestion disorder, a varix, an abscess, a fistula, a cancer, and an infection.

As used here, the term "locally administering" refers to administration at or within close proximity to a vascular condition in a subject. For example, upon cessation of the administration, a shear-thinning composition provided herein will remain substantially localized at the site of the administration. In some embodiments, a shear-thinning composition provided herein is locally administered at the site of a vascular condition (e.g., administered within abscess, administered within a tumor, administered within an aneurysm, and the like).

The present invention provides shear-thinning compositions. The expression "shear-thinning" or "shear-thinning behavior", refers to a decrease in viscosity (i.e., increasing flow rate) of a composition with increasing application of shear stress. For example, a shear-thinning composition (i.e. a composition exhibiting shear-thinning behavior) can exhibit a decrease in viscosity (i.e. increase in flow) upon application of an increasing rate of shear stress. As shown in Reference Example <NUM>, shear-thinning behavior was not observed in the individual components of exemplary compositions (e.g., gelatin, 9NC0), but was observed in the compositions upon combination of the components (e.g., a composition comprising both gelatin and silicate nanoparticles (e.g., 9NC75)).

As used herein, the term "gelatin", alone or in combination with other terms, refers to a mixture of proteins and peptides derived from the partial hydrolysis or denaturing of collagen. The intermolecular, intramolecular, and hydrogen bonds which stabilize collagen proteins and peptides are broken down to form gelatin, for example, by acid hydrolysis, alkali hydrolysis, or enzymatic hydrolysis. In some embodiments, the gelatin may be derived from a mammalian source. Examples of such gelatins include, but are not limited to porcine gelatin (e.g., type-A porcine gelatin, gelatin derived from porcine skin, gelatin derived from porcine bones, and the like), bovine gelatin (e.g., gelatin derived from bovine skin, type B bovine gelatin, gelatin derived from bovine bones, and the like), and equine gelatin.

In some embodiments, the shear-thinning compositions provided herein comprise a gelatin derivative selected from methacrylated gelatin (GelMA), acrylated gelatin or thiolated gelatin. As used here, the term "gelatin derivative", alone or in combination with other terms, refers to gelatin that has been reacted with various types of reagents to functionalize the gelatin (e.g., methacrylated gelatin (GelMA), acrylated gelatin, thiolated gelatin). For example, derivatives of gelatin may be prepared by reacting gelatin with an acid anhydride or acid chloride, including, but not limited to, phthalic anhydride, maleic anhydride, succinic anhydride, benzoic anhydride, isatoic anhydride, methacrylic anhydride, <NUM>,<NUM>-dibromo phthalic anhydride, benzoyl chloride, p-nitro benzoyl chloride, <NUM>-hydroxy-<NUM>-naphthoyl chloride, o-hydroxy benzoyl chloride, phthalyl chloride, and ethyl chlorocarbonate. In some embodiments, the shear-thinning compositions provided herein comprise methacrylated gelatin (GelMA), acrylated gelatin, or thiolated gelatin.

In some embodiments, the shear-thinning compositions provided herein comprise about <NUM> percent to about <NUM> percent by weight of gelatin or a derivative thereof, for example, from about <NUM> percent to about <NUM> percent by weight, about <NUM> percent to about <NUM> percent by weight, about <NUM> percent to about <NUM> percent by weight, about <NUM> percent to about <NUM> percent by weight, about <NUM> percent to about <NUM> percent by weight, about <NUM> percent to about <NUM> percent by weight, or from about <NUM> percent to about <NUM> percent by weight.

As used herein, the term "silicate nanoparticles", used alone or in combination with other terms, refers to silicate layered clays. Example silicate layered clays include, but are not limited to, laponite, montmorillonite, saponite, hectorite, kaolinite, palygorskite, and sepiolite. Silicate nanoparticles can be prepared, for example, by dialysis and similar purification techniques known in the art to remove any impurities. In some embodiments, overall charge of the silicate nanoparticles is negative. In some embodiments, the silicate nanoparticles are from about <NUM> to about <NUM> in diameter, for example, from about <NUM> to about <NUM> in diameter, from about <NUM> to about <NUM> in diameter, or from about <NUM> to about <NUM> in diameter. In some embodiments, the silicate nanoparticles are from about <NUM> to about <NUM> in thickness. In some embodiments, the silicate nanoparticles are about <NUM> in thickness.

In some embodiments, the silicate nanoparticles comprise silicate nanoplatelets. As used herein, the term "silicate nanoplatelets", used alone or in combination with other terms, refers to silicate layered clays characterized by a discotic charge distribution on the surface. Silicate nanoplatelets can be prepared, for example, by dispersion and sonication in an aqueous solution. In some embodiments, the silicate nanoplatelets comprise a positively charged edge and a negatively charged surface. In some embodiments, the overall charge of the silicate nanoplatelets is negative. In some embodiments, the silicate nanoplatelets are from about <NUM> to about <NUM> in diameter, for example, from about <NUM> to about <NUM> in diameter, from about <NUM> to about <NUM> in diameter, or from about <NUM> to about <NUM> in diameter. In some embodiments, the silicate nanoplatelets are from about <NUM> to about <NUM> in thickness. In some embodiments, the silicate nanoplatelets are about <NUM> in thickness.

In some embodiments, the shear-thinning compositions provided herein comprise about <NUM> percent to about <NUM> percent by weight of the silicate nanoparticles, for example, about <NUM> percent to about <NUM> percent by weight, about <NUM> percent to about <NUM> percent by weight, about <NUM> percent to about <NUM> percent by weight, about <NUM> percent to about <NUM> percent by weight, or about <NUM> percent to about <NUM> percent by weight.

In some embodiments, the ratio of silicate nanoparticles to gelatin or a derivative thereof, is from about <NUM> to about <NUM>, for example, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM> or about <NUM>. In some embodiments, the ratio of gelatin or a derivative thereof to silicate nanoparticles is from about <NUM> to about <NUM>, for example, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM> or about <NUM>.

In some embodiments, the shear-thinning compositions provided herein comprise about <NUM> percent to about <NUM> percent by weight of gelatin or a derivative thereof and silicate nanoparticles together, for example, from about <NUM> percent to about <NUM> percent by weight, from about <NUM> percent to about <NUM> percent by weight, from about <NUM> percent to about <NUM> percent by weight, from about <NUM> percent to about <NUM> percent by weight, from about <NUM> percent to about <NUM> percent by weight, about <NUM> percent to about <NUM> percent by weight, from about <NUM> percent to about <NUM> percent by weight, about <NUM> percent to about <NUM> percent by weight, or about <NUM> percent to about <NUM> percent by weight. In some embodiments, the shear-thinning compositions provided herein comprise about <NUM> percent by weight gelatin and about <NUM> percent by weight silicate nanoparticles, about <NUM> percent by weight gelatin and about <NUM> percent by weight silicate nanoparticles, about <NUM> percent by weight gelatin and about <NUM> percent by weight silicate nanoparticles, about <NUM> percent by weight gelatin and about <NUM> percent by weight silicate nanoparticles, about <NUM> percent by weight gelatin and about <NUM> percent by weight silicate nanoparticles, about <NUM> percent by weight gelatin and about <NUM> percent by weight silicate nanoparticles, about <NUM> percent by weight gelatin and about <NUM> percent by weight silicate nanoparticles, about <NUM> percent by weight gelatin and about <NUM> percent by weight silicate nanoparticles, or about <NUM> percent by weight gelatin and about <NUM> percent by weight silicate nanoparticles.

In some embodiments, a shear-thinning composition provided herein is a physically crosslinked gel comprising gelatin or a derivative thereof, and silicate nanoparticles. In some embodiments, the shear-thinning composition is a physically crosslinked hydrogel comprising gelatin or a derivative thereof, and silicate nanoparticles.

According to the invention, the shear-thinning composition provided herein further comprises water, for example, ultra-pure water (e.g., Milli-Q) or buffered water (e.g., phosphate buffered saline). In some embodiments, the shear-thinning composition comprises from about <NUM> percent to about <NUM> percent by weight of water, for example, from about <NUM> percent to about <NUM> percent by weight, about <NUM> percent to about <NUM> percent by weight, about <NUM> percent to about <NUM> percent by weight, about <NUM> percent to about <NUM> percent by weight, about <NUM> percent to about <NUM> percent by weight, or about <NUM> percent to about <NUM> percent by weight.

In some embodiments, the shear-thinning composition comprises, for example, about <NUM> percent by weight gelatin, about <NUM> percent by weight silicate nanoparticles, and about <NUM> percent by weight water, about <NUM> percent by weight gelatin, about <NUM> percent by weight silicate nanoparticles, and about <NUM> percent by weight water, about <NUM> percent by weight gelatin, about <NUM> percent by weight silicate nanoparticles, and about <NUM> percent by weight water, about <NUM> percent by weight gelatin, about <NUM> percent by weight silicate nanoparticles, and about <NUM> percent by weight water, about <NUM> percent by weight gelatin, about <NUM> percent by weight silicate nanoparticles, and about <NUM> percent by weight water, about <NUM> percent by weight gelatin, about <NUM> percent by weight silicate nanoparticles, and about <NUM> percent by weight water, about <NUM> percent by weight gelatin, about <NUM> percent by weight silicate nanoparticles, and about <NUM> percent by weight water, about <NUM> percent by weight gelatin, about <NUM> percent by weight silicate nanoparticles, and about <NUM> percent by weight water, or about <NUM> percent by weight gelatin, about <NUM> percent by weight silicate nanoparticles, and about <NUM> percent by weight water.

In some embodiments, a shear-thinning composition provided herein is self-healing. As used herein, the expression "self-healing", used alone or in combination with other terms, refers to recovery of the elastic gel strength of a composition upon removal of a stress. In some aspects, a self-healing composition may recover elastic gel strength from about <NUM> seconds to about <NUM> minute after removal of a stress, for example, from <NUM> seconds to <NUM>. , from <NUM> seconds to <NUM> seconds, from <NUM> seconds to <NUM> minute, from <NUM> seconds to <NUM> seconds, from <NUM> seconds to <NUM> seconds, from <NUM> seconds to <NUM> seconds, from <NUM> seconds to <NUM> seconds, from <NUM> seconds to <NUM> seconds, from <NUM> seconds to <NUM> minute, from <NUM> seconds to <NUM> seconds, from <NUM> seconds to <NUM> seconds, from <NUM> seconds to <NUM> seconds, from <NUM> seconds to <NUM> minute, from <NUM> seconds to <NUM> seconds, from <NUM> seconds to <NUM> seconds, from <NUM> seconds to <NUM> seconds, or from about <NUM> seconds to <NUM> minute.

In some embodiments, a shear-thinning composition provided herein flows upon application of a pressure greater than the yield stress, for example, application of a pressure about <NUM>% greater, about <NUM>% greater, about <NUM>% greater, about <NUM>% greater, about <NUM>% greater about <NUM>% greater, about <NUM>% greater, about <NUM>% greater, about <NUM>% greater, or about <NUM>% greater than the yield stress. In some embodiments, the yield stress of the shear-thinning composition is from about <NUM> Pa to about <NUM> Pa, for example, from about <NUM> Pa to about <NUM> Pa, from about <NUM> Pa to about <NUM> Pa, from about <NUM> Pa to about <NUM> Pa, from about <NUM> Pa to about <NUM> Pa, from about <NUM> Pa to about <NUM> Pa, or from about <NUM> Pa to about <NUM> Pa.

In some embodiments, shear-thinning compositions provided herein are useful for accelerating blood clotting (e.g., reducing blood clotting time). Prior to the application of a shear-thinning composition, an active bleeding site may be characterized as bleeding at a rate from about <NUM>/min to about <NUM>/min, for example, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, or <NUM>/min to <NUM>/min.

In some embodiments, the active bleeding is an internal active bleeding. An internal active bleeding site may be characterized by a rate of blood flow from <NUM>/min to <NUM>/min, for example, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, <NUM>/min to <NUM>/min, or <NUM>/min to <NUM>/min.

In some embodiments, a shear-thinning composition provided herein can be prepared according to procedures provided in <CIT>.

In some embodiments, a shear-thinning composition provided herein comprises chitosan. In some embodiments, the shear-thinning composition comprises about <NUM> to about <NUM>% chitosan, for example, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, about <NUM> to about <NUM>%, or about <NUM> to about <NUM>% chitosan. In some embodiments, the composition comprises about <NUM> to about <NUM>% chitosan. In some embodiments, the composition comprises about <NUM>% chitosan.

The present invention provides shear-thinning composition for use in treating cancer.

In some embodiments, the present invention provides a shear-thinning composition for use in treating a hemorrhage, where the hemorrhage is an internal hemorrhage associated with a cancer.

In some embodiments, hemorrhage is associated with a medical disorder, wherein the medical disorder is cancer. In some embodiments, the hemorrhage is associated with a cancer, wherein the cancer is selected from the group consisting of breast cancer, prostate cancer, benign prostatic hyperplasia, esophageal cancer, liver cancer, colon cancer, endometrial cancer, brain cancer, bladder cancer, cancer of the uterus, fibroid (leiomyoma) uterus, cancer of the ovary, lung cancer, sarcoma, bone cancer, pancreatic cancer, renal cancer, and stomach cancer. In some embodiments, the cancer is selected from the group consisting of gastric cancer, stomach cancer, liver cancer, and lung cancer. In some embodiment, the cancer is gastric cancer or stomach cancer.

In some embodiments, the treating comprises locally administering the shear-thinning composition at the site of the hemorrhage. In some embodiments, the treating comprises locally administering the shear-thinning composition at the site of the hemorrhage in an amount effective to reduce or stop bleeding of the hemorrhage. In some embodiments, the administration is endovascular or percutaneous administration.

In some embodiments, the shear-thinning composition is administered to reduce or stop bleeding in a subject wherein the subject has been treated with one or more anti-coagulants, one or more anti-platelet agents, or a combination thereof (e.g., prior to administration of the shear-thinning composition). In some embodiments, the subject has been treated with one or more anti-coagulants, one or more anti-platelet agents, or a combination thereof, less than about <NUM> hours prior to administering the shear-thinning composition, for example, less than about <NUM> hours, less than about <NUM> hours, less than about <NUM> hours, less than about <NUM> hours, less than about <NUM> hours, less than about <NUM> hours, less than about <NUM> hours, less than about <NUM> hours, or less than about <NUM> hour prior to administering the shear-thinning composition. In a subject having been treated with one or more anti-coagulants, one or more anti-platelet agents, or a combination thereof, the standard of therapy, which can include using coil embolization, may be ineffective, as the standard of therapy requires that the subject have the ability to form a blood clot to occlude a vessel to prevent bleeding. The shear-thinning compositions provided herein can be used to occlude anticoagulated blood vessels, as the composition does not require the subject to form a blood clot to occlude the blood vessel.

In some embodiments, the shear-thinning composition is administered to reduce or stop bleeding in a subject, wherein the subject has been treated with:.

In some embodiments, the bleeding comprises active bleeding. In some embodiments, the bleeding comprises active bleeding after the subject has been treated with the standard embolization therapy (e.g., coil embolization).

In some embodiments, the shear-thinning composition is administered to stop bleeding in a subject wherein the subject has been treated with one or more anti-coagulants. Example anti-coagulants include, but are not limited to, warfarin, apixaban, dabigatran, rivaroxaban, edoxaban, dalteparin, desirudin, enoxaparin, fondaparinux, and heparin. In some embodiments, the anti-coagulant is selected from the group consisting of warfarin, apixaban, dabigatran, rivaroxaban, and edoxaban.

In some embodiments, the shear-thinning composition is administered to reduce or stop bleeding in a subject wherein the subject has been treated with one or more anti-platelet agents. Example anti-platelet agents include, but are not limited to, anagrelide, aspirin, cilostazol, clopidogrel, dipyridamole, prasugrel, ticagrelor, ticlopidine, and vorapaxar. In some embodiments, the anti-platelet agent is selected from the group consisting of anagrelide, aspirin, cilostazol, clopidogrel, dipyridamole, prasugrel, ticagrelor, ticlopidine, and vorapaxar.

One or more additional agents are used in combination with the compositions provided herein for the use in treating a vascular disorder or condition in a subject in need thereof, where the vascular disorder or condition is a cancer. According to the invention, the additional agent is a therapeutic agent or a diagnostic agent. In some embodiments, the additional agent is a therapeutic agent. In some embodiments, the additional agent is a diagnostic agent. According to the invention, defined by the appended set of claims, the present invention refers to a shear-thinning composition for use in treating a vascular disorder or condition in a subject in need thereof, where the vascular disorder or condition is a cancer, comprising administering to the subject a shear-thinning composition provided herein and an additional therapeutic agent. According to the invention defined by the appended set of claims, the present invention refers to a shear-thinning composition for use in treating a vascular disorder or condition in a subject in need thereof, where the vascular disorder or condition is a cancer, comprising administering to the subject a shear-thinning composition provided herein and one or more additional diagnostic agents. According to the invention defined by the appended set of claims, the present invention refers to a shear-thinning composition for use in treating a vascular disorder or condition in a subject in need thereof, where the vascular disorder or condition is a cancer, comprising administering to the subject a shear-thinning composition provided herein and an additional diagnostic agent.

According to the invention, at least one of the additional agents is preloaded onto at least one surface of the shear-thinning composition prior to the treating, or at least one of the additional agents is preloaded into the shear-thinning composition prior to the treating. In some embodiments, at least one of the additional agents is preloaded onto at least one surface of the shear-thinning composition and at least one of the additional agents is preloaded into the shear-thinning composition prior to the treating. In some embodiments, at least one of the additional agents is a therapeutic agent.

In some embodiments, each of the one or more additional therapeutic agents is a chemotherapeutic agent.

Example chemotherapeutics include, but are not limited to, proteosome inhibitors (e.g., bortezomib), thalidomide, revlimid, and DNA-damaging agents such as melphalan, doxorubicin, cyclophosphamide, vincristine, etoposide, carmustine, and the like.

One or more of the following agents may be used in combination with the compositions provided herein and are presented as a non-limiting list: a cytostatic agent, cisplatin, doxorubicin, taxol, etoposide, irinotecan, topotecan, paclitaxel, docetaxel, epothilones, tamoxifen, <NUM>-fluorouracil, methotrexate, temozolomide, cyclophosphamide, SCH <NUM>, R115777, L778,<NUM>, BMS <NUM>, gefitinib, erlotinib hydrochloride, antibodies to EGFR, imatinib mesylate, intron, ara-C, gemcitabine, uracil mustard, chlormethine, ifosfamide, melphalan, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, floxuridine, cytarabine, <NUM>-mercaptopurine, <NUM>-thioguanine, fludarabine phosphate, oxaliplatin, folinic acid, pentostatin, vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, teniposide, 17α-ethinylestradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, dromostanolone propionate, testolactone, megestrol acetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianisene, hydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesteroneacetate, leuprolide, flutamide, toremifene, goserelin, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane, mitoxantrone, levamisole, vinorelbine, anastrazole, letrozole, capecitabine, reloxafine, hexamethylmelamine, bevacizumab, Bexxar®, Velcade®, Zevalin®, Trisenox®, Xeloda®, vinorelbine, porfimer, Erbitux®, liposomal, thiotepa, altretamine, melphalan, trastuzumab, fulvestrant, exemestane, ifosfamide, rituximab, C225, alemtuzumab, clofarabine, cladribine, aphidicolin, sunitinib, dasatinib, tezacitabine, Sml1, triapine, didox, Trimidox®, amidox, <NUM>-AP, MDL-<NUM>,<NUM>, bendamustine, ofatumumab, and GS-<NUM> (also known as CAL-<NUM>).

In some embodiments, each of the one or more diagnostic agents is independently selected from the group consisting of a magnetic agent, an electrical agent, or any combination thereof.

Example magnetic agents include, but are not limited to, magnetic resonance imaging agents, iron oxide nanoparticles, zinc oxide nanoparticles, magnesium oxide particles, metallic particles for contrast agents, and polymeric nanoparticles containing metallic nanoparticles.

Example electrical agents include, but are not limited to, metallic nanoparticles, graphene / graphene oxide, and carbon nanotubes.

The present invention, as defined by the appended set of claims, provides a shear-thinning composition for use in treating cancer in a subject in need thereof. In some embodiments, the cancer is selected from the group consisting of breast cancer, prostate cancer, benign prostatic hyperplasia, esophageal cancer, liver cancer, colon cancer, endometrial cancer, brain cancer, bladder cancer, cancer of the uterus, fibroid (leiomyoma) uterus, cancer of the ovary, lung cancer, sarcoma, bone cancer, pancreatic cancer, renal cancer, and stomach cancer. In some embodiments, the cancer comprises a solid tumor. In some embodiments, the additional agent is a diagnostic agent. In some embodiments, the additional agent is a therapeutic agent. In some embodiments, the additional therapeutic agent is a sensitizing drug to adjuvant therapy (e.g., a sensitizing drug to a chemotherapy or radiation therapy). Example sensitizing drugs include, but are not limited to, misonidazole, bromodeoxyuridine, cisplatin, <NUM>-fluorouracil [<NUM>-FU], taxanes, tirapazamine, mitomycin C, and paclitaxel.

In some embodiments, the additional therapeutic agent is a chemotherapeutic agent. In some embodiments, the additional therapeutic agent is released from the shear-thinning composition upon contacting the cancer with the shear-thinning composition.

The compositions provided herein can be administered by a variety of routes, depending upon the area to be treated. In some embodiments, the shear-thinning compositions are administered by parenteral administration. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal intramuscular, injection, or infusion; or intracranial, (e.g., intrathecal or intraventricular administration). Parenteral administration can be in the form of a single bolus dose, or may be, for example, by a continuous perfusion pump. In some embodiments, the administering comprises injecting the shear-thinning composition. In some embodiments, the administering comprises an image guided endovascular procedure or an image guided percutaneous procedure where computed tomography, fluoroscopy or ultrasound imaging is used to deliver the composition. In some embodiments, the administering comprises injecting the shear-thinning composition into the vascular system of a subject. In some embodiments, the administering comprises injecting the shear-thinning composition into a cancer of the subject. In some embodiments, the cancer is a solid tumor.

In some embodiments, the shear-thinning composition is administered to displace a solid organ. In some embodiments, the shear-thinning composition is administered to displace a solid organ during a surgical procedure (e.g., a biopsy procedure, an ablation procedure, and the like). In some embodiments, the surgical procedure further comprises imaging the subject (e.g., a tissue, cell, or bone of the subject) with an appropriate imaging technique. Example imaging techniques include, but are not limited to, computed tomography, ultrasound, and flurosocopy imaging techniques. In some embodiments, the surgical procedure is a biopsy procedure. For example, the shear-thinning composition can be used to displace a solid organ (e.g., an adrenal gland) to provide safe needle access for a tissue biopsy (e.g., a lymph node biopsy). In some embodiments, the surgical procedure is an ablation procedure. In some embodiments, the ablation procedure is an organ ablation procedure or a tumor ablation procedure. Without being bound by any theory, a shear-thinning composition as provided herein can function as a "heat-sink" to protect adjacent tissues when solid tissue is thermally ablated. For example, the biomaterial can be injected in between a kidney and the large bowel to allow safe ablation of kidney tumor-protecting the bowel from potential thermal injury.

In some embodiments, the administering is performed using a catheter or a syringe. In some embodiments, the administering is performed using a catheter. In some embodiments, the shear-thinning composition is preloaded onto at least one surface of a medical device prior to the treating. In some embodiments, the shear-thinning composition is preloaded into a medical device prior to the treating. In some embodiments, the shear-thinning composition is preloaded into a catheter prior to the treating. In some embodiments, the shear-thinning composition is preloaded into a syringe prior to the treating.

The amount of the composition administered to a subject will vary depending upon the composition being administered, the purpose of the administration, the state of the subject, the manner of administration, and the like. In therapeutic applications, compositions can be administered to a subject to cure or at least partially arrest the symptoms and its complications. Effective doses will depend on the necessary treatment as well as by the judgment of the attending clinician depending upon factors such as the type and severity of the complication (e.g., aneurysm, abscess, GI bleed, and the like), the age, weight and general condition of the subject, and the like.

The present application further provides a kit comprising a shear-thinning composition provided herein and one or more catheters. The kit is useful in the treatment of a vascular disorder or condition associated with a cancer.

In some embodiments, the kit comprises one or more components of a shear-thinning composition provided herein and one or more catheters. For example, the components can be separately packaged or contained.

In some embodiments, the kit comprises a pharmaceutically acceptable amount of the shear-thinning composition preloaded onto one or more of the catheters. In some embodiments, the kit comprises a pharmaceutically acceptable amount of the shear-thinning composition preloaded into one or more of the catheters.

In some embodiments, the kit comprises a shear-thinning composition provided herein and one or more sterile syringes. In some embodiments, the shear-thinning composition is preloaded into one or more of the sterile syringes.

Instructions, either as inserts or as labels, indicating quantities of the composition to be administered, guidelines for administration, and/or guidelines for mixing components of the composition, can also be included in a kit provided herein. In some embodiments, the instructions further comprise instructions for performing one or more of the methods provided herein. In some embodiments, the instructions further comprise instructions for quantities of the composition to be administered, guidelines for administration, and/or guidelines for mixing components of the composition.

In some embodiments, the kit further comprises a chemotherapeutic agent.

The kits provided herein can further include, if desired, one or more conventional pharmaceutical kit components, such as, for example, containers with one or more pharmaceutically acceptable carriers.

The invention will be described in greater detail by way of specific examples.

Physical mixtures of gelatin and silicate nanoplatelets were used to formulate the compositions disclosed herein. Synthetic silicate nanoplatelets (Laponite XLG) were purchased from Southern Clay Products, Inc. (Louisville, KY). Type-A porcine skin gelatin was obtained from Sigma Aldrich (Milwaukee, WI).

As a general procedure, silicate nanoplatelets were exfoliated in ultrapure (Milli-Q) water using a vortexer to enhance the surface area available for interaction with gelatin. Next, a gelatin stock, heated to liquefy the solution, was vigorously mixed with the exfoliated silicate at room temperature. Vigorous agitation was necessary to prevent clumping of the nanoplatelets during gelation; however, the nanoplatelets were stably dispersed after the gel had set.

The compositions of the examples were prepared with total solid concentrations (e.g., gelatin + silicate nanoplatelets) of <NUM>, <NUM>, and <NUM> weight percent. The ratio of solid components used in the compositions provided herein (e.g. gelatin:nanoplatelet and chitosan: nanoplatelet) is from about <NUM>:<NUM> to about <NUM>:<NUM>. Shear-thinning compositions provided herein are abbreviated by XNCY where X represents the percent of the shear-thinning composition by weight made up of solid materials (e.g., gelatin, silicate nanoplatelets, and chitosan) and Y represents the percent of the solid material that is silicate nanoplatelets. In the absence of silicate nanoplatelets, the gelatin solution was a viscous liquid at <NUM>. Upon mixing silicate nanoplatelets with gelatin, the composition gelled in about <NUM> minute.

Oscillatory shear rheology was performed over a temperature range from <NUM> to <NUM>, mimicking common environmental and physiological temperatures that the compositions may be exposed during sample preparation and in vivo applications.

Small Angle X-Ray Scattering (SAXS) was performed at the National Synchrotron Light Source (NSLS) at Brookhaven National Laboratory at beamline X27C. Samples were placed in a <NUM> thick washer and sealed between Kapton tape. Samples were equilibrated at <NUM> and <NUM> for <NUM> minutes prior to data collection. Scattering patterns were collected for <NUM> seconds per sample. Radial integration of the two dimensional scattering pattern was performed to yield a one dimensional scattering curve, which was corrected for empty cell and dark field scattering. Thin disk form factor model fitting was performed in MatLab using a nonlinear fit algorithm to fit the radius and a Gaussian distribution for polydispersity of the composition.

An Anton Paar MCR <NUM> rheometer was used for mechanical testing. A <NUM> diameter parallel plate geometry with a gap height of <NUM> was used for temperature sweeps and mineral oil was placed around the circumference of the plate to prevent evaporation of water from the composition for all tests. Compositions were equilibrated for <NUM> minutes prior to testing, followed by a <NUM> minute steady shear at <NUM>-<NUM>. <NUM> of equilibrium time was sufficient for the viscosity to return to a higher plateau value, after which point testing was initiated. Frequency and shear rate sweeps were performed at <NUM> and <NUM>, with frequencies from <NUM> - <NUM> at <NUM>% strain and shear rates from <NUM> to <NUM>-<NUM> with <NUM> points/decade. Frequency sweeps were performed with a cone geometry (<NUM> diameter, <NUM>° angle, <NUM> truncation gap). Stress-controlled temperature sweeps were performed from <NUM>-<NUM> at <NUM> Pa stress and <NUM>. All other tests were performed at <NUM>. Oscillatory stress sweeps were performed from <NUM>-<NUM> Pa at <NUM>. Strain Sweeps were performed from <NUM>-<NUM>% at <NUM>. Recovery testing was conducted at <NUM> by applying <NUM>% strain, a value outside of the linear viscoelastic range, followed by <NUM>% strain for <NUM> minutes to monitor gel recovery. Interfacial strength was also measured by applying a linearly increasing strain to a system of a composition and coagulated blood. Shear stress was measured until <NUM>,<NUM>% strain. The maximum stress attained was used as a measure of the strength of the clot system.

Zeta potentials of gelatin and silicate nanoplatelets were determined in ultrapure water (Milli-Q) and phosphate buffered saline (PBS), pH <NUM> (Invitrogen) using a <NUM> laser in a Malvern ZEN3600 (Malvern Instruments, UK). Silicate nanoplatelets were dissolved with vigorous agitation (vortexing) while gelatin was dissolved with stirring at <NUM>.

Viscosity vs shear rate curves were plotted on a log-log plot and fitted using the nonlinear least-squares regression algorithm in MATLAB. The linear region of the curves was fit with the fluid power law, Equation <NUM>: <MAT> where η is the viscosity, ηc is the viscosity at <NUM>-<NUM>, γ̇ is the shear rate and n is the power. Values of n < <NUM> suggest a shear-thinning material.

Blood was mixed with <NUM>% <NUM>-M CaCl<NUM> in MilliQ deionized (DI) water and pipetted to mix. <NUM>µL of whole blood was added to successive wells of a <NUM> well plate that were either uncoated, coated with a shear-thinning composition, or containing a <NUM> section of an embolic coil. After selected times, a well was washed with <NUM> % saline solution and the liquid aspirated to leave only coagulated blood. Well plate images were taken using a Zeiss Axio Zoom V16 stereomicroscope.

Human plasma was separated from citrated whole blood by gravity settling of the red blood cells. Plasma was used to incubate weighed shear-thinning composition (<NUM>µL) in <NUM> Eppendorf tubes (VWR) at <NUM>. Over <NUM>, the plasma was removed, the remaining shear-thinning composition weighed, and replaced with fresh plasma.

The injectability of the material was analyzed using a mechanical tester (Instron Model <NUM>). The shear-thinning composition was added to a <NUM>-mL plastic syringes (ID=<NUM>, BD) and injected through medical catheters (<NUM>-French Beacon®, pediatric pigtail flush, and <NUM>-French Beacon®, multipurpose A, Cook Incorporated) or needles (<NUM> and <NUM>, Becton-Dickinson), using standard luer-lock fittings. The syringe plunger was depressed using an upper compressive platen and the housing of the syringe or catheter was fitted into a lower tensile grip to prevent movement during the experiment. The injection rate was controlled by changing the cross speed of the compression platen to achieve the desired flow rates. The force on the plunger was measured with a <NUM> N load cell and recorded using Bluehill <NUM> software.

Citrated human whole blood (Research Blood Components, Brighton, MA) was used for all blood related testing.

Hemolysis testing was performed according to protocols described in <NPL>. Briefly, citrated whole blood was diluted 50X into <NUM>% saline solutions. The shear-thinning composition was flattened in <NUM>-mL Eppendorf tubes in a swinging bucket centrifuge. Equal volumes of diluted blood and either the shear-thinning composition, saline (negative control) or DI water (positive control) were incubated at <NUM> for <NUM> under agitation in a shaker incubator (<NUM> rpm, Labline Instruments). Samples were centrifuged (<NUM> rpm, Labnet) and the supernatant was transferred into wells of a <NUM>-well plate. Percent hemolysis was calculated according to the Equation <NUM>: <MAT> where Asample is the absorbance at <NUM> of the shear-thinning composition containing supernatant, Aneg is the absorbance of the saline diluted blood and Apos is the absorbance of the DI water diluted blood.

Shear-thinning compositions were mixed with contrast dye (Visipaque™) at a ratio of <NUM>:<NUM> Visipaque™:water and sterilization was performed as described herein. Following induction of anesthesia, mice were placed in a supine position over a warming platform to maintain a core temperature of <NUM>. Laser Doppler perfusion imaging of the hind limbs was performed to obtain per-injection baseline scans. A <NUM>-<NUM> incision was made longitudinally on the anterior thigh of one hind limb. The vessels in the thigh were exposed under a surgical microscope using a combination of sharp and blunt dissection to expose the femoral artery and vein. The femoral artery segment distal to the inguinal ligament was mobilized and freed of its surrounding tissue. Two <NUM>-<NUM> silk sutures were passed distally and proximally underneath the artery to allow for gentle manipulation of the artery by lifting and aligning the vessel for injection. The artery was injected distally with a shear-thinning composition (6NC75) using <NUM> gauge sterile syringe needles. After injection, the incision was closed with a <NUM> polypropylene sutures and the mouse remained on the warming table for <NUM> under general anesthesia followed by post shear-thinning composition injection laser Doppler perfusion imaging. The perfusion scanning of the injected hind limb was compare to the non-injected contralateral hind limb to confirm ischemia. After laser Doppler imaging was completed, animals were euthanized and the carcasses were placed inside a non-opaque container for contrast-enhanced micro computed tomography scanning (µCT) imaging.

µCT imaging was completed to produce high-resolution three-dimensional images constructed of two-dimensional trans-axial projections of the euthanized mouse hind limb to detect the radiodense contrast-containing shear-thinning composition casting of the vasculature <NUM> after injection using Nikon XT H <NUM> (µCT) and Bruker SkyScan <NUM> imaging scanners. The parameters used in the a Nikon platform were <NUM> kV, <NUM> W, <NUM> projections, <NUM> frames per projection, and <NUM> exposure and for the SkyScan system were 70kV, <NUM>µA, <NUM> projections, and <NUM> frames per projection. The Nikon specimens were reconstructed with Nikon's CT Agent and CT Pro 3D software, and viewed in VGStudio MAX Version <NUM>. The SkyScan specimens were reconstructed with SkyScan's NRecon software, and viewed in their CTAn Version <NUM> software.

Female Yorkshire swine (Sus scrofa domestica; n = <NUM>; weight, <NUM> to <NUM>; Tufts Veterinary School, North Grafton, MA) were allowed to acclimate for at least <NUM> d in a cage; the night before procedure, food was withheld but water was provided ad libitum. For the procedure, all swine received tiletamine-zolazepam (<NUM>/kg intramuscularly; Telazol, Zoetis, Florham Park, NJ) and atropine (<NUM>/kg IM) as pre-induction medication and then were placed supine on the operating table. After anesthesia induction (<NUM>/kg intravenously; propofol), all swine were intubated (internal diameter of endotracheal tube, <NUM> to <NUM>) and ventilated mechanically (Evita <NUM>, Dräger, Lübeck, Germany).

During the procedure, electrocardiogram, transcutaneous oxyhemoglobin saturation (SpO2), end-tidal CO<NUM> concentration, inspired oxygen fraction and core temperature were monitored. A bolus of propofol (<NUM>/kg IV) was used to maintain general anesthesia. Following anesthesia, access to the carotid artery or to the common femoral vein was obtained using ultrasound and C-Arm fluoroscopy (Siemens). The access needle and wire were exchanged for a <NUM> French catheter (Cook Medical). Using a glidewire, the target vessel was selected and embolized using a sterilized shear-thinning composition (6NC75) as described herein. Following the procedure, the catheter was removed and hemostasis at the puncture site was obtained using manual compression for up to <NUM> minutes. The animals were subsequently recovered and placed back into their cages.

Two-tailed Student's t test were performed for experiments with <NUM> groups. One-way analysis of variance (ANOVA) with Tukey post hoc test (p<<NUM> was defined as significant) were performed on experiments with more than <NUM> test groups. Standard deviation was the measure of uncertainty in all data. All statistical analysis and graphing were performed with the GraphPad Prism <NUM> software.

Stock solutions of <NUM>% (w/w) gelatin and <NUM>, <NUM>, or <NUM> % (w/w) silicate nanoplatelets were prepared in water. Milli-Q water was heated to <NUM> to dissolve gelatin and <NUM> water was used for nanoplatelet stock solutions to slow gelation and allow for full dissolution of nanoplatelet particles prior to gelling. The nanoplatelet gels were kept at room temperature to fully hydrate until a clear gel formed. The compositions were again heated and vortexed at <NUM> rpm for <NUM> minutes to achieve the appropriate solid concentration and nanoplatelet loading. Once formed, the compositions were stored at <NUM>.

Composition 9NC0 had a gel-sol transition temperature of <NUM>, too low for application as a hemostat. However, the addition of silicate nanoplatelets to gelatin improved the thermal stability, increasing the sol-gel transition to above <NUM> for compositions having total solids concentrations of <NUM> weight percent or greater. In contrast, composition having a total solid concentration of <NUM> weight percent were not solid within the experimentally observed temperature range. Physiological stability was observed for all compositions having a total solids concentration of <NUM> weight percent. Physiological stability was also observed for compositions 6NC50, 6NC75, and 6NC100. Weight percentages of example compositions is shown below in Table <NUM>.

Composition samples were placed in <NUM> Eppendorf tubes and weighed. Each sample was centrifuged in a swinging bucket rotor centrifuge to obtain a flat interface. Each sample was then soaked in phosphate buffered saline (PBS, pH <NUM>; Invitrogen), stored at <NUM>. At set times, the PBS was removed, and the composition was reweighed. The change in weight was recorded up to <NUM> hours after initial soaking. PBS was replaced after each weighing.

Scattering measurements of gelatin-silicate nanoparticle compositions suggested the presence of disk shaped particles, indicating that clay particles remain exfoliated in the composition. Small Angle X-ray Scattering (SAXS) intensity curves of the compositions exhibited a power law decay with an exponent of -<NUM> at high q, the scattering vector, characteristic of disk-shaped scatterers. The scattering intensity from 9NC75 can be fit with a thin disk model with a radius of <NUM> ± <NUM>, in agreement with the reported size of the silicate nanoplatelets. The SAXS intensity curves suggest that scattering was produced from individual nanoplatelets dispersed within the gelatin and not aggregates of nanoplatelets.

Storage modulus (G') and loss modulus (G") of gelatin and gelatin-silicate compositions with total solid concentrations of <NUM>, <NUM>, and <NUM> weight percent were monitored from <NUM> to <NUM>. Gelatin (NC0) was observed to flow at all solid concentrations above <NUM>. The addition of silicates improved the thermal stability of the composition network. All temperature sweeps were performed at <NUM> Pa stress and <NUM> and are shown in <FIG>.

Physiological stability was determined by measuring the weight of compositions with total solid concentration of <NUM>, <NUM>, and <NUM> weight percent stored in phosphate buffered solution (PBS) at <NUM>. Gelatin (NC0) immediately dissolved in PBS, while compositions having a total solid concentration of <NUM> and <NUM> weight percent maintained their structural integrity throughout a <NUM> hour test.

Silicate addition to gelatin modulated the rheological response of the compositions, resulting in a shear-thinning behavior observed at <NUM>. Preliminary investigations using a <NUM>-gauge needle indicated that all silicate-containing compositions could be injected and form self-supporting structures, suggesting the presence of a yield stress and recovery potential. Linear oscillatory shear rheology showed that the crossover frequency was below <NUM> for 9NC75 and 9NC100, maintaining solid-like (G'>G") properties over the tested frequency range. Strain sweeps were performed at <NUM> and indicated a crossover point when tan(δ) = <NUM> that decreased with increasing silicate loading, as shown in <FIG>. Frequency sweeps were performed at <NUM>. Compositions 9NC25, 9NC50, 9NC75, and 9NC100 exhibited increased moduli for compositions with higher nanoplatelet loading, as shown in <FIG>. Oscillatory strain into the nonlinear regime illustrated yielding behavior, an important parameter for designing hydrogels for minimally invasive therapies. Yield stress of gels as a function of nanoplatelet loading and total solid weight percent (<NUM>%, <NUM>% and <NUM>% solid) is shown in <FIG>. In oscillatory shear rate sweeps, the yield stress was defined as a <NUM>% departure of the stress from the initial linearity on a stress-strain plot. Tests were performed at <NUM>, where gelatin readily flows and lacks a yield stress. An increase in the silicate concentrations from <NUM>% (9NC0) to <NUM>% (9NC100) increased the yield stress from <NUM> Pa to <NUM> Pa, as shown in <FIG>. A yield stress was observed in 9NC100 but not in 9NC0, suggesting that the yield stress behavior was derived from the presence of the dispersed nanoplatelets in the hydrogel composition, consistent with the known shear-thinning capability of nanoplatelets. Because increasing the concentration of gelatin reduces the yield stress, the presence of higher concentrations of gelatin eased delivery of the composition by injection.

Recovery of the elastic gel strength in less than <NUM> seconds was observed in compositions for nanoplatelet loadings greater than <NUM>% (9NC50, 9NC75, and 9NC100). Recovery was tested by straining above the crossover point, observed from strain sweeps, to break the network, resulting in G''> G', followed by removal of the strain. Such rapid self-healing after the removal of stress can prevent material flow after application to a wound site. This provides a significant advantage over self-assembling peptides, which risk being washed away because they have relatively long self-healing times after the deformation of physically crosslinked networks. Results of the composition moduli indicate rapid recovery of the storage modulus after repeated application of high oscillatory strain amplitudes, suggesting rapid recovery of the physically crosslinked networks. After four cycles of high and low oscillatory strain, the modulus observed for composition 9NC50 during large amplitude strain oscillations was <NUM>% lower than the initial modulus. At higher silicate loading (9NC75 and 9NC100), the moduli were <NUM>% and <NUM>% lower compared to the initial values. Extended monitoring indicated that after <NUM> seconds the moduli reached asymptotic values, indicating completion of the healing process. Aging of composition 9NC100 was observed when samples were monitored over a period of hours at <NUM>% strain, <NUM>. The effects of aging were able to be countered by application of high shear rates (<NUM>-<NUM>) prior to testing, which returned moduli to their initial, non-aged values.

Solutions of silicate nanoplatelets possessed a zeta potential of -<NUM> mV, whereas gelatin solutions had a zeta potential of <NUM> mV. Because the two components had opposite charges, electrostatic interactions between silicate and gelatin were expected. This was also supported by earlier findings which showed that strong interactions between montmorillonite (another type of silicate clay) and gelatin can function to increase the sol-gel transition temperature of the composite. Zeta potential measurements suggest that electrostatic interactions between nanoplatelets and gelatin contributed to the observed increase in the thermal stability.

Using an in vitro aneurysm model (<FIG>), shear-thinning compositions were injected using a <NUM> French catheter (<FIG>). As shown in <FIG>, approximately <NUM> cc of the composition was injected during real-time fluoroscopy guidance. Subsequent contrast injection demonstrates absence of the wide neck aneurysm sac. As shown in <FIG>, to simulate gastrointestinal bleeding and its treatment, a long segment main branch received an injection of the shear-thinning composition (segment in between dotted lines). Subsequent contrast injection demonstrates exclusion of the injected segment. A unique property of this composition is that hemostasis is not coagulation dependent. The shear-thinning composition produces a cast of the injected vessel or aneurysm preventing flow into this segment. In modern treatments, coil embolization are generally performed; these coils rely on adequate coagulation for hemostasis. There is significant clinical failure of coil embolization cases often seen in coagulopathy patients; such failure would be a non-issue when using shear-thinning compositions.

In a further example, anticoagulated whole blood (citrated, Research Blood Components) was flowed through a PDMS tube (ID: <NUM>, wall modulus: approximately <NUM> kPa) with a syringe pump, with the pressure being monitored upstream (<FIG>). A <NUM>-French (ID: <NUM>) catheter was fed through the tubing to inject coils or shear-thinning composition directly into the PDMS mimic vessel. The catheter was removed, flow was started, and the pressure was measured until movement of the embolic agents in the tube. <FIG> shows pressure curves over time, which show that the control and coil samples are similar, reproducing the clinical scenario. The shear-thinning composition, however, requires significantly higher pressures to displace even when anticoagulated blood is used (<FIG>).

Shear-thinning compositions were characterized utilizing rheology to measure the mechanical properties of the shear-thinning composition. It can be used to determine the viscosity of materials under a variety of conditions such as changing temperature and applied torques, which can model the process of injection through a syringe, catheter, or needle. As shown in <FIG>, the shear-thinning composition was subjected to multiple injection rates (termed shear rates in rheological terms) and temperatures while measuring viscosity. Higher shear rates reflect a faster rate of injection and lower viscosity values reflect a material that is easier to flow (e.g. water has a lower viscosity than honey). In shear-thinning materials, once the viscosity begins to decrease, the material is "shear-thinning" and begins to flow. As the percent of silicate nanoplatelet is increased, the maximum viscosity attained prior to flowing (when the viscosity begins to decrease) is increased as shown in <FIG>. After waiting <NUM> minutes, the material is still able to be injected, summarized for multiple compositions in <FIG> (right trace) and <FIG> (right trace), with open symbols for the initial peak viscosity and closed symbols representing the peak viscosity after <NUM> minutes. The ability to pause and re-inject an embolic material is unique to shear-thinning compositions and something that is not possible today with metal coils or commercial embolics (see e.g., Onyx embolic agents). As shown in <FIG>, the temperature dependence is shown to be statistically insignificant when higher nanoplatelet concentrations are used, for initial peak viscosities <FIG> (left trace) and peak viscosities after <NUM> minutes <FIG> (right trace). Indeed, the temperature stability of the shear-thinning compositions is highlighted when the stiffness is monitored over a range of temperatures, from <NUM> to <NUM> (<FIG>). At higher nanoplatelet compositions (e.g., <FIG>), there is independence in mechanical properties on the applied temperature.

Longer term monitoring of shear-thinning composition injectability shows a statistically insignificant change in the peak viscosity over the span of <NUM> hour after the injectability was initially measured, as shown in <FIG>. This suggests that the shear-thinning composition is not hardening after equilibrating over <NUM> hour, highlighting the ability to keep the embolic in a catheter for the extent of a <NUM> hour procedure. This is a unique property of shear-thinning compositions; during embolization procedures, for example, the operator can stop and reassess progress and then continue safely with the procedure. In contrast, there is minimal time to pause when performing coil embolization to avoid occluding a microcatheter with thrombus and there is no time to pause and assess progress using Onyx embolic agents to avoid cementing the catheter to the embolic agent.

The stiffness of the shear-thinning composition is modified from an elastic solid to a viscous liquid as the deformation of the shear-thinning composition is changed. However, it is able to recover its stiffness after multiple cycles of high and low deformation without weakening, as shown in <FIG>. This highlights the rapid recovery of the shear-thinning composition stiffness after high deformation that is experienced during injection.

Using a syringe pump and system of valves and pressure, the pressure needed to inject the shear-thinning composition through catheters and needles can be measured. The injection pressure is dependent on how quickly the shear-thinning composition is injected, as shown in <FIG>. Compared to standard liquids injected through catheters, (e.g., saline solutions and contrast dye), the shear-thinning compositions require more force to inject, as shown in <FIG> but all forces needed are below the maximum functional pressure of many catheters (approximately <NUM> MPa).

The time it commonly takes for blood to clot outside of the body is <NUM>-<NUM> minutes. One example of a shear-thinning composition contains silicate nanoplatelets, which accelerate the speed of blood clotting. The utilization of clays (e.g., silicate nanoplatelets) to impact blood clotting has already been shown to work in topical hemostats such as QuikClot. Compared to controls, increased shear-thinning composition concentrations accelerate the clotting time to <NUM>-<NUM> minutes, as shown in <FIG>. These results are similar to the currently used fibered metallic coils, as shown in <FIG>. These properties, for example, will allow for a clot to fill an aneurysm sac and produce a cast of the sac. The strength of the clot can improve the stability of the embolic.

The interactions of blood with the shear-thinning composition are characterized by the level of red blood cell lysis, termed hemolysis. Stiffer shear-thinning compositions have lower hemolysis percent, as shown in <FIG>. Additionally, incubation of shear-thinning compositions in blood plasma showed no significant change in the shear-thinning composition mass over the span of <NUM> hours, as shown in <FIG>.

Viscosities of shear-thinning compositions from <NUM>-<NUM> wt. % were tested for their injectability by monitoring viscosity as injection rate (shear rate) was increased by rheology, as shown in <FIG>. Using the correlation between flow rate and shear rate (injection rate), flow rates were converted to shear rates, as shown below in Table <NUM> and the resulting viscosity observed at this shear rate was determined from the viscosity curves as shown below in Table <NUM>. During flow, the observed viscosities of these shear-thinning compositions are comparable to castor oil or corn syrup, highly viscous but able to flow when subjected to sufficient forces.

Ongoing tests are being performed within microfluidic flow systems. From these tests, the shear-thinning composition performance under high pressure and flow can be analyzed for stability and disruption or degradation of the shear-thinning composition. Models that can replicate vessel occlusion, as shown in <FIG> and aneurysm embolization, as shown in <FIG> have been developed from simple PDMS microchannels. The shear-thinning composition (dyed blue) is stable in the microfluidic system with flowing water (dyed green) at <NUM>, as shown in <FIG>.

As shown in <FIG>, a shear-thinning-Chitosan composition (<NUM>% chitosan) produced approximately <NUM> of bacteria free zone when the composition is placed onto a confluent Staphylococcus aureus plate.

These experiments were performed to demonstrate that, following intravascular shear-thinning composition injection, the composition remains within the injected site and takes the shape of the vasculature without fragmentation despite blood pressures that exceed physiologic human systolic pressures and heart rates ><NUM>. The right groin was exposed revealing the femoral vasculature, shown in <FIG>, and approximately <NUM>µL of the <NUM>% shear-thinning composition formulation was injected, as shown in <FIG>. The black arrow indicates the vessel puncture site; slight blanching of the vessel can be seen suggesting proximal and distal flow of the shear-thinning composition producing an occlusive cast. Similarly, the left flank was exposed revealing the left renal vasculature; <NUM>µL of the shear-thinning composition was again injected (not shown). Approximately <NUM> hours later, an ultrahigh resolution CT imaging was performed (<NUM> x <NUM> slice thickness reconstructed at <NUM> x <NUM>, <NUM> mA, KV <NUM>, rotation time <NUM> sec, Pitch <NUM>; <NUM> mGy, DLP <NUM> mGycm); the composition is indicated by the arrow in the coronal and axial views, as shown in <FIG>. In each case, the shear-thinning composition successfully produced a cast of the vasculature without any evidence of fragmentation and non-target embolization. The shear-thinning composition was mixed with Visipaque contrast solution to help visualize the composition during CT imaging. No evidence of non-target contrast enhancement was identified in these state-of-the-art ultrahigh resolution CT imaging. Additional images are shown in <FIG>.

Creep tests were performed on an Anton Paar MCR702 SingleDrive system with a disposable aluminum cone geometry (<NUM> diameter, <NUM>° angle, <NUM> truncation gap) and a sandblasted lower plate. Tests were performed at <NUM> and samples were sealed with mineral oil to prevent sample drying. Shear-thinning composition 6NC75 was loaded and creep was measured by applying stresses (σ) of <NUM>, <NUM>, and <NUM> Pa for <NUM>, followed by relaxation for <NUM> and monitoring the resulting strain (ε). Creep compliance was calculated according to Equation <NUM> from the <NUM> Pa creep curve and used to extrapolate the strain expected at <NUM>, <NUM>, <NUM>, and <NUM> Pa (<NUM>-<NUM> dynes/cm<NUM>), shear stress values experienced along the wall of vasculature (see e.g., <NPL>).

<FIG> show creep and creep recovery of a representative intravascular shear-thinning composition, 6NC75 (<FIG>) and compliance curves from creep data (<FIG>). The compliance curve derived from the <NUM> Pa creep curve (J<NUM> Pa(t)) was used to extrapolate the creep response at lower stress values (<NUM>-<NUM> Pa) that are experienced along a vasculature wall (<NUM>-<NUM> Pa) as shown in <FIG>. Expected creep caused by physiologically relevant stresses is below <NUM>% strain and, therefore, is likely not a source of hydrogel deformation following delivery into the vasculature. Almost complete recovery after shear was observed, suggesting minimal permanent deformation in the shear-thinning composition.

Simulated Body Fluid (SBF; <NUM> sodium chloride, <NUM> sodium bicarbonate, <NUM> potassium chloride, <NUM> potassium phosphate dibasic, <NUM> magnesium chloride hexahydrate, <NUM> <NUM> hydrochloric acid, <NUM> calcium chloride, <NUM> sodium sulfate, <NUM> Tris per <NUM> SBF) was made with <NUM> % (w/w) glycerol to simulate whole blood (see e.g., <NPL>). Porcine aorta and abdominal vasculature was acquired from Research <NUM>, Inc. Vasculature including the aortic bifurcation and iliac arteries was excised and zip-tied to the tubing of a digital pressure gauge (PASPORT Dual Pressure Sensor). The vessels were submerged in a Simulated Body Fluid (SBF)-filled container and SBF was flowed through the vessels with a syringe pump, with the pressure being monitored upstream. Lastly, a <NUM>-French (Internal Diameter = <NUM>) catheter was fed through the tubing to inject the shear-thinning composition directly into the vessel. The catheter was removed and the pressure was measured until ejection of the shear-thinning composition plug from the vessel. 18A-18E show results of the ex vivo occlusion studies using porine iliac arteries and simulated body fluid to mimic whole blood.

Shear-thinning composition samples were transferred into syringes and the bottom sealed with a luer lock compatible syringe cap. Syringes were centrifuged at <NUM> rpm in a swinging bucket rotor (plunger removed) to remove any bubbles and compact the shear-thinning composition. Plungers were reinserted and filled, capped syringes were placed in a plastic bag, in a glass jar filled with ice to prevent excessive heat from damaging the shear-thinning composition. The jar was placed in the irradiation chamber of a Gammacell 220E irradiator and irradiated for <NUM>, resulting in a dosage of <NUM> kGy (<NUM> gray min-<NUM>). Shear rate sweep of the shear-thinning composition 6NC75 before and after gamma irradiation showed no significant change in the shear stress profile, as shown in <FIG>. Clotting time reduction of blood in contact with the shear-thinning composition was not significantly changed by gamma irradiation of the shear-thinning composition for sterilization, and addition of contrast dye (Visipaque™).

Shear-thinning composition 6NC75 was inoculated with <NUM><NUM> CFU mL-<NUM> BL21 Escherichia coli (E. coli) (see e.g., <NPL>; and <NPL>), according to a standard colony forming unit (CFU) calculation. CFU concentrations of overnight cultures were enumerated by plating serial dilutions of the culture onto LB agar plates without antibiotic. coli was mixed with the shear-thinning composition and separated into <NUM> parts: one that was placed in a sterile tube as the positive control and the other half prepared for γ-sterilization as described above. After irradiation, sterile pipette tips were used to sample the shear-thinning compositions and placed into culture tubes containing LB media. Tubes were incubated overnight (<NUM>) and OD<NUM> was measured. Serial dilutions of the overnight culture were also made in saline solution to perform the standard CFU calculation. <NUM>µL of each dilution was added to a LB agar plate without antibiotic and spread down the plate by tilting. The number of colonies formed at the first dilution containing between <NUM> and <NUM> colonies was used for the calculation of CFU concentration according to Equation <NUM>: <MAT> where n is the dilution used for counting (e.g. <NUM> for a <NUM> fold dilution). Samples that generated no colonies from the initial undiluted overnight culture were considered as being less than <NUM><NUM> CFU mL-<NUM>. OD<NUM> of LB media incubated in shear-thinning composition-coated pipette tips for <NUM> showed no increase in optical density of 6NC75 inoculated with E. coli, as shown in <FIG>.

<FIG> show a visual comparison of agar plates treated with dilutions of overnight cultures containing unsterilized inoculated shear-thinning compositions (<FIG>), γ-sterilized inoculated shear-thinning compositions (<FIG>), sterile pipette tips (negative control, <FIG>), and a sterile Tornado embolization coil (<FIG>).

Shear-thinning compositions 6NC50 and 6NC75 were injected into the vessel or aneurysm of a clinical vasculature embolization model. The system was perfused manually with contrast dye (Ultravist®, Bayer) and imaged under fluoroscopy (Siemens; Artis Zeego) before and after injection of the shear-thinning composition. The shear-thinning compositions occluded the entire model vessel, delivered simply from a preloaded shear-thinning composition-filled syringe with no other necessary embolic preparation.

Shear-thinning compositions were prepared with multiple ratios of iodinated contrast dye (Visipaque™):water and added into syringes that were centrifuged to remove any bubbles in the syringe. The syringes were placed in a head and body modular phantom (Gammex 461A) and images were obtained on the same dual-energy CT scanner. Results of the phantom imaging studies are shown in <FIG>.

Pig vessels were fixed in formalin (Sigma, HT <NUM>-<NUM>, St. Louis, MO) for overnight at room temperature. Paraffin embedded blocks were sectioned at <NUM> and mounted on positively charged slide glass (Fisher Scientific, #<NUM>-<NUM>-<NUM>, Pittsburgh, PA). Slides were baked at <NUM> for <NUM>, deparaffinized (<NUM> x <NUM>% Xylene, <NUM> x <NUM>% ethanol, <NUM> x <NUM>% ethanol, <NUM> x <NUM>% ethanol, <NUM> x <NUM>% ethanol, <NUM> each) and rehydrated in double-distilled water for <NUM>.

Slides were stained in Gill's #<NUM> Hematoxylin solution (Sigma, GHS-<NUM>, St. Louis, MO) for <NUM> and washed in running water for <NUM>. Slides were then stained in Eosin Y (Sigma, HT110116, St. Louis, MO) for <NUM> and washed for <NUM>. Slides were dehydrated <NUM> x in <NUM>% ethanol, <NUM> x80 % ethanol, <NUM> x in <NUM>% ethanol, <NUM> x <NUM>% ethanol for <NUM>, respectively. Next, the slides were dried and mounted with permanent mounting solution (Histo Mount Solution, #<NUM>, Life Technologies, Thermo-Fisher Scientific, Grand Island, NY).

Collagen in the vessels were stained according to Mason's Trichrome Stain Kit (Sigma HT1079, St. Louis, MO). Slides were stained in freshly prepared Weigert's iron Hematoxylin solution (Sigma HT15-1KT, St. Louis, MO) for <NUM> in a humid chamber and washed in running tap water for <NUM>. Then the slides were stained in Biehrich Scarlet Acid Fuchsin solution for <NUM> and rinsed in phosphomolybdic-phosphotungstic acid solution for <NUM>. Aniline blue solution was applied for <NUM> and washed in tap water for <NUM>. Slides were dried at <NUM> for <NUM> and mounted with permanent mounting solution (Histo Mount Solution, #<NUM>, Life Technologies, Thermo-Fisher Scientific, Grand Island, NY). Nuclei are stained in black, cytoplasm in red and collagen in blue in the tissue. Representative images of histology staining of shear-thinning composition occluded and patent vessels are shown in <FIG>.

Antigen retrieval was performed in <NUM> sodium citrate, pH <NUM> solution (Life Technologies #<NUM>, Thermo-Fisher Scientific, Grand Island, NY). Vessel tissues were permeabilized in <NUM>% Triton X-<NUM> (Sigma T8532, St. Louis, MO) in PBS and endogenous peroxidase activity was quenched in <NUM>% H<NUM>O<NUM> in <NUM>% methanol for <NUM> at room temperature, respectively. Endogenous biotin activity was blocked by Avidin-Biotin Blocking Kit (Life Technologies, #<NUM>, Thermo-Fisher Scientific, Grand Island, NY). Non-specific protein was blocked in <NUM>% goat serum in PBS for <NUM> at room temperature. Polyclonal rabbit anti proliferating cell nuclear antigen (PCNA, Santa Cruz, #SC7907, <NUM>:<NUM>, Santa Cruz, CA), CD68 (Aviva, ARP63008_P050, <NUM>:<NUM>, San Diego, CA) and MPO (AbCam, #<NUM>, <NUM>:<NUM>, Cambridge, MA) were incubated in % goat serum for overnight at <NUM> in a humid chamber. Biotinylated goat anti-rabbit second antibody (Vector, BA4001, Burlingame, CA) was applied for <NUM> at room temperature. Signal was amplified with Avidin-Biotin-Complex solution (Vectastain Elite ABC kit, Vector Laboratories PK6100, Burlingame, CA). Permanent insoluble dark brown color (<NUM>, <NUM>'-diaminobenzidine, DAB substrate, Vector Laboratories SK4100, Burlingame, CA) was developed under a light microscope. Tissue sections were counterstained with Gill's #<NUM> Hematoxylin (Sigma, GHS-<NUM>, St. Louis, MO) for <NUM> and dehydrated and then permanently mounted.

Residual shear-thinning composition was present in the vessel with infiltrating inflammatory cells evident in the vessel lumen surrounding the remaining biomaterial. Immunohistochemistry for myeloperoxidase (MPO) revealed MPO-positive cells infiltrating the vessel luminal area. MPO is known to be expressed primarily by neutrophils of the myeloid cell lineage but can also be present in monocytes and macrophages (see e.g., <NPL>). There was negligible MPO staining observed in the vessel wall, highlighting that inflammation was confined to the lumen. Macrophages, as indicated by CD68 staining, were predominately expressed surrounding the remaining shear-thinning composition, as shown in <FIG>, second row), suggesting that there was ongoing degradation and clearance of residual shear-thinning composition inside the occluded vessel by phagocytic cells. In addition, there was active cellular proliferative activity, as determined by PCNA staining. The PCNA staining localized primarily to the areas of residual shear-thinning composition, suggesting that there is continued remodeling that parallels shear-thinning composition clearance within the occluded vessel lumen.

H&E histology samples were imaged using a Zeiss Axio Zoom V16 stereomicroscope and the image files imported into ImageJ. Regions containing shear-thinning composition or unfilled with connective tissue within the intraluminal region of the vessel were outlined using the polygon tool and the area was measured (shear-thinning composition area). Next, the area of the intraluminal region of the vessel was measured (Vessel Area) and the percent vessel occluded was calculated according to Equation <NUM>.

<FIG> shows the replacement of shear-thinning composition with connective tissue in the vessel which was quantified by measuring the amount of intraluminal area filled with connective tissue from H&E histology images over time, reflecting the percentage of vessel remodeled. As shown in <FIG>, when the remodeled vessel percent was plotted against the histology slide's location proximal or distal to shear-thinning composition injection, higher levels of remodeling were noted proximal to the shear-thinning composition injection which then decreased as the sampled tissue became distal to the injection. As shown in <FIG>, the spatial dependence of vessel occlusion was noted in histology slides sampled proximal and distal to the site of embolization.

Clearance of the shear-thinning composition and remodeling of the embolized vessel was dependent on temporal and spatial factors (<FIG>). Temporally, histological sections from later time points were more advanced in their clearance of the shear-thinning composition and subsequent vessel remodeling. An increase in the percent of the vessel remodeled with connective tissue was observed, as the shear-thinning composition was cleared from the site (<FIG>). Spatially, comparing proximal and distal histologic cross sections of the embolized vessel, a trend toward decreased connective tissue presence was observed. There was near <NUM>% connective tissue occupying the vessel at proximal locations and a decreased presence of connective tissue in more distal sections (<FIG>). The most distal section showed an increase in the presence of connective tissues relative to its adjacent more proximal sections, likely due to its proximity to the blood stream and the ability for phagocytic cells to interact with the shear-thinning composition at this extreme distal location during remodeling. The high percentage of connective tissues present at both the proximal and distal sites of occlusion result in an embolized site with minimal likelihood for subsequent fragmentation or recanalization.

Porcine models were further tested where a shear-thinning composition (6NC75) was delivered to target vessels via standard clinical percutaneous catheterization techniques and tools. Because fragmentation and displacement of the shear-thinning composition following embolization of a target vessel is an undesired outcome, initial experiments were designed to assess stability of the injected shear-thinning composition in the arterial vasculature (<FIG>).

Female Yorkshire pigs were anesthetized, intubated, and monitored throughout the procedure as described herein. Under ultrasound guidance, the right carotid artery was accessed using a standard <NUM> needle and wire. Under fluoroscopic guidance, a <NUM>-French Cobra <NUM> catheter (Cook Medical) and a <NUM>" angled glidewire (Terumo) were brought to the infrarenal aorta and digital subtraction angiography (DSA) was performed demonstrating the lumbar arteries and the iliac arteries. The lumbar artery was catheterized and successfully embolized within seconds following the injection of approximately <NUM> of gamma-irradiated shear-thinning composition (6NC75) from a premade <NUM>-mL syringe (<FIG> shows a magnified view of the shear-thinning composition edge with patent proximal lumbar artery and aorta.

Since this shear-thinning composition remained in place and did not migrate over a period of <NUM>, the left external iliac artery (EIA) was embolized next. The same catheter was brought just distal to the internal iliac artery origin and approximately <NUM> of the shear-thinning composition was injected (<FIG>). Flow immediately ceased without any evidence for displacement or fragmentation; the shear-thinning composition remained at the injected site. This was a notable outcome because EIA is at high flow (approximately <NUM>/s) with a diameter of <NUM>, similar to the human iliac arteries; when compared to the clinical scenario, coil embolization of this high flow, large-diameter artery would be at high risk for coil displacement, which was not the case for the shear-thinning composition.

Next, the durability of shear-thinning composition embolization in the forelimb venous vasculature was tested. Forelimb veins were chosen because any fragmentation or displacement of the shear-thinning composition would lead to pulmonary embolism, an objective assessment of performance that was tested using computed tomography (CT) imaging. Using a <NUM>-French Davis catheter (Cook Medical), various centrally positioned forelimb veins were embolized in three pigs using <NUM>-<NUM> of the gamma-sterilized shear-thinning composition 6NC75 (<FIG>). These pigs survived <NUM>, <NUM>, and <NUM> days. Just prior to necropsy, CT imaging was performed using a <NUM> slice dual-energy scanner (Definition Flash, Siemens) before and after intravenous administration of iodinated contrast media (Visipaque™). CT imaging revealed no evidence of artifact from the shear-thinning composition that would obscure the region of embolization. CT imaging also revealed that there was no evidence of any pulmonary embolism as indicated by the highly sensitive iodine maps of the lungs up to <NUM> days post-embolization.

Claim 1:
A shear-thinning composition comprising a therapeutic agent or a diagnostic agent, gelatin or a derivative thereof, silicate nanoparticles, and water, for use in treating cancer;
wherein the therapeutic agent is selected from the group consisting of a chemotherapeutic agent and a sensitizing drug to an adjuvant therapy and the diagnostic agent is selected from a magnetic agent and an electrical agent;
wherein the therapeutic agent or diagnostic agent is:
(a) preloaded onto at least one surface of the shear-thinning composition prior to the treating; or
(b) preloaded into the shear-thinning composition prior to the treating,
wherein the gelatin derivative is methacrylated gelatin, acrylated gelatin, or thiolated gelatin.