Biological dressing containing glycosylglycerol as well as preparation method and application of biological dressing

Abstract

The invention relates to a biological dressing containing glycosylglycerol and a preparation method and application thereof. The biological dressing comprises a base material layer and a functional coating loaded on the base material layer, wherein the functional coating comprises glycosylglycerol (GG), a biocompatible high polymer material and a pH responsive dye; the pH responsive dye can generate macroscopic color change when the pH is increased due to wound infection; the functional coating further comprises mussel mucin and / or epidermal growth factor (EGF), and the functional coating and GG form a sequential controlled release system. The preparation method comprises the step of forming the functional coating through a visible light-initiated'one-step 'cross-linking load or a coaxial electrostatic spinning technology. The dressing disclosed by the invention not only has excellent moisturizing and healing promoting performance, but also can realize early intelligent early warning (pH response) of wound infection and on-demand release (enzyme response) of drugs, and can be used for synergistically resisting bacteria and inflammation, remarkably accelerating the healing process of chronic wound difficult to heal and reducing scar formation.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medical devices and biomaterials, in particular to a glyceryl glucoside-containing biological dressing and its preparation method and application, specifically to a novel intelligent responsive biological dressing, in particular to a glyceryl glucoside-containing biological dressing with infection intelligent early warning, time-controlled release and enzyme-responsive degradation functions, its preparation method and its application in wound repair and nursing. BACKGROUND

[0002] Wound healing is a complex and highly coordinated biological process, including the inflammatory phase, the proliferative phase and the remodeling phase. An ideal wound dressing should be able to simulate the barrier function of healthy skin and provide a moist, breathable, sterile and healing-promoting microenvironment for the wound. Traditional dressings such as gauze and cotton pads, although have certain absorption of exudate and protection of wound, are easy to adhere to the new granulation tissue, and replacement will cause secondary damage, and poor moisture retention is not conducive to cell migration and epithelialization.

[0003] With the development of material science, new functional dressings such as hydrocolloid, foam, hydrogel and alginate dressing have emerged, which have significantly improved in terms of moisture retention and absorption of exudate. However, for chronic non-healing wounds such as diabetic foot ulcers, pressure sores, venous ulcers, etc., existing dressings are often still insufficient. These wounds often have problems such as blood circulation disorders, persistent inflammation, high proteolytic enzyme activity and impaired cell function, and there is an urgent need for "intelligent" dressings with active healing-promoting function.

[0004] Glyceryl glucoside (GG) is a naturally occurring glycoside compound composed of one molecule of glycerol and one molecule of glucose connected by a glycosidic bond. Recent studies have found that GG has excellent moisturizing properties and cell protection effects. It can up-regulate the expression of aquaporin 3 (AQP3) and promote the uptake of water and glycerol by skin keratinocytes and fibroblasts, thereby enhancing skin barrier function and improving skin hydration. In addition, studies have shown that GG can reduce UV-induced cell damage and has certain anti-inflammatory and antioxidant potential. However, the application of GG in the field of wound dressings to actively intervene and accelerate the wound healing process using its unique biological functions has not been systematically reported and mature products have not been developed.

[0005] There are some attempts in the prior art to add moisturizers or active ingredients to dressings. For example, patent CN116473897B discloses a hyaluronic acid-containing liquid dressing and its preparation method and application, which relates to the field of daily chemical technology. The present application uses wheat germ oil, dendrobium water extract and chamomile ethanol extract as moisturizers on the basis of hyaluronic acid, fully utilizes the properties of functional ingredients such as hyaluronic acid, wheat germ oil, dendrobium polysaccharide and chamomile flavonoids, on the one hand, promotes skin regeneration and repair, maintains moisture in the skin, plays a role in moisturizing and preventing skin aging, on the other hand, forms a closed layer on the skin to prevent water loss, increases the moisturizing effect, and through the hydration capacity of functional ingredients, improves the effect of moisture absorption and moisture retention, further enhances the moisturizing effect, plays a role in moisturizing through endogenous and exogenous mechanisms, thereby achieving ideal moisturizing effect, and can be used for moisturizing products. Although its moisturizing effect is good, the promotion of cell migration is limited.

[0006] For example, patent CN118121751A discloses a growth factor-containing hydrogel wound dressing and its preparation method, which belongs to the technical field of medical dressings. The dressing contains 1.5-4 parts of gel matrix, 0.5-4 parts of chitosan, 0.5-4 parts of trehalose, 0.0001-0.001 parts of tannic acid, 1-30 parts of glycerol, 10-30 mg / mL of epidermal growth factor, and the balance is water. A preparation method of the chitosan hydrogel wound dressing containing growth factors, characterized in that it comprises the following steps: step S1: dissolve glycerol in deionized water to obtain a glycerol-water mixed solution; step S2: dissolve chitosan in the glycerol-water mixed solution; step S3: add trehalose and tannic acid and mix thoroughly; step S4: add gel matrix and mix thoroughly; step S5: add epidermal growth factor and mix thoroughly; step S6: add triethanolamine to adjust the pH to 6.0-7.0, stand at room temperature to eliminate bubbles, sterilize and pack, and the growth factor-containing hydrogel dressing is obtained. It is a growth factor-containing dressing, which has significant healing-promoting effect, but the release of growth factors cannot be controlled, is easily degraded by wound proteases, and the growth factors are expensive, have poor stability, are easily inactivated, and have potential safety risks.

[0007] GG is a natural, safe, stable and relatively low-cost active ingredient. If it can be combined with a suitable dressing substrate, a new type of biological dressing can be developed, which is expected to significantly improve the healing performance of the dressing under the premise of safety, and fill the market gap.

[0008] Therefore, there is an urgent need in the art to develop a "unified" smart dressing which can not only monitor the wound state (such as infection) in real time, but also intelligently release different functional active ingredients (such as activating cells for moisture preservation first, then promoting proliferation, and simultaneously resisting bacteria and inflammation) according to the changes in the wound microenvironment, and its structure can guide tissue regeneration. This is still a technical problem that has not been solved in the art, and has great clinical needs and market prospects. SUMMARY

[0009] In view of the problems in the prior art, such as single function of wound dressing, lack of intelligent responsiveness, low utilization rate of active ingredients, limited treatment effect on complex chronic wounds, etc., the present application aims to provide a novel multifunctional intelligent responsive biological dressing. The dressing not only provides excellent physical protection and moisturizing effect, but also realizes: 1) early and naked-eye visible warning of wound infection; 2) time sequence controlled release and on-demand release of various active ingredients such as moisturizing, antibacterial and growth promoting; 3) active guidance of tissue regeneration through biomimetic structure.

[0010] The primary object of the present application is to provide a glycerol glucoside (GG) containing biological dressing. Through the synergistic effect of GG and biocompatible polymer material, the dressing can provide an ideal moist environment for the wound and effectively promote cell migration and proliferation.

[0011] Another object of the present application is to provide a preparation method of the above-mentioned biological dressing. The method is simple in process, mild in conditions, easy to scale up, and can effectively maintain the biological activity of GG.

[0012] Still another object of the present application is to provide the use of the above-mentioned biological dressing in the preparation of medical devices for promoting wound (especially chronic wound) healing, moisturizing and reducing scar formation.

[0013] The above objects of the present application are achieved by the following technical solutions:

[0014] In a first aspect, the present application provides a glycerol glucoside containing biological dressing, comprising a substrate layer and a functional coating layer loaded on the substrate layer, wherein the functional coating layer comprises glycerol glucoside (GG), biocompatible polymer material and a pH responsive dye; the pH responsive dye can cause naked-eye visible color change when the wound infection causes the pH to rise; the mass of the glycerol glucoside accounts for 0.1% to 10% of the total mass of the biological dressing.

[0015] Preferably, the functional coating layer further comprises mussel myoglobin and / or epidermal growth factor (EGF). The mass ratio of the mussel myoglobin to the glycerol glucoside is (1:10) to (1:1), and the two components synergistically play the roles of moisturizing, antibacterial and healing promotion. The concentration of the epidermal growth factor is 1-100 μg / g of the dressing, which is used to precisely promote cell proliferation.

[0016] Preferably, the pH-responsive dye is one of bromothymol blue (color change range: yellow 6.0- blue 7.6), phenol red or alizarin red S. The biocompatible polymer material is a light-crosslinkable or enzyme-responsive material, such as one of methacrylated gelatin (GelMA), sodium alginate-calcium ion network, or hyaluronic acid-dopamine complex.

[0017] Preferably, the substrate layer is a bacterial cellulose (BC) membrane, or a polycaprolactone (PCL) / gelatin (Gelatin) fiber membrane with anisotropic pores prepared by electrospinning technology. The BC membrane provides extreme moisturizing and air permeability, and the anisotropic fiber membrane can guide the directional migration of cells.

[0018] More preferably, the functional coating is a time-controlled release system, in which glycerol glucoside and mussel myoglobin are the rapid release phase, with a release rate of more than 70% within 24 hours, to quickly create a humid environment and inhibit early infection; and epidermal growth factor is the slow release phase, with a cumulative release rate of not more than 50% within 7 days, to achieve long-term proliferation promotion.

[0019] Further, the functional coating is also designed to be responsive to matrix metalloproteinases (MMPs). When the concentration of MMPs at the wound site abnormally increases, the specific peptide cross-linking bond in the functional coating breaks, causing the coating to degrade and accelerate the release of the loaded drug components, thereby achieving on-demand drug delivery.

[0020] In a second aspect, the present application provides two innovative methods for preparing the above-mentioned biological dressing:

[0021] Method one: "one-step method" using visible light-induced crosslinking, which is mild and efficient, and can well protect the active ingredients. The steps include preparing a prepolymer solution containing a photoinitiator, a polymer, GG, a dye, mussel myoglobin, etc., and then instantaneously curing and forming a functional coating after coating and visible light irradiation.

[0022] S1. Preparation of prepolymer solution: dissolve the photoinitiator LAP, methacrylated gelatin, glycerol glucoside, pH-responsive dye, and mussel myoglobin in PBS, stir uniformly and avoid light;

[0023] S2. Coating and crosslinking: coat the prepolymer solution on the substrate layer, and crosslink and cure under visible light irradiation at a wavelength of 405 nm for 30-60 seconds to form a functional coating;

[0024] S3. Post-processing: after cutting, packaging, and irradiation sterilization, the biological dressing is obtained.

[0025] Method two: using coaxial electrospinning technology, the substrate and functional layer integrated product can be prepared, and the time sequence control release of core-sheath structure can be realized. PCL solution is used as the shell layer, and gelatin, GG and EGF aqueous solution are used as the core layer to perform coaxial electrospinning, and a film is directly formed.

[0026] A1. Prepare the shell layer solution: dissolve polycaprolactone in an organic solvent to form a uniform solution;

[0027] A2. Prepare the core layer solution: dissolve gelatin, glycerol glucoside and epidermal growth factor in deionized water to form a uniform solution;

[0028] A3. Coaxial electrospinning: using the shell layer solution as the outer shell and the core layer solution as the core material, coaxial electrospinning is performed to directly form a fiber film of the substrate layer and the functional coating integrated on the receiving device;

[0029] A4. Post-processing: after the fiber film is crosslinked and solidified, it is cut, packaged and sterilized to obtain the biological dressing.

[0030] In a third aspect, the present application provides the use of the above-mentioned biological dressing in the preparation of a medical device for promoting wound healing (especially chronic infected wounds), intelligently warning infection and reducing scar formation. The wound includes but is not limited to surgical incisions, burns, scalds, abrasions, cuts, diabetic foot ulcers, pressure sores, venous ulcers, etc. It is particularly suitable for chronic difficult-to-heal wounds that require high moisture and active repair.

[0031] In summary, compared with the prior art, the present application has at least one of the following beneficial technical effects:

[0032] 1. The present application discloses a biological dressing containing glycerol glucoside and its preparation method and application. The biological dressing is composed of a substrate layer and a functional coating layer. The substrate layer provides physical support and release carrier, and can firmly adhere to the wound surface. The functional coating layer loads glycerol glucoside and biocompatible polymer materials to promote wound healing, moisturizing and reduce scar formation. Glycerol glucoside as an active ingredient has good moisturizing performance and tissue repair function, which helps to maintain a moist wound environment and accelerate the wound healing process. Biocompatible polymer materials act as carriers, which not only can stably package glycerol glucoside, but also can provide certain adhesion, sustained release or mechanical support properties according to the selected materials. Among them, the mixture of hyaluronic acid and sodium alginate as a high polymer material can balance between moisturizing and promoting healing by adjusting the proportion, and its good biocompatibility and adhesion help to improve the use effect of the biological dressing. In summary, the biological dressing realizes the effects of promoting wound healing, maintaining a moist environment and reducing scar formation through the synergistic effect of each component, and provides an efficient and safe wound care solution.

[0033] 2. The present application creatively introduces glycerol glucoside (GG) into biological dressings. GG not only provides and maintains a moist healing environment for wounds through its strong moisturizing effect, but also actively promotes the migration and proliferation of keratinocytes and fibroblasts by up-regulating the expression of aquaporin AQP3, thereby accelerating the formation of granulation tissue and the process of epithelialization, and significantly shortening the wound healing time. This is particularly important for chronic wounds with impaired cell function. The biocompatible polymers in the functional coating (such as hyaluronic acid, sodium alginate) have extremely strong water-locking ability, which synergizes with the moisturizing effect of GG to maintain wound moisture for a long time without immersion. At the same time, the porous or gel-like coating formed allows gas exchange, maintains wound ventilation, and avoids the risk of anaerobic bacteria breeding.

[0034] 3. The present application has high safety and biocompatibility: GG is a naturally derived ingredient, safe and non-toxic, with extremely low irritation. The selected polymer materials are also clinically proven biocompatible materials. The entire dressing does not contain toxic and harmful substances, is safe and reliable to use, and reduces the risk of allergies and adverse reactions. The present application overcomes the shortcomings of existing active dressings: compared with dressings containing growth factors, GG is chemically stable, not easily inactivated, resistant to storage, and low in cost, making it easier to achieve industrialized production and clinical application.

[0035] 4. The present application creatively integrates three intelligent functions of pH-responsive infection early warning, enzyme-responsive drug on-demand release, and time-controlled release of active ingredients into one. The dressing can change from "passive covering" to "active management", and clinicians and patients can visually judge the infection situation through color change, intervene in time, and avoid the aggravation of infection. The present application also has multiple active ingredients synergistic effect: by introducing mussel mucin, not only the antibacterial effect is enhanced, and the cytotoxicity of silver ions is avoided, but also its own adhesion-promoting and healing-promoting properties produce unexpected synergistic effects with the moisturizing property of GG and the proliferation-promoting property of EGF (see Example 4), significantly surpassing the effect of single component or simple mixture.

[0036] 5. The present application adopts innovative process to protect activity: "one-step method" visible light crosslinking and "coaxial electrospinning" technology, which avoids the damage of high temperature, organic solvent or strong chemical crosslinking agent to sensitive active substances such as growth factors, GG, greatly improves the loading rate and survival rate of active ingredients, and the process is simple and efficient, suitable for industrialization. Biomimetic structure guides repair: using bacterial cellulose or anisotropic electrospinning fiber membrane as the substrate, not only provides excellent physical properties, its nanofiber structure can better simulate the extracellular matrix (ECM), provide a climbing scaffold for cells, actively guide cell directional migration and ordered proliferation, and help reduce scar formation. Solve clinical pain points: the dressing is especially suitable for complex and refractory chronic wounds, which can effectively deal with the core problems such as susceptibility to infection, poor healing environment, and low cell function, and has great clinical value and market prospect.

[0037] In summary, the present application provides a novel component, unique structure, powerful and intelligent and efficient biological dressing by cross-fusion of multiple technologies, solves many bottleneck problems in the prior art, and has outstanding substantial characteristics and significant progress. BRIEF DESCRIPTION OF DRAWINGS

[0038] Figure 1 is a real photo of the glycerol glucoside-containing intelligent biological dressing prepared in Example 1 of the present application.

[0039] Figure 2 is a structural schematic diagram of the biological dressing described in the present application (the functional coating is loaded on the substrate layer).

[0040] Figure 3 is a process flow diagram for preparing the functional coating by visible light crosslinking "one-step method" in Example 1 of the present application.

[0041] Figure 4 is a process flow diagram for preparing an integrated dressing by coaxial electrospinning technology in Example 2 of the present application.

[0042] Reference signs: 1, substrate layer; 2, functional coating. DETAILED DESCRIPTION

[0043] In order to make the purpose, technical scheme and advantages of the present application clearer and more apparent, the present application will be further described in detail below in combination with the drawings and examples. It should be understood that the specific examples described herein are only used to explain the present application and do not limit the present application. Based on the examples in the present application, all other examples obtained by those of ordinary skill in the art without making creative efforts fall within the scope of the present application.

[0044] The test methods used in the following examples are conventional methods unless otherwise specified; the materials, reagents, etc. used are commercially available reagents and materials unless otherwise specified.

[0045] Reference Figure 1 and Figure 2 The present application relates to the technical field of medical devices, and specifically discloses a glycerol glucoside-containing intelligent response biological dressing and a preparation method and application thereof. The biological dressing comprises a substrate layer 1 and a functional coating 2 loaded thereon, the functional coating 2 comprises glycerol glucoside (GG), a biocompatible polymer material and a pH-responsive dye; the pH-responsive dye can undergo a macroscopically visible color change when the pH is increased due to wound infection; the mass of the glycerol glucoside accounts for 0.1% to 10% of the total mass of the biological dressing. Preferably, the functional coating 2 further comprises mussel myoglobin and / or epidermal growth factor (EGF) and constitutes a time-controlled release system with the GG.

[0046] The preparation method comprises forming the functional coating 2 by visible light-induced “one-step” cross-linking loading or coaxial electrospinning technology. The dressing of the present application not only has excellent moisturizing and healing properties, but also can realize early intelligent early warning (pH response) of wound infection and on-demand release of drugs (enzyme response), and can synergistically achieve antibacterial and anti-inflammatory effects, significantly accelerating the healing process of chronic non-healing wounds and reducing scar formation.

[0047] Example 1: Intelligent response hydrogel dressing containing GG (visible light cross-linking method)

[0048] Raw material preparation:

[0049] Substrate layer 1: bacterial cellulose (BC) membrane (10 cm x 10 cm, thickness 0.2 mm), which is soaked and washed with ultrapure water until neutral and then used.

[0050] Biocompatible polymer: methacrylated gelatin (GelMA, degree of substitution ~ 70%).

[0051] Active ingredients: glycerol glucoside (GG, purity > 98%), mussel myoglobin (MAP, purity > 90%), recombinant human epidermal growth factor (rhEGF).

[0052] Intelligent response ingredients: bromothymol blue (pH-responsive dye), matrix metalloproteinase 2 (MMP-2)-responsive polypeptide cross-linking agent (sequence: GPLGIAGQ).

[0053] Photoinitiator: lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP).

[0054] Solvent: phosphate buffer (PBS, 0.01 M, pH 7.4).

[0055] Reference Figure 3 Preparation step:

[0056] S1. Preparation of pre-polymer solution: 0.1 g LAP, 5.0 g GelMA, 2.0 g GG, 0.2 g MAP (GG:MAP = 10:1), 10 mg bromothymol blue, 5 mg MMP-2 responsive peptide were weighed and dissolved in 92.695 g PBS. The solution was stirred at 300 rpm under 37℃ and dark condition for 2 hours until all components were completely dissolved, obtaining a uniform pre-polymer solution with a light blue color. Finally, 1.0 mg rhEGF (final concentration 10 μg / g pre-polymer solution) was added under ice bath operation, and the solution was gently shaken to mix well to avoid air bubbles.

[0057] S2. Coating and crosslinking: The BC membrane was laid in a polytetrafluoroethylene mold. The above-mentioned pre-polymer solution was carefully poured onto the BC membrane, and the coating thickness was controlled to be about 1.0 mm. Then, the mold was moved under the blue light curing box with a wavelength of 405 nm, and the irradiation intensity was 10 mW / cm2. The pre-polymer solution was crosslinked and cured under irradiation for 45 seconds, and a stable, light yellow intelligent hydrogel functional coating 2 was formed on the surface of the BC membrane.

[0058] S3. Post-processing: The prepared composite dressing was taken out with sterile tweezers, cut into a size of 5 cm x 5 cm, and vacuum packaged in an aluminum foil bag. Finally, the finished product intelligent biological dressing was obtained by irradiation sterilization (absorbed dose 25 kGy) using a cobalt-60 source. It was calculated that GG accounted for about 1.8% of the total mass of the dressing.

[0059] Performance test:

[0060] pH response: The dressing pieces were immersed in pH 6.0 and pH 7.8 buffer solutions, respectively. As shown in Table 1, the dressing remained light yellow at pH 6.0; at pH 7.8, the color changed to obvious blue within 1 minute, which was clearly visible to the naked eye, indicating that it had good infection warning ability.

[0061] Table 1

[0062]

[0063] Enzyme-responsive drug release: The dressing was soaked in PBS containing MMP-2 (100 ng / mL) (simulating an infected environment) and shaken at 37℃. The release rate of GG was detected by HPLC. As shown in Table 2, compared with the control group without MMP-2, the release rate of GG in the experimental group was significantly accelerated, proving that it had the function of enzyme-responsive on-demand drug release.

[0064] Table 2

[0065]

[0066] Time-controlled release: Release experiment was performed in PBS (without enzyme). The results are shown in Table 3, the release rates of GG and MAP were both over 75% within 24 hours, while the cumulative release rate of rhEGF was only 42% within 7 days, successfully achieving time-controlled release with fast and slow combination.

[0067] Table 3

[0068]

[0069] Example 2: Core-sheath fiber membrane dressing containing GG (coaxial electrospinning method)

[0070] Raw material preparation:

[0071] Sheath material: Polycaprolactone (PCL, Mn = 80,000).

[0072] Core material: Gelatin (from bovine hide, Type B).

[0073] Active ingredient: Glycerol glucoside (GG, purity > 98%), recombinant human epidermal growth factor (rhEGF).

[0074] Solvent: Hexafluoroisopropanol (HFIP, for sheath), aqueous acetic acid solution (2% v / v, for core).

[0075] Reference Figure 4 Preparation steps:

[0076] A1. Prepare the sheath solution: weigh 1.2 g of PCL into 8.8 g of HFIP, magnetically stir for 6 hours to obtain a uniform and transparent sheath solution with a mass concentration of 12%.

[0077] A2. Prepare the core solution: weigh 0.8 g of gelatin and 0.4 g of GG into 8.8 g of 2% aqueous acetic acid solution, stir in a 40°C water bath until completely dissolved to obtain a uniform solution. After cooling to room temperature, add 0.04 mg of rhEGF (final concentration 4 μg / g of core solution) and mix gently.

[0078] A3. Coaxial electrospinning:

[0079] Using a coaxial electrospinning device, the sheath solution and the core solution were injected into two independent syringe pumps respectively.

[0080] The inner diameter of the coaxial needle (core) was 0.4 mm, and the outer diameter (sheath) was 0.8 mm.

[0081] Process parameters: shell flow rate 0.8 mL / h, core flow rate 0.4 mL / h; applied voltage 18 kV; receiving distance 15 cm; drum receiver rotation speed 800 rpm.

[0082] Under the above parameters, continuous spinning was carried out for 4 hours, and a uniform PCL / gelatin core-sheath fiber membrane with anisotropic channels was formed directly on the receiving drum. Among them, PCL is a sheath, which plays a protective and slow-release role; gelatin, GG and EGF are cores, which are responsible for active release.

[0083] A4. Post-processing: the fiber membrane was taken off from the receiver and placed in a dryer to crosslink with glutaraldehyde vapor for 6 hours to stabilize the gelatin. Then it was repeatedly washed with deionized water to remove residual acetic acid and crosslinking agent. Finally, it was freeze-dried, cut (5 cm x 5 cm), packaged and EO sterilized to obtain the finished integrated fiber dressing.

[0084] Performance test:

[0085] Morphology characterization: scanning electron microscopy (SEM) showed that the fiber surface was smooth, uniform in thickness, and had no beading phenomenon. Transmission electron microscopy (TEM) confirmed the clear core-sheath structure.

[0086] Cell orientation: fibroblasts were inoculated on the fiber membrane for culture. The cells elongated and arranged along the orientation direction of the fiber, indicating that the anisotropic structure could effectively guide the directional migration of cells.

[0087] Example 3: Animal experiment to verify the healing promotion and antibacterial effect

[0088] Model establishment: healthy SD rats were selected, and a full-thickness skin defect wound with a diameter of 1.5 cm was made on their back, and S. aureus was inoculated to establish an infection model.

[0089] Grouping treatment: the rats were randomly divided into 4 groups:

[0090] Model group: the wound was covered with vaseline gauze.

[0091] Control dressing group: covered with blank hydrogel dressing (containing only GelMA) without GG and MAP.

[0092] GG dressing group: covered with hydrogel dressing containing only GG (Example 1 formula without MAP and EGF).

[0093] Inventive group: covered with the complete smart dressing prepared in Example 1.

[0094] Observation and detection:

[0095] Infection early warning observation: daily observation after operation. The invented dressing turned from yellow to blue within 24-48 hours in the infection model group, while the other groups had no color change, confirming the effectiveness of its early warning function.

[0096] Wound healing rate: photograph and calculate the wound healing rate on the 3rd, 7th, 10th, and 14th day after operation. The results are shown in Table 4. The healing rate of the invented group was always the fastest, and the healing rate on the 14th day was significantly higher than that of the other groups (p<0.01).

[0097] Table 4

[0098]

[0099]

[0100] Bacterial load analysis: wound tissue was taken on the 7th day after operation for colony counting. The bacterial load (CFU / g of tissue) of the invented group was significantly lower than that of the model group, the control dressing group, and the GG dressing group, proving the strong synergistic antibacterial effect of MAP and GG.

[0101] Histological analysis: wound tissue was taken on the 14th day for H&E staining and Masson staining. The invented group showed the most complete epithelialization, the richest granulation tissue, and the most orderly collagen arrangement, with the lightest scar.

[0102] Example 4: Verification experiment of multifunctional synergistic effect

[0103] Grouping:

[0104] Group A: dressing without any active ingredients (blank control, only GelMA hydrogel)

[0105] Group B: dressing containing only GG (2%)

[0106] Group C: dressing containing GG (2%) + mussel myoglobin (0.2%)

[0107] Group D: dressing containing GG (2%) + EGF (10 μg / g)

[0108] Group E: invented dressing (containing GG (2%) + MAP (0.2%) + EGF (10 μg / g) + pH dye)

[0109] Performance test:

[0110] Antibacterial experiment: against S. aureus and E. coli. The antibacterial circle diameter and antibacterial rate of Group C (GG+MAP) were significantly larger than those of Group B (only GG) and Group A (p<0.05), indicating that GG and MAP have a synergistic antibacterial effect. Group E has the best effect because it contains all the ingredients.

[0111] Cell migration and proliferation:

[0112] The cell scratch healing rate of group E was significantly higher than that of any other single or double component group (p<0.01).

[0113] After 3 days of culture, the cell proliferation rate (OD value) of group E was significantly higher than that of other groups (p<0.01).

[0114] It is proved that the multi-component time-controlled release produces a synergistic healing-promoting effect of "1+1+1>3".

[0115] Inflammation factor detection: The levels of TNF-α and IL-6 in the cell culture supernatant were detected by ELISA. The levels of inflammatory factors in groups C and E were significantly lower than those in other groups, indicating that MAP has good anti-inflammatory effect and cooperates with GG to create a more healing microenvironment.

[0116] Comparative example 1: dressing without pH-responsive dye

[0117] Except that bromothiol is not added in step S1, the remaining steps are exactly the same as example 1, and a dressing without intelligent early warning function is prepared for comparison and verification of the necessity of pH-responsive function.

[0118] Conclusion: The above examples fully prove the effectiveness, creativity and significant progress of the present application. By introducing pH-responsive dyes, enzyme-responsive peptides, mussel mucin, EGF and other active ingredients, and using advanced "one-step" photo-crosslinking and coaxial electrospinning technology, a new type of multifunctional biological dressing is successfully prepared, which integrates intelligent early warning, time-controlled release, on-demand drug delivery, synergistic antibacterial and anti-inflammatory, guided tissue regeneration and other functions. Its outstanding performance, especially the rapid healing ability and high-efficiency antibacterial activity shown on the infected chronic wound model, is not a simple superposition of existing technologies, but produces an unexpected synergistic effect, solving the major problem faced in clinical practice.

[0119] The above is only the preferred specific embodiment of the present application, but the protection scope of the present application is not limited thereto, any change or replacement easily thought of by those skilled in the art within the technical range disclosed by the present application should be covered within the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. A biological dressing containing glyceryl glucoside, characterized in that, The dressing includes a substrate layer (1) and a functional coating (2) loaded on the substrate layer (1). The functional coating (2) contains glycerol glucoside, a biocompatible polymer, and a pH-responsive dye. The pH-responsive dye is capable of undergoing a visible color change when the pH increases due to wound infection. The glycerol glucoside accounts for 0.1% to 10% of the total mass of the biological dressing.

2. The biological dressing containing glycerol glucoside according to claim 1, characterized in that, The functional coating (2) further comprises mussel adhesive protein and / or epidermal growth factor; the mass ratio of mussel adhesive protein to glycerol glucoside is (1:10) to (1:1); the concentration of epidermal growth factor is 1-100 μg / g dressing.

3. A biological dressing containing glycerol glucoside according to claim 1 or 2, characterized in that, The pH-responsive dye is one of bromothymol blue, phenol red, or alizarin red S; the biocompatible polymer material is one of methacrylamide gelatin, sodium alginate-calcium ion network, or hyaluronic acid-dopamine complex.

4. The biological dressing containing glycerol glucoside according to claim 1, characterized in that, The substrate layer (1) is a bacterial cellulose membrane or a polycaprolactone / gelatin fiber membrane with anisotropic channels prepared by electrospinning technology.

5. The biological dressing containing glycerol glucoside according to claim 2, characterized in that, The functional coating (2) is a time-controlled release system, wherein glycerol glucoside and mussel adhesive protein are the rapid release phases, with a release rate of over 70% within 24 hours; and epidermal growth factor is the sustained release phase, with a cumulative release rate of no more than 50% within 7 days.

6. The biological dressing according to claim 1, characterized in that, The functional coating (2) is also designed to respond to matrix metalloproteinases (MMPs); when the concentration of MMPs increases, the functional coating (2) degrades, accelerating the release of the loaded drug components.

7. A method for preparing a glycerol glucoside-containing biological dressing as described in any one of claims 1-6, characterized in that, A one-step method using visible light-induced cross-linking is employed. S1. Preparation of prepolymer solution: Dissolve photoinitiator LAP, methacrylamide gelatin, glyceryl glucoside, pH-responsive dye, and mussel adhesive protein in PBS, stir well and protect from light; S2. Coating and crosslinking: The prepolymer liquid is coated onto the substrate layer (1) and crosslinked and cured for 30-60 seconds under visible light with a wavelength of 405nm to form a functional coating (2); S3. Post-processing: After cutting, packaging, and irradiation sterilization, the biological dressing is obtained.

8. A method for preparing a glycerol glucoside-containing biological dressing as described in any one of claims 1-6, characterized in that, Coaxial electrospinning technology is used: A1. Preparation of shell solution: Dissolve polycaprolactone in an organic solvent to form a homogeneous solution; A2. Preparation of the core layer solution: Dissolve gelatin, glyceryl glucoside, and epidermal growth factor in deionized water to form a homogeneous solution; A3. Coaxial electrospinning: using the shell solution as the outer shell and the core solution as the core material, coaxial electrospinning is performed to directly form a fiber membrane integrating the substrate layer (1) and the functional coating (2) on the receiving device. A4. Post-processing: After the fiber membrane is cross-linked and cured, it is cut, packaged, and sterilized to obtain the biological dressing.

9. The method according to claim 7 or 8, characterized in that, The method further includes the step of microencapsulating the pH-responsive dye before adding it to adjust its response sensitivity.

10. The use of the glycerol glucoside-containing biological dressing as described in any one of claims 1-6 in the preparation of medical devices for promoting wound healing, providing intelligent infection early warning, and reducing scar formation.

Images