A liquid functional dressing containing camellia oil extract for difficult-to-heal wounds and its preparation method

Abstract

The present invention discloses a preparation method of a liquid functional dressing containing camellia oil extract for difficult-to-heal wounds, which comprises the following components by weight: 3.5-3.9 parts of camellia oil extract, 18-22 parts of sodium alginate, 3-5 parts of tara gum, and 0.4-0.7 parts of gallic acid. The camellia oil extract of the present invention can extract the effective components of camellia oil with antibacterial and wound granulation tissue regeneration-promoting effects through the treatment of adding catechin pulsed direct current electric field and composite enzyme treatment, achieving the effect of promoting the healing of difficult-to-heal wounds.

Description

Technical Field

[0001] The present invention relates to the field of liquid functional dressings, and particularly to a preparation method of a liquid functional dressing containing camellia oil extract for difficult-to-heal wounds. Background Art

[0002] A difficult-to-heal wound is defined by the International Wound Healing Society as a wound that cannot achieve anatomical and functional complete healing through a normal, orderly, and timely repair process. A chronic non-healing wound (ulcer) on the body surface refers to a wound on the body surface formed by various reasons that has not healed after more than 1 to 3 months (the time is not completely absolute) of treatment and shows no tendency to heal. From a pathophysiological perspective, a non-healing wound that cannot be histologically repaired regularly and orderly after skin tissue defect is called a difficult-to-heal wound.

[0003] There are various etiologies for difficult-to-heal wounds, including but not limited to: 1. Various traumas such as burns, scalds, and car accidents; 2. Pressure injuries; 3. Vascular diseases such as varicose veins and vasculitis; 4. Metabolic diseases such as diabetes; 5. Immune diseases such as gout; 6. Radiation ulcers; 7. Infectious ulcers, especially those caused by various infections, such as infections caused by Mycobacterium tuberculosis; 8. Implantation of artificial materials, and it is very common that the exposure of various artificial materials such as titanium mesh after implantation into the human body leads to the formation of difficult-to-heal wounds.

[0004] Camellia oil is also known as camellia seed oil. Camellia oil is rich in polyphenols, camellia saponins, phytosterols, flavonoids and some other unique active ingredients. Camellia oil contains a relatively high content of oleic acid and rich active ingredients, so it has good affinity with the skin, strong permeability, is easy to be absorbed by the skin, and has functions such as antioxidant, anti-aging, refreshing, and moisturizing. The active ingredients of camellia oil not only have strong antioxidant effects, but also can inhibit a variety of bacteria, fungi, and yeasts, especially have strong inhibitory ability against common pathogenic bacteria such as Staphylococcus aureus and Escherichia coli. Camellia saponins have a broad-spectrum antibacterial effect, especially have a very strong inhibitory effect on Escherichia coli, Staphylococcus aureus, and yeasts. Phytosterols are a kind of plant active ingredient that is structurally similar to animal cholesterol but has completely different functions. It can scavenge free radicals, have antioxidant effects, and has a unique anti-inflammatory effect, and has good inhibitory effects on Escherichia coli, Staphylococcus aureus, etc.

[0005] Therefore, applying camellia oil extract to the dressing for difficult-to-heal wounds can accelerate the healing of difficult-to-heal wounds. Summary of the Invention

[0006] In view of this, the present invention proposes a preparation method of a liquid functional dressing containing camellia oil extract for difficult-to-heal wounds to solve the above problems.

[0007] A liquid functional dressing containing camellia oil extract for difficult-to-heal wounds, comprising the following components by weight: 3.5 - 3.9 parts of camellia oil extract, 18 - 22 parts of sodium alginate, 3 - 5 parts of Caesalpinia spinosa gum, and 0.4 - 0.7 parts of gallic acid.

[0008] Further, the preparation method of the camellia oil extract includes the following steps:

[0009] (1) Add catechin to camellia oil and treat it under pulsed direct current electric field for 8 - 12 min;

[0010] (2) Mix and soak the camellia oil after the treatment in step (1) with a complex enzyme and perform microwave treatment;

[0011] (3) Subject the camellia oil after the treatment in step (2) to high-temperature and high-pressure treatment to obtain the camellia oil extract.

[0012] Further, in step (1), the mass ratio of catechin to camellia oil is 3 - 5:100.

[0013] Further, in step (1), the pulsed direct current electric field parameters are: peak voltage 25 - 28 V, pulse frequency 2.3 - 2.7 kHz, and duty cycle 0.6 - 0.8.

[0014] Further, in step (2), the complex enzyme is composed of pectinase, partial glyceride lipase, and cellulase with a mass ratio of 1:2 - 4:1.5 - 1.9, and the mass ratio of the complex enzyme to the camellia oil after the treatment in step (1) is 0.01 - 0.03:1.

[0015] Further, in step (2), the soaking temperature is 35 - 45 °C, the soaking time is 2 - 4 h, and the microwave treatment is performed at 80 - 90 °C with 400 - 500 W for 20 - 30 min.

[0016] Further, in step (3), the high-temperature and high-pressure treatment is carried out at 150 - 200 Pa with a reaction temperature of 170 - 200 °C for 70 - 90 min.

[0017] Further, the preparation method of the liquid functional dressing includes the following steps:

[0018] S1: Add sodium alginate, Caesalpinia spinosa gum, and gallic acid to water to obtain a mixed solution 1;

[0019] S2: Add the camellia oil extract to the mixed solution 1 to obtain the liquid functional dressing.

[0020] Further, in the step S1, the amount of water is 1.2 - 1.5 times the total mass of sodium alginate, Caesalpinia spinosa gum, and gallic acid. Sodium alginate, Caesalpinia spinosa gum, and gallic acid are added to water and stirred at 150 - 180 r / min for 20 - 30 min. After stirring, it is left to stand for 3 - 5 h.

[0021] Further, in the step S2, the camellia oil extract is added to the mixed solution 1 and heated to 30 - 50 °C, and stirred at 100 - 120 r / min for 10 - 15 min. After stirring, it is heated to 70 - 80 °C and left to stand for 60 - 100 min.

[0022] Advantages of the present invention:

[0023] The liquid functional dressing of the present invention is composed of camellia oil extract, sodium alginate, Caesalpinia spinosa gum, and gallic acid. Sodium alginate, Caesalpinia spinosa gum, and gallic acid in the present invention are used as excipients, which can improve the absorption capacity of the dressing for wound exudate, maintain a dry wound surface environment, and avoid bacterial reproduction. The extraction method of the camellia oil extract in the present invention can extract the effective components in camellia oil with antibacterial and wound granulation tissue regeneration promotion effects. The liquid functional dressing of the present invention has good anti-inflammatory effects and the function of improving the proliferation activity of fibroblasts, enabling the wound to quickly transition from the inflammatory stage to the granulation tissue proliferation stage, and further achieving the effect of promoting the healing of difficult-to-heal wounds. Specific embodiments

[0024] To better understand the technical content of the present invention, specific embodiments are provided below to further illustrate the present invention.

[0025] Example 1

[0026] Experimental group 1

[0027] (1) Catechin and camellia oil are mixed according to a mass ratio of 3:100 and processed at a peak voltage of 25 V, a pulse frequency of 2.3 kHz, and a duty cycle of 0.6 for 8 min;

[0028] (2) A composite enzyme composed of pectinase, partial glyceride lipase, and cellulase according to a mass ratio of 1:2:1.5 is prepared. The camellia oil processed in step (1) and the composite enzyme are mixed and soaked at a temperature of 35 °C for 2 h. After soaking, it is processed at 400 W at 80 °C for 20 min;

[0029] (3) The camellia oil processed in step (2) is reacted at 170 °C under 150 Pa for 70 min to obtain the camellia oil extract.

[0030] Experimental group 2

[0031] (1) Mix catechin and chaulmoogra oil according to a mass ratio of 5:100, and process for 12 min at a peak voltage of 28 V, a pulse frequency of 2.7 kHz, and a duty cycle of 0.8;

[0032] (2) Compose a composite enzyme from pectinase, partial glyceride lipase, and cellulase according to a mass ratio of 1:4:1.9. Mix the chaulmoogra oil processed in step (1) with the composite enzyme, soak at a temperature of 45 °C for 4 h, and then process at 90 °C with 500 W for 30 min;

[0033] (3) React the chaulmoogra oil processed in step (2) at 200 °C for 90 min under 200 Pa to obtain a chaulmoogra oil extract.

[0034] Experimental group 3

[0035] (1) Mix catechin and chaulmoogra oil according to a mass ratio of 3 - 5:100, and process for 10 min at a peak voltage of 27 V, a pulse frequency of 2.5 kHz, and a duty cycle of 0.67;

[0036] (2) Compose a composite enzyme from pectinase, partial glyceride lipase, and cellulase according to a mass ratio of 1:3:1.7. Mix the chaulmoogra oil processed in step (1) with the composite enzyme, soak at a temperature of 40 °C for 3 h, and then process at 85 °C with 450 W for 25 min;

[0037] (3) React the chaulmoogra oil processed in step (2) at 190 °C for 80 min under 180 Pa to obtain a chaulmoogra oil extract.

[0038] Experimental group 4

[0039] (1) Compose a composite enzyme from pectinase, partial glyceride lipase, and cellulase according to a mass ratio of 1:3:1.7. Mix the chaulmoogra oil with the composite enzyme, soak at a temperature of 40 °C for 3 h, and then process at 85 °C with 450 W for 25 min;

[0040] (2) React the chaulmoogra oil processed in step (1) at 190 °C for 80 min under 180 Pa to obtain a chaulmoogra oil extract.

[0041] Experimental group 5

[0042] (1) Mix catechin and chaulmoogra oil according to a mass ratio of 3 - 5:100, and process for 10 min at a peak voltage of 27 V, a pulse frequency of 2.5 kHz, and a duty cycle of 0.67;

[0043] (2) A composite enzyme is composed of pectinase, protease and cellulase in a mass ratio of 1:1. After the treatment in step (1), the camellia oleifera oil and the composite enzyme are mixed and soaked at a temperature of 40°C for 3 hours, and then treated at 450 W at 85°C for 25 minutes after soaking;

[0044] (3) The camellia oleifera oil treated in step (2) is reacted at 190°C for 80 minutes under 180 Pa to obtain a camellia oleifera oil extract.

[0045] The antibacterial test is carried out on the camellia oleifera oil extract prepared by Experimental Group 3 - 5.

[0046] The test method uses the paper disk agar diffusion method to detect the antibacterial effect, and the test bacteria are Staphylococcus aureus, Candida albicans, and Pseudomonas aeruginosa.

[0047] The preparation method of the antibacterial tablets is as follows: in the experimental group, 0.5 mL of the camellia oleifera oil extract solution (the mass ratio of camellia oleifera oil extract to water is 1:5) is dropped at the center of different filter papers respectively, and after drying, it is reserved for use. The control group is to drop 0.5 mL of camellia oleifera oil (untreated camellia oleifera oil) at the center of different filter papers.

[0048] The preparation method of the inoculated plates: After Staphylococcus aureus, Candida albicans, and Pseudomonas aeruginosa are cultured overnight using a nutrient medium, the cultures of Staphylococcus aureus, Candida albicans, and Pseudomonas aeruginosa are diluted to 10 6 cfu / mL with phosphate buffer saline, and then inoculated onto nutrient agar plates respectively by the plate coating method, and left to dry at room temperature for 5 minutes.

[0049] The determination of the antibacterial effect: The antibacterial tablets and the inoculated plates are attached, and the attached inoculated plates are placed in an incubator at 37°C for 24 hours respectively. The antibacterial diameter (mm) is measured using a vernier caliper. Each experimental group is repeated three times, and the average value is taken.

[0050] Table 1 Antibacterial effect test results (mm)

[0051] Name Staphylococcus aureus Candida albicans Pseudomonas aeruginosa Experimental group 3 21.47 17.66 25.93 Experimental group 4 17.64 13.72 20.52 Experimental group 5 19.96 12.31 23.68 Control group 12.62 7.96 14.34

[0052] The test results show that the camellia oleifera oil extract prepared by the present invention has good antibacterial effects. In Experimental Group 4, catechin was not used for treatment, resulting in the oxidation of camellia oleifera oil during the high - temperature heating process and the decline of the finished product effect. In the present invention, the combination of catechin and pulsed direct current electric field treatment can prevent the high - temperature oxidation of camellia oleifera oil and is beneficial to the leaching of active ingredients. In Experimental Group 5, the composition of the composite enzyme was changed, resulting in the decline of the enzymatic hydrolysis effect. The purpose of using the composite enzyme for enzymatic hydrolysis in the present invention is to extract the active ingredients of camellia oleifera oil with antibacterial properties and promoting wound granulation tissue regeneration.

[0053] The following examples use the camellia oil extract prepared in experimental group 3 to prepare a liquid functional dressing.

[0054] Example 2

[0055] (1) Weigh the following components by weight: 3.5 parts of camellia oil extract, 18 parts of sodium alginate, 3 parts of Caesalpinia spinosa gum, and 0.4 part of gallic acid.

[0056] (2) Add sodium alginate, Caesalpinia spinosa gum, and gallic acid to water and stir at 150 r / min for 20 min. After stirring, let it stand for 3 h. The amount of water used is 1.2 times the total mass of sodium alginate, Caesalpinia spinosa gum, and gallic acid to obtain mixed solution 1.

[0057] (3) Add the camellia oil extract to mixed solution 1, heat it to 30 °C, and stir at 100 r / min for 10 min. After stirring, heat it to 70 °C and let it stand for 60 min to obtain the liquid functional dressing.

[0058] Example 3

[0059] (1) Weigh the following components by weight: 3.9 parts of camellia oil extract, 22 parts of sodium alginate, 5 parts of Caesalpinia spinosa gum, and 0.7 part of gallic acid.

[0060] (2) Add sodium alginate, Caesalpinia spinosa gum, and gallic acid to water and stir at 180 r / min for 30 min. After stirring, let it stand for 5 h. The amount of water used is 1.5 times the total mass of sodium alginate, Caesalpinia spinosa gum, and gallic acid to obtain mixed solution 1.

[0061] (3) Add the camellia oil extract to mixed solution 1, heat it to 50 °C, and stir at 120 r / min for 15 min. After stirring, heat it to 80 °C and let it stand for 100 min to obtain the liquid functional dressing.

[0062] Example 4

[0063] (1) Weigh the following components by weight: 3.7 parts of camellia oil extract, 20 parts of sodium alginate, 4 parts of Caesalpinia spinosa gum, and 0.55 part of gallic acid.

[0064] (2) Add sodium alginate, Caesalpinia spinosa gum, and gallic acid to water and stir at 160 r / min for 25 min. After stirring, let it stand for 4 h. The amount of water used is 1.3 times the total mass of sodium alginate, Caesalpinia spinosa gum, and gallic acid to obtain mixed solution 1.

[0065] (3) Add the camellia oil extract to mixed solution 1, heat it to 40 °C, and stir at 110 r / min for 15 min. After stirring, heat it to 75 °C and let it stand for 80 min to obtain the liquid functional dressing.

[0066] Test Example 1 Cell Viability Test

[0067] The test was carried out using mouse fibroblast L929 as the raw material. The detection method was MTT colorimetry, and the detection item was cell proliferation viability.

[0068] Experimental procedure: Inoculate at an inoculation density of 1×10 4 cells / mL in a 96-well plate and culture. Incubate at 37°C, in an atmosphere of 5% CO2 and humidity above 90% for 12 h. Add 10 μL of the liquid functional dressing prepared in Examples 2-4, and culture for 24 and 48 h respectively. After the culture is completed, add 10 μL of MTT solution and continue to culture for 4 h. Discard the culture supernatant. Add 100 μL of DMSO solution to each well, shake for 10 min, and use an enzyme-linked immunosorbent assay (ELISA) reader to measure and record the photometric value at a wavelength of 570 nm. Repeat the measurement three times.

[0069] The addition of cell growth culture medium was used as the blank group, the non-addition of the liquid functional dressing was used as the control group, and Examples 2-5 were used as the test groups.

[0070] Cell viability = (test group - blank group) / (control group - blank group) × 100%

[0071] Table 2 Experimental results of cell viability test

[0072]

[0073]

[0074] The experimental results show that the liquid functional dressing prepared by the present invention can improve the proliferation viability of fibroblasts, promote the growth of wound surface cells, and further promote the growth of granulation tissue.

[0075] Test Example 2 Animal Experiment

[0076] Six 8-10-week-old male Sprague-Dawley (SD) rats were used for the test. The experimental material was the liquid functional dressing of Example 4.

[0077] 1. Establishment of type II diabetic rat model

[0078] Inject streptozotocin (60 mg / kg) into the rats until the fasting blood glucose level of the rats exceeds 17 mM, which is regarded as successful establishment of the model.

[0079] 2. Experimental procedure

[0080] Anesthetize the rats, remove the hair on the back of the rats, make a wound with a diameter of 10 mm on the back of the rats using a skin drill, apply the liquid functional dressing to the wound, and the application amount is 0.5 g. After 5 days, take the wound skin to detect the inflammatory factors (TNF-α and IL-6) of the wound. The detection method is an ELISA kit.

[0081] Table 3 Detection Results of Inflammatory Factors

[0082] Name TNF-α (pg / ml) IL-6 (pg / ml) Example 4 16.8 11.5

[0083] The test results show that the liquid functional dressing of the present invention can effectively inhibit the growth of inflammatory factors, enable the wound to quickly transition from the inflammatory stage to the granulation tissue proliferation stage, and further achieve the effect of promoting the healing of difficult-to-heal wounds.

[0084] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims

1. A liquid functional dressing containing camellia oil extract for non-healing wounds, characterized in that, It comprises the following components by weight parts: 3.5 - 3.9 parts of camellia oil extract, 18 - 22 parts of sodium alginate, 3 - 5 parts of Caesalpinia spinosa gum, and 0.4 - 0.7 parts of gallic acid; The camellia oil extract is prepared by the following method: (1) Catechin is added to camellia oil, and the mass ratio of catechin to camellia oil is 3 - 5:

100. It is treated for 8 - 12 min under pulsed direct current electric field, with a peak voltage of 25 - 28 V, a pulse frequency of 2.3 - 2.7 kHz, and a duty cycle of 0.6 - 0.8; (2) The camellia oil after being treated in step (1) is mixed and soaked with a composite enzyme, and then microwave - treated. The composite enzyme is composed of pectinase, partial glyceride lipase, and cellulase with a mass ratio of 1:2 - 4:1.5 - 1.

9. The mass ratio of the composite enzyme to the camellia oil after being treated in step (1) is 0.01 - 0.03:

1. The soaking temperature is 35 - 45 °C, and the soaking time is 2 - 4 h. The microwave treatment is carried out at 80 - 90 °C with 400 - 500 W for 20 - 30 min; (3) The camellia oil after being treated in step (2) is subjected to high - temperature and high - pressure treatment, and the reaction is carried out at 170 - 200 °C for 70 - 90 min.

2. The liquid functional dressing containing camellia oil extract for difficult-to-heal wounds according to claim 1, wherein The preparation method of the liquid functional dressing comprises the following steps: S1: Sodium alginate, Caesalpinia spinosa gum, and gallic acid are added to water to obtain a mixed solution 1; S2: The camellia oil extract is added to the mixed solution 1 to obtain the liquid functional dressing.

3. The liquid functional dressing containing the extract of chaulmoogra oil for difficult-to-heal wounds according to claim 2, characterized in that In the step S1, the amount of water used is 1.2 - 1.5 times the total mass of sodium alginate, Caesalpinia spinosa gum, and gallic acid. Sodium alginate, Caesalpinia spinosa gum, and gallic acid are added to water and stirred at 150 - 180 r / min for 20 - 30 min, and after stirring, it is left to stand for 3 - 5 h.

4. The liquid functional dressing containing the extract of Camellia oleifera Abel oil for difficult-to-heal wounds according to claim 2, characterized in that, In the step S2, the camellia oil extract is added to the mixed solution 1, heated to 30 - 50 °C, and stirred at 100 - 120 r / min for 10 - 15 min. After stirring, it is heated to 70 - 80 °C and left to stand for 60 - 100 min.

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