Solvatochromic indicators for detecting infection

Abstract

A solvatochromic indicator for detecting the presence of bacterial infection is disclosed. The indicator is made of sugar beet (Beta vulgaris) and optionally further comprises turmeric (Curcuma longa), isopropyl alcohol, and any combination thereof, and undergoes a detectable color change in the presence of infection. A filament treated with a solvatochromic indicator is further provided, where the detectable color change of the indicator is based on the pH of the tissue having the bacterial infection. TIFF2024514717000002.tif83134

Description

[Technical field]

[0001] Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 63 / 173,611, filed April 12, 2021, the contents of which are incorporated by reference in their entirety herein.

[0002] FIELD OF THEINVENTION The present invention relates to compositions and methods for efficiently detecting pH changes using solvatochromic indicators. In a specific embodiment, the present invention also relates to efficiently detecting infection at a surgical site based on visual detection of pH changes. [Background technology]

[0003] background Surgical site infections are one of the most common causes of hospital-acquired or nosocomial infections. These infections are generally caused by gram-positive bacterial species such as Staphylococcus aureus and Streptococcus pyogenes, and gram-negative bacteria such as Pseudomonas aeruginosa and Escherichia coli. Clinical symptoms may include localized redness, pain, wound drainage, delayed healing, swelling, and fever.

[0004] Millions of surgical procedures are performed in inpatient hospitals, outpatient affiliated hospitals, or free-standing settings. Surgical site infections can occur in up to 5% of these surgical procedures, and infections are associated with significant morbidity and mortality. Moreover, it often leads to increased hospital stays and additional costs due to prolonged antibiotic therapy.

[0005] Core guidelines for surgical procedures often include recommendations for preventing surgical site infections that are often generalized across surgical procedures. Among these recommendations, administration of preoperative antimicrobials before skin incision, thorough cleaning of the skin before and after surgery, and wound monitoring are the most commonly used methods. Additionally, hospitals around the world have adjusted their surgical protocols in an attempt to reduce the likelihood of surgical site infections. However, protocols for checking incisions for obvious signs of infection have remained unchanged, despite the fact that it is very difficult to proceed once signs of infection have occurred, and therefore why surgical site infections are so dangerous. If a surgical site infection is detected early, the wound can be treated with antibiotics as opposed to the need for additional surgery. These concerns apply broadly to all types of wound sites, especially in developing countries where medical care can be difficult to obtain.

[0006] For this purpose, there is a need for a method for efficiently detecting bacterial infection in both clinical and non-clinical environments.The disclosed compositions, methods and uses provide for efficiently detecting pH changes in medical, biomedical, veterinary or clinical environments, or any other environments that need to be kept free of bacteria.Detecting pH changes can be applied to visually detect infection in wound sites, such as surgical sites.Disclosed herein is a novel method for detecting pH changes, and therefore bacterial infection, in any environment.

[0007] Additionally, there is a need for inexpensive, easily accessible tools to diagnose infection in developing countries, where people may not be able to see a doctor, and the use of simple, widely applicable compositions to indicate infection would allow a person to know that medical attention is needed. Summary of the Invention

[0008] In one aspect, the present disclosure provides a composition comprising at least one extract from sugar beet (Beta vulgaris) bulb, at least one extract from turmeric (Curcuma longa) root, and at least 10% alcohol.

[0009] In one aspect, the present disclosure provides a composition comprising a solvatochromic indicator, wherein the solvatochromic indicator is a pH responsive visual indicator, and wherein the composition further comprises at least one extract from turmeric root.

[0010] In one aspect, the disclosure provides a method comprising preparing a first extract from at least one sugar beet bulb; preparing a second extract from at least one turmeric root; and combining the first and second extracts in a solution comprising alcohol. [Brief description of the drawings]

[0011] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0012] The present aspects are illustrated by the figures of the accompanying drawings, which may not necessarily be to scale, and in which like references indicate similar elements.

[0013] [Figure 1] FIG. 1A shows the scale of sugar beet extract over a range of pH conditions. FIG. 1B shows a set of polyester cotton blend sutures treated with sugar beet extract. From left to right, the sutures are subjected to a pH 5 solution, a no solution control, and a pH 9 solution. The image shows the sutures after 5 minutes have elapsed. FIG. 1C shows the same set of three sutures as seen in FIG. 1B, except that approximately 60 hours have elapsed. The sutures treated with sugar beet extract and pH 9 solution are visually different from the control and pH 5 treatments. [Diagram 2] Figure 2A depicts a set of swabs. Each swab was treated with 1 ml of sugar beet bulb extract. Figure 2B depicts the same set of swabs 1 hour after application of the following solutions; from left to right: a control with nothing added, a pH 5 solution, a pH 9 solution, distilled water, 91% alcohol, and a pH 9 solution followed by application of 1 ml of water. [Diagram 3] The antibacterial properties of the mixture containing the extract from sugar beet bulb are demonstrated. Figure 3A shows normal growth of E. coli when grown in a Petri dish at 37°C for 12 hours. Figure 3B shows the same Petri dish as Figure 3A, except that a filament containing an extract from sugar beet bulb has been overlaid on this portion of the dish, and no E. coli growth is visible. Figure 3C is a photograph taken after the filament has been removed, and again, no E. coli growth is visible in or around the area contacted by the filament. [Figure 4-1] Figure 4A is an exemplary illustration of the chemical structure of Parylene C that can be used as a thin film coating compound with the solvatochromic indicators of the present invention. Figure 4B is an exemplary illustration of the steps in a coating process for a thin film coating on Parylene C. (VSi Parylene "Coating and Deposition Process" https: / / vsiparylene.com / about-parylene / #deposition-process). [Figure 4-2] See description of Figure 4-1. [Diagram 5]Figures 5A-C show three sets of sutures, each set containing three sutures containing a mixture of sugar beet-derived extract and turmeric-derived extract in 10% alcohol and coated with Parylene C. Figures 5D-F show the same three sets of sutures shown in Figures 5A-C, except that Figure 5D was treated with a pH 5 solution and Figure 5F was treated with a pH 9 solution. Figure 5E has no solution added and represents a negative control. Figures 5G-I show the same set of sutures as Figures 5D-F 10 minutes after application of the above mentioned solutions. The color change in response to a basic pH of 9 is evident compared to Figures 5G-H. [Figure 6] Figure 6A shows two tubes of purple cabbage extract in a solution of pH 5 (left, labeled C5) and a solution of pH 9 (right, labeled C9). Figure 6B shows two tubes of equal amounts of purple cabbage extract and sugar beet extract in a solution of pH 5 (left, labeled BC5) and a solution of pH 9 (right, labeled BC9). Figure 6C shows two tubes of sugar beet extract in a solution of pH 5 (right, labeled B5) and a solution of pH 9 (left, labeled B9), shown as a positive control. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] Detailed Description of the Invention Unless otherwise defined, all technical and scientific terms used have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. Where a term is provided in the singular form, the inventors also contemplate aspects of this disclosure described by the plural form of that term. In the event of a discrepancy between terms and definitions used in references incorporated by reference, the terms used in this application shall have the definitions given herein. Other technical terms used have their ordinary meaning in the art in which they are used, as exemplified by various art-specific dictionaries, such as "The American Heritage® Science Dictionary" (Editors of the American Heritage Dictionaries, 2011, Houghton Mifflin Harcourt, Boston and New York), "McGraw-Hill Dictionary of Scientific and Technical Terms" (6th edition, 2002, McGraw-Hill, New York), or "Oxford Dictionary of Biology" (6th edition, 2008, Oxford University Press, Oxford and New York).

[0015] Any references cited herein, including, for example, all patents, published patent applications, and non-patent publications, are hereby incorporated by reference in their entirety.

[0016] When a grouping of options is presented, any and all combinations of the members that make up that grouping of options are specifically contemplated. For example, if an item is selected from the group consisting of A, B, C, and D, we specifically contemplate each option individually (e.g., A alone, B alone, etc.), as well as combinations such as A, B, and D; A and C; B and C. The term "and / or" when used in a list of two or more items means any one of the listed items alone or in combination with any one or more of the other listed items. For example, the phrase "A and / or B" is intended to mean either or both of A and B, i.e., A alone, B alone, or a combination of A and B. The phrase "A, B, and / or C" is intended to mean A alone, B alone, C alone, a combination of A and B, a combination of A and C, a combination of B and C, or a combination of A, B, and C.

[0017] When a range of numerical values ​​is provided herein, the range is understood to include the endpoints of the range, as well as any number between the defined endpoints of the range. For example, "between 1 and 10" includes any number between 1 and 10, as well as the number 1 and the number 10.

[0018] When the term "about" is used in reference to a numerical value, it is understood to mean plus or minus 10%. For example, "about 100" includes 90 to 110.

[0019] As used herein, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" can include a plurality of compounds, including mixtures thereof.

[0020] As used herein, betaine, also known as betaine anhydrous or trimethylglycine, is a neutral compound with a positively charged cationic functional group. Betaine occurs naturally in the human body and can be found in foods such as beets and spinach. In some embodiments, betaine is obtained from at least one beet (sugar beet). In further embodiments, betaine is obtained from a source other than sugar beet. In further embodiments, betaine is used as a solvatochromic indicator at wound or surgical sites to diagnose bacterial infection.

[0021] As used herein, pH is defined as a measure of the acidity and alkalinity of a solution, it is a number on a scale where a value of 7 represents neutrality, lower numbers indicate increasing acidity, and higher numbers indicate increasing alkalinity, with each unit of change representing a ten-fold or logarithmic change in acidity or alkalinity. pH can also be defined as the negative logarithm of the effective hydrogen ion concentration or hydrogen ion activity in gram equivalents per liter of solution. pH values ​​below 7 are also known as acidic pH scores, and pH values ​​above 7 are also known as basic pH scores.

[0022] Healthy human skin is naturally acidic, typically having a pH of 5. Wounds and surgical sites can become infected with microorganisms or bacteria, and these infections can potentially cause illness or death in patients. Infected wounds have a pH of 9 or higher. The present invention provides an apparatus and method for detecting infection in surgical sites using a medical device in combination with a color-changing indicator.

[0023] As used herein, solvatochromic indicator refers to a compound that can be used to access the polarity of a solvent by UV, visible, or near infrared spectroscopy, without limitation. See, for example, Reichardt, Chem. Rev., 1994, pp. 2319-2358. In some embodiments, the solvatochromic indicator provides a visual indication of a change in pH when exposed to a material that has a basic pH compared to the previous environment of the solvatochromic indicator. In further embodiments, the visual indication of the change in pH is a color change of the composition that includes the solvatochromic indicator. In another embodiment, the solvatochromic indicator shows a visual indication of a change in pH when applied to a wound site with bacterial infection. In further embodiments, the solvatochromic indicator that includes betaine darkens when exposed to a basic pH compared to the neutral pH that occurs when the indicator is applied to a site with bacterial infection. In further embodiments, the solvatochromic indicator comprising an extract of sugar beet bulb darkens when exposed to a basic pH compared to the neutral pH that occurs when the indicator is applied to a site with a bacterial infection, hi further embodiments, the solvatochromic indicator comprising an extract of sugar beet bulb changes from red to purple when exposed to a basic pH compared to the neutral or acidic pH that occurs when the indicator is applied to a site with a bacterial infection.

[0024] As used herein, "extract" refers to a composition, solid, liquid, or a combination of both, obtained from the disruption of cell walls and cell membranes. Molecules within cell walls and cell membranes can be extracted by any means commonly known in the art. In some embodiments, extracts are prepared by routine processes such as blending, grinding, crushing, beating, scraping, or any combination thereof, and are materials that contain the molecules desired to be extracted. In some embodiments, extracts are prepared through techniques commonly practiced in the art, such as sonication. It is envisioned that any equivalent method known in the art can be used to prepare extracts for use in the disclosed methods and compositions. Further processing of the extract to remove impurities is also envisioned, including but not limited to any type of size exclusion method.

[0025] In one aspect, a solvatochromic pH indicator comprising an extract from the bulb of the sugar beet plant is used to dye sutures or wound dressings. The sutures or wound dressings comprising the solvatochromic pH indicator comprising sugar beet undergo a color change in the presence of a basic pH. In a further aspect, the solvatochromic pH indicator marks a basic pH environment via a purple color. Bacterial infections are known to create basic pH conditions at an infected wound site. Thus, the compositions disclosed herein comprising the solvatochromic pH indicator comprising sugar beet provide a novel method for detecting bacterial infection at a wound site.

[0026] The bulb of the sugar beet plant contains the chemical betaine, which is a solvatochromic indicator. One example of a solvatochromic indicator containing betaine is Reichardt's dye. In one aspect, the indicator of the present invention undergoes a visible color change when the pH becomes more basic, which occurs when bacteria are present and actively metabolizing. In a further aspect, the color change is to purple in a basic pH environment. In another aspect, the solvatochromic indicator of the present invention can contain turmeric in the form of an extract obtained from the root of the plant. In a further aspect, the turmeric extract is mixed into a solution containing the solvatochromic indicator. In another aspect, the sugar beet is mixed in a solution containing isopropyl or any other alcohol. In further aspects, the solution containing the solvatochromic indicator comprises about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or at least 20% alcohol. In further aspects, the solution containing the solvatochromic indicator comprises at least 13 percent alcohol. In further aspects, the solution containing the solvatochromic indicator comprises at least 15 percent alcohol. In further aspects, the solution containing the solvatochromic indicator comprises at least 20 percent alcohol. In further aspects, the solution containing the solvatochromic indicator comprises at least 7 percent alcohol. In further aspects, the solution containing the solvatochromic indicator comprises at least 5 percent alcohol. In further aspects, the solution containing the solvatochromic indicator comprises at least 3 percent alcohol. In another aspect, the alcohol is diluted in water in the final solution.

[0027] In an embodiment, the solvatochromic indicator comprises an equal amount of sugar beet extract and an equal amount of turmeric extract. In an embodiment, the solvatochromic indicator comprises two parts sugar beet extract and one part turmeric extract. In an embodiment, the solvatochromic indicator comprises one part sugar beet extract and two parts turmeric extract. In an embodiment, the solvatochromic indicator comprises two parts sugar beet extract and three parts turmeric extract.

[0028] As used herein, filament refers to any material, natural or synthetic, suitable for absorbing or coating a composition containing a solvatochromic indicator, without limitation. In one aspect, the solvatochromic indicator comprises betaine. In a preferred embodiment, the betaine is present in an extract from sugar beet bulb. In one embodiment, the filament can be porous or non-porous. In a further embodiment, the filament is suitable for use in medical procedures. In a further embodiment, the filament is suitable for use as a wound suture. In another embodiment, the filament is suitable for use as a wound dressing. Non-limiting examples of filaments include gauze, medical swabs, cotton, prolene, silk, polyester, catgut, polyglactin, polyglycol, poliglecaprone, polydioxanone, polypropylene, polyamide, and mixtures thereof. In a further embodiment, the filament is composed of synthetic fibers. In a further embodiment, the filament is composed of natural fibers.

[0029] In embodiments, filaments, such as sutures or wound dressings, may be saturated, infused, coated, or otherwise treated with the solvatochromic indicators of the present invention. In one aspect, the solvatochromic indicator comprises betaine. In a preferred embodiment, the betaine is present in an extract from sugar beet bulbs. In a further aspect, a method is provided in which a solution comprising a solvatochromic indicator is added to a suture or wound dressing prior to application to a wound. In another aspect, a method is provided in which a solution comprising a solvatochromic indicator is added to a suture or wound dressing after application to a wound. In each of these embodiments, the color change brought about by the pH change caused by the presence of bacteria is significant enough to be detected by the human eye alone.

[0030] As used herein, a wound site is a place on the human or animal body where the surface of the skin is broken or abraded.In one aspect, a surgical site is a wound site.Without being limited, a wound can be a laceration, a cut, a deep cut, a tear, a hole, an incision, or a lesion.In a further aspect, a wound site can be any environment that is prone to bacterial growth and / or infection.

[0031] As used herein, a surgical site is defined in this disclosure as a particular part of the body, either human or animal, where surgery has been performed. A surgical site may also include a site on the body where other open wounds, such as lacerations, cuts, gashes, tears, holes, incisions, or lesions, have occurred. Surgical site infections may occur at any of these sites and may take a long time to detect using traditional methods.

[0032] Bacteria frequently associated with wound site infections include, by way of non-limiting example, Staphylococcus subspecies (ssp.), Streptococcus subspecies, Pseudomonas subspecies, Escherichia subspecies, Enterococcus subspecies, Proteus subspecies, Serratia subspecies, Enterobacter subspecies, Acetinobacter subspecies, and Klebsiella subspecies. Additional bacteria frequently associated with wound site infections include, but are not limited to, Streptococcus pyogenes, Pseudomonas aeruginosa, Enterococcus faecalis, Proteus mirabilis, Serratia marcescens, Enterobacter clocae, Acetinobacter anitratus, Klebsiella pneumoniae, Escherichia coli, Staphyloccus aureus, and coagulase-negative Staphylococci.

[0033] As used herein, suture refers to a medical device (such as present at a surgical or wound site) that is commonly used to close incisions, punctures, lacerations, and the like by holding body tissue together after surgery. A suture is typically a thread connected to a needle that is sewn through open body tissue. The suture is secured using a surgical knot. In an embodiment, the suture is saturated with the solvatochromic indicator of the present disclosure. In another embodiment, the suture is coated with the solvatochromic indicator of the present disclosure. In one aspect, the solvatochromic indicator comprises betaine. In a preferred embodiment, the betaine is present in an extract from sugar beet bulb.

[0034] Wound dressings are commonly used to enclose or contain wounds or surgical sites. Non-limiting examples of wound dressings include gauze sponges, gauze bandages, non-adhesive pads, non-adhesive wound dressings, foam dressings, calcium alginate, hydrogel dressings, hydrocolloids, alginates, composites, and transparent dressings. In one embodiment, any solvatochromic indicator of the present disclosure can be used in conjunction with any wound dressing. In one aspect, the solvatochromic indicator comprises betaine. In a preferred embodiment, betaine is present in an extract from sugar beet bulb.

[0035] As used herein, thin film coating refers to depositing a substance on a material that includes any of the solvatochromic indicators of the present disclosure. In one aspect, the solvatochromic indicator includes betaine. In a preferred embodiment, betaine is present in an extract from sugar beet bulb. In a further embodiment, the thin film coating allows the interaction of the external environment with the solvatochromic indicators of the present disclosure. In one embodiment, the thin film coating of the present invention is a thin film coating of Parylene C or any suitable material with similar properties. The thin film coating can be coated on any material that includes any of the solvatochromic indicators of the present disclosure. The thin film coating can be deposited on any material that includes any of the solvatochromic indicators of the present disclosure.

[0036] In one aspect, the disclosure provides a method comprising contacting a wound with a filament comprising at least one solvatochromic indicator; detecting a change in pH based on a visual change of the at least one solvatochromic indicator; and diagnosing a bacterial infection based on the visual change. In a further embodiment, the at least one solvatochromic indicator is an extract from at least one sugar beet bulb. In a further embodiment, the solution comprises an extract from at least one turmeric root. In a further embodiment, the solution comprises about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or at least 20% alcohol. In one aspect, the solution further comprises at least 13 percent alcohol. In one aspect, the solution further comprises at least 15 percent alcohol. In some aspects, the solution further comprises at least 20 percent alcohol. In some aspects, the solution further comprises at least 7 percent alcohol. In some aspects, the solution further comprises at least 5 percent alcohol. In some aspects, the solution further comprises at least 3 percent alcohol. In another aspect, the alcohol is diluted in water in the final solution. In another embodiment, the solvatochromic indicator is betaine. In another embodiment, the wound is contacted with a filament in the form of a suture. In another embodiment, the wound is contacted with a filament in the form of a wound dressing. In a further embodiment, the filament is coated with a thin film coating.

[0037] In one aspect, the disclosure provides a method comprising contacting a wound with a filament; applying a solution comprising at least one solvatochromic indicator to the filament; detecting a change in pH based on a visual change of the at least one solvatochromic indicator; and diagnosing a bacterial infection based on the visual change. In a further embodiment, the at least one solvatochromic indicator is an extract from at least one sugar beet bulb. In a further embodiment, the solution comprises an extract from at least one turmeric root. In a further embodiment, the solution comprises 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or at least 20% alcohol. In one aspect, the solution further comprises at least 13 percent alcohol. In one aspect, the solution further comprises at least 15 percent alcohol. In some aspects, the solution further comprises at least 20 percent alcohol. In some aspects, the solution further comprises at least 7 percent alcohol. In some aspects, the solution further comprises at least 5 percent alcohol. In some aspects, the solution further comprises at least 3 percent alcohol. In another aspect, the alcohol is diluted in water in the final solution. In another embodiment, the solvatochromic indicator is betaine. In another embodiment, the wound is contacted with a filament in the form of a suture. In another embodiment, the wound is contacted with a filament in the form of a wound dressing. In a further embodiment, the filament is coated with a thin film coating.

[0038] In one aspect, the disclosure provides a method comprising preparing a first extract from at least one sugar beet bulb; preparing a second extract from at least one turmeric root; and combining the first and second extracts in a solution comprising alcohol. In one embodiment, the solution comprises about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or at least 20% alcohol. In a further aspect, the solution further comprises at least 13 percent alcohol. In a further aspect, the solution further comprises at least 15 percent alcohol. In a further aspect, the solution further comprises at least 20 percent alcohol. In a further aspect, the solution further comprises at least 7 percent alcohol. In a further aspect, the solution further comprises at least 5 percent alcohol. In a further aspect, the solution further comprises at least 3 percent alcohol. In another aspect, the alcohol is diluted in water in the final solution. In another embodiment, the method includes applying the solution to the filament. In a further embodiment, the filament is in a form selected from the group consisting of a suture, a swab, a gauze sponge, a gauze bandage, a non-adhesive pad, and a non-adhesive wound dressing. In another embodiment, the method includes encapsulating the filament in a thin film coating. In a further embodiment, the thin film coating is a coating of Parylene C. In a further embodiment, the first extract from at least one sugar beet bulb and the second extract from at least one turmeric root are prepared using sonication or an equivalent method to rupture cell walls and cell membranes.

[0039] In one aspect, the present disclosure provides a composition for use in detecting bacterial infection at a wound site, comprising a solvatochromic indicator, wherein the solvatochromic indicator is a pH-responsive visual indicator.In another aspect, the solvatochromic indicator comprises at least one extract from sugar beet bulb.In another embodiment, the solvatochromic indicator comprises at least one extract from turmeric root.In a further embodiment, the solvatochromic indicator comprises at least one extract from sugar beet bulb and at least one extract from turmeric root.In one embodiment, the solvatochromic indicator comprises about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or at least 20% alcohol. In some aspects, the solvatochromic indicator comprises at least 13 percent alcohol. In some aspects, the solvatochromic indicator comprises at least 15 percent alcohol. In some aspects, the solvatochromic indicator comprises at least 20 percent alcohol. In some aspects, the solvatochromic indicator comprises at least 7 percent alcohol. In some aspects, the solvatochromic indicator comprises at least 5 percent alcohol. In some aspects, the solvatochromic indicator comprises at least 3 percent alcohol. In another aspect, the alcohol is diluted in water in the final solution. In another aspect, the alcohol is isopropyl alcohol.

[0040] In one aspect, the present disclosure provides a composition for use in detecting bacterial infection at a surgical site, comprising a solvatochromic indicator, wherein the solvatochromic indicator is a pH-responsive visual indicator. In another aspect, the solvatochromic indicator comprises betaine. In another embodiment, the solvatochromic indicator comprises at least one extract from turmeric root. In a further embodiment, the solvatochromic indicator comprises betaine and at least one extract from turmeric root. In another embodiment, the solvatochromic indicator comprises about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or at least 20% alcohol. In one aspect, the solvatochromic indicator comprises at least 13 percent alcohol. In some aspects, the solvatochromic indicator comprises at least 15 percent alcohol. In some aspects, the solvatochromic indicator comprises at least 20 percent alcohol. In some aspects, the solvatochromic indicator comprises at least 7 percent alcohol. In some aspects, the solvatochromic indicator comprises at least 5 percent alcohol. In some aspects, the solvatochromic indicator comprises at least 3 percent alcohol. In another aspect, the alcohol is diluted in water in the final solution. In another aspect, the alcohol is isopropyl alcohol.

[0041] In one aspect, the present disclosure provides a composition comprising a solvatochromic indicator and a filament. In one aspect, the solvatochromic indicator comprises betaine. In a preferred embodiment, the betaine is present in an extract from sugar beet bulb. In one embodiment, the filament is selected from the group consisting of cotton, proline, silk, polyester, catgut, polyglactin, polyglycol, poliglecaprone, polydioxanone, polypropylene, and polyamide. In another embodiment, the filament comprises a thin film coating. In a further embodiment, the thin film coating comprises parylene C.

[0042] In one aspect, the present disclosure provides a composition comprising a solvatochromic indicator for use at or on a wound site. In one aspect, the solvatochromic indicator comprises betaine. In a preferred embodiment, the betaine is present in an extract from sugar beet bulb. In one embodiment, the wound site is on a human body. In another embodiment, the wound site is on an animal body. In a further embodiment, the animal is selected from the group consisting of a dog, a cat, a cow, a sheep, a goat, a llama, a pig, a horse, and a donkey.

[0043] It will be understood by those skilled in the art that the compositions and methods provided herein can be applied to applications related to any field not disclosed herein, but where there is a known need for detection of bacterial growth. For example, the compositions and methods provided herein can be adapted for use in the food industry in detecting bacterial growth on food or preparation surfaces. In some aspects, use in the food industry is envisioned in a non-limiting manner in professional kitchens, industrial kitchens, personal kitchens, food storage facilities, food storage refrigerators, food storage freezers, and the like. Additional applications are also envisioned in the biomedical industry, where there is a known need for detection of bacterial growth. For example, the use of catheters is common in many medical fields, namely nephrology, and there is a need for compositions and methods for the detection of bacteria. Catheters incorporating the disclosed compositions can meet these needs. Furthermore, it is envisioned that additional agents can be added to the compositions of the present application that provide binding affinity to materials that are not porous, or that otherwise show poor uptake of the disclosed solvatochromic indicator without the additional agent. In this manner, solid surfaces can be treated with solvatochromic indicators and coated with thin film coatings for use in detecting bacterial growth as indicated by a change in pH.

[0044] The following exemplary, non-limiting embodiments are contemplated.

[0045] Aspect 1: A composition comprising at least one extract from sugar beet bulb, at least one extract from turmeric root, and at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or at least 20% alcohol.

[0046] Aspect 2: 2. The composition of embodiment 1, further comprising a filament.

[0047] Aspect 3: The composition of embodiment 1 or 2, wherein the filament is in the form of a suture.

[0048] Aspect 4: The composition of any one of the preceding aspects, further comprising a thin film coating encapsulating the filaments.

[0049] Aspect 5: Aspects 5. The composition of any one of aspects 1-4, wherein the thin film coating comprises Parylene C.

[0050] Aspect 6: 1. A composition for use in detecting a bacterial infection at a wound site comprising a solvatochromic indicator, the solvatochromic indicator is a pH responsive visual indicator; The composition further comprises at least one extract from turmeric root. composition.

[0051] Aspect 7: The composition of embodiment 6, wherein the solvatochromic indicator comprises a betaine.

[0052] Aspect 8: The composition of embodiment 6 or 7, wherein the solvatochromic indicator comprises an extract from at least one sugar beet bulb.

[0053] Aspect 9: 9. The composition of any one of aspects 6-8, further comprising at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or at least 20% alcohol.

[0054] Aspect 10: The composition of any one of aspects 6-9, further comprising a filament.

[0055] Aspect 11: The composition of any one of aspects 6-10, wherein the filament is in a form selected from the group consisting of sutures, gauze, gauze sponges, gauze bandages, non-adhesive pads, and non-adhesive wound dressings.

[0056] Aspect 12: 12. The composition of any one of aspects 6-11, further comprising a thin film coating encapsulating the filaments.

[0057] Aspect 13: Aspect 13. The composition of any one of aspects 6-12, wherein the thin film coating comprises Parylene C.

[0058] Aspect 14: Preparing a first extract from at least one sugar beet bulb; Preparing a second extract from at least one turmeric root; and combining said first and second extracts in a solution comprising an alcohol. A method comprising:

[0059] Aspect 15: The method of embodiment 14, further comprising applying the solution to the filament.

[0060] Aspect 16: 16. The method of embodiment 14 or 15, further comprising encapsulating the filaments in a thin film coating.

[0061] Aspect 17: The method of any one of aspects 14-16, wherein the thin film coating comprises Parylene C.

[0062] Aspect 18: The method of any one of aspects 14-17, wherein the filament is in a form selected from the group consisting of a suture, a gauze sponge, a gauze bandage, a non-adhesive pad, and a non-adhesive wound dressing.

[0063] Aspect 19: 19. The method according to any one of aspects 14 to 18, wherein the first and second extracts are prepared by sonication or an equivalent process.

[0064] Aspect 20: 20. The method of any one of aspects 14-19, wherein the solution comprises a concentration of alcohol of at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or at least 20%.

[0065] Aspect 21: 1. A composition for use in detecting a bacterial infection at a wound site comprising a solvatochromic indicator, The solvatochromic indicator comprises a pH responsive visual indicator; composition.

[0066] Aspect 22: A composition for use in detecting bacterial infection at a wound site comprising at least one extract from sugar beet bulb.

[0067] Aspect 23: A composition for use in detecting pH changes at a wound site comprising at least one extract from sugar beet bulb and at least one extract from turmeric root.

[0068] Aspect 24: 1. A composition for use in detecting pH changes at a wound site comprising at least one extract from sugar beet bulb, at least one extract from turmeric root, and isopropyl alcohol.

[0069] Aspect 25: 1. A composition for use in detecting pH changes at a wound site comprising at least one extract from sugar beet bulb, at least one extract from turmeric root, and at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or at least 20% alcohol.

[0070] Aspect 26: 1. A composition for use in detecting pH changes at a wound site comprising a filament, at least one extract from sugar beet bulb, and at least one extract from turmeric root.

[0071] Aspect 27: 1. A composition for use in detecting pH changes at a wound site comprising a filament, at least one extract from sugar beet bulb, at least one extract from turmeric root, and isopropyl alcohol.

[0072] Aspect 28: 1. A composition for use in detecting pH changes at a wound site comprising a filament, at least one extract from sugar beet bulb, at least one extract from turmeric root, and at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or at least 20% alcohol.

[0073] Aspect 29: A composition for use in detecting bacterial infection at a wound site comprising betaine.

[0074] Aspect 30: 1. A composition for use in detecting bacterial infection at a wound site comprising betaine and at least one extract from turmeric root.

[0075] Aspect 31: A composition for use in detecting pH changes at a wound site comprising a filament and a betaine.

[0076] Aspect 32: 1. A composition for use in detecting pH changes at a wound site comprising a filament, betaine, and at least one extract from turmeric root.

[0077] Aspect 33: 1. A composition comprising a filament, betaine, at least one extract from turmeric root, and isopropyl alcohol for use in detecting pH changes at a wound site.

[0078] Aspect 34: 1. A composition comprising a filament, betaine, at least one extract from turmeric root, and at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or at least 20% alcohol for use in detecting pH changes at a wound site.

[0079] Aspect 35: 1. A composition comprising a filament, betaine, and at least one extract from turmeric root for use in detecting pH changes at a wound site.

[0080] Aspect 36: The composition of any one of aspects 21-35, wherein the filaments are selected from the group consisting of cotton, prolene, silk, polyester, catgut, polyglactin, polyglycol, poliglecaprone, polydioxanone, polypropylene, and polyamide.

[0081] Aspect 37: The composition of any one of aspects 21-36, wherein the filaments further comprise a thin film coating.

[0082] Aspect 38: Aspects 21-37, wherein the thin film coating comprises Parylene C.

[0083] Aspect 39: The composition of any one of aspects 23-28, 30, and 32-35, wherein the at least one extract from turmeric root constitutes at least about 10%, 20%, 30%, 40%, 50%, or 60% of the composition.

[0084] Aspect 40: The composition of any one of aspects 22-28 and 32-35, wherein an extract from at least one sugar beet bulb constitutes at least about 10%, 20%, 30%, 40%, 50%, or 60% of the composition.

[0085] Aspect 41: The composition according to any one of aspects 1 to 13 and 21 to 40, which is capable of detecting a bacterial infection on a wound site on the human body.

[0086] Aspect 42: The composition according to any one of aspects 1 to 13 and 21 to 40, which is capable of detecting a bacterial infection on a wound site on an animal body.

[0087] Aspect 43: The composition of embodiment 42, wherein the animal is selected from the group consisting of dogs, cats, cows, sheep, goats, llamas, pigs, horses, and donkeys.

[0088] Aspect 44: applying to the wound site a filament comprising a solution comprising betaine, at least one extract from turmeric root, and at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or at least 20% alcohol; visually detecting a color change of the color indicator; and A process for diagnosing the presence or absence of infection A method comprising: A color change occurs when the color indicator comes into contact with a basic pH. method.

[0089] Aspect 45: applying a wound dressing to the wound site; applying a solution comprising betaine to the wound dressing; visually detecting a color change of the color indicator; and A process for diagnosing the presence or absence of infection A method comprising: A color change occurs when the color indicator comes into contact with a basic pH. method.

[0090] Aspect 46: 31. The method of embodiment 29 or 30, wherein the solution further comprises at least one extract from turmeric root and at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or at least 20% alcohol.

[0091] Aspect 47: 28. The method of embodiment 27, wherein the infection is selected from the group consisting of Staphylococcus spp., Streptococcus spp., Pseudomonas spp., Escherichia spp., Enterococcus spp., Proteus spp., Serratia spp., Enterobacter spp., Acetinobacter spp., and Klebsiella spp.

[0092] Aspect 48: The method according to any one of aspects 14 to 20 and 44 to 47, which is carried out on an animal.

[0093] Aspect 49: The method of embodiment 48, wherein the animal is selected from the group consisting of dogs, cats, cows, sheep, goats, llamas, pigs, horses, and donkeys.

[0094] Aspect 50: The composition according to any one of aspects 1 to 13 and 21 to 40, which can be used in the food service industry.

[0095] Aspect 51: The composition according to any one of aspects 1 to 13 and 21 to 40, which is applied to a catheter.

[0096] Aspect 52: 1. A composition comprising a solvatochromic indicator, the solvatochromic indicator is a pH responsive visual indicator; The composition further comprises at least one extract from turmeric root. composition.

[0097] Aspect 53: 53. The composition of embodiment 52, wherein the solvatochromic indicator is a betaine.

[0098] Aspect 54: 54. The composition of claim 52 or 53, wherein the solvatochromic indicator is an extract from at least one sugar beet bulb.

[0099] Aspect 55: 55. The composition of any one of aspects 52-54, further comprising at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or at least 20% alcohol.

[0100] Aspect 56: 56. The composition of any one of aspects 52-55, further comprising a filament.

[0101] Aspect 57: 57. The composition of any one of aspects 52-56, wherein the filament is in a form selected from the group consisting of sutures, gauze, gauze sponges, gauze bandages, non-adhesive pads, and non-adhesive wound dressings.

[0102] Aspect 58: 58. The composition of any one of aspects 52-57, further comprising a thin film coating encapsulating the filaments.

[0103] Aspect 59: Aspects 52-58, wherein the thin film coating comprises Parylene C.

[0104] Having now generally described the present disclosure, the same will be more readily understood by reference to the following examples, which are provided by way of illustration and are not intended to be limiting of the disclosure, unless otherwise specified. EXAMPLES

[0105] Example 1: Determine viable materials that change color with pH The bulb of at least one sugar beet plant is cut into pieces of approximately 200 g. The pieces are added to 400 milliliters (ml) of distilled water in a beaker and boiled at 400° C. A polyester cotton blend filament material is cut into 2 meter long strings and stirred into the boiling mixture and boiled for at least 30 minutes with frequent stirring. After boiling, the filaments are removed and dried on a non-absorbent surface. This process is repeated with various types of materials including yarn, polyester, cotton, vicryl, and monocryl.

[0106] After drying, each of the filament materials is treated with a solution that is either pH 5, pH 7 (neutral, control), or pH 9 to determine the intensity of the visual response during the solvatochromic reaction. The solution appears red at pH 5 and purple at pH 9. See FIG. 1A. The material treated with the pH 9 solution showed a color response that clearly distinguished between pH 9 and pH 5. The polyester cotton blend showed the most robust color response. See FIG. 1B and C. The yarn (75% viscose and 25% polyester) also showed a robust color change at pH 9. The color change response of the sugar beet extract is further tested by adding the extract to a cotton swab and testing against pH 5 and pH 9 solutions with the addition of water and 91% isopropyl alcohol as well. See FIG. 2. The addition of 91% isopropyl alcohol does not affect the color change response and aids in the solubility of the extract containing solution. The addition of extra water may reduce the color change response. See FIG. 2.

[0107] Example 2: Preparation of sugar beet extract A sugar beet extract is made by cutting 250 grams of sugar beet bulbs into small cubes. The sugar beet cubes are added to 600 ml of distilled water and boiled at 400° C. for at least 45 minutes. The mixture is removed from the heat, cooled, and then blended in an emulsion blender. Alternatively, the mixture can be sonicated or subjected to an equivalent treatment. The mixture is then returned to the heat and boiled for at least 30 minutes. The mixture is then removed from the heat and cooled. 100 ml of 91% isopropyl alcohol is added and the mixture is poured into a strainer to remove any remaining solid material. The filtration step can be repeated as necessary until all solid material is removed.

[0108] Example 3: Preparation of turmeric extract An extract of turmeric is made by cutting 500 grams of turmeric root into small cubes. The turmeric cubes are added to 1200 ml of distilled water and boiled at 400°C for at least 45 minutes. The mixture is removed from the heat, cooled, and then blended in an emulsion blender. Alternatively, the mixture can be sonicated or subjected to an equivalent treatment. The mixture is then returned to the heat and boiled for at least 30 minutes. The mixture is then removed from the heat and cooled. 100 ml of 91% isopropyl alcohol is added and the mixture is poured into a strainer to remove any remaining solid material. The filtration step can be repeated as necessary until all solid material has been removed.

[0109] Example 4: Preparation of Solvatochromic Mixtures Sugar beet extract and turmeric extract are prepared as described in Examples 2 and 3, respectively. A solvatochromic mixture containing approximately 10% isopropyl alcohol is prepared by mixing equal amounts of sugar beet extract and turmeric extract. The solvatochromic mixture can be used in a similar manner as the sugar beet extract described in Example 1.

[0110] Example 5: Solvatochromic mixtures contain antibacterial properties To determine whether the solvatochromic mixtures of Examples 1 and 2 have antibacterial properties, sugar beet bulb extracts were exposed to E. coli in culture. Petri dishes containing LB agar or similar are prepared using standard procedures. The petri dishes are inoculated with E. coli using standard laboratory procedures. Filament material is treated with the solvatochromic mixtures of Examples 1, 2, 3, or 4 and boiled together to sterilize. After a short cooling period, the filament material is placed on one half of the petri plate, the other half being a positive growth control. The plate is incubated at 37° C. for 12 hours. After incubation, the positive growth control side of the petri plate shows normal growth, while the side treated with the solvatochromic mixture shows no growth, confirming the antibacterial properties of the solvatochromic mixture. See FIG. 3.

[0111] Example 6: Development of materials containing solvatochromic mixtures Any form of sterile filament can be used to create a material containing a solvatochromic indicator that is useful for diagnosing bacterial infection in a biomedical environment or any other environment where bacterial growth is a problem, such as the food service industry.

[0112] For example, any material suitable for sutures can be used to make sutures containing a solvatochromic mixture. The filament material is added to a tube containing 35 ml of the solvatochromic mixture of Example 4. The material is immersed in the solvatochromic mixture at room temperature for 48 hours. After 48 hours, the material is removed from the solvatochromic mixture and allowed to dry on a non-absorbent surface.

[0113] Example 7: Thin film coating of treated filaments Materials made with the solvatochromic mixture in Example 5 can be treated with a thin film coating to confine the solvatochromic mixture while allowing interaction with the external environment. For example, a thin film coating of Parylene C (FIG. 4A) has been shown to work well with the solvatochromic mixture of the present disclosure, confine the solvatochromic mixture while being permeable enough to allow detection of pH in the external environment. Briefly, any deposition process can be used to apply the thin film coating. Here, the suture treated with the solvatochromic mixture is coated using a VSi Parylene C deposition process (FIG. 4B, VSi Parylene, Broomfield Colorado, 80021). Parylene C is deposited using a liquid-promoted adhesion protocol and subsequent coating as shown in FIG. 4B.

[0114] Parylene C treated sutures are sewn into a material that mimics skin. Three sets of sutures are set up, one treated with pH 5 solution, one treated with pH 9 solution, and one control. See Figure 5A-C. A solution of appropriate pH is added to the sutures as time zero, the control is untreated. See Figure 5D-F. After 10 minutes, a color change reaction can be seen in response to the pH 9 solution, while the pH 5 solution does not cause a color change (compare Figure 5G and 5I). Note that Parylene C still allows absorption of the pH 9 solution that has diffused underneath the artificial membrane. This demonstrates that the thin film coating does not affect uptake into the suture and subsequent color change (see subtle color change in the control stitch, Figure 5H).

[0115] Example 8: Identification of additional solvatochromic indicators useful for diagnosing bacterial infections Any material with solvatochromic properties that allow for a visible color change reaction in response to a basic pH environment can be used to create a solvatochromic indicator for use in the methods disclosed herein. For example, purple cabbage (Brassica oleracea, which has purple or red leaves), also known as red cabbage or blaukraut, has suitable properties and has been shown to cause a strong color change in response to a pH 9 environment. See Figure 6A.

[0116] 250 grams of purple cabbage leaves are cut into small strips and added to 600 ml of 400° C. distilled water and boiled for at least 45 minutes. The strips are removed from the heat and allowed to cool for 10 minutes. After the cooling period, an emulsion blender is used to disrupt the cell membranes. Sonication or any equivalent procedure can also be used to create the purple cabbage leaf extract. The mixture is then returned to the heat and boiled for an additional 30 minutes. After cooling for at least 10 minutes, 100 ml of 91% isopropyl alcohol is added to the solution. The mixture is poured into a strainer to remove any remaining solid material. The filtration step can be repeated as necessary until all solid material has been removed. The cabbage extract can be used alone or in combination with the sugar beet extract. See Figures 6A and 6B.

Claims

1. A composition for detecting bacterial infection comprising betaine, at least one extract derived from turmeric (Curcuma longa) root, and at least 10% alcohol, the composition being indicative of a bacterial infection.

2. The composition of claim 1, further comprising a filament.

3. 3. The composition of claim 2, wherein the filament is in a form selected from the group consisting of sutures, gauze, gauze sponges, gauze bandages, non-adherent pads, and non-adherent wound dressings.

4. 3. The composition of claim 2, further comprising a thin film coating encapsulating the filaments.

5. 5. The composition of claim 4, wherein the thin film coating comprises Parylene C.

6. 1. A composition for detecting a bacterial infection comprising a solvatochromic indicator, the solvatochromic indicator is a pH responsive visual indicator; the composition further comprises at least one extract from turmeric root; and The composition may be capable of indicating a bacterial infection. The composition.

7. 7. The composition of claim 6, wherein the solvatochromic indicator comprises a betaine.

8. 7. The composition of claim 6, wherein the solvatochromic indicator comprises an extract from at least one sugar beet (Beta vulgaris) bulb.

9. 7. The composition of claim 6, further comprising filaments.

10. 10. The composition of claim 9, wherein the filament is in a form selected from the group consisting of sutures, gauze, gauze sponges, gauze bandages, non-adherent pads, and non-adherent wound dressings.

11. 10. The composition of claim 9, further comprising a thin film coating encapsulating the filaments.

12. 12. The composition of claim 11, wherein the thin film coating comprises Parylene C.

13. Preparing a first extract from at least one sugar beet bulb; preparing a second extract from at least one turmeric root; combining the first extract and the second extract in a solution comprising alcohol; applying the solution to the filaments; and contacting the wound site with the filament; A method comprising:

14. The method of claim 13 , further comprising encapsulating the filament in a thin film coating.

15. The method of claim 14 , wherein the thin film coating comprises Parylene C.

16. 14. The method of claim 13, wherein the filament is in a form selected from the group consisting of a suture, a gauze sponge, a gauze bandage, a non-adherent pad, and a non-adherent wound dressing.

17. 14. The method of claim 13, wherein the first extract and the second extract are prepared by sonication or an equivalent process.

18. The composition of claim 1, wherein the detection of a bacterial infection includes detection of a bacterial infection at a wound site.

19. The composition of claim 6, wherein the detection of a bacterial infection includes detection of a bacterial infection at a wound site.

20. The method of claim 13, further comprising detecting a bacterial infection.

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