ENHANCED WOUND HEALING USING A NANOEMULGEL FORMULATION AND METHOD THEREOF

Abstract

Disclosed herein is an enhanced wound healing using a nanoemulgel formulation and method thereof (100) that comprises asiatic acid, solubilized in a mixture of virgin coconut oil and isopropyl myristate in a 1:1 ratio to form an organic phase, a surfactant mixture, comprising tween 20 and span 80 in a 4:1 ratio, incorporated into the organic phase to create a stable emulsion, an aqueous phase, into which the organic phase and surfactant mixture are transferred and sonicated to form a nano emulsion with reduced globule size and enhanced surface area, chitosan, prepared as a 0.1% solution in distilled water containing 0.25% glacial acetic acid, added dropwise to the nano emulsion under continuous stirring to form a chitosan-coated nano emulsion, carbopol 934, incorporated into the chitosan-coated nano emulsion to convert it into a nanoemulgel with enhanced viscosity and a gel-like consistency suitable for wound application.

Specification

Description:FIELD OF DISCLOSURE
[0001] The present disclosure generally relates to the field of pharmaceutical compositions, more specifically, relates an enhanced wound healing using a nanoemulgel formulation and method thereof.
BACKGROUND OF THE DISCLOSURE
[0002] The present formulation is utilizing nanoemulgel technology to deliver active ingredients, allowing deeper penetration into the wound site. This feature ensures that the ingredients such as Asiatic acid, chitosan, and virgin coconut oil are reaching deeper layers of tissue, promoting faster and more efficient healing compared to traditional surface-level treatments. This depth of penetration is particularly beneficial in treating chronic or deeper wounds.
[0003] The combination of Asiatic acid, chitosan, and virgin coconut oil is providing a synergistic effect that enhances wound healing. Asiatic acid stimulates collagen production, chitosan promotes cellular regeneration, and virgin coconut oil offers anti-inflammatory and antimicrobial properties. This unique formulation offers comprehensive wound care by accelerating the healing process, reducing inflammation, and minimizing the risk of infection.
[0004] The nanoemulgel system is offering better stability and prolonged release of active ingredients, which improves the efficiency of the treatment. This sustained release ensures continuous delivery of therapeutic agents over time, reducing the frequency of application and improving patient compliance. The formulation is also characterized by excellent spreadability and skin adherence, making it easy to apply on various wound types.
[0005] Current wound care products such as ointments and creams are typically limited in their ability to penetrate beyond the surface layers of the skin. These formulations are often only treating superficial wounds effectively, leaving deeper or chronic wounds untreated. As a result, these products are ineffective for more severe wounds, requiring more frequent and prolonged applications.
[0006] Many existing products are containing synthetic chemicals that can cause irritation, allergic reactions, or other adverse effects, especially in sensitive skin types. The use of preservatives, artificial fragrances, and harsh chemicals is compromising the comfort and recovery of patients. This irritation not only delays healing but also increases the risk of complications.
[0007] Traditional wound healing products are often lacking strong antimicrobial properties, making wounds more vulnerable to infections. This limitation is especially problematic in cases of diabetic ulcers or compromised immune systems, where infection control is critical. Without sufficient antimicrobial action, these products are unable to offer comprehensive wound care, leading to slower healing and increased medical complications
[0008] Thus, in light of the above-stated discussion, there exists a need for an enhanced wound healing using a nanoemulgel formulation and method thereof.
SUMMARY OF THE DISCLOSURE
[0009] The following is a summary description of illustrative embodiments of the invention. It is provided as a preface to assist those skilled in the art to more rapidly assimilate the detailed design discussion which ensues and is not intended in any way to limit the scope of the claims which are appended hereto in order to particularly point out the invention.
[0010] According to illustrative embodiments, the present disclosure focuses on a method for enhancing wound healing using a nanoemulgel formulation which overcomes the above-mentioned disadvantages or provide the users with a useful or commercial choice.
[0011] An objective of the present disclosure is to provide a nanoemulgel formulation that enhances the bioavailability and therapeutic efficacy of Asiatic acid, chitosan, and virgin coconut oil for promoting faster wound healing.
[0012] Another objective of the present disclosure is to develop a stable nanoemulgel that ensures prolonged release of active ingredients, offering sustained therapeutic effects over an extended period of time.
[0013] Another objective of the present disclosure is to promote deeper penetration of active ingredients into the skin layers to target deeper wounds and facilitate faster tissue regeneration.
[0014] Another objective of the present disclosure is to utilize the anti-inflammatory properties of virgin coconut oil and chitosan to reduce swelling, redness, and irritation in wounded skin areas.
[0015] Another objective of the present disclosure is to enhance collagen synthesis and cellular proliferation through the inclusion of Asiatic acid, accelerating wound closure and tissue repair.
[0016] Another objective of the present disclosure is to improve the antimicrobial properties of the wound treatment by leveraging chitosan's antibacterial effects, reducing the risk of infections during the healing process.
[0017] Another objective of the present disclosure is to offer a formulation with high spreadability and skin adherence, ensuring easy application and effective coverage of wounds for various wound types and sizes.
[0018] Another objective of the present disclosure is to provide a natural and biocompatible wound treatment option, reducing the risk of irritation, allergic reactions, and other adverse effects that are commonly associated with synthetic chemicals in conventional products.
[0019] Yet another objective of the present disclosure is to minimize the frequency of application required by offering a formulation with controlled release properties, improving patient compliance and convenience.
[0020] Yet another objective of the present disclosure is to optimize the nanoemulgel formulation using advanced techniques like ultrasonication and design expert software, ensuring precise control over particle size and distribution for maximum therapeutic benefits.
[0021] In light of the above, in one aspect of the present disclosure, an enhanced wound healing using a nanoemulgel formulation is disclosed herein. The formulation comprises asiatic acid, solubilized in a mixture of virgin coconut oil and isopropyl myristate in a 1:1 ratio to form an organic phase. The formulation includes a surfactant mixture, comprising tween 20 and span 80 in a 4:1 ratio, incorporated into the organic phase to create a stable emulsion. The formulation also includes an aqueous phase, into which the organic phase and surfactant mixture are transferred and sonicated to form a nano emulsion with reduced globule size and enhanced surface area. The formulation also includes chitosan, prepared as a 0.1% solution in distilled water containing 0.25% glacial acetic acid, added dropwise to the nano emulsion under continuous stirring to form a chitosan-coated nano emulsion. The formulation also includes carbopol 934, incorporated into the chitosan-coated nano emulsion to convert it into a nanoemulgel with enhanced viscosity and a gel-like consistency suitable for wound application.
[0022] In one embodiment, the nano emulsion has a globule size ranging between 100 nm to 200 nm.
[0023] In one embodiment, the chitosan-coated nano emulsion has a polydispersity index (PDI) less than 0.3.
[0024] In one embodiment, the asiatic acid is present in a concentration of 0.5% to 2.0% by weight.
[0025] In one embodiment, the virgin coconut oil and isopropyl myristate in the organic phase are in a 1:1 ratio.
[0026] In one embodiment, the nanoemulgel formed by incorporating Carbopol 934 has a viscosity ranging from 2,000 to 10,000.
[0027] In one embodiment, the surfactant mixture of Tween 20 and Span 80 is used in a 4:1 ratio.
[0028] In one embodiment, the chitosan coating is applied as a 0.1% solution prepared in distilled water with 0.25% glacial acetic acid.
[0029] In one embodiment, the nanoemulgel exhibits a pH range between 5.5 and 7.0.
[0030] In light of the above, in one aspect of the present disclosure, a method for preparing an enhanced wound healing nanoemulgel formulation is disclosed herein. The method comprising solubilizing asiatic acid in an organic phase comprising virgin coconut oil and isopropyl myristate to create a homogenous solution. The method includes incorporating a surfactant mixture comprising tween 20 and span 80 into the organic phase to form an emulsion. The method also includes transferring the organic phase and surfactant mixture into an aqueous phase and sonicating the mixture to produce a nano emulsion. The method also includes preparing a 0.1% chitosan solution in distilled water containing 0.25% glacial acetic acid, and adding the chitosan solution dropwise to the nano emulsion under continuous stirring to coat the nano emulsion particles. The method also includes incorporating Carbopol 934 into the chitosan-coated nano emulsion to convert the nano emulsion into a nanoemulgel.
[0031] These and other advantages will be apparent from the present application of the embodiments described herein.
[0032] The preceding is a simplified summary to provide an understanding of some embodiments of the present invention. This summary is neither an extensive nor exhaustive overview of the present invention and its various embodiments. The summary presents selected concepts of the embodiments of the present invention in a simplified form as an introduction to the more detailed description presented below. As will be appreciated, other embodiments of the present invention are possible utilizing, alone or in combination, one or more of the features set forth above or described in detail below.
[0033] These elements, together with the other aspects of the present disclosure and various features are pointed out with particularity in the claims annexed hereto and form a part of the present disclosure. For a better understanding of the present disclosure, its operating advantages, and the specified object attained by its uses, reference should be made to the accompanying drawings and descriptive matter in which there are illustrated exemplary embodiments of the present disclosure.
BRIEF DESCRIPTION OF THE DRAWINGS
[0034] To describe the technical solutions in the embodiments of the present disclosure or in the prior art more clearly, the following briefly describes the accompanying drawings required for describing the embodiments or the prior art. Apparently, the accompanying drawings in the following description merely show some embodiments of the present disclosure, and a person of ordinary skill in the art can derive other implementations from these accompanying drawings without creative efforts. All of the embodiments or the implementations shall fall within the protection scope of the present disclosure.
[0035] The advantages and features of the present disclosure will become better understood with reference to the following detailed description taken in conjunction with the accompanying drawing, in which:
[0036] FIG. 1 illustrates a block diagram of an enhanced wound healing using a nanoemulgel formulation, in accordance with an exemplary embodiment of the present disclosure;
[0037] FIG. 2 illustrates a flowchart of a method for preparing an enhanced wound healing nanoemulgel formulation, in accordance with an exemplary embodiment of the present disclosure;
[0038] FIG. 3 illustrates a perspective view of graphical abstract of the work depicting the preparation process, in accordance with an exemplary embodiment of the present disclosure;
[0039] FIG. 4 illustrates a perspective view of the cumulative drug release of CS-ASA-NEG vs Marketed product. The data indicate that CS-ASA-NEG had better in-vitro release profile compared to the marketed product. The data represent Mean±S.D, in accordance with an exemplary embodiment of the present disclosure;
[0040] FIG. 5A illustrates a perspective view of the cytotoxic effect (0-1μg/ml) for 24hrs on HEK293 cells, in accordance with an exemplary embodiment of the present disclosure;
[0041] FIG. 5B illustrates a perspective view of the cytotoxic effect of 0-1μg/ml) for 24hrs on RAW 264.7) without, in accordance with an exemplary embodiment of the present disclosure;
[0042] FIG. 5C illustrates a perspective view of the LPS, in accordance with an exemplary embodiment of the present disclosure;
[0043] FIG. 5D illustrates a perspective view of the cell morphology observed under a phase contrast Microscope (Leica Software), in accordance with an exemplary embodiment of the present disclosure;
[0044] FIG. 6A illustrates a perspective view of the micrographs of cell migration observed at 5X magnification (Phase Contrast inverted microscope-Leica), in accordance with an exemplary embodiment of the present disclosure;
[0045] FIG. 6B illustrates a perspective view of the rates of cell migration were calculated from the area of wound coverage over a period of 24 h relative to the scarification area at 0 h.. Performed on in-vitro cell lines (RAW264.7). Data presented are in mean± standard deviation. Significance was measured using one-way ANOVA. #p<0.05, ##p<0.01 and ###p<0.001 vs control group. *p<0.05 and **p<0.01 and ***p<0.001 vs LPS-control group, in accordance with an exemplary embodiment of the present disclosure;
[0046] FIG. 7A illustrates a perspective view of the in-vivo skin irritation studies in Wistar rats, in accordance with an exemplary embodiment of the present disclosure;
[0047] FIG. 7B illustrates a perspective view of the in-vivo wound contraction, in accordance with an exemplary embodiment of the present disclosure;
[0048] FIG. 7C illustrates a perspective view of the excision model wound healing for Control, Standard, NEG and CS-ASA-NEG treated Wistar rats, in accordance with an exemplary embodiment of the present disclosure; and
[0049] FIG. 8 illustrates a perspective view of the histopathology of the healed skin after 21 days of treatment, in accordance with an exemplary embodiment of the present disclosure.
[0050] Like reference, numerals refer to like parts throughout the description of several views of the drawing.
[0051] The enhanced wound healing using a nanoemulgel formulation and method thereof is illustrated in the accompanying drawings, which like reference letters indicate corresponding parts in the various figures. It should be noted that the accompanying figure is intended to present illustrations of exemplary embodiments of the present disclosure. This figure is not intended to limit the scope of the present disclosure. It should also be noted that the accompanying figure is not necessarily drawn to scale.
DETAILED DESCRIPTION OF THE DISCLOSURE
[0052] The following is a detailed description of embodiments of the disclosure depicted in the accompanying drawings. The embodiments are in such detail as to communicate the disclosure. However, the amount of detail offered is not intended to limit the anticipated variations of embodiments; on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the present disclosure.
[0053] In the following description, numerous specific details are set forth in order to provide a thorough understanding of the embodiments of the present disclosure. It may be apparent to one skilled in the art that embodiments of the present disclosure may be practiced without some of these specific details.
[0054] Various terms as used herein are shown below. To the extent a term is used, it should be given the broadest definition persons in the pertinent art have given that term as reflected in printed publications and issued patents at the time of filing.
[0055] The terms "a" and "an" herein do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced items.
[0056] The terms "having", "comprising", "including", and variations thereof signify the presence of a component.
[0057] Referring now to FIG. 1 to FIG. 8 to describe various exemplary embodiments of the present disclosure. FIG. 1 illustrates a flowchart of an enhanced wound healing using a nanoemulgel formulation 100, in accordance with an exemplary embodiment of the present disclosure.
[0058] The formulation 100 may include asiatic acid, solubilized in a mixture of virgin coconut oil and isopropyl myristate in a 1:1 ratio to form an organic phase, a surfactant mixture, comprising tween 20 and span 80 in a 4:1 ratio, incorporated into the organic phase to create a stable emulsion, an aqueous phase, into which the organic phase and surfactant mixture are transferred and sonicated to form a nano emulsion with reduced globule size and enhanced surface area, chitosan, prepared as a 0.1% solution in distilled water containing 0.25% glacial acetic acid, added dropwise to the nano emulsion under continuous stirring to form a chitosan-coated nano emulsion, carbopol 934, incorporated into the chitosan-coated nano emulsion to convert it into a nanoemulgel with enhanced viscosity and a gel-like consistency suitable for wound application.
[0059] The nanoemulsion has a globule size ranging between 100 nm to 200 nm.
[0060] The chitosan-coated nanoemulsion has a polydispersity index (PDI) less than 0.3.
[0061] The asiatic acid is present in a concentration of 0.5% to 2.0% by weight.
[0062] The virgin coconut oil and isopropyl myristate in the organic phase are in a 1:1 ratio.
[0063] The nanoemulgel formed by incorporating Carbopol 934 has a viscosity ranging from 2,000 to 10,000.
[0064] The surfactant mixture of Tween 20 and Span 80 is used in a 4:1 ratio.
[0065] The chitosan coating is applied as a 0.1% solution prepared in distilled water with 0.25% glacial acetic acid.
[0066] The nanoemulgel exhibits a pH range between 5.5 and 7.0.
[0067] The method may include solubilizing asiatic acid in an organic phase comprising virgin coconut oil and isopropyl myristate to create a homogenous solution, incorporating a surfactant mixture comprising tween 20 and span 80 into the organic phase to form an emulsion, transferring the organic phase and surfactant mixture into an aqueous phase and sonicating the mixture to produce a nano emulsion, preparing a 0.1% chitosan solution in distilled water containing 0.25% glacial acetic acid, and adding the chitosan solution dropwise to the nano emulsion under continuous stirring to coat the nano emulsion particles, incorporating Carbopol 934 into the chitosan-coated nano emulsion to convert the nano emulsion into a nanoemulgel.
[0068] At 102, asiatic acid, solubilized in a mixture of virgin coconut oil and isopropyl myristate in a 1:1 ratio to form an organic phase.
[0069] At 104, a surfactant mixture, comprising tween 20 and span 80 in a 4:1 ratio, incorporated into the organic phase to create a stable emulsion.
[0070] At 106, an aqueous phase, into which the organic phase and surfactant mixture are transferred and sonicated to form a nano emulsion with reduced globule size and enhanced surface area.
[0071] At 108, chitosan, prepared as a 0.1% solution in distilled water containing 0.25% glacial acetic acid, added dropwise to the nano emulsion under continuous stirring to form a chitosan-coated nano emulsion.
[0072] At 110, carbopol 934, incorporated into the chitosan-coated nano emulsion to convert it into a nanoemulgel with enhanced viscosity and a gel-like consistency suitable for wound application.
[0073] FIG. 2 illustrates a flowchart of a method for preparing an enhanced wound healing nanoemulgel formulation, in accordance with an exemplary embodiment of the present disclosure.
[0074] At 202, solubilize asiatic acid in an organic phase comprising virgin coconut oil and isopropyl myristate to create a homogenous solution.
[0075] At 204, incorporate a surfactant mixture comprising tween 20 and span 80 into the organic phase to form an emulsion.
[0076] At 206, transfer the organic phase and surfactant mixture into an aqueous phase and sonicating the mixture to produce a nano emulsion.
[0077] At 208, prepare a 0.1% chitosan solution in distilled water containing 0.25% glacial acetic acid, and adding the chitosan solution dropwise to the nano emulsion under continuous stirring to coat the nano emulsion particles.
[0078] At 210, incorporate Carbopol 934 into the chitosan-coated nano emulsion to convert the nano emulsion into a nanoemulgel.
[0079] FIG. 3 illustrates a perspective view of graphical abstract of the work depicting the preparation process, in accordance with an exemplary embodiment of the present disclosure.
[0080] At 302, the chitosan-coated asiatic acid-loaded nanoemulgel formulation (cs-asa-neg) nanoemulgel is prepared by combining virgin coconut oil and isopropyl myristate with a surfactant mixture and transferring it to an aqueous phase. Ultrasonication is used to create a nano emulsion, which is then coated with chitosan and mixed with Carbopol 934 to achieve the desired gel-like consistency. This process results in a stable and effective nanoemulgel formulation for wound healing.
[0081] At 304, the organic phase is a crucial component of the nanoemulgel formulation. It consists of a mixture of virgin coconut oil and isopropyl myristate. These oils provide a hydrophobic environment that facilitates the solubilization of Asiatic acid, a lipophilic compound. The combination of these oils also contributes to the overall stability and biocompatibility of the nanoemulgel.
[0082] At 306, the aqueous phase serves as the continuous medium in the nanoemulgel formulation. It is typically composed of water or a water-based solvent. The aqueous phase provides a suitable environment for the dispersion of the nano emulsion droplets and ensures the overall stability and biocompatibility of the formulation. The choice of aqueous phase can influence factors such as drug solubility, viscosity, and the overall properties of the nanoemulgel.
[0083] At 308, the crude emulsion is the initial stage in the preparation of the nanoemulgel. It involves combining the organic phase, consisting of virgin coconut oil and isopropyl myristate, with the surfactant mixture of Tween 20 and Span 80. This mixture is then transferred to the aqueous phase and subjected to mechanical agitation, such as propeller mixing, followed by ultrasonication. This process results in the formation of a coarse emulsion with larger droplets, which serves as the starting point for further refinement and optimization.
[0084] At 310, the nano emulsion is a key component of the formulation. It is a stable dispersion of oil droplets in water, with droplet sizes in the nanometre range. The small size of the droplets enhances the surface area, improving drug delivery and absorption. Nano emulsions are also known for their increased stability, transparency, and tenable rheological properties, making them ideal for topical applications like wound healing.
[0085] At 312, ultrasonication is a process that involves the application of high-frequency sound waves to a liquid or suspension. In the context of nano emulsion formation, ultrasonication is used to break down larger droplets into smaller, more uniform droplets. The high-frequency sound waves create cavitation bubbles that collapse, generating intense shear forces that disrupt the droplets. This process helps to reduce the droplet size and improve the stability of the nano emulsion.
[0086] At 314, the optimized nano emulsion (ASA-NE) is combined with a 0.1% chitosan solution prepared in distilled water containing glacial acetic acid. This mixture is then stirred overnight to allow the chitosan to coat the nano emulsion droplets. The chitosan coating enhances the stability of the nanoemulgel and provides additional functional properties, such as improved biocompatibility and controlled drug release. This step is crucial in transforming the nano emulsion into a nanoemulgel suitable for topical application.
[0087] At 316, the chitosan-coated nanoemulgel is the final product of the formulation process. The chitosan coating provides additional stability to the nano emulsion, preventing aggregation of the droplets and ensuring a longer shelf life. Chitosan is a natural biopolymer with excellent biocompatibility and mucoadhesive properties, which enhance the adherence of the nanoemulgel to the wound site, promoting effective drug delivery and retention.
[0088] At 318, chitosan-coated asiatic acid-loaded nanoemulgel formulation (cs-asa-ne) is the abbreviation for the chitosan-coated asiatic acid-loaded nanoemulgel formulation (cs-asa-ne). This final product incorporates the beneficial properties of Asiatic acid, chitosan, and the nanoemulgel delivery system. chitosan-coated asiatic acid-loaded nanoemulgel formulation (cs-asa-ne) is designed to provide enhanced wound healing by delivering Asiatic acid to the wound site, promoting collagen synthesis, and reducing inflammation. The chitosan coating ensures controlled release and improved stability, making chitosan-coated asiatic acid-loaded nanoemulgel formulation (cs-asa-ne) a promising candidate for wound care applications.
[0089] At 320, Carbopol is a polymer that is added to the nanoemulgel formulation to increase its viscosity and provide a gel-like consistency. This gel-like consistency improves the adherence of the formulation to the wound site, ensuring prolonged contact with the affected area and enhanced drug delivery. The addition of 1% carbopol helps to optimize the rheological properties of the nanoemulgel, making it suitable for topical application and preventing premature drainage from the wound.
[0090] At 322, Triethanolamine (TEA) is a common pH adjuster used in formulations to neutralize the acidic nature of the solution. In this case, TEA is added dropwise to the nanoemulgel formulation to adjust the pH to the desired range, typically between 5.5 and 7.0. This pH adjustment is important for ensuring the stability, biocompatibility, and effectiveness of the nanoemulgel. By carefully controlling the pH, the formulation can be optimized for topical application and minimize any potential irritation to the wound site.
[0091] At 324, chitosan-coated asiatic acid-loaded nanoemulgel formulation (cs-asa-neg) is the final product of the nanoemulgel formulation. It incorporates the beneficial properties of Asiatic acid, chitosan, and the nanoemulgel delivery system. Chitosan-coated asiatic acid-loaded nanoemulgel formulation (cs-asa-neg) is designed to provide enhanced wound healing by delivering Asiatic acid to the wound site, promoting collagen synthesis, and reducing inflammation. The chitosan coating ensures controlled release and improved stability, making chitosan-coated asiatic acid-loaded nanoemulgel formulation (cs-asa-neg) a promising candidate for wound care applications.
[0092] At 326, In-vitro analysis involves studying the properties and behaviour of the nanoemulgel formulation under controlled laboratory conditions. This includes evaluating its drug release profile, ex vivo permeation through skin or other tissues, and cytotoxicity assessment. By conducting in-vitro studies, researchers can gain insights into the formulation's characteristics and predict its potential performance in vivo. These experiments help to optimize the formulation and ensure its safety and efficacy before proceeding to animal or clinical trials.
[0093] At 328, Chitosan is a natural biopolymer derived from chitin, a component of the exoskeletons of crustaceans and insects. It is a biodegradable and biocompatible material with several beneficial properties, including antibacterial activity, haemostatic properties, and the ability to promote tissue regeneration. In the nanoemulgel formulation, chitosan serves as a coating for the nanoemulsion droplets, enhancing their stability and providing additional functional benefits for wound healing.
[0094] At 330, Asiatic acid is a bioactive compound extracted from the plant centella asiatica, commonly known as gotu kola. It possesses potent anti-inflammatory, antioxidant, and wound healing properties. Asiatic acid stimulates collagen synthesis, promotes angiogenesis, and inhibits inflammation, all of which are essential for effective wound healing. Its inclusion in the nanoemulgel formulation enhances the overall therapeutic efficacy and accelerates the healing process.
[0095] At 332, A cytotoxicity assay is a laboratory technique used to assess the toxicity of a substance, such as a drug or chemical, on cells. It involves exposing cells to the substance and measuring their viability or response. In the case of the nanoemulgel formulation, cytotoxicity assays can be conducted to determine its potential toxicity to cells and ensure its safety for topical application. This information is crucial for evaluating the biocompatibility of the formulation and identifying any potential adverse effects.
[0096] At 334, A cell migration assay is a laboratory technique used to study the movement of cells in response to various stimuli. It can provide insights into the wound healing process, as cell migration is essential for tissue repair and regeneration. In the context of the nanoemulgel formulation, cell migration assays can be conducted to evaluate its ability to promote cell movement and facilitate wound closure. These assays can help to assess the formulation's efficacy in stimulating tissue repair and regeneration.
[0097] At 336, In vivo evaluation involves assessing the efficacy and safety of the nanoemulgel formulation in a living organism, such as an animal model. This type of evaluation provides valuable information on the formulation's performance in a more complex and realistic environment. In vivo studies can assess wound healing rates, inflammation levels, and any potential adverse effects. By conducting in vivo experiments, researchers can validate the findings from in vitro studies and evaluate the clinical relevance of the nanoemulgel formulation for wound healing applications.
[0098] At 338, A skin irritation study is conducted to assess the potential of a substance to cause irritation or damage to the skin. In the case of the nanoemulgel formulation, this study is essential to ensure its safety for topical application. The study involves applying the formulation to a specific area of the skin and monitoring for any signs of irritation, such as redness, swelling, or pain. By evaluating the skin irritation potential, researchers can determine the safety of the nanoemulgel for use in wound healing applications.
[0099] At 340, A wound contraction study is conducted to assess the ability of a wound to close and heal over time. This is a key indicator of effective wound healing. In vivo studies involving animal models or human subjects are typically used to evaluate wound contraction rates. The study involves monitoring the size of the wound over time and comparing it to a control group. By measuring the rate of wound contraction, researchers can assess the efficacy of the nanoemulgel formulation in promoting tissue repair and regeneration.
[0100] At 342, A histopathological study involves examining tissue samples under a microscope to assess their structure and cellular composition. In the context of wound healing, histopathological analysis can provide valuable insights into the healing process, including the formation of granulation tissue, collagen deposition, and the presence of inflammatory cells. By examining tissue samples from treated and control groups, researchers can compare the healing response and evaluate the efficacy of the nanoemulgel formulation in promoting tissue regeneration and preventing complications.
[0101] FIG. 4 illustrates a perspective view of the cumulative drug release of CS-ASA-NEG vs Marketed product. The data indicate that CS-ASA-NEG had better in-vitro release profile compared to the marketed product. The data represent Mean±S.D, in accordance with an exemplary embodiment of the present disclosure.
[0102] FIG. 5A illustrates a perspective view of the cytotoxic effect (0-1μg/ml) for 24hrs on HEK293 cells, in accordance with an exemplary embodiment of the present disclosure.
[0103] FIG. 5B illustrates a perspective view of the cytotoxic effect of 0-1μg/ml) for 24hrs on RAW 264.7) without, in accordance with an exemplary embodiment of the present disclosure.
[0104] FIG. 5C illustrates a perspective view of the LPS, in accordance with an exemplary embodiment of the present disclosure.
[0105] FIG. 5D illustrates a perspective view of the cell morphology observed under a phase contrast Microscope (Leica Software), in accordance with an exemplary embodiment of the present disclosure.
[0106] FIG. 6A illustrates a perspective view of the micrographs of cell migration observed at 5X magnification (Phase Contrast inverted microscope-Leica), in accordance with an exemplary embodiment of the present disclosure.
[0107] FIG. 6B illustrates a perspective view of the rates of cell migration were calculated from the area of wound coverage over a period of 24 h relative to the scarification area at 0h.. Performed on in-vitro cell lines (RAW264.7). Data presented are in mean± standard deviation. Significance was measured using one-way ANOVA. #p<0.05, ##p<0.01 and ###p<0.001 vs control group. *p<0.05 and **p<0.01 and ***p<0.001 vs LPS-control group, in accordance with an exemplary embodiment of the present disclosure.
[0108] FIG. 7A illustrates a perspective view of the in-vivo skin irritation studies in Wistar rats, in accordance with an exemplary embodiment of the present disclosure.
[0109] FIG. 7B illustrates a perspective view of the in-vivo wound contraction, in accordance with an exemplary embodiment of the present disclosure.
[0110] FIG. 7C illustrates a perspective view of the excision model wound healing for Control, Standard, NEG and CS-ASA-NEG treated Wistar rats, in accordance with an exemplary embodiment of the present disclosure.
[0111] FIG. 8 illustrates a perspective view of the histopathology of the healed skin after 21 days of treatment, in accordance with an exemplary embodiment of the present disclosure.
[0112] While the invention has been described in connection with what is presently considered to be the most practical and various embodiments, it will be understood that the invention is not to be limited to the disclosed embodiments, but on the contrary, is intended to cover various modifications and equivalent arrangements included within the scope of the appended claims. A person of ordinary skill in the art may be aware that, in combination with the examples described in the embodiments disclosed in this specification, units and algorithm steps may be implemented by electronic hardware, computer software, or a combination thereof.
[0113] The foregoing descriptions of specific embodiments of the present disclosure have been presented for purposes of illustration and description. They are not intended to be exhaustive or to limit the present disclosure to the precise forms disclosed, and many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described to best explain the principles of the present disclosure and its practical application, and to thereby enable others skilled in the art to best utilize the present disclosure and various embodiments with various modifications as are suited to the particular use contemplated. It is understood that various omissions and substitutions of equivalents are contemplated as circumstances may suggest or render expedient, but such omissions and substitutions are intended to cover the application or implementation without departing from the scope of the present disclosure.
[0114] Disjunctive language such as the phrase "at least one of X, Y, Z," unless specifically stated otherwise, is otherwise understood with the context as used in general to present that an item, term, etc., may be either X, Y, or Z, or any combination thereof (e.g., X, Y, and/or Z). Thus, such disjunctive language is not generally intended to, and should not, imply that certain embodiments require at least one of X, at least one of Y, or at least one of Z to each be present.
[0115] In a case that no conflict occurs, the embodiments in the present disclosure and the features in the embodiments may be mutually combined. The foregoing descriptions are merely specific implementations of the present disclosure, but are not intended to limit the protection scope of the present disclosure. Any variation or replacement readily figured out by a person skilled in the art within the technical scope disclosed in the present disclosure shall fall within the protection scope of the present disclosure. Therefore, the protection scope of the present disclosure shall be subject to the protection scope of the claims.
, Claims:I/We Claim:
1. An enhanced wound healing using a nanoemulgel formulation and method thereof, the method (100) comprising:
asiatic acid, solubilized in a mixture of virgin coconut oil and isopropyl myristate in a 1:1 ratio to form an organic phase;
a surfactant mixture, comprising tween 20 and span 80 in a 4:1 ratio, incorporated into the organic phase to create a stable emulsion;
an aqueous phase, into which the organic phase and surfactant mixture are transferred and sonicated to form a nano emulsion with reduced globule size and enhanced surface area;
chitosan, prepared as a 0.1% solution in distilled water containing 0.25% glacial acetic acid, added dropwise to the nano emulsion under continuous stirring to form a chitosan-coated nano emulsion;
carbopol 934, incorporated into the chitosan-coated nano emulsion to convert it into a nanoemulgel with enhanced viscosity and a gel-like consistency suitable for wound application.
2. The formulation (100) as claimed in claim 1, wherein the nano emulsion has a globule size ranging between 100 nm to 200 nm.
3. The formulation (100) as claimed in claim 1, wherein the chitosan-coated nano emulsion has a polydispersity index (PDI) less than 0.3.
4. The formulation (100) as claimed in claim 1, wherein the Asiatic acid is present in a concentration of 0.5% to 2.0% by weight.
5. The formulation (100) as claimed in claim 1, wherein the virgin coconut oil and isopropyl myristate in the organic phase are in a 1:1 ratio.
6. The formulation (100) as claimed in claim 1, wherein the nanoemulgel formed by incorporating Carbopol 934 has a viscosity ranging from 2,000 to 10,000.
7. The formulation (100) as claimed in claim 1, wherein the surfactant mixture of Tween 20 and Span 80 is used in a 4:1 ratio.
8. The formulation (100) claimed in claim 1, wherein the chitosan coating is applied as a 0.1% solution prepared in distilled water with 0.25% glacial acetic acid.
9. The formulation (100) as claimed in claim 1, wherein the nanoemulgel exhibits a pH range between 5.5 and 7.0.
10. A method for preparing an enhanced wound healing nanoemulgel formulation (100), the method (100) comprising:
solubilizing asiatic acid in an organic phase comprising virgin coconut oil and isopropyl myristate to create a homogenous solution;
incorporating a surfactant mixture comprising tween 20 and span 80 into the organic phase to form an emulsion;
transferring the organic phase and surfactant mixture into an aqueous phase and sonicating the mixture to produce a nano emulsion;
preparing a 0.1% chitosan solution in distilled water containing 0.25% glacial acetic acid, and adding the chitosan solution dropwise to the nano emulsion under continuous stirring to coat the nano emulsion particles;
incorporating Carbopol 934 into the chitosan-coated nano emulsion to convert the nano emulsion into a nanoemulgel;

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