Wound dressing composition containing fucoidan and olive leaf extract

Abstract

The present invention relates to a wound dressing composition that has a skin wound healing effect by promoting the growth of epidermal basal layer stem cells. The present invention provides a wound dressing composition containing 60-70 wt% glycerin, 28-39 wt% honey, 0.2-3.0 wt% fucoidan, 0.2-3.0 wt% olive leaf extract, and 0.1-0.3 wt% xanthan gum.

Description

[Technical Field]

[0001] The present invention relates to a wound dressing composition that has a skin wound healing effect by promoting the growth of stem cells in the basal layer of the epidermis of the skin. [Background technology]

[0002] Korean Patent No. 10-2088767 is an invention related to a "composition for increasing the biological activity of stem cells using mixture 4F," which provides a composition for inhibiting aging, promoting proliferation, or inducing differentiation of stem cells, containing fucoidan, tauroursodeoxycholic acid, oleuropein, and vascular endothelial growth factor as active ingredients.

[0003] The "stem cells using Mixture 4F" shown in the Korean registered patent not only acquire undifferentiated stemness but also have the effect of improving cell proliferation and migration ability. After transplantation of stem cells into the body, it can improve cell survival and engraftment rate and enhance vascular and tissue regeneration ability, so it can be used in a variety of fields such as stem cell differentiation and the prevention or treatment of ischemic diseases.

[0004] Meanwhile, human skin tissue is composed of the epidermis, dermis, and hypodermis. The epidermis can be divided into the stratum corneum, which is made up of dead cells, and the living epidermis, which can be divided into four layers (stratified epithelial tissue): the stratum lucidum, stratum granulosum, stratum spinosum, and stratum basale.

[0005] When the skin is injured, stem cells are supplied to the wound from the basal layer, and the wound is healed through the division and growth of the stem cells. Therefore, utilizing Korean Patent No. 10-2088767 and related research results, we have developed a wound dressing composition that can achieve skin wound healing effects by promoting the growth of stem cells in the basal layer. Summary of the Invention [Problem to be solved by the invention]

[0006] The present invention has been made in consideration of the above circumstances, and its object is to provide a wound dressing composition that uses fucoidan and olive leaf extract and has the effect of healing skin wounds by promoting the growth of stem cells in the basal layer of the epidermis. [Means for solving the problem]

[0007] The present invention provides a wound dressing composition comprising 60 to 70 wt% of glycerin, 28 to 39 wt% of honey, 0.2 to 3.0 wt% of fucoidan, 0.2 to 3.0 wt% of olive leaf extract, and 0.1 to 0.3 wt% of xanthan gum.

[0008] The fucoidan can be obtained by mixing dried mekabu powder or extract with purified water, centrifuging the mixture, filtering, and recovering it. The olive leaf extract can be obtained by filtering powder made from crushed olive leaves, or by sterilizing a liquid extracted from olive leaves by heating it at 120 to 125°C for 15 to 18 minutes.

[0009] Depending on the content of the xanthan gum, the viscosity of the wound dressing composition can be adjusted within the range of 500 to 10,000 mPa·s. [Effects of the Invention]

[0010] The present invention provides a wound dressing composition comprising the components thereof and the optimal content ratio thereof.

[0011] The organic effects of 1) the exudate drainage effect due to the osmotic pressure of glycerin, 2) the moisturizing effect of honey and the antibacterial and antimicrobial effects due to the action of putrefaction-preventing enzymes, 3) the growth promotion effect of basal layer stem cells due to fucoidan, and 4) the antioxidant, anti-inflammatory, antibacterial and inflammation-reducing effects of olive leaf extract, can provide a healing effect on skin wounds, and xanthan gum provides the appropriate viscosity to maintain a film on the wound area. [Brief explanation of the drawings]

[0012] [Figure 1] 1 is a graph showing the results of measuring the cytotoxicity of fucoidan and olive leaf extract (oleuropein) at different concentrations against human dermal fibroblasts and epidermal keratinocytes. [Figure 2] 1 shows SEM photographs and a graph showing the results of measuring the wound healing effects of fucoidan and olive leaf extract (oleuropein) at different concentrations on human dermal fibroblasts. [Figure 3] 1 shows SEM photographs and a graph showing the results of measuring the wound healing effects of fucoidan and olive leaf extract (oleuropein) at different concentrations on human epidermal keratinocytes. [Figure 4] 1 is a graph showing the wound recovery rate by date for the control group and the test group. [Figure 5] 1 is a graph showing the recovery rate by date for the test group compared to the control group. [Figure 6] Photographs showing the wound healing status of the control group and the test group on different days. DETAILED DESCRIPTION OF THE INVENTION

[0013] It contains 60 to 70 wt% of glycerin, 28 to 39 wt% of honey, 0.2 to 3.0 wt% of fucoidan, 0.2 to 3.0 wt% of olive leaf extract, and 0.1 to 0.3 wt% of xanthan gum,

[0014] The fucoidan is obtained by mixing a powder or extract of dried Mekabu seaweed with purified water, centrifuging the mixture, filtering, and recovering the mixture.

[0015] The wound dressing composition is characterized in that the olive leaf extract is obtained by sterilizing a liquid obtained by filtering powder obtained by crushing olive leaves or a liquid extracted from olive leaves by heating it at 120 to 125°C for 15 to 18 minutes. [Mode for Carrying Out the Invention]

[0016] The present invention relates to a wound dressing composition that utilizes substances known to promote skin regeneration when the skin barrier is destroyed by wounds, burns, ulcers, bruises, post-surgical wounds, childbirth, chronic wounds, dermatitis, etc., and that prevents wound infection and exhibits antioxidant effects using substances that are relatively easy to obtain and whose safety has already been assured.

[0017] Stem cells are cells that have the ability to self-renew and differentiate into two or more cell types. The proliferative potential of basal layer stem cells maintains the homeostasis of stratified epithelial tissues. Therefore, when the skin is injured, stem cells are supplied to the wound from the basal layer at the epidermal-dermal junction, and the wound is healed through the division and growth of stem cells.

[0018] The present invention applies fucoidan and olive leaf extract, which are culture medium compositions for stem cell therapeutic agents studied in Korean Patent Registration No. 10-2088767, to a wound dressing composition.

[0019] However, when the skin is injured, the supply and proliferation of stem cells from the basal layer may be restricted by exudates, microorganisms, and other contaminants at the wound site. Therefore, it is important to use a wound dressing to clean the wound site, relieve the heat and pain caused by the wound, and ensure that the wound dressing has a certain viscosity so that it does not drip easily when applied to the wound site.

[0020] The present invention was arrived at in consideration of the above matters, and provides a wound dressing composition comprising: 60-70 wt% glycerin, 28-39 wt% honey, 0.2-3.0 wt% fucoidan, 0.2-3.0 wt% olive leaf extract, and 0.1-0.3 wt% xanthan gum.

[0021] The wound dressing composition provided by the present invention is based on glycerin and honey, contains fucoidan and olive leaf extract as active ingredients for medicinal effects, and contains xanthan gum as a thickener.

[0022] Glycerin (C3H8O3) is a non-irritating substance that is widely used as a main ingredient in enemas, lubricants, creams, eye drops, injections, etc., and is also widely used as an ingredient in wound dressings.

[0023] Glycerin has a high osmotic pressure, which causes cells in the body to contract, allowing exudates to be expelled from the inside of the wound to the outside. At the same time, microorganisms and other contaminants in the wound are also separated and expelled to the outside, and glycerin also prevents the exudates, microorganisms and other contaminants from re-entering the body. This cleanses the inside of the wound and promotes wound healing.

[0024] Furthermore, because glycerin has a high binding power, when applied to a wound, it does not penetrate into the body tissue but remains outside the wound. When this glycerin is mixed with xanthan gum (described later) to increase its viscosity, it forms a film on the wound.

[0025] The wound dressing composition of the present invention contains 60 to 70 wt % of the glycerin.

[0026] Honey is a viscous liquid produced by bees after ingesting sucrose secreted from nectar glands of flowers. The sucrose is broken down into fructose and glucose by bee enzymes. Honey does not spoil due to the osmotic effect of its high sugar content and the presence of several anti-spoilage enzymes in nectar. Therefore, when honey is applied to a wound on the skin, the action of the anti-spoilage enzymes and osmotic effect described above causes the moisture in the bacteria to move toward the honey, thereby performing an antibacterial function that causes the bacteria that have lost their moisture to wither and die.

[0027] Honey also has a skin soothing function, forming a protective film on the skin to reduce heat and pain caused by wounds, and a moisturizing function, which keeps the skin moist and relieves discomfort such as heat, itching, and dryness, and softens the skin.

[0028] The wound dressing composition of the present invention contains 28 to 39 wt % of the honey.

[0029] Fucoidan is a substance that gives brown algae flexibility, protecting them from strong currents. Fucoidan, extracted from brown algae, is a polysaccharide formed by the combination of a monosaccharide called fucose and a sulfate group, and is known for its anticoagulant, antitumor, antihypertensive, antihypertensive, hepatocyte growth factor (HGF) production induction, antihyperglycemic, immune cell regulating, antiallergic, and antiviral effects.

[0030] Fucoidan contains a large amount of highly hydrophilic sulfate groups, which has a high skin regenerative power. It supports and activates the proliferative potential of basal layer stem cells, contributing to the maintenance of homeostasis of stratified epithelial tissues. In other words, it promotes the division and growth of stem cells in the basal layer at the epidermis-dermis junction, thereby facilitating wound healing.

[0031] The fucoidan may have a structure such as Chemical Structure 1 below.

[0032] [Chemical structure 1] TIFF2025539175000002.tif69130

[0033] In the present invention, dried wakame powder or extract is mixed with purified water, centrifuged three times, and then filtered to recover fucoidan, which has a molecular weight in the range of 1,000 to 2,000,000 Da. The wound dressing composition of the present invention contains 0.2 to 3.0 wt% of the fucoidan.

[0034] Olive leaf extract is a component of oleuropein extracted from olive leaves. It has excellent antioxidant, anti-inflammatory, and antibacterial effects, and is also effective in alleviating inflammation, making it widely used as a skin protectant and natural antibiotic. Oleuropein is a generic term for polyphenols present in olive fruit and olive leaves, and olive leaf extract contains a higher amount of oleuropein than olive oil. Oleuropein, the olive leaf extract, may have the following chemical structure:

[0035] [Chemical structure 2] TIFF2025539175000003.tif43120

[0036] In the present invention, the wound healing effect is maximized by sterilizing the filtered solution of crushed olive leaves or the solution extracted from olive leaves by heating it at 120-125°C for 15-18 minutes.

[0037] The wound dressing composition of the present invention contains 0.2 to 3.0 wt % of the olive leaf extract.

[0038] Xanthan gum is a polymeric polysaccharide obtained by purely cultivating and fermenting carbohydrates using the fungus Xanthomonas campestris. The gum is then purified, dried, and ground in isopropyl alcohol. It is a mixture of glucose, mannose, and sodium, potassium, and calcium salts of glucuronic acid. As an extracellular polysaccharide, its main backbone is beta-1,4-glucose (cellulose), with a triose sugar (mannose-glucuronic acid-mannose) attached to every two glucose residues via a side chain. The mannose attached to the glucose is acetylated. Xanthan gum is used as a food additive, stabilizer, thickener, binder, emulsifier, caking agent, and foaming agent.

[0039] The viscosity of the wound dressing composition of the present invention varies depending on the concentration and content of glycerin and the content of honey. However, by adjusting the amount of xanthan gum added within the range of 0.1 to 0.3 wt%, the viscosity of the wound dressing composition can be adjusted within the range of 500 to 10,000 mPa·s.

[0040] The wound dressing composition described above can be used as a medicine, a quasi-drug, or a cosmetic composition by stirring at a speed of 80 to 300 rpm for 24 to 120 hours to achieve homogenous mixing. Depending on the application, the wound dressing composition can be adjusted to a pH range of 5 to 9 and can be made translucent or opaque.

[0041] The wound dressing composition can be commercialized by pouring about 0.1 to 500 cc into a container, and the wound dressing composition in the container can be sterilized by irradiating it with gamma rays.

[0042] The following describes the results of independent tests using human dermal fibroblasts and keratinocytes.

[0043] 1. Measurement of cytotoxicity in human dermal fibroblasts and epidermal keratinocytes To confirm the effect of the mixture of fucoidan and olive leaf extract (oleuropein) on the proliferation of human dermal fibroblasts (adult HDFa cells) and human epidermal keratinocytes (HaCaT cells), an MTT assay was performed. Human dermal fibroblasts and epidermal keratinocytes were plated at 1–3 × 10 cells / well in 96-well plates. 3 After incubating for 24 hours, a 1000x stock solution of a standard mixture of fucoidan and olive leaf extract (oleuropein) in a 1:1 weight ratio was diluted with sterile distilled water to prepare working solutions of 50x, 25x, 10x, 1x, and 0.1x concentrations. After incubation, MTT solution was added and incubated, followed by measuring the absorbance at 540nm.

[0044] As a result, when the survival rate (proliferation rate) of human dermal fibroblasts in the negative control group, in which no mixture was added, was taken as 100%, the survival rate (proliferation rate) of human dermal fibroblasts after 24 hours of treatment with each concentration of the mixture was 98.38% at 0.1x concentration, 98.10% at 1x concentration, 88.40% at 10x concentration, and 84.17% at 25x concentration, confirming that there was no significant difference in cytotoxicity. For human epidermal keratinocytes, the survival rate was 98.96% at 0.1x concentration, 97.65% at 1x concentration, 99.42% at 10x concentration, 93.79% at 25x concentration, and 86.36% at 50x concentration, confirming that there was no significant difference in cytotoxicity (see Figure 1).

[0045] 2. Measurement of the wound healing efficacy of human dermal fibroblasts and epidermal keratinocytes To confirm the effect of the mixture of fucoidan and olive leaf extract (oleuropein) on wounds in human dermal fibroblasts and epidermal keratinocytes, a wound healing assay was performed. 5After distributing cells / well and culturing for 24 hours, horizontal scratches were made using a yellow tip and the cells were washed twice with medium. A 1,000x stock solution of a standard mixture of fucoidan and olive leaf extract (oleuropein) at a weight ratio of 1:1 was individually diluted with sterile distilled water to create working solutions of 50x, 25x, 10x, 1x, and 0.1x concentrations. These working solutions were then added to the wells and cultured for 24 hours. After culturing, the surface area of ​​the horizontal scratches was measured using Image J software and photographed under a microscope at 0 and 24 hours.

[0046] As a result, when treated with a mixture of fucoidan and olive leaf extract (oleuropein), the scratch area of ​​human dermal fibroblasts was measured at 76.6% (23.4% damage recovery rate) in the negative control group, while after 24 hours it was confirmed to be 18.8% (81.2% damage recovery rate) at 0.1x concentration, 25.0% (75% damage recovery rate) at 1x concentration, and 34.3% (65.7% damage recovery rate) at 10x concentration. It is understood that the efficacy of wound recovery is maximized when the mixture is used at low concentrations (see Figure 2).

[0047] The scratch area of ​​human epidermal keratinocytes was 39.8% (damage recovery rate 60.2%) in the negative control group, but after 24 hours it was 5.6% (damage recovery rate 94.4%) at 0.1x concentration, 8.6% (damage recovery rate 91.4%) at 1x concentration, and 11.7% (damage recovery rate 88.3%) at 10x concentration. Again, it appears that the wound recovery effect is even greater when the mixed substance is at a lower concentration (see Figure 3).

[0048] Therefore, in the present invention, small amounts of fucoidan and olive leaf extract are added within the range of 0.2 to 3.0 wt% and 0.2 to 3.0 wt%, respectively, of the entire wound dressing.

[0049] The following example compositions were arbitrarily selected within the range of 60-70 wt% glycerin, 28-39 wt% honey, 0.2-3.0 wt% fucoidan, 0.2-3.0 wt% olive leaf extract, and 0.1-0.3 wt% xanthan gum (midpoint of each component, 65 wt% glycerin, 33 wt% honey, 0.9 wt% fucoidan, 0.9 wt% olive leaf extract, and 0.2 wt% xanthan gum). The details of the toxicity tests conducted by an accredited testing organization are shown in Table 1 below.

[0050] [Table 1]

[0051] It was confirmed that the contents of each test and the compositions of the examples of the present invention were completely free of any harmful effects, and the results of each test are as follows:

[0052] 1. Test 1: Skin sensitization test using guinea pigs This test was conducted to evaluate the presence or absence of skin sensitization that occurs after administering the test substance (the composition of the Examples of the present invention) to test animals (guinea pigs, female).

[0053] The negative control substance and the test substance were administered to 5 or 10 animals in each group in the primary intradermal induction, secondary topical induction, and tertiary induction tests, and mortality, symptoms, and body weight were observed and measured during the test period. In addition, skin reactions at all application sites were evaluated 24±2 hours and 48±2 hours after removal of the tertiary patch.

[0054] As a result of the test, none of the test animals administered with the negative control substance solution or the test substance solution died during the test period, and no symptoms, abnormal behavior, or changes in body weight that could be attributed to the administered substance were observed. No erythema or swelling was observed at the challenge site in any of the animals, and the Magnusson & Kligman Scale Grade was 0. In conclusion, the test substance was evaluated as not having skin sensitizing properties.

[0055] [Table 2]

[0056] [Table 3]

[0057] [Table 4]

[0058] 2. Test 2: Single-dose acute systemic toxicity test in mice The eluate of the test substance and the negative control substance in this test were the same as those in "Test 1" above.

[0059] This test was conducted to evaluate the potential for toxic reactions after a single intraperitoneal administration of the test substance to test animals (male SPF (Specific Pathogen Free) mice), and the negative control substance and the test substance eluate were administered in a single dose to five mice per test group. Mortality, symptoms, and body weight were observed and measured during the test period, and a macroscopic necropsy examination was performed on the day of necropsy.

[0060] As a result of the test, none of the test animals administered the negative control substance or the test substance died during the test period. In all groups, no abnormal symptoms or abnormal behaviors that could be attributed to the administered substance were observed, and no changes in body weight that could be attributed to the administered substance were also observed. No abnormal findings that could be attributed to the administered substance were also observed upon gross dissection.

[0061] 3. Test 3: Cytotoxicity test using L929 cells This test is intended to confirm the presence or absence of cytotoxicity of the test substance using L929 cells.

[0062] L929 cells were seeded into a 6-well plate and cultured for 24±2 hours in a 37±1°C, 5±1% CO2 incubator. After dispensing and solidifying agar-containing culture medium (prepared by adding approximately 50 ml of FBS (Fetal Bovine Serum) and 10 ml of penicillin-streptomycin to 440 ml of MEM (Minimum Essential)), the cells were stained with neutral red.

[0063] The negative control material was a 1.0 cm RM-C (High density polyethylene film) selected according to ISO 10993. 2 The positive control material was RM-B (0.25% ZDBC polyurethane film) selected according to ISO 10993, 1.0 cm thick. 2 The test substance was a composition of the example of the present invention cut into 1.0 cm pieces and sterilized at 121±2°C for 20 minutes to prepare three pieces. 2 Three 0.45 μm filters were prepared by thoroughly soaking the filter.

[0064] The negative control substance, positive control substance, and test substance prepared as described above were applied to stained L929 cells and observed. The negative control group showed no change in cell shape beneath or around the sample, and no lysis was observed. Furthermore, there were no areas of decolorization, so the cytotoxicity rating was 0. The positive control group showed rounded cells, and the lysis and decolorization areas extended up to 0.4 cm around the sample, resulting in a cytotoxicity rating of 3. The test group showed cell abnormalities and decolorization limited to the area directly beneath the sample, resulting in a cytotoxicity rating of 2.

[0065] When the results of the negative and positive control groups were confirmed, the test process was deemed to be appropriate. Based on the combined test results, the cytotoxicity level of the test substance was determined to be Grade 2, indicating that it was not cytotoxic.

[0066] [Table 5]

[0067] 4. Test 4: Intradermal reaction test using rabbits (direct contact) The eluate of the test substance and the negative control substance in this test were the same as those in "Test 1" above.

[0068] This test was designed to evaluate the potential for irritation that a test substance may induce when its solution was administered intradermally (intradermally) to female rabbits (strain: New Zealand White (NZW), SPF).

[0069] The test substance was dissolved in saline (SC) and cottonseed oil (CSO) and the solution was administered to three rabbits. On the left side of the back of the depilated rabbits, 0.2 ml of the test substance solution was administered intradermally at 5 sites for each dissolution medium (SC, CSO) for a total of 10 sites, centered around the spine, and on the right side, 0.2 ml of the negative control substance was administered at 5 sites for each substance (SC, CSO) for a total of 10 sites.

[0070] During the test period, the mortality, symptoms, and body weight of the test animals were observed and measured.

[0071] As a result of the test, no animals died during the test period, and none of the animals showed any specific symptoms that could be attributed to the administered substance. There was also no change in body weight that could be attributed to the administered substance.

[0072] The intradermal reaction at the administration sites of the test substance eluate and negative control substance was observed, and the intradermal reaction score at each administration site was calculated 24±2 hours, 48±2 hours, and 72±2 hours after administration according to the observation criteria. The difference in the average score at the administration site for each eluate was less than 1.0, and when the eluate was the same, there was no difference between the test substance and the negative control substance.

[0073] No specific irritation was observed at the site of administration of the test substance, and therefore the test substance is not expected to cause a local reaction within the rabbit skin.

[0074] [Table 6]

[0075] [Table 7]

[0076] 5. Test 5: Pyrogenicity test using rabbits This study was designed to evaluate the potential for pyrogenic reactions after a single administration of the eluate of the test substance into the ear vein of female rabbits (strain: New Zealand White (NZW), SPF) to verify the biological safety of the test substance. SC was used as the elution vehicle.

[0077] Before administration of the test substance solution, the control body temperature was measured, and the test substance solution was administered once to the auricular vein of three rabbits at a volume of 10 ml / kg. Test groups were assigned so that the difference in control body temperature between individual test rabbits was less than 1.0°C, and individuals with a control body temperature of more than 39.8°C were excluded from the test.

[0078] One hour after the administration of the test substance, body temperature was measured five times at 30-minute intervals to determine the difference from the control body temperature, while observing the mortality rate and symptoms during the test period.

[0079] As a result of the test, none of the test animals administered with the solution of the test substance showed an increase in body temperature of 0.5°C or more compared to the control body temperature, and the test substance was determined to be non-pyrogenic (two of the three rabbits showed no increase in body temperature, while the body temperature of one rabbit increased by a maximum of 0.2°C). Furthermore, none of the test animals died during the test period, and no animals showing abnormal symptoms were observed.

[0080] 6. Test 6: Skin irritation test using rabbits (direct contact) This test was designed to evaluate the potential for irritation that a test substance may induce when it comes into direct contact with the skin of female rabbits (strain: New Zealand White (NZW), SPF).

[0081] The test substance and negative control substance were applied to the backs of hairless rabbits. According to ISO 10993-23, the test substance was applied directly to the skin by cutting a 5.0 x 5.0 cm sterile gauze pad into four equal pieces to make four 2.5 x 2.5 cm sterile gauze pads, which were then stacked 12 times. 0.5 ml of the test substance was instilled into the skin using a syringe. The negative control substance was also applied according to ISO 10993-23, by cutting a 5.0 x 5.0 cm sterile gauze pad into four equal pieces to make four 2.5 x 2.5 cm sterile gauze pads, which were then stacked 12 times. 0.5 ml of sterile saline (SC) was instilled into the skin using a syringe.

[0082] The application site was wrapped with appropriate pressure to avoid abdominal irritation and secured with a bandage to prevent the test substance and negative control substance from being lost. 4 to 24 hours after application, the test substance and negative control substance were removed from the application site.

[0083] Mortality, symptoms, and body weight were observed and measured during the test period. Skin reactions at the application site were observed 1±0.1 h, 24±2 h, 48±2 h, and 72±2 h after removal of the applied substance according to the observation criteria, and a skin irritation score was calculated.

[0084] As a result of the test, no animals died during the test period, and none of the animals showed any abnormal symptoms or changes in body weight that could be attributed to the administered substance.

[0085] The primary irritation index (PII) at the application sites of the test substance and negative control substance was 0.0 (Negligible). In conclusion, it is determined that the test substance does not cause skin irritation in rabbit skin.

[0086] [Table 8]

[0087] 7. Test 7: Endotoxin test using lysate reagent (colorimetric method) This test is intended to quantitatively confirm the toxicity (endotoxin) of the test substance using a lysate reagent.

[0088] Endotoxin-free Reagent Water (LRW, LAL Reagent Water) was selected as the dissolution medium for the test substance in accordance with USP40 and USP43, and dissolution was performed under the conditions in Table 9 below in accordance with ISO10993-12.

[0089] [Table 9]

[0090] The eluate of the test substance was applied to a PTS cartridge and measured with a PTS Reader. The channel information of the PTS cartridge is as follows:

[0091] [Table 10]

[0092] The endotoxin standard value of the eluate is determined as follows using the endotoxin standard value (20 EU / device) of the test substance.

[0093] ERL (Endotoxin Release Limit) =K×N / V=20EU×1 / 40ml=0.5EU / ml

[0094] K: the Endotoxin limit per device(20EU)

[0095] N:the number of tested devices

[0096] V:the total volume of water used to extract the endotoxin from the devices tested

[0097] Using the endotoxin standard value of the eluate calculated as above, the maximum effective dilution ratio is determined as follows.

[0098] Maximum effective dilution factor = ERL × concentration of test substance / λ (most sensitive sensitivity of PTS cartridge) = 0.5EU / ml × 1 / 0.005EU / ml = 100 times

[0099] The most sensitive PTS cartridge: 0.005EU / ml

[0100] Concentration of test substance: 1 (100% eluate)

[0101] The test groups are as follows:

[0102] [Table 11]

[0103] Group A was applied to channels 1 and 3 of the PTS cartridge, and group B was applied to channels 2 and 4 of the PTS cartridge to measure endotoxin recovery and confirm reaction interference factors. Crns and group D confirmed the information contained in the PTS cartridge by confirming the reliability of the calibration curve and LRW.

[0104] After preparing the eluate of the test substance as a test solution, the pH was measured using pH-indicator paper (MERCK). The pH of the test solution was found to be 6.0 to 7.0. 25 μl of the test solution was applied to each of the four channels of the PTS cartridge. The absorbance at 395 nm was measured using a PTS Reader at a temperature of 37 ± 1°C.

[0105] As a result of the test, the Rxn time CV between the sample and spike was less than 25%, demonstrating the suitability of the test, and the endotoxin recovery rate was within the range of 50-200%, indicating that there were no interfering factors in the test solution, and the endotoxin concentration of the test solution was less than 0.5EU / ml. Therefore, the endotoxin concentration of the eluate of the test substance used in the test under these test conditions was determined to be less than 0.5EU / ml.

[0106] [Table 12]

[0107] Meanwhile, the viscosity of the wound dressing composition must be 500 to 10,000 mPa·s (Ministry of Food and Drug Safety Notification No. 2023-2, General Test Methods of the Korean Pharmacopoeia, 57. Viscosity Measurement Method).

[0108] The viscosity of the test example of the composition of the present invention was measured to be 5,420 mPa·s, which was within the standard range.

[0109] [Table 13]

[0110] The following are the results of a wound induction test on test animals, which was conducted to comparatively evaluate the healing ability of the wound dressing composition of the present invention.

[0111] Test animals (specific pathogen-free (SPF) mice) were anesthetized (using isoflurane respiratory anesthesia) and their back skin was disinfected. A wound was then induced by removing skin tissue using a biopsy punch (5 mm diameter). The natural healing process was observed for a control group, and the wound dressing composition (test substance) of the present invention was applied and the healing process was observed for a test group. The test substance was applied to sterile gauze (3 cm x 5 cm) (0.2 g per application) and evenly applied to the wound and surrounding skin. An adhesive band (3 cm x 10 cm) was cut to completely wrap the abdomen and back of the test animal, and the gauze was placed in contact with the wound to secure the abdomen and back of the test animal.

[0112] The wound area was measured once a day from the first day to the tenth day of the test, and the wound recovery rate by day and the recovery rate of the test group relative to the control group were calculated based on the measurements.

[0113] To measure the wound area, the gauze and bandage are cut and removed with sterilized surgical scissors. To measure the exact size of the wound area (lesion site), the scab is removed with sterile tweezers. Bleeding is stopped for 1 minute with sterile gauze. The wound area is calculated based on the long axis and shortened length of the wound lesion measured using Vernier calipers.

[0114] The components of the wound dressing composition applied to the test group are as shown in Table 14 below, and were applied once a day to each wound lesion and the skin surrounding the lesion in the manner described above.

[0115] [Table 14]

[0116] [Figure 4] is a graph showing the wound healing rate by date for the control group and the test group, [Figure] is a graph showing the healing rate by date for the test group compared to the control group, and [Figure 6] is a photograph showing the wound healing status by date for the control group and the test group.

[0117] In Figure 4, the test group showed a faster wound healing rate up to day 5 compared to the control group, and after day 5, the healing rates of the test and control groups were almost the same. In Figure 5, the healing rate of the test group compared to the control group increased rapidly until day 4, and the difference in healing rate gradually decreased from day 4 onwards. Therefore, it can be seen that the test group contributed to faster wound healing.

[0118] Although the present invention has been described in connection with the test examples as described above, various modifications and variations are possible within the scope of the gist of the present invention, and the present invention can be used in various fields. Therefore, the scope of the claims of the present invention includes modifications and variations that fall within the true scope of the previous invention. [Industrial Applicability]

[0119] The present invention can be used as a wound dressing for wound and burn healing and scar amelioration on human or animal skin.

Claims

1. 60 to 70 wt% glycerin, Honey 28-39wt% Fucoidan 0.2 to 3.0 wt% Olive leaf extract 0.2-3.0 wt% 0.1 to 0.3 wt% xanthan gum, 1. A wound dressing composition comprising:

2. 2. The wound dressing composition according to claim 1, wherein the fucoidan is obtained by mixing a powder or extract of dried Mekabu seaweed with purified water, centrifuging the mixture, filtering, and recovering the mixture.

3. 2. The wound dressing composition according to claim 1, wherein the olive leaf extract is obtained by filtering a powder obtained by crushing olive leaves or by sterilizing a liquid extracted from olive leaves by heating it at 120 to 125°C for 15 to 18 minutes.

4. 4. The wound dressing composition according to claim 1, wherein the viscosity is adjusted to a range of 500 to 10,000 mPa·s by adjusting the content of the xanthan gum.

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