TW201208717A - Wound dressing containing citrus extract and fabrication method thereof - Google Patents

Abstract

Immersing or wet spinning is used to fabricate wound dressing containing citrus extract, and thus the wound recovering effect of the wound dressing can be increased.

Description

201208717 Six invention description: The technical field to which the invention belongs: The present invention relates to a wound containing a tangerine extract of an orange extract, and in particular relates to [prior art] the current global wound dressing market The vast majority. The recovery has the following stages: ·, the inflammatory period of physical trauma and the long skin remodeling period. At different times; need = the same function of the material to help them recover. Therefore, the recruitment of two = can be elastically covered in the wound on the wound, the injury of the material, the isolation of the microbial infection wound, the degree of dryness of the following functions, the degree of pain that provides the wound energy = ^ control of the wound ^ m (four) material 'and Reducing injuries There are currently some studies in progress to identify related biological materials for use in wound dressings. [Summary of the Invention] Therefore, one aspect of the present invention is a wound dressing and a method for manufacturing the same, to increase the effect of the citrus extract. The wound healing material promotes the wound healing = the implementation state The wound dressing adjusted orange extract containing the citrus extract is further prepared by using the fiber obtained by wet spinning to contain the citrus extract: 201208717 According to another embodiment, the wound dressing containing the phase orange extract may also be the following method The sponge-like wound dressing is firstly immersed in a solution containing citrus extract, and then freeze-dried to obtain a wound dressing containing citrus extract. The citrus extract wound dressing prepared by the above method, The weight ratio of the citrus extract/applied material contained therein is 0.18-0.72. It can be seen from the above embodiment that the wound dressing containing the citrus extract can be obtained by a simple method. The wound dressing containing the citrus extract prepared by the above method has a fairly good wound healing effect. [Embodiment] = The above provides a kind of citrus extract-containing wound And dressing materials

Ik method. It is only necessary to use a very simple method of extracting wound dressings. Moreover, it is said that each of the citrus (four)/reported "wound soil containing citrus extract" has a very good wound healing effect. In the following example, the dry medicinal extract contains the citrus extract. The manufacturing method of the material. These technical details are required for easy understanding of the described embodiments. At the same time:;: wide or components, only the sound of the 4', known, ., σ structure content. The heart, 'type in the pattern, please, in order to properly simplify the 'oxygen species' in the septic tissue, causing the sputum, the scorpion-/the oxygen-producing species (reactive) b cover + . st test). The above reactive oxygen species are, for example, force (. her tlve state oxime), hydrogen peroxide base owned =,), single lane (〇〇H), superoxide radical (. (3⁄43⁄4] 201208717 and hydrogen peroxide ( H2〇2), etc. In the early stage of injury, low concentrations of reactive oxygen species can help wound healing by attracting and activating macrophage cells (macr〇phage). However, when the oxidative stress is long-term or large, Let macrophages secrete a large amount of cytokine, which produces chronic inflammation, which causes the injured tissue to begin anti-oxidation defense mechanism, thus slowing the tissue repair rate and slowing the wound recovery. If the oxidative stress is more serious, Causes permanent damage to skin aging, apoptosis, and skin necrosis. Therefore, in order to treat wounds, in addition to substances that promote cell proliferation, some antioxidants are needed to promote wound healing. The results of the study 'discovered that citrus peel extract has at least the effect of promoting cell proliferation and angiogenesis (analysis of sweet orange peel formation) (Lai Hongliang et al; Taiwan citrus industry development "Special issue", W〇2005/105127) 'Citrus fruit extract also has the effect of promoting cell growth. In addition, 'citrus extract is also rich in flavonoids, cartenoids, vitamin c, Vitamin B1, B2, etc., all of which have anti-oxidant effects and can alleviate inflammation. The above-mentioned Kluj-song orange extract has both multiple functions of promoting cell proliferation and growth, promoting angiogenesis and antioxidant activity. In the following embodiments of the present invention, a biological material is used as a substrate, and an orange extract is added to be applied to the manufacture of a wound dressing. At the same time, the wound dressing to which the citrus extract is added is tested for wound healing after use. Effects of citrus extracts Common citrus fruits are citrus reticulata, citrus sinensis, citrus tankan Hayata and kumquat (marumikumquat), where citrus fruits are taken. Fruit s] 201208717 and the peel were separately extracted with a juice machine and the money was extracted with ethanol as a solvent. After the stroke liquid was dried in the air extractor cabinet, The remaining solid is the citrus extract. The vitamin C content of a quantitative citrus extract, also known as ascorbic acid, is a good reduction d & can be reduced to Γ, so it can be used The acid-depleting (κι〇3) method for the shoulder-shoulder J firstly causes the gorging acid to be unloaded and decomposed (10) in an acidic environment to produce an iodine molecule (〗 2) and then reacts the iodine molecule with vitamin C. After C has been fully reacted, excess iodine molecules will bind to the starch indicator to produce a blue-black complex that is known to reach the end of the titration. At this time, the amount of vitamin C can be estimated by the amount of potassium iodate, and the results are shown in Table 1. Table 1: Vitamin C of each citrus extract contains strontium. -------- Citrus extract vitamin C per gram of extract contains orange peel 0.2257 orange fruit 0.4264 willow peel 0.3762 willow fruit 0.5204 sea pear skin "——___ --- 0.2508 pear fruit 0.4076 Kumquat skin 0.1693 Kumquat fruit 1^ 0.2257 201208717 Quantitative citrus extract and other carotenoid content of carotenoids are determined using Siikran DERE (§tikran DERE, Tohit GTJNE§, Ridvan SIVACI Turk. J. Bot. , 22, (1998), 13-18.) The method published was measured. Siikran's method requires first measuring the absorbance at 470 nm, 653 nm, and 666 nm for samples of different carotenoids, ie, A470, A653, and A666. Then, the chlorophyll a content (Ca) and the chlorophyll b content (Cb) were calculated according to the following two equations (1) and (2), respectively.

Ca = 15·65χΑ666 - 7.34〇xA653 (1)

Cb = 27.05χΑ653 - 11.21χΑ666 (2) Finally, using the chlorophyll a content (Ca) and the chlorophyll b content (Cb), the total carotenoid content (Cx+c) is calculated by the following equation (3). Establish a concentration standard curve.

Cx+c = 1000χΑ470 - 2.86〇xCa - 129.2><Cb/245 (3) Then 'measure the values of A470, A653 and A666 of the sample to be tested, and convert it by the concentration standard curve of carotenoid The content of carotenoids in the sample to be tested. The carotenoid content of each citrus extract is shown in Table 2. Carotenoid content - the content of carotene in the body (Yang) 201208717 Orange skin... 4.45 Orange fruit 18.82 Liuding skin 6.08 Liuding fruit 11.78 Sea pear skin ------------ 7.26 Sea Pear fruit " -------- 15.50 gold skin ------------ 6.24 Kumquat fruit 7.34 In vitro cell proliferation experiments Cell proliferation is associated with wound healing rate. Therefore, the activity of promoting various cell proliferation of various citrus extracts was first measured. Two, five mouse fibroblasts (fibroblast ' L929) were implanted in the cell culture dish. After 3-6 hours of cell culture, various citrus extracts of various concentrations were added. The cells were further cultured for 72 hours, and the growth of each plate was observed. The total number of cells in each plate was counted, and the total number of cells obtained φ is shown in Table 3. The control group to which no orange extract was added was cultured for 72 hours in the same cell culture. The total number of cells observed in the control group was 17,500. Table 2. After 72 hours of incubation with different citrus extracts, the total number of cells obtained was observed. Total dry weight (Mg) of citrus extract per plate 30 60 120 240 480 960 1920 3840 7680 Orange peel 12600 22000 26800 32500 25000 17500 15000 13500 9900 fs ] 8 201208717 Orange fruit 9500 14000 18000 20000 57500 37500 34000 18500 13000 Liudingpi 12600 14000 18000 20000 35000 35000 26000 20000 14000 Liuding fruit 14000 19000 22500 27500 50000 30000 23000 16000 13000 Sea pear skin 14500 19000 21000 25000 40000 30000 25000 22000 16500 Sea pear fruit 12600 15000 20500 22500 50000 35000 27500 22500 16000 Golden orange peel 18000 22000 26100 30000 37500 40000 34500 27500 16500 Kumquat fruit 12600 15000 22500 27500 42500 35000 31000 28100 18500 It can be seen from the results in Table 3 that the best effect on promoting cell proliferation is the addition of orange fruit extract with a total dry weight of 480 /zg, after cultivation. The total number of cells was 3.29 times the total number of cells in the control group. Next, the total lyophilized fruit extract and the pear fruit extract were added at a total dry weight of 480 />tg, and the total number of cells after the culture was 2.86 times of the total number of control cells. From the above, it is understood that the citrus extract has an effect of promoting cell proliferation by extracts of orange fruit, lycopene fruit and sea pear fruit. However, instead of adding as much as possible, add the right amount of citrus extract. Antioxidant activity test Antioxidant activity is associated with slowing down inflammation, so the antioxidant activity of various orange extracts is then determined. The antioxidant activity of various citrus extracts is measured by the DPPH (11-Diphenyl-2-Picrylhydrazyl) free radical scavenging ability. DPPH is a very stable free radical with a strong absorbance at 517 nm. When the antioxidant capacity of the analyte is better, the more hydrogen atoms can be transferred to the DPPH, the DPPH is lowered at the absorbance of 517 nm 201208717. The above principle can be expressed by the following chemical formula.

Therefore, the method for testing the antioxidant activity of various citrus extracts is to determine the DPPH absorbance of various citrus extracts before and after reaction with DPPH for 30 minutes, and to reflect the antioxidant activity of various citrus extracts by the magnitude (%) of decrease in absorbance. active. The standard test sample used here is BHA (butylated hydroxyanisole), which has a scavenging ability (i.e., antioxidant power) of 640 / xg DPPH free radical of 92.52%. The series of DPPH free radical scavenging abilities of various citrus extracts are shown in Table 4. Table 4: Removal of 640 /ig DPPH free radicals by various citrus extracts • Ability (ie antioxidant capacity, in %). Citrus extract total dry weight 240 gg 480 /xg 960 /xg orange peel 88.05 88.34 88.92 orange fruit 87.76 89.50 92.71 willow skin 87,46 88.63 84.26 willow fruit 86.59 87.46 86.59 sea pear skin 86.59 83.09 86.59 sea pear fruit 88.34 89.2 90.96 Gold file leather 84.55 86.01 84.s 201208717

Tianyi Si 1 夭 子 果实 果实 果实 果实 果实 果实 果实 果实 果实 果实 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 960 〉 The oxidizing power is also quite good, with 89.5 〇〇/0 and 89 ^ σ A divided by about 90% of DPPH free radicals. · °’ is able to clear

Preparation of wound dressings containing sweet extracts - soaking method Main: The steps of the soaking method are as follows. First take a sponge or non-woven cloth soak = orange extract ^, the time is about U hours. The fabric with the citrus extract is then dried by cold-bead drying to obtain a wound dressing containing the citrus extract. The material of the above sponge or non-woven fabric may be, for example, chitosan, acacia, collagen egg, or any combination thereof. These materials are more or less biocompatible, protect wounds and promote wound healing, and have been applied to medical materials such as dressings and hemostatic materials. Combined with the citrus extract, it will further enhance the wound healing material to promote wound healing. The following is a method for preparing a chitosan-containing material containing a citrus extract by taking a chitosan nonwoven fabric and a sponge as an example. In the preparation of chitosan nonwoven fabrics, chitosan fibers are first prepared in a viable wet spinning manner. The method may be, for example, first preparing a 5 wt% chitosan solution (prepared by 2 wt G/0 acetic acid aqueous solution), and then using a mixed solution of ethanol and water containing 5 wt% NaOH (EtOH: H20=1:1) Ratio) 201208717 As a molding liquid for chitosan fibers. Next, the obtained chitosan fiber was dried in a vacuum oven, and the dried chitosan fiber was immersed in the orange extract for about 2 hours. Finally, the chitosan fiber containing the citrus extract is dried by the cold; east drying method to obtain the chitosan fiber containing the orange extract. Next, after the obtained citrus extract-containing chitosan fiber is sufficiently washed with water, the chitosan fiber is cut into a 38 mm staple fiber, and then the chitosan non-woven fabric is produced. A non-woven fabric composed of chitosan fibers containing citrus extract. In the preparation of chitosan sponge, a solution containing 2 wt% of chitosan and a bridging agent are poured into a forming box and subjected to a bridging reaction to form a cake. Next, freeze-drying was carried out to make the cake into a sponge. Then, it was heated at 80 - 1 ° C for 1 hour, and the inside of the chitosan sponge was further subjected to a bridging reaction. The chitosan is then soaked in the citrus extract, and after it has sufficiently absorbed the citrus extract, it is freeze-dried again to obtain a chitosan sponge containing the citrus extract. Preparation of wound dressings containing citrus extracts - Wet spinning The main steps of the wet spinning process are as follows. A quantitative citrus extract is first added to the solution of the wet-spun raw material, and then the wet-spun raw material solution is injected into the molding liquid through the spinning nozzle to directly form the fiber containing the orange extract. Then, vacuum drying is carried out to obtain a fiber containing a citrus extract. The above wet-spun raw material may be, for example, chitosan, sodium alginate, collagen, gelatin or any combination of the above. Hereinafter, a method for preparing chitosan fibers containing citrus extracts will be described by taking chitosan as an example. The chitosan fibers are first prepared in a viable wet spinning manner. Method Example 12 201208717 It is possible to prepare a 5 wt% solution of chitosan (modified with 2 wt% aqueous acetic acid), and then add an ethanol solution of citrus extract. Then, a mixed solution of ethanol and water containing 5 wt% of NaOH (Et〇H: H2〇 = 3:8 by volume) was used as a forming liquid of chitosan fibers. Here, since the citrus extract is extracted by ethanol, the ethanol content in the forming liquid is reduced to reduce the loss of the orange extract. Next, the resulting chitosan fiber containing the citrus extract was dried in a vacuum oven. Next, the obtained citrus extract-containing chitosan ruthenium fiber was sufficiently washed with water, and then the fiber was cut into staple fibers of 38 mm. Further, a chitosan non-woven fabric is produced to obtain a chitosan non-woven fabric containing a citrus extract. The retention rate of the citrus extract in the raw material of the dressing material can be compared in the subsequent animal experiments. It can be estimated that the chitosan-containing citrus extract prepared by the above three methods can retain the citrus extract. What is the ratio of the objects? The method for analyzing the content of the citrus extract containing the citrus extract material is as follows. First, the weight of the dressing is first weighed and then the above-mentioned wound dressing containing the citrus extract is dissolved with 〇5 to 10% by weight of acetic acid. Next, using the carotenoids contained in the citrus extract as an indicator compound, by measuring the carotenoid content and the data in Table 2 above, it is possible to estimate the amount of the citrus extract remaining in the dressing. The citrus extract content values of the respective dressings obtained by taking the best-performing orange fruit extract and the chitosan raw material as an example are listed in Table 5.

Table 5: Retention rate of citrus extract of chitosan-coated material U 13 201208717 Type of perforated material A. Fiber B. Fiber --- C. Sea brocade (soaked) Chitosan-coated material weight (g) 0.061 0.124 0.147 Orange fruit extract 10.579 2.189 6.566* Addition amount (mg) Orange fruit extract 4.361 2.136 5.307 Retention amount (mg) ---- Retention rate (%) 41.22 97.59 80.82 -------·*·*· * The initial addition amount and target content of the citrus extract in the material of the dressing are determined according to the retention rate of Table 5, and the amount of the orange extract extracted from various chitosan materials is determined. If the amount of citrus extract added in the in vitro cell proliferation assay is shown in Table 3, the amount of citrus extract to be added to the raw material can be estimated based on the retention rate in Table 5 above and the method described below. As a result of the in vitro cell proliferation assay, the total amount of citrus extract added to a plate of 2,500 cells is shown in Table 3. In addition, when the cells are closely arranged, the density is about 38,000 cells/cm 2 , so it is possible to estimate the amount of the corresponding orange stalk required for the wound per square centimeter of the size of the wound. The calculation formula is as follows: Dry weight of citrus extract (Table 3 data) X 38000/2500 = dry weight of the orange stalk required per square centimeter of wound. Next, calculate the dry weight of the amount of material required per square centimeter of wound. For what purpose, obtain the desired weight ratio of citrus extract dry weight / dry weight of the dressing. Finally, by using the data of 201208717 five, the data of the desired orange extract in the raw material is reversed: = . The calculation results obtained by the above method are listed in Table 6 below. ##表^,· The initial addition amount of the extract of the thumb orange in the raw material and the target of the citrus extract in the dressing, „ 知3 $ 'The value is calculated (dry weight of citrus extract / C. Sponge corresponding to the citrus corresponding to the citrus extract of Table III (soaking) --- The target content of the extract is added (Mg) 0.029 0.023 30 0.058 0.045 60 0.116 0.090 120 0.232 0.180 240 0.464 0.360 480 0.968 0.720 960 1.856 1.440 1920 3.712 2.880 3840 7.424 5.760 7680 (wet visit) 0.057 In Table 6, the calculation of the content of citrus extract in raw materials is more complicated, as explained below. In the wet spinning method, the dry weight of citrus extract and several The weight of the poly-dry weight is calculated from the weight of the citrus extract and chitosan in the chitosan solution (ie, the wet-spun raw material). In the soaking method, the dry weight of the citrus extract and the chitin The weight of the dry weight of the polysaccharide is calculated by the dry weight of the orange extract in the absorbed extract and the dry weight of the fiber or the brocade immersed in the citrus extract. From the results of Tables 3 and 6, it can be seen that In the material (citrus extract dry 15 201208717 / dry weight of the material) The target ratio is 0.360 - 0.720, which is better for cell proliferation. Evaluating the cell safety of the citrus extract-containing material After obtaining the citrus-containing extract material as described above, the next step Cell safety assessment will be performed first. The evaluation method is as follows: First, subculture of L929 cells, 2χ105 cells in each well of the cell culture plate, continue to culture at 37 ° C and 5% CO 2 . 24 hours. Transfer the cells to MEM medium, pass UV lamp, add commercially available gauze after sterilization or chitosan with citrus extract, continue at 37 ° C and 5% C02 After culturing for 24 hours, a neutral red dye (chemical name 3-amino-6-methylamino-2-mercaptophenazine hydrochloride) was added, and after reacting with the cells for 1 hour, the neutral red was removed and photographed. The number of living cells. Neutral red dye will stain the living cells in red, while dead cells will not be stained. The experimental results are commercially available gauze and chitosan dressings containing citrus extracts (wet spinning and soaking) ) have no cells Toxicity. Animal experiments Next, animal experiments were conducted to test the effect of promoting the healing of wounds containing citrus extracts. The method of animal experiments was as follows. Healthy rats were selected and shaved in the back of the rats after anesthesia shaving. Take two 2x2 cm2 wounds. Cover the wound with a size of 2x2 cm2 (about 0.08g weight) and then bandage. After one, two and three weeks of the experiment, the rats are anesthetized and dismantled. Bandage the bandage. After the sample dressing was removed, the wound was washed with physiological saline to observe the healing of the wound. Here, the experimental results of the cell proliferation test and the antioxidant power of the orange fruit are shown in Table 7. Wound Healing Day 7 Day 14 Day 21 Day 21 Commercial Gamble 17.5 21.3 23.3 - David (Wet Spin) 23.0 33.5 43.8 Chitin Glycoside Containing Orange Fruit Extract - '''''---1 «1« #As Wet Spinning* * 43.5 76.5 82.0 Soaking * * * 26.8 67.5 81.3 Yangshoukou ©槚-day η observation of the injury)/Original wound area X 100% Hazelnut fruit extract dried cardinal The dry weight of the glycan is about 0.36. The orange fruit extract dry butter (4) has a dry weight of about 0.36. According to the preliminary results, it is known that for the effect of promoting wound healing, the chitosan fiber of the fiber is almost exclusively chitosan. It can be known from the above disclosure that only the benefit is obtained: the wound dressing containing the citrus extract is obtained. . And 'can ride into the general biocompatible record ^ mouth;;!: fruit clustering, sodium alginate, collagen, gelatin) promotion [sl 17 201208717 although the invention has been disclosed in the above embodiments, but The present invention is not limited to the scope of the invention, and the scope of protection of the present invention is defined by the scope of the appended claims. Prevail. [Simple description of the diagram]

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Claims (1)

201208717 VII. Patent Application L A wound dressing containing citrus extract, comprising: a dressing material, which is chitosan, sodium alginate, gelatin, gelatin or any combination thereof; and, a citrus The extract, the citrus extract / the weight ratio of the medicinal material is 0.18 - 0·72 〇 L L. The wound dressing according to claim 1, wherein the weight ratio of the citrus extract / ° hai material is 0.36 - 0.72. A wound dressing according to claim 1, wherein the citrus extract is derived from an extract of orange fruit, willow fruit or sea pear fruit. 1 . A method for producing a citrus extract wound dressing, the method comprising: immersing a dressing material in a solution of a citrus extract, the soaking time is about θ 4 Α the citrus extract absorbed by the money material Dry weight 7 The dry weight ratio of the dry material is 0.18-0.72; and the citrus extract is dried by freeze drying. Method 2: a method for producing a wound dressing comprising an orange extract, the method comprising: adding an orange extract to a wet-spun raw material solution t; performing wet spinning to obtain a fiber containing a citrus extract; and Roll drying method to dry the citrus-containing extract ^ Wei ^ The fiber Wei] 19 201208717 The weight ratio of the dry weight of the citrus extract / the dry weight of the fiber is 0.18-0.72. 6. The method of claim 4 or 5, wherein the fiber comprises a weight ratio of the dry weight of the citrus extract to the dry weight of the fiber of from 0.36 to 0.72. The manufacturing method according to claim 4, wherein the material of the dressing material is chitosan, sodium alginate, collagen, gelatin or any combination thereof. 8. The method of manufacture according to claim 4, wherein the citrus extract is derived from an extract of orange fruit, willow fruit or sea pear fruit. 20 201208717 IV. Designation of Representative Representatives: (1) The representative representative of the case is: (No). (2) A brief description of the symbol of the representative figure: None 5. If there is a chemical formula in this case, please disclose the chemical formula that best shows the characteristics of the invention: