A medical nanocellulose gel cold compress patch and a preparation method thereof

Abstract

The application discloses a medical nanocellulose gel cold compress patch and a preparation method thereof. The cold compress patch comprises a cold compress part, wherein the cold compress part is sequentially provided with a film layer, a temperature insulation layer, a nanocellulose gel layer and a backing layer from top to bottom. An anti-diffusion frame is arranged at the outer ring of the cold compress part, which can effectively prevent the essence liquid in the cold compress part from diffusing to a non-cold compress part, thereby avoiding waste of the essence liquid. A sticky layer is fixed to the lower part of both sides of the anti-diffusion frame, and the sticky layer has a mesh structure and a hexagonal mesh hole, thereby ensuring the air permeability of the cold compress patch during use. A release paper is attached to the bottom end of the sticky layer. During use, the release paper is torn off, the cold compress part is aligned with a part to be cold compressed, and the sticky layer is pressed to be attached to the skin. The nanocellulose gel layer comprises a nanocellulose matrix and an essence liquid. The cold compress patch is a natural biological component, and due to the particularity of the nanocellulose structure, the skin can be deeply moisturized, thereby meeting the requirements of people in modern society for skin care.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to a medical nanocellulose gel cold compress and its preparation method. Background Technology

[0002] As people's living standards improve, they are paying more and more attention to skin care. Post-operative wounds or those resulting from cosmetic procedures often require physical cooling to aid healing. Traditional cold compresses use alcohol or ice, while existing cold compresses use synthetic hydrogels containing chemical ingredients instead of traditional gauze. Traditional cold compresses have a shorter duration of effect and are inconvenient to carry. The sharp edges of ice cubes can also damage the wound. Existing hydrogel cold compresses often contain chemical ingredients, which are not conducive to wound healing and can irritate the skin. They are not suitable for sensitive skin. Furthermore, when cold compresses need to be applied for a longer period, traditional cold compresses have poor moisturizing properties and may actually absorb moisture from the skin if left on for too long, which is not conducive to wound healing or skin care. Summary of the Invention

[0003] Therefore, it is necessary to provide a medical nanocellulose gel cold compress and its preparation method. The cold compress is a natural biological component, and due to the special structure of nanocellulose, it can deeply moisturize the skin, refresh the skin, and meet the skin care requirements of people in modern society.

[0004] To achieve the above objectives, the specific technical solution of the present invention is as follows:

[0005] Firstly, a medical nanofiber cellulose gel cooling patch is provided, comprising a cooling patch, which, from top to bottom, consists of a thin film layer, a heat-insulating layer, a nanofiber cellulose gel layer, and a backing layer. An anti-diffusion frame is provided around the outer edge of the cooling patch to effectively prevent the essence in the cooling patch from diffusing to non-cooling areas, thus avoiding waste. Adhesive layers are fixed to both sides of the lower part of the anti-diffusion frame. The adhesive layers have a mesh structure with hexagonal mesh openings to ensure breathability during use. Release paper is adhered to the bottom of the adhesive layers. In use, the release paper is peeled off, the cooling patch is aligned with the area requiring cooling, and the adhesive layers are pressed to adhere it to the skin.

[0006] Preferably, the nanocellulose gel layer comprises a nanocellulose gel matrix and an essence, wherein the components of the essence and their mass percentages are as follows: glycerin 2-3%, propylene glycol 0.6-1%, butylene glycol 0.6-1%, polyglutamic acid 0.01-2%, grape seed extract 0.01-2%, Bletilla striata extract 0.01-2%, Dendrobium officinale extract 0.01-2%, Spirulina extract 0.01-2%, Isatis indigotica root extract 0.01-2%, Paeonia suffruticosa root bark extract 0.01-2%, Paeonia lactiflora extract 0.01-2%, phenoxyethanol ≤0.03%, and the balance is water.

[0007] Glycerin is used as a dispersant, water, propylene glycol and butylene glycol are used as solvents, polyglutamic acid is used as a moisturizer, grape seed extract, Bletilla striata extract, Dendrobium officinale extract, Spirulina extract, Isatis indigotica root extract, Paeonia suffruticosa root bark extract and Paeonia lactiflora extract are used as skin conditioning agents, and phenoxyethanol is used as a preservative.

[0008] Grape seed extract has high antioxidant properties, which can delay aging, whiten and moisturize, remove blemishes and acne scars, promote scar healing, and effectively reduce inflammation and allergies; Bletilla striata extract can astringe and stop bleeding, reduce swelling and promote tissue regeneration, and also whiten and remove blemishes; Dendrobium officinale extract has highly effective anti-aging and moisturizing abilities; Spirulina extract can not only promote skin metabolism and reduce scars, but also inhibit bacterial growth and has good antibacterial ability; Isatis indigotica root extract is bitter and cold in nature, and can reduce inflammation and pain, disperse blood stasis and stop bleeding. As a medicinal ingredient in cold compresses, it can assist wound healing while relieving pain and inflammation; Paeonia suffruticosa root bark extract is bitter and slightly cold in nature, and can promote blood circulation, cool blood and remove blood stasis. When combined with Paeonia lactiflora extract, it enhances its effects of promoting blood circulation and dispersing blood stasis.

[0009] Preferably, the nanocellulose gel matrix is ​​biosynthesized by *Acetobacter xylinum* and has a three-dimensional mesh structure.

[0010] Furthermore, a method for preparing the medical nanocellulose gel cold compress is provided, characterized by comprising the following steps:

[0011] 1) Revive the bacterial strain;

[0012] 2) Preserve the bacterial strain;

[0013] 3) A nanocellulose gel matrix was prepared by culturing *Acetobacter xylodis* strains;

[0014] 4) Purify the nanocellulose gel matrix;

[0015] 5) The nanocellulose gel matrix obtained in step 4) was photographed and subjected to electron microscopy, and the results are as follows: Figure 4As shown, Figure (a) is a photograph of the morphology of the nanocellulose gel matrix, Figure (b) is a surface electron microscope image of the nanocellulose gel matrix, and Figure (c) is a cross-sectional electron microscope image of the nanocellulose gel matrix. Figure 4 The morphology and nanofiber structure of the nanocellulose gel matrix can be observed. Its fiber structure is a three-dimensional mesh structure, which gives the nanocellulose gel matrix excellent mechanical properties. The synthesis of the nanocellulose gel matrix involves bacteria synthesizing it layer by layer at the gas-liquid interface. Therefore, the fibers are tightly bound in the interlacing direction, and the tightness between layers contributes to the fiber interlacing direction. Figure 5 As shown in Figure (a) is a photograph of the nanocellulose gel matrix, Figure (b) is an electron microscope image of the fiber interlacing surface, and Figure (c) is an electron microscope image of the layered surface), this makes the mechanical properties of the nanocellulose gel matrix anisotropic.

[0016] 6) Cut the nanocellulose gel matrix obtained in step 4) into shape according to the use requirements, and then immerse it in the pre-prepared essence to fully absorb the essence and obtain the nanocellulose gel layer (3).

[0017] 7) The sample obtained in step 6) is subjected to quality inspection. After passing the inspection, the nanocellulose gel layer (3) is pasted, assembled, connected and packaged with the film layer (1), the heat insulation layer (2), the backing layer (4), the anti-diffusion frame (5), the adhesive layer (6) and the release paper (7).

[0018] Preferably, step 1) of reviving the bacterial strain includes: placing the bottle containing the *Acetobacter xylose* strain in a sterile laminar flow hood, wiping the bottle mouth with a cotton ball soaked in medical alcohol, then heating it with the outer flame of an alcohol lamp, adding sterile distilled water to the top of the heated bottle to crack the glass, removing the cracked top of the bottle with tweezers, using a pipette to draw 0.3-0.5 mL of liquid culture medium, adding it to the bottle, gently shaking to prepare a suspension of the lyophilized bacterial cells, transferring the suspension to a slant of a solid culture medium, culturing at 26-30°C for 36 hours, and storing the revived *Acetobacter xylose* in a refrigerator at 4°C.

[0019] Preferably, step 2) of preserving the bacterial strain includes:

[0020] Short-term preservation (three months) method: Under aseptic conditions, use an inoculation needle to gently brush the bacterial culture onto a slant solid medium in a "Z" pattern (20ml test tube, slant about 2 / 3 height). Incubate in a constant temperature incubator at 30℃ for 48 hours. If new colonies are observed to grow on the slant, store the bacterial culture on the slant in a refrigerator at 4℃. This is a short-term preservation method, and nanocellulose needs to be transferred every three months.

[0021] Long-term preservation method (one to twenty years): Under aseptic conditions, inoculate the bacterial strain into a sterile culture medium containing 20% ​​glycerol and store it in an ultra-low temperature freezer at -70°C. It can be preserved for one to twenty years.

[0022] Preferably, step 3) of preparing the nanocellulose gel matrix includes: dispensing the prepared *Acetobacter xylinum* strain culture medium into suitable containers, including 250ml culture flasks, various square and rectangular culture dishes, sealing the containers with filter paper, sealing film, or gauze, and then sterilizing them at 121°C for 20 minutes under high temperature and high pressure. After cooling overnight, in a sterile laminar flow hood, inoculate the bacterial strain into the culture medium at an inoculation rate of 8-10%. After inoculation, shake the container to distribute the bacterial strain evenly and place it in a 30°C incubator for static incubation. For cell culture plates: after 48 hours of bacterial culture, transfer the culture medium to the wells of the cell culture plate using a pipette in a laminar flow hood. The incubation time varies from 3 to 20 days depending on the required thickness.

[0023] Preferably, step 4) purifying the nanocellulose gel matrix includes: soaking the nanocellulose gel matrix in distilled water for 2 days, then boiling it in a 0.1 mol / L NaOH solution for 30 minutes, then rinsing the boiled matrix until neutral, and storing it in distilled water at 4°C for later use.

[0024] Preferably, step 5) involves photographing and electron microscopy to obtain a three-dimensional mesh structure of the nanocellulose gel matrix.

[0025] Furthermore, the application of the medical nanocellulose gel cold compress in the preparation of postoperative or cosmetic medical products is also provided.

[0026] Based on the above technical solution, the present invention has the following beneficial effects:

[0027] 1. Nanocellulose gel matrix is ​​a natural hydrogel produced and secreted by microorganisms. It has the same molecular structure as plant cellulose, but has properties that plant cellulose does not have. The cellulose secreted by microorganisms has high purity and does not contain impurities such as hemicellulose and lignin; it has high degree of polymerization (DP=4000-10000), high degree of hydration (>99%), and high degree of crystallinity (60-80%).

[0028] 2. The high moisturizing properties of the nanocellulose gel matrix solve the problem of membrane material drying out due to prolonged cold compresses;

[0029] 3. Compared with the production of plant cellulose, the synthetic cellulose produced by microbial fermentation has the following advantages: 1) The production of bacterial cellulose is not affected by region and climate; 2) The productivity and quality of bacterial cellulose produced by microorganisms are controllable; 3) Cellulose synthesized by microorganisms does not contain impurities such as hemicellulose and lignin, and does not require purification; 4) Microorganisms can obtain functionalized cellulose through genetic modification; 5) Industrial and agricultural waste can be used for the microbial fermentation production of cellulose.

[0030] 4. The nanocellulose gel membrane can absorb the moisture of the essence. When applied cold, the essence seeps out from the membrane and is absorbed by the skin. The membrane begins to thin and gradually tightens laterally. The nanostructure substrate makes the release of the essence a penetrating delivery process from the inside out, which can deeply moisturize the skin and refresh it.

[0031] 5. The addition of various Chinese herbal medicine ingredients to the skin conditioning agent promotes wound healing and also provides analgesic and soothing effects, improving the comfort of using the cold compress.

[0032] 6. The addition of skin conditioning agents enables the product to repair the skin barrier and promote scar repair while providing cold compresses, thus meeting people's needs for beauty and allowing the invention to be used for post-operative repair in medical aesthetics.

[0033] The medical cold compress of this invention is convenient to use, has a good moisturizing effect, and can be used for a long time without drying out the film material or absorbing moisture from the skin. In addition, the essence of this invention contains natural skin conditioning agents, which are safe, non-toxic and non-irritating, and can be used with confidence by sensitive skin. It has good biocompatibility, and its essence contains anti-inflammatory and blood-activating ingredients, which can help wound healing. It also has the effects of soothing the skin, cooling blood and relieving pain. It can quickly repair the skin barrier and promote scar repair, satisfying the needs of wound healing while also maintaining the skin, thus meeting people's needs for cold compress beauty function. Attached Figure Description

[0034] Figure 1 A schematic diagram of the structure of the medical nanocellulose gel cold compress patch of the present invention;

[0035] Figure 2 Schematic diagram of the adhesive layer structure;

[0036] Figure 3 Nanocellulose gel matrices obtained by culturing in different containers;

[0037] Figure 4 Photographs and electron microscope images of the nanocellulose gel matrix: Figure (a) is a morphological photograph, Figure (b) is a surface electron microscope image, and Figure (c) is a cross-sectional electron microscope image.

[0038] Figure 5The fiber interlacing pattern of the nanocellulose gel matrix is ​​shown in Figure (a), which is a photograph of the nanocellulose gel matrix; Figure (b) is an electron microscope image of the fiber interlacing surface; and Figure (c) is an electron microscope image of the layered surface.

[0039] In the diagram: 1. Thin film layer; 2. Thermal insulation layer; 3. Nanocellulose gel layer; 4. Backing layer; 5. Anti-diffusion frame; 6. Adhesive layer; 7. Release paper. Detailed Implementation

[0040] The following embodiments are used to illustrate the present invention, but are not intended to limit the scope of the invention. Unless otherwise specified, the technical means used in the embodiments are conventional means well known to those skilled in the art.

[0041] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0042] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0043] Example 1

[0044] like Figure 1 As shown, the medical nanocellulose gel cold compress of the present invention includes a cold compress part, wherein the cold compress part is provided with a thin film layer 1, a heat insulation layer 2, a nanocellulose gel layer 3 and a backing layer 4 in sequence from top to bottom. An anti-diffusion frame 5 is provided on the outer ring of the cold compress part, and an adhesive layer 6 is fixed on both sides of the lower part of the anti-diffusion frame 5. Figure 2 As shown, the adhesive layer 6 has a mesh structure with hexagonal mesh openings to ensure breathability during the use of the cold compress. Release paper 7 is attached to the bottom of the adhesive layer 6.

[0045] When using, peel off the release paper 7, align the cooling compress with the area that needs cooling, and press the adhesive layer 6 to stick it to the skin.

[0046] The nanocellulose gel layer comprises a nanocellulose gel matrix and an essence, the composition of which is as follows:

[0047] Glycerin 2%, Propylene glycol 0.6%, Butylene glycol 0.6%, Polyglutamic acid 0.01%, Grape seed extract 0.01%, Bletilla striata extract 0.01%, Dendrobium officinale extract 0.01%, Spirulina extract 0.01%, Isatis indigotica root extract 0.01%, Paeonia suffruticosa root bark extract 0.01%, Paeonia lactiflora extract 0.01%, Phenoxyethanol 0.001%, Balance: Water.

[0048] The nanocellulose gel matrix was biosynthesized by *Acetobacter xylinum*.

[0049] The preparation method of medical nanofiber cellulose gel cold compress is as follows:

[0050] 1. Resuscitation of microbial strains

[0051] Place the bottle containing the *Acetobacter xylose* strain in a sterile laminar flow hood. Wipe the bottle opening with a cotton ball soaked in medical alcohol, then heat it with the outer flame of an alcohol lamp. Add sterile distilled water to the heated bottle top until the glass cracks. Remove the cracked bottle top with tweezers. Use a pipette to add 0.3-0.5 mL of liquid culture medium to the bottle, gently agitate, and prepare a suspension of lyophilized bacteria. Transfer the suspension to a slant of solid culture medium and incubate at 26-30°C for 36 hours. Store the revived *Acetobacter xylose* in a refrigerator at 4°C.

[0052] 2. Preservation of microbial strains

[0053] Short-term preservation (three months) method: Under aseptic conditions, use an inoculation needle to gently brush the bacterial culture onto a slant solid culture medium in a "Z" pattern (20ml test tube, slant about 2 / 3 full). Incubate at 30℃ for 48 hours. If new colonies are observed growing on the slant, store the cultured bacterial culture at 4℃. This is a short-term preservation method, and it needs to be transferred to nanocellulose every three months.

[0054] Long-term preservation method (one to twenty years): Under aseptic conditions, inoculate the bacterial strain into a sterile culture medium containing 20% ​​glycerol and store it in an ultra-low temperature freezer at -70°C. It can be preserved for one to twenty years.

[0055] 3. Preparation of nanocellulose gel matrix

[0056] Dispense the prepared culture medium into suitable containers, including 250ml culture flasks and various square and rectangular culture dishes. Seal the containers using filter paper, film, or gauze, then autoclave at 121°C for 20 minutes. Allow to cool overnight. In a sterile laminar flow hood, inoculate the bacterial culture into the culture medium at an inoculation rate of 8-10%. After inoculation, shake to distribute the bacteria evenly and incubate at 30°C. For cell culture plates: After 48 hours of bacterial culture, transfer the culture medium to the wells of a cell culture plate using a pipette in a laminar flow hood. The incubation time varies from 3 to 20 days depending on the desired thickness.

[0057] 4. Purification treatment of nanocellulose gel matrix

[0058] The nanocellulose gel matrix was soaked in distilled water for 2 days, then boiled in a 0.1 mol / L NaOH solution for 30 minutes. The boiled membrane was then rinsed until neutral and stored in distilled water at 4°C for later use. Nanocellulose gel matrices obtained from different container cultures were shown below. Figure 3 As shown.

[0059] 5. The nanocellulose gel matrix obtained in step 4 was photographed and subjected to electron microscopy. The results are as follows: Figure 4 As shown, Figure (a) is a photograph of the morphology of the nanocellulose gel matrix, Figure (b) is a surface electron microscope image of the nanocellulose gel matrix, and Figure (c) is a cross-sectional electron microscope image of the nanocellulose gel matrix. Figure 4 The morphology and nanofiber structure of the nanocellulose gel matrix can be observed. Its fiber structure is a three-dimensional mesh structure, which gives the nanocellulose gel matrix excellent mechanical properties. The synthesis of the nanocellulose gel matrix involves bacteria synthesizing it layer by layer at the gas-liquid interface. Therefore, the fibers are tightly bound in the interlacing direction, and the tightness between layers contributes to the fiber interlacing direction. Figure 5 As shown in Figure (a) is a photograph of the nanocellulose gel matrix, Figure (b) is an electron microscope image of the interwoven fiber surface, and Figure (c) is an electron microscope image of the layered surface), this makes the mechanical properties of the nanocellulose gel matrix anisotropic.

[0060] 6. Cut the nanocellulose gel matrix obtained in step 4 into the shape required for use, and then immerse it in the pre-prepared essence to fully absorb the essence, thus obtaining nanocellulose gel layer 3.

[0061] 7. Perform quality inspection on the sample obtained in step 6. After the inspection is qualified, attach, assemble, connect and package the nanocellulose gel layer 3 with the film layer 1, the heat insulation layer 2, the backing layer 4, the anti-diffusion frame 5, the adhesive layer 6 and the release paper 7.

[0062] Example 2

[0063] like Figure 1 As shown, the medical nanocellulose gel cold compress of the present invention includes a cold compress part, wherein the cold compress part is provided with a thin film layer 1, a heat insulation layer 2, a nanocellulose gel layer 3 and a backing layer 4 in sequence from top to bottom. An anti-diffusion frame 5 is provided on the outer ring of the cold compress part, and an adhesive layer 6 is fixed on both sides of the lower part of the anti-diffusion frame 5. Figure 2 As shown, the adhesive layer 6 has a mesh structure with hexagonal mesh openings to ensure breathability during the use of the cold compress. Release paper 7 is attached to the bottom of the adhesive layer 6.

[0064] When using, peel off the release paper 7, align the cooling compress with the area that needs cooling, and press the adhesive layer 6 to stick it to the skin.

[0065] The nanocellulose gel layer comprises a nanocellulose gel matrix and an essence, the composition of which is as follows:

[0066] Glycerin 3%, Propylene glycol 1%, Butylene glycol 1%, Polyglutamic acid 2%, Grape seed extract 2%, Bletilla striata extract 2%, Dendrobium officinale extract 2%, Spirulina extract 2%, Isatis indigotica root extract 2%, Paeonia suffruticosa root bark extract 2%, Paeonia lactiflora extract 2%, Phenoxyethanol 0.03%, Balance: Water.

[0067] The nanocellulose gel matrix was biosynthesized by *Acetobacter xylinum*.

[0068] The preparation method of the medical nanocellulose gel cold compress is the same as that in Example 1.

[0069] Example 3

[0070] like Figure 1 As shown, the medical nanocellulose gel cold compress of the present invention includes a cold compress part, wherein the cold compress part is provided with a thin film layer 1, a heat insulation layer 2, a nanocellulose gel layer 3 and a backing layer 4 in sequence from top to bottom. An anti-diffusion frame 5 is provided on the outer ring of the cold compress part, and an adhesive layer 6 is fixed on both sides of the lower part of the anti-diffusion frame 5. Figure 2 As shown, the adhesive layer 6 has a mesh structure with hexagonal mesh openings to ensure breathability during the use of the cold compress. Release paper 7 is attached to the bottom of the adhesive layer 6.

[0071] When using, peel off the release paper 7, align the cooling compress with the area that needs cooling, and press the adhesive layer 6 to stick it to the skin.

[0072] The nanocellulose gel layer comprises a nanocellulose gel matrix and an essence, the composition of which is as follows:

[0073] Glycerin 2.5%, Propylene glycol 0.8%, Butylene glycol 0.8%, Polyglutamic acid 1%, Grape seed extract 1%, Bletilla striata extract 1%, Dendrobium officinale extract 1%, Spirulina extract 1%, Isatis indigotica root extract 1%, Paeonia suffruticosa root bark extract 1%, Paeonia lactiflora extract 1%, Phenoxyethanol 0.02%, Balance: Water.

[0074] The nanocellulose gel matrix was biosynthesized by *Acetobacter xylinum*.

[0075] The preparation method of the medical nanocellulose gel cold compress is the same as that in Example 1.

[0076] Example 4

[0077] like Figure 1 As shown, the medical nanocellulose gel cold compress of the present invention includes a cold compress part, wherein the cold compress part is provided with a thin film layer 1, a heat insulation layer 2, a nanocellulose gel layer 3 and a backing layer 4 in sequence from top to bottom. An anti-diffusion frame 5 is provided on the outer ring of the cold compress part, and an adhesive layer 6 is fixed on both sides of the lower part of the anti-diffusion frame 5. Figure 2As shown, the adhesive layer 6 has a mesh structure with hexagonal mesh openings to ensure breathability during the use of the cold compress. Release paper 7 is attached to the bottom of the adhesive layer 6.

[0078] When using, peel off the release paper 7, align the cooling compress with the area that needs cooling, and press the adhesive layer 6 to stick it to the skin.

[0079] The nanocellulose gel layer comprises a nanocellulose gel matrix and an essence, the composition of which is as follows:

[0080] Glycerin 3%, Propylene glycol 0.8%, Butylene glycol 0.8%, Polyglutamic acid 1%, Grape seed extract 2%, Bletilla striata extract 2%, Dendrobium officinale extract 2%, Spirulina extract 2%, Isatis indigotica root extract 1%, Paeonia suffruticosa root bark extract 1%, Paeonia lactiflora extract 1%, Phenoxyethanol 0.02%, Balance: Water.

[0081] The nanocellulose gel matrix was biosynthesized by *Acetobacter xylinum*.

[0082] The preparation method of the medical nanocellulose gel cold compress is the same as that in Example 1.

[0083] Example 5

[0084] like Figure 1 As shown, the medical nanocellulose gel cold compress of the present invention includes a cold compress part, wherein the cold compress part is provided with a thin film layer 1, a heat insulation layer 2, a nanocellulose gel layer 3 and a backing layer 4 in sequence from top to bottom. An anti-diffusion frame 5 is provided on the outer ring of the cold compress part, and an adhesive layer 6 is fixed on both sides of the lower part of the anti-diffusion frame 5. Figure 2 As shown, the adhesive layer 6 has a mesh structure with hexagonal mesh openings to ensure breathability during the use of the cold compress. Release paper 7 is attached to the bottom of the adhesive layer 6.

[0085] When using, peel off the release paper 7, align the cooling compress with the area that needs cooling, and press the adhesive layer 6 to stick it to the skin.

[0086] The nanocellulose gel layer comprises a nanocellulose gel matrix and an essence, the composition of which is as follows:

[0087] Glycerin 2%, Propylene glycol 1%, Butylene glycol 1%, Polyglutamic acid 2%, Grape seed extract 1%, Bletilla striata extract 1%, Dendrobium officinale extract 1%, Spirulina extract 1%, Isatis indigotica root extract 2%, Paeonia suffruticosa root bark extract 2%, Paeonia lactiflora extract 2%, Phenoxyethanol 0.02%, Balance: Water.

[0088] The nanocellulose gel matrix was biosynthesized by *Acetobacter xylinum*.

[0089] The preparation method of the medical nanocellulose gel cold compress is the same as that in Example 1.

[0090] Test case

[0091] 1. Take 50 mg of each of the five nanocellulose gel matrix samples prepared in Examples 1-5 and place them in five conical flasks. Add a 100 mg / L methylene blue solution to each flask, mix thoroughly with the aid of a water bath shaker, and conduct adsorption experiments at room temperature. After a certain reaction time (0 h, 2 h, 6 h, 9 h, 12 h, 24 h), the supernatant was centrifuged and the absorbance was measured at a wavelength of 664 nm. The results are shown in Table 1.

[0092] Table 1. UV absorbance values ​​of methylene blue solutions treated at different time periods

[0093]

[0094]

[0095] As can be seen from Table 1, the absorbance of the methylene blue solution treated with the nanocellulose gel matrix prepared in Examples 1-5 decreased significantly within 24 hours, proving that the nanocellulose gel matrix prepared in Examples 1-5 has high adsorption performance, enabling it to absorb the essence well.

[0096] 2. Antibacterial effect experiment

[0097] Several circular sterile filter paper discs, 3 mm thick and 6 mm in diameter, were immersed in sterilized nanocellulose gel layer sample extracts, physiological saline, and penicillin for 30 min respectively. 100 μL of a bacterial suspension containing approximately 0.5 Mcf of Staphylococcus aureus was evenly spread on the surface of culture medium plates. Six filter paper discs were then placed on each plate with a certain distance between them using sterile forceps. All operations were performed on a clean bench. The plates were then incubated at 37°C for 24 h, and the diameter of the inhibition zone of the circular filter paper discs was measured. Filter paper discs numbered 1, 2, and 3 were immersed in penicillin; those numbered 4, 5, 6, 7, and 8 were immersed in the sample extracts of Examples 1, 2, 3, 4, and 5 respectively; and those numbered 9, 10, and 11 were immersed in physiological saline. The results are shown in Table 2.

[0098] Table 2. Results of antibacterial test

[0099] Filter paper number 1 2 3 4 5 6 7 8 9 10 11 Diameter of the inhibition zone (mm) 57 57 56 16.5 18.5 17 17.5 18 6 6.5 6

[0100] As shown in Table 2, the diameter of the inhibition zone of the filter paper soaked in the sample extract was significantly larger than that of the filter paper soaked in physiological saline, but smaller than that of the filter paper soaked in penicillin. This indicates that Examples 1, 2, 3, 4 and 5 all had a more significant inhibitory effect on Staphylococcus aureus than physiological saline, but their inhibitory effect was not as strong as that of penicillin.

[0101] 3. Toxicity testing

[0102] HaCat cells in the logarithmic growth phase were selected, seeded in 96-well plates, and cultured for 24 hours. 100 μL of pre-prepared essence solution was added to each well. The composition of the essence solution was as shown in Examples 1-5. Six replicates were set up for each group. A negative control group was also set up. 100 μL of cell culture medium was added to the negative control group. After culturing for another 24 hours, WST-1 assay was performed. The results of the percentage of viable cells in each component are shown in Table 3.

[0103] Table 3. Percentage of HaCat active cells

[0104]

[0105] As can be seen from Table 3, there was no significant difference in the survival rate of HaCat cells in the components with added essence and the control group. This indicates that the essence prepared in Examples 1-5 has no significant effect on cell growth and reproduction, and can be considered as having no cytotoxicity. At the same time, it can be demonstrated that the essence has good biocompatibility.

[0106] 4. Animal experiments

[0107] Six healthy SD rats were anesthetized by intraperitoneal injection of pentobarbital 30 mg / kg. A circular full-thickness skin excision with a diameter of 3 cm was surgically removed from one side of the spine on the back, reaching the fascia. The symmetrical skin on the other side of the spine served as a normal control. Hemostasis was then performed on the wounds. The rats were numbered, and samples of the cold compresses from Examples 1-5 were applied to the wound surfaces of five rats numbered 1-5, respectively. The remaining rat, numbered 6, which was not treated with the cold compress, was treated with topical saline solution as a control group. After the six rats regained consciousness, they were housed individually. The dressings were changed every other day at the same dosage until healing. The healing status of each rat was observed on days 3, 10, 15, 20, and 25. The results are shown in Table 4.

[0108] Table 4. Wound healing status in rats

[0109]

[0110]

[0111] As can be seen from Table 4, Examples 1-5 of the present invention all have significant effects in promoting wound healing and have excellent moisturizing and repairing effects on skin tissue, which can significantly reduce scars.

[0112] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A medical nanofiber cellulose gel cooling patch, comprising a cooling compress portion, characterized in that, The cold compress part is provided with a thin film layer (1), a heat insulation layer (2), a nanocellulose gel layer (3) and a backing layer (4) from top to bottom. The outer ring of the cold compress part is provided with an anti-diffusion frame (5). The anti-diffusion frame (5) can effectively prevent the essence in the cold compress part from diffusing to the non-cold compress part. Adhesive layers (6) are fixed on both sides of the lower part of the anti-diffusion frame (5). The adhesive layer (6) has a mesh structure with hexagonal mesh holes. Release paper (7) is attached to the bottom of the adhesive layer (6). The nanocellulose gel layer (3) comprises a nanocellulose gel matrix and an essence, wherein the components of the essence and their mass percentages are as follows: glycerol 2-3%, propylene glycol 0.6-1%, butylene glycol 0.6-1%, polyglutamic acid 0.01-2%, grape seed extract 0.01-2%, Bletilla striata extract 0.01-2%, Dendrobium officinale extract 0.01-2%, Spirulina extract 0.01-2%, Isatis indigotica root extract 0.01-2%, Paeonia suffruticosa root bark extract 0.01-2%, Paeonia lactiflora extract 0.01-2%, phenoxyethanol ≤0.03%, and the remainder is water; The nanocellulose gel matrix is ​​biosynthesized by *Acetobacter xylodis* and has a three-dimensional mesh structure.

2. The preparation method of the medical nanocellulose gel cold compress according to claim 1, characterized in that, Includes the following steps: 1) Revive the bacterial strain; 2) Preserve the bacterial strain; 3) A nanocellulose gel matrix was prepared by culturing *Acetobacter xylodis* strains; 4) Purify the nanocellulose gel matrix; 5) The nanocellulose gel matrix obtained in step 4) was photographed and subjected to electron microscopy; 6) Cut the nanocellulose gel matrix obtained in step 4) into shape according to the use requirements, and then immerse it in the pre-prepared essence to fully absorb the essence and obtain the nanocellulose gel layer (3). 7) The sample obtained in step 6) is subjected to quality inspection. After passing the inspection, the nanocellulose gel layer (3) is pasted, assembled, connected and packaged with the film layer (1), the heat insulation layer (2), the backing layer (4), the anti-diffusion frame (5), the adhesive layer (6) and the release paper (7).

3. The method according to claim 2, characterized in that, Step 1) The steps for reviving the bacterial strain include: placing the bottle containing the *Acetobacter xylose* strain in a sterile laminar flow hood, wiping the bottle mouth with a cotton ball soaked in medical alcohol, then heating it with the outer flame of an alcohol lamp, adding sterile distilled water to the top of the heated bottle to crack the glass, removing the cracked top of the bottle with tweezers, using a pipette to draw 0.3-0.5 mL of liquid culture medium and adding it to the bottle, gently shaking to prepare a suspension of the lyophilized bacterial cells, transferring the suspension to a slant of solid culture medium, culturing at 26-30°C for 36 hours, and storing the revived *Acetobacter xylose* in a refrigerator at 4°C.

4. The method according to claim 2, characterized in that, Step 2) The steps for preserving the bacterial strain include: Short-term preservation method: Under aseptic conditions, use an inoculation needle to gently brush the bacterial culture onto a slant solid medium in a "Z" pattern. Incubate at 30°C for 48 hours. If new colonies are observed to grow on the slant, store the bacterial culture in a 4°C freezer. This is a short-term preservation method, and nanocellulose needs to be transferred every three months. Long-term preservation method: Under aseptic conditions, inoculate the bacterial culture into a sterile medium containing 20% ​​glycerol and store in a -70°C ultra-low temperature freezer. It can be preserved for one to twenty years.

5. The method according to claim 2, characterized in that, Step 3) The steps for preparing the nanocellulose gel matrix include: dispensing the prepared *Acetobacter xylinum* strain culture medium into suitable containers, including 250ml culture flasks, various square and rectangular culture dishes, sealing the containers with filter paper, film, or gauze, and then autoclaving at 121°C for 20 minutes, cooling overnight, and inoculating the bacteria into the culture medium in a sterile laminar flow hood at an inoculation rate of 8-10%. After inoculation, shake to distribute the bacteria evenly, and place in a 30°C incubator for static incubation. For cell culture plates: after 48 hours of bacterial culture, transfer the culture medium to the wells of the cell culture plate using a pipette in a laminar flow hood. The incubation time varies from 3 to 20 days depending on the required thickness.

6. The method according to claim 2, characterized in that, Step 4) The purification steps of the nanocellulose gel matrix include: soaking the nanocellulose gel matrix in distilled water for 2 days, then boiling it in a 0.1 mol / L NaOH solution for 30 minutes, then rinsing the boiled matrix until neutral, and storing it in distilled water at 4°C for later use.

7. The method according to claim 2, characterized in that, Step 5) Photographs and electron microscopy revealed that the fibrous structure of the nanocellulose gel matrix is ​​a three-dimensional mesh structure.

8. The application of the medical nanocellulose gel cold compress according to claim 1 in the preparation of postoperative or medical aesthetic products.

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