CN114668898A - Application of fat emulsion in preparation of wound repair material - Google Patents
Application of fat emulsion in preparation of wound repair material Download PDFInfo
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- CN114668898A CN114668898A CN202210301994.5A CN202210301994A CN114668898A CN 114668898 A CN114668898 A CN 114668898A CN 202210301994 A CN202210301994 A CN 202210301994A CN 114668898 A CN114668898 A CN 114668898A
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- Prior art keywords
- fat emulsion
- wound
- wound repair
- fat
- fat milk
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Images
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/60—Materials for use in artificial skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3637—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the origin of the biological material other than human or animal, e.g. plant extracts, algae
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
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Abstract
The invention belongs to the technical field of wound repair, and particularly relates to application of fat emulsion in preparation of a wound repair material. The fat milk has good anti-inflammatory effect in vitro and in vivo, and can obviously reduce the expression of proinflammatory cytokines TNF-alpha and IL-1 beta; meanwhile, the fat emulsion can also promote angiogenesis and cell migration, acts on each period of wound healing, and has the obvious effect of promoting wound repair; the wound repair dressing prepared based on the fat emulsion is applied to a diabetic wound mouse model, so that the blood vessel density of the wound can be increased, the healing rate of the wound is accelerated, and tissue epithelialization and more collagen deposition formation are promoted.
Description
Technical Field
The invention belongs to the technical field of wound repair. More particularly, it relates to the use of fat emulsions in the preparation of wound repair materials.
Background
The skin is the largest organ of the human body and is a protective barrier isolating the interior of the body from contact with the outside of the environment. When the skin is damaged and not repaired in time due to daily life or other diseases, the wound surface is easily infected and develops into a chronic wound, such as Diabetic Foot Ulcer (DFU), which is a common diabetic syndrome and even endangers life in severe cases, and amputation is needed.
Research shows that wound healing mainly comprises four stages of hemostasis coagulation stage, inflammation stage, proliferation stage and tissue remodeling stage: in the hemostasis and blood coagulation period, platelets are activated to secrete related factors to generate a series of cascade reactions, so that blood is promoted to be coagulated into thrombus, the blood coagulation effect is achieved, and meanwhile, the organism is stimulated to generate inflammatory reaction and the like; in the inflammatory phase, inflammatory cells such as neutrophils and macrophages engulf tissue fragments and kill pollutants in the environment such as invading bacteria, and simultaneously secrete various cytokines to stimulate fibroblast division, collagen synthesis and angiogenesis; during the proliferation period, the fibroblasts generate various extracellular matrix components such as collagen, fibronectin and the like, synthesize granulation tissues, differentiate into myofibroblasts at the same time, and secrete related proteins to promote wound contraction; in the tissue remodeling stage, collagen, fibronectin and the like are continuously synthesized and decomposed under the co-regulation of relevant cytokines and cells to reach a balance for continuous remodeling.
In order to promote the rapid progress of the healing of the wound in each period, various wound repairing materials are researched in the prior art, for example, Chinese patent application discloses an organic-inorganic hybrid dressing for wound repair, the dressing is prepared by modifying silicon dioxide with fucoidan under a 3-glycidyl ether oxypropyl trimethoxy silane coupling agent, has good biocompatibility and no cytotoxicity, can adsorb inflammatory factors, promotes cell migration and angiogenesis, and promotes the healing of chronic wounds; however, the preparation method of the material has multiple steps and complex operation, and the application of the material is greatly limited.
Disclosure of Invention
The invention aims to overcome the defects and shortcomings of multiple steps and complex operation of the existing preparation method of the wound repair material and provide the application of the existing fat emulsion product clinically applied to the preparation of the wound repair material.
The invention aims to provide a wound repair dressing.
Another object of the present invention is to provide a method for preparing the wound repair dressing.
The above purpose of the invention is realized by the following technical scheme:
the inventor finds in practice that the fat milk has good anti-inflammatory action in vitro and in vivo, and can obviously reduce the expression of proinflammatory cytokines TNF-alpha and IL-1 beta; meanwhile, the fat emulsion can also promote angiogenesis and cell migration, acts on each period of wound healing, and has a remarkable effect of promoting wound repair.
The invention therefore claims the use of fat emulsions for the preparation of wound repair materials.
Preferably, the fat milk comprises 50-200 mg/mL of soybean oil and C6～2425-100 mg/mL of medium-chain triglyceride and 6-12 mg/mL of lecithin.
More preferably, C is6～24The medium chain triglyceride is selected from one or two of caprylic triglyceride and capric triglyceride.
In addition, the invention also provides a wound repair dressing which comprises the fat emulsion and the carrier matrix.
The wound repairing dressing prepared by the invention has better repairing effect on wounds such as burn, crush wound, impact wound, open wound and the like; experiments prove that the wound repair dressing prepared based on the fat emulsion is applied to an in-vivo diabetes wound mouse model, so that the blood vessel density of a mouse chronic wound can be increased, the wound healing rate is accelerated, tissue epithelization and more collagen deposition are promoted, and the wound repair healing is accelerated.
Preferably, the fat milk comprises 50-200 mg/mL of soybean oil and C6～2425-100 mg/mL of medium-chain triglyceride and 6-12 mg/mL of lecithin.
Further, the carrier matrix is selected from one or more of medical gauze, medical sponge, medical foam and hydrogel.
Preferably, the hydrogel is a physical gel or a chemical gel. The physical gel can be gelatin, agar and the like, and the chemical gel can be polyacrylamide-polyacrylic acid hydrogel, alginic acid-chitosan hydrogel and the like. More preferably, the hydrogel is methacrylic anhydrified hyaluronic acid.
Furthermore, the mass ratio of the fat emulsion to the carrier matrix is 1: (2-2X 10)6). PreferablyThe mass ratio of the fat emulsion to the carrier matrix is 1: (200 to 20000).
In addition, the invention also provides a preparation method of the wound repair dressing, which comprises the following steps:
diluting the fat emulsion in a biocompatible solvent to ensure that the concentration of the fat emulsion is 0.01-10000 mug/mL, and loading the obtained fat emulsion on a carrier matrix to obtain the fat emulsion.
Further, the biocompatible solvent is water, PBS buffer or liquid medium.
Preferably, the concentration of the fat milk is 1-1000 mug/mL, preferably, the concentration of the fat milk is 10-100 mug/mL, and more preferably, the concentration of the fat milk is 100 mug/mL.
Preferably, the pH value of the PBS buffer solution is 6.5-7.5.
Preferably, the liquid culture medium is selected from one or more of a DMEM medium, a MEM medium, an RPMI 1640 medium, an F12 medium or an IMDM medium.
The invention has the following beneficial effects:
the invention provides the application of the fat milk in preparing a wound repair material, the fat milk has good anti-inflammatory action in vitro and in vivo, and can obviously reduce the expression of proinflammatory cytokines TNF-alpha and IL-1 beta; meanwhile, the fat emulsion can also promote angiogenesis and cell migration, acts on each period of wound healing, and has the obvious effect of promoting wound repair; the wound repair dressing prepared based on the fat emulsion is applied to a diabetic wound mouse model, so that the blood vessel density of the wound can be increased, the healing rate of the wound is accelerated, and tissue epithelialization and more collagen deposition formation are promoted.
In addition, the fat milk is approved by the FDA to be clinically used for providing nutrition for patients who can not obtain enough nutrition in the gastrointestinal tract, the safety is guaranteed, and the fat milk is simple in preparation method, green, safe, low in cost, easy to implement and large-scale in application.
Drawings
FIG. 1 is a graph showing DLS results obtained by examining the particle size of fat milk in application example 1.
Fig. 2 is a TEM image of the morphology of fat milk in application example 2.
FIG. 3 is a fluorescence microscope photograph showing the cell compatibility of fat milk detected at different concentrations in application example 3.
FIG. 4 is a graph of data obtained by examining the effect of fat milk at various concentrations on the proliferation rate of HUVEC in application example 3.
FIG. 5 is a graph showing the data of the effect of fat milk of different concentrations on the transcriptional level of inflammatory cytokine Tnf-alpha gene in application example 4.
FIG. 6 is a graph showing the data of the effect of fat milk of different concentrations on the transcription level of the inflammatory cytokine Il-1. beta. gene in application example 4.
FIG. 7 is a micrograph showing the effect of fat emulsion of different concentrations on the vascularizing ability of cells in the normal environment of application example 5.
FIG. 8 is a graph showing the quantitative results of the influence of fat milk of different concentrations on the cell vascularizing ability in the normal environment in application example 5.
FIG. 9 is a graph showing the data of the quantitative results of the influence of fat milk on the cell vascularization ability in the inflammatory environment in application example 5.
FIG. 10 is a micrograph showing the effect of different concentrations of fat milk on cell migration in application example 6.
Fig. 11 is a graph showing the results of wound repair of diabetic rats on the back of the wound surface by the fat milk-based wound repair dressing in application example 7.
Fig. 12 is a graph showing the results of immunofluorescence staining, HE staining and Masson staining of the wound repair dressing based on fat emulsion prepared in application example 7 on the repair effect of rat skin back wound tissue.
Detailed Description
The invention is further described with reference to the drawings and the following detailed description, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
Fat emulsion (250mL fat emulsion containing 12.5-50 g of soybean oil, C)6～246.25-25 g of medium-chain triglyceride, 1.5-3 g of lecithin): purchased at the seventh subsidiary hospital of Zhongshan university.
The whole culture medium: DMEM medium with 10% FBS + 1% double antibody was purchased from cantonen, inc.
Unless otherwise indicated, reagents and materials used in the following examples are commercially available.
Example 1 preparation of a fat milk-based wound repair dressing
A preparation method of a wound repair dressing based on fat milk comprises the following steps:
s1, diluting fat emulsion in a PBS (pH 7.4) buffer solution to obtain a fat emulsion solution with the concentration of 100 mu g/mL;
s2, uniformly mixing 1mL of the fat emulsion solution obtained in the step S1 with 20mg of methacrylic acid anhydridized hyaluronic acid, and carrying out ultraviolet irradiation for 30S to obtain the wound repair dressing.
Example 2 preparation of a fat milk-based wound repair dressing
A preparation method of a wound repair dressing based on fat milk comprises the following steps:
s1, diluting fat emulsion in a PBS (pH 7.4) buffer solution to obtain a fat emulsion solution with the concentration of 100 mu g/mL;
s2, absorbing the fat emulsion solution into gauze to soak the gauze, and obtaining the wound repair dressing.
Example 3 preparation of a fat milk-based wound repair dressing
A preparation method of a wound repair dressing based on fat milk comprises the following steps:
s1, diluting fat emulsion in a PBS (pH 7.4) buffer solution to obtain a fat emulsion solution with the concentration of 100 mu g/mL;
s2, absorbing the fat emulsion solution into the sponge to soak the sponge, and obtaining the wound repair dressing.
Example 4 preparation of a fat milk-based wound repair dressing
A preparation method of a wound repair dressing based on fat emulsion comprises the following steps:
s1, diluting fat emulsion in a PBS (pH 7.4) buffer solution to obtain a fat emulsion solution with the concentration of 100 mu g/mL;
s2, absorbing the fat emulsion solution into foam to soak the foam, and obtaining the wound repair dressing.
Example 5 preparation of a fat milk-based wound repair dressing
A preparation method of a wound repair dressing based on fat milk comprises the following steps:
s1, weighing 100mg of grade A acrylic acid anhydridized hyaluronic acid matrix (HA-MA), dissolving in 5g of PBS buffer solution (pH 7.4), and stirring for 2 hours;
s2, sucking the fat emulsion and mixing the fat emulsion with an HA-MA matrix to enable the concentration of the fat emulsion in the matrix to be 100 mu g/mL, and irradiating the fat emulsion for 30s to form gel through ultraviolet light to obtain the wound repair dressing.
Application example 1 DLS test of particle size of fat emulsion
1. Experimental methods
Diluting fat emulsion with PBS buffer solution (pH 7.4) to make its concentration be 100 μ g/mL, mixing well to obtain fat emulsion solution; sucking 1mL of fat emulsion solution into a cuvette; and (5) placing the cuvette in a nanometer particle size analyzer for detection.
2. Results of the experiment
The results of DLS particle size measurement of fat milk are shown in FIG. 1, and it can be seen from the figure that the size range of fat milk is 130-590 nm, and the average size is 267 nm.
Application example 2 appearance of fat milk was observed by TEM
1. Experimental methods
Diluting fat emulsion with PBS buffer solution (pH 7.4) to make its concentration be 100 μ g/mL, mixing well to obtain fat emulsion solution; taking out the TEM copper mesh by using a pair of tweezers, and placing the TEM copper mesh under a heat lamp; sucking the diluted fat emulsion solution by a 10-microliter gun head, dripping about 5 microliter of the fat emulsion solution on a copper mesh, turning on a heat lamp, heating for about 20 minutes, and allowing the copper mesh to be basically completely dried; and taking out the copper mesh, placing the copper mesh in the air, air-drying for 30 minutes, and placing the copper mesh in a transmission electron microscope to observe the appearance of the fat emulsion.
2. Results of the experiment
The appearance of the fat emulsion observed by TEM is shown in FIG. 2, and it can be seen that the fat emulsion has uniform spherical appearance.
Application example 3 cytocompatibility test of fat milk
1. Experimental methods
(a) Inoculating Human Umbilical Vein Endothelial Cells (HUVEC) in 24 and 96-well plates respectively, and adding a whole culture medium overnight for wall adhesion;
(b) removing the old culture medium, adding 5 kinds of fat milk-containing whole culture medium with different concentrations (10. mu.g/mL, 50. mu.g/mL, 100. mu.g/mL, 300. mu.g/mL and 500. mu.g/mL) into the experimental group, and adding only whole culture medium into the negative control group;
(c) respectively co-culturing for 1, 2 and 3 days, removing the old culture medium, adding 200 μ L of DMEM medium containing calcein and Propidium Iodide (PI) into each well of a 24-well plate, incubating for 10 minutes, and placing the 24-well plate in an inverted fluorescence microscope to observe the proliferation condition of cells; adding 20 mu L of MTT solution into each hole of a 96-hole plate, incubating for 4h, removing culture solution, adding 100 mu L of DMSO into each hole to dissolve crystals, oscillating for 8-10 min by a shaking table, placing the hole plate on an enzyme labeling instrument, and detecting the light absorption value by adopting the wavelength of 490 nm.
2. Results of the experiment
Results of the effect of fat milk at different concentrations on the HUVEC proliferation rate are shown in FIGS. 3 and 4, when the concentration of the fat milk is lower than or equal to 100 mu g/mL, basically no dead cells are observed, the proliferation rate of the cells is basically consistent with that of the negative control group, and the normal growth of the cells is not affected. However, when the fat milk concentration is 300. mu.g/mL or more, more dead cells are observed in FIG. 3, and the cell proliferation rate in FIG. 4 is significantly suppressed.
Application example 4 Effect of fat milk on the transcriptional levels of inflammatory cytokines Tnf-alpha and Il-1 beta genes
1. Experimental methods
(a) Inoculating RAW264.7 macrophage in a 12-well plate, and adding a full culture medium overnight for wall adhesion;
(b) removing the old culture medium, adding a DMEM culture medium into the negative group, adding DMEM culture media containing 10 mu g/mL and 100 mu g/mL fat emulsion solutions into the negative experiment group respectively, and incubating for 24 h; adding DMEM culture medium containing lipopolysaccharide (LPS, 100ng/mL) into the positive group, and adding DMEM culture medium containing 10 mug/mL and 100 mug/mL fat emulsion solution and LPS into the positive experiment group respectively;
(c) after LPS induces inflammation for 2h, removing a culture medium containing LPS, washing cells for 2 times by using PBS buffer solution (pH 7.4), adding a DMEM culture medium into a positive group, adding DMEM culture media containing 10 mu g/mL and 100 mu g/mL fat emulsion solutions into a positive experiment group respectively, and continuing to incubate for 6 h;
(d) removing the culture medium, washing the cells for 2 times by PBS, adding 1mL Trizol solution into each sample, blowing and beating uniformly, and collecting in a 1.5mL EP tube to stand for 10min at room temperature;
(e) adding 200 μ L chloroform into each tube, shaking vigorously for 15s, standing at room temperature for 5min, 4 deg.C, 14000g, and centrifuging for 15 min; taking 200-400 mu L of supernatant in a new EP tube, adding 0.5mL of isopropanol into each tube, standing at room temperature for 10min, keeping the temperature at 4 ℃, keeping the temperature at 14000g, and centrifuging for 15 min; discarding the supernatant, adding ice-precooled 75% ethanol (DEPC water), washing for 2 times, 12000g, and centrifuging for 5 min; discarding the supernatant, naturally drying at room temperature for 5-10 min, and dissolving RNA by using 20 mu LDEPC water;
(e) detecting the purity and concentration of RNA by using a nucleic acid protein detector;
(f) taking 1 mu g of RNA per 20 mu L of system for reverse transcription;
(g) and (3) carrying out qRT-PCR reaction by using a PCR instrument, and detecting the transcription levels of inflammatory cytokines Tnf-alpha and Il-1 beta genes.
2. Results of the experiment
The results of the effect of the fat emulsion solution on the transcription levels of inflammatory cytokines Tnf-alpha and Il-1 beta genes are shown in fig. 5 and 6. As can be seen, after the RAW264.7 macrophage is incubated in different concentrations of fat milk for 24h, compared with a negative group, Tnf-alpha and Il-1 beta transcription levels are not different; after LPS induction, the positive group can see that the gene transcription levels of inflammatory cytokines Tnf-alpha and Il-1 beta are obviously up-regulated, but the gene transcription levels of Tnf-alpha and Il-1 beta are obviously reduced after macrophages are treated by 100 mu g/mL fat emulsion solution.
The results show that the fat emulsion does not stimulate cells to generate inflammatory reaction and has good anti-inflammatory effect.
Application example 5 Effect of fat milk on cell vascularization
1. Experimental methods
(a) Thawing matrigel at 4 ℃, and adding 10 mu Lmatrigel on an ibidi 96-well plate after the matrigel is thawed;
(b) placing the 96-well plate in an incubator at 37 ℃ for solidification for 30 min;
(c) meanwhile, 5000/50mL of HUVEC cell suspension was prepared;
(d) after 30min, 50 μ L HUVEC cell suspension was added to each well;
(e) after the cells are completely settled on matrigel, gently sucking the culture solution away, adding a DMEM culture medium into a control group, and adding DMEM culture media containing 10 microgram/mL and 100 microgram/mL fat milk into 10FE and 100FE respectively; adding DMEM culture medium after RAW264.7 macrophage is induced by LPS for 24h into a positive group (LPS), and respectively adding DMEM culture medium which contains 10 mu g/mL and 100 mu g/mL fat milk and is induced by LPS for 24h into 10FE + and 100FE + groups;
(f) the tube formation was observed after 2h, 6h and 8h and photographed.
2. Results of the experiment
The results of the vascularization ability of the fat emulsion are shown in fig. 7 and 8, and it can be seen from the figures that the fat emulsion not only can promote the formation of the lumen structure, but also can promote the formation of more pipelines; compared with a control group, the fat emulsion can form an obvious lumen after 2 hours, and can form a tubular structure earlier; and along with the migration of time, the tubular structure becomes more and more obvious after 4h, and the number of pipelines becomes more and more. The above results indicate that fat milk can promote the formation of the lumen structure and the formation of the number of channels, and shows concentration dependence.
The data in FIG. 9 is obtained by analysis and calculation of imageJ software, and it can be seen that when HUVEC cells are in an inflammatory environment, the fat milk can still improve the hemangioblast effect of the cells.
Application example 6 Effect of fat milk on HUVEC migration Capacity
1. Experimental methods
(a) Planking, 6-well plates (25X 10 holes per well)4) When HUVEC cells are full;
(b) marking 3 straight lines on each pore plate by using a 200uL gun head, removing old liquid, gently cleaning the pore plates for 1-2 times by using PBS (pH 7.4) buffer solution, adding a DMEM culture medium into a negative group, and respectively adding DMEM culture media containing 10 mu g/mL and 100 mu g/mL fat milk into a negative experiment group; adding a DMEM culture medium after the LPS induces the macrophages for 24 hours into the positive group, and respectively adding DMEM culture medium after the LPS induces the macrophages for 24 hours, wherein the DMEM culture medium contains 10 mu g/mL of fat milk and 100 mu g/mL of fat milk; taking a picture, and then putting the picture back to the incubator for continuous culture;
(c) and observing the migration condition of the scratches every 12h, photographing, and finishing the experiment after the migration of a certain group is finished.
2. Results of the experiment
The effect of fat milk on HUVEC cell migration ability is shown in FIG. 10, and it can be seen from the figure that 10. mu.g/mL and 100. mu.g/mL fat milk both promote HUVEC cell migration and exhibit concentration-dependent characteristics. Meanwhile, in an inflammatory environment, the effect of the fat milk on promoting the migration of HUVEC is more remarkable, especially for a fat milk (100FE + group) solution of 100 mug/mL. After 72h, the HUVEC cells in the 100FE + group completely migrated, and the migration rate reached 100%. This result indicates that fat milk promotes the migration of HUVECs both in normal culture environments and in inflammatory environments.
Application example 7 Effect of fat milk-based wound repair dressing on wound repair in diabetic mice
1. Experimental methods
(1) Skin wound modeling
(a) Selecting male C57 mice with the weight of 20-22 g as study objects, and injecting streptozotocin (150mg kg) into the abdominal cavity of all the mice after fasting for 12h-1) Induction of type I diabetes;
(b) after one week, after fasting the mice for 12h, detecting the blood sugar level of the mice, and if the fasting blood sugar exceeds 16.7mM, considering that the type I diabetes mellitus of the mice is successfully modeled;
(c) randomly dividing mice successfully modeled into 2 groups, injecting barbital sodium into the abdominal cavity to anaesthetize the mice, shaving the backs of the mice, and then cutting 5mm round skins on the backs of the mice in a full-layer manner;
(2) medicine applying on wound
The wounds of the control group were treated with HA-MA hydrogel, and the wounds of the experimental group were treated with the wound repair dressing prepared in example 5, and were applied daily and photographed.
2. Results of the experiment
The wound area size, immunofluorescence staining, HE staining and Masson staining results of the wound repair dressing based on the fat emulsion, which is prepared by the invention, on the influence of the wound repair dressing on the skin back wound tissue repair of the diabetic mouse are shown in FIGS. 11 and 12.
Fig. 11 shows that the wound repair dressing based on fat milk can effectively accelerate the time for wound healing, and the skin wound of the diabetic mouse is basically completely healed 11 days after the operation, but obvious wounds exist in the control group. FIG. 12IL-1 β immunofluorescent staining shows that a fat milk-based wound repair dressing can reduce inflammatory response in wounds and promote wound healing; PDGF-B immunofluorescent staining shows that the wound repair dressing based on fat emulsion can promote healing tissues to form richer blood vessels; as can be seen from the HE and Masson dyeing results, compared with the control group, the wound repairing dressing group based on fat emulsion has the advantages of higher wound granulation tissue degree, abundant vascularity, abundant hair follicles and complete epithelization of the wound.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. Use of fat emulsion in the preparation of a wound repair material.
2. The use of claim 1, wherein the fat emulsion comprises 50-200 mg/mL soybean oil, C6～2425-100 mg/mL of medium-chain triglyceride and 6-12 mg/mL of lecithin.
3. The use according to claim 2, wherein C is6～24The medium chain triglyceride is selected from one or two of caprylic triglyceride and capric triglyceride.
4. A wound repair dressing comprising a fat emulsion and a carrier matrix.
5. The wound repair dressing of claim 4, wherein the fat emulsion comprises 50-200 mg/mL soybean oil, C6～2425-100 mg/mL of medium-chain triglyceride and 6-12 mg/mL of lecithin.
6. The wound repair dressing according to claim 4, wherein the carrier matrix is selected from one or more of medical gauze, medical sponge, medical foam, and hydrogel.
7. The wound repair dressing of claim 6, wherein the hydrogel is a physical gel or a chemical gel.
8. The wound repair dressing of any one of claims 4-7, wherein the mass ratio of the fat emulsion to the carrier matrix is 1: (2-2X 10)6)。
9. A method of preparing a wound repair dressing according to any one of claims 4 to 8, comprising the steps of:
diluting the fat emulsion in a biocompatible solvent to ensure that the concentration of the fat emulsion is 0.01-10000 mug/mL, and loading the obtained fat emulsion on a carrier matrix to obtain the fat emulsion.
10. The method of claim 9, wherein the biocompatible solvent is selected from the group consisting of water, PBS buffer, and liquid medium.
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