A composition and its preparation method and application

Abstract

The present invention provides a composition, preparation method, and application thereof. The composition comprises cellulose and an active substance. The active substance has a zeta potential or charge property opposite to that of the cellulose, with the absolute value of the active substance's zeta potential being 10-100 mV, and the sum of the absolute values of the zeta potentials of the cellulose and the active substance being 25-200 mV. Cellulose can form strong electrostatic adsorption and / or hydrogen bonding with the active substance, as well as physical encapsulation, stabilizing the spatial structure of the active substance. The composition can be used in pharmaceuticals for repairing damaged skin, healing wounds, and treating skin diseases.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a composition, a preparation method and an application thereof. Background Art

[0002] Active substances include peptides and collagen. Peptides can be divided into monopeptides, oligopeptides, and polypeptides based on molecular weight. Collagen is a biopolymer and the primary component of animal connective tissue. It is also the most abundant and widely distributed functional protein in mammals, accounting for 25%-30% of the total protein content, and even exceeding 80% in some organisms. It is a family of proteins with multiple encoding genes that can form a variety of collagen molecules.

[0003] In existing technologies, most skin active ingredients are reducing agents and are therefore unstable. Many active substances are stable only within a specific pH range. If the pH value of the system exceeds the active substance's stable range, it may lead to decomposition or deterioration of the active substance. Temperature and light exposure are also important factors affecting the stability of active substances. Increased temperature accelerates chemical reaction rates, thereby accelerating the degradation of active substances. Ultraviolet radiation can cause photodegradation reactions, leading to decomposition or deterioration of active substances.

[0004] The molecular structure of an active substance is a key factor in determining its activity. For example, collagen's triple helix structure is the foundation of its biological activity, enabling it to possess properties such as high tensile strength, biodegradability, and cell growth promotion. If collagen loses its triple helix structure, its biological activity disappears. However, ensuring the activity of active ingredients in pharmaceuticals remains a challenge and presents several limitations. Summary of the Invention

[0005] In order to solve the above technical problems, the present invention provides a composition comprising cellulose and an active substance, wherein the Zeta potential or charging property of the active substance is opposite to that of the cellulose.

[0006] The inventors have discovered that by controlling the Zeta potential or charge properties of the active substance to be opposite to the Zeta potential or charge properties of the cellulose, the cellulose has appropriate flexibility / rigidity and network structure, can form strong electrostatic adsorption and / or hydrogen bonding and physical encapsulation with the active substance, stabilize the spatial structure of the active substance, achieve a sustained release effect, and can be used in medicines, etc.

[0007] In some embodiments, the cellulose forms a complex or composite with the active substance.

[0008] In some embodiments, the absolute value of the zeta potential of the cellulose is 15-100 mV, preferably 20-60 mV, and more preferably 30-50 mV.

[0009] In some embodiments, the absolute value of the Zeta potential of the active substance is 10-100 mV, preferably 20-50 mV, and more preferably 30-40 mV.

[0010] In some embodiments, the sum of the absolute values of the zeta potential of the cellulose and the active substance is 25-200 mV, preferably 60-90 mV.

[0011] In some embodiments, the cellulose is nanocellulose, which has a diameter of less than 100 nanometers, preferably less than 50 nanometers, and more preferably 5-20 nanometers;

[0012] In some embodiments, the cellulose is nanocellulose, and its aspect ratio is greater than 50, preferably greater than 100, and more preferably greater than 200.

[0013] In some embodiments, the cellulose is modified cellulose, which is modified by converting the hydroxyl groups in the cellulose into groups with positive or negative charges; preferably, the modified cellulose is obtained by replacing the hydroxyl groups in the cellulose with carboxymethyl, carboxyl, phosphoric acid or sulfonic acid groups to obtain a modified cellulose with a negative charge; and the modified cellulose is obtained by replacing the hydroxyl groups in the cellulose with quaternary ammonium groups to obtain a modified cellulose with a positive charge.

[0014] In some embodiments, the active substance is selected from bioactive proteins such as keratin, silk fibroin, collagen or cell growth factors; bioactive peptides, including polypeptides, oligopeptides, cyclic peptides according to molecular weight and structure, and signal peptides, neurotransmitter inhibitory peptides, carrier peptides or enzyme inhibitory peptides according to function.

[0015] In some embodiments, the Zeta potential of the cellulose and the active substance is controlled by adjusting the pH values of the cellulose and the active substance before mixing, and then the two are mixed to form a complex or a composite. In this way, the cellulose and the active substance can be tightly bound together, and even if the system pH value of the formed complex or composite is subsequently adjusted, the tight bond between the cellulose and the active substance will not be destroyed.

[0016] In some embodiments, the zeta potential of the cellulose and the active substance is controlled separately by adjusting the pH value, and the pH is adjusted in the range of 3-11, for example, the pH of the cellulose and the active substance is adjusted to 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11.

[0017] In some embodiments, in the composition, the mass ratio of the cellulose to the active substance is 1:100-1:10000, for example, 1:100, 1:500, 1:1000, 1:3000, 1:5000, 1:7000, 1:10000.

[0018] In some embodiments, the composition further comprises cellulase, preferably the cellulase comprises at least one of endo-enzyme, exo-enzyme and glucosidase.

[0019] In some embodiments, in the composition, the added amount of the cellulase is 100-500 U / g, based on the absolute dry mass of cellulose.

[0020] In some embodiments, in the composition, the filter paper enzyme (FPA) activity of the cellulase is 30-60 U / mg, preferably 40-50 U / mg, such as 45 U / mg.

[0021] In some embodiments, the surface adsorption rate of the cellulose in the composition to the active substance is: the adsorption rate after 5 hours of adsorption is 60% or more, preferably the adsorption rate is 70% or more, more preferably the adsorption rate is 90% or more; or the adsorption rate after 10 hours of adsorption is 85% or more, preferably the adsorption rate is 95% or more, more preferably the adsorption rate is 98% or more.

[0022] The present invention also provides a method for preparing the above-mentioned composition, which comprises:

[0023] (S1) preparing a solution of the active substance;

[0024] (S2) adding the cellulose solution to (S1) and mixing; preferably, mixing is performed by high-speed vortex oscillation;

[0025] And also includes optional steps (S3), (S4):

[0026] (S3) preparing a cellulase solution and refrigerating it for later use;

[0027] (S4) mixing the cellulase solution in step (S3) and the solution of the composition obtained in step (S2); preferably, mixing is performed by high-speed vortex oscillation.

[0028] In some embodiments, when the cellulose is modified cellulose, the method for preparing the composition further comprises preparing the modified cellulose:

[0029] (SS1) converts the hydroxyl groups on the cellulose surface into groups with positive or negative charges;

[0030] and further comprising the optional step (SS2):

[0031] (SS2) high pressure homogenization treatment performed after step (SS1);

[0032] Preferably, in the step (SS1), the hydroxyl groups in the cellulose are replaced with carboxymethyl, carboxyl, phosphoric acid, or sulfonic acid groups to obtain negatively charged cellulose; and the hydroxyl groups in the cellulose are replaced with quaternary ammonium groups to obtain positively charged cellulose.

[0033] In some embodiments, the concentration of the active substance solution in step (S1) is 0.01-25 wt %, preferably 0.1-5 wt %, such as 1.0 wt %;

[0034] In some embodiments, the step (S1) further includes adjusting the pH of the solution of the active substance after preparing it, and the pH is adjusted in the range of 3-11. The reagents used to adjust the pH include acid solutions, alkaline solutions or buffer solutions, commonly used acid solutions such as hydrochloric acid, sulfuric acid, acetic acid, etc., commonly used alkaline solutions such as sodium hydroxide, sodium bicarbonate, concentrated ammonia solution, etc., and commonly used buffer solutions such as phosphate buffer, borate buffer, boric acid buffer, etc.

[0035] In some embodiments, the concentration of the cellulose solution in step (S2) is 0.01-25 wt %, preferably 0.1-5 wt %, such as 1.0 wt %;

[0036] In some embodiments, the volume ratio of the active substance solution to the cellulose solution in step (S2) is 1:0.1-100, for example, a volume ratio of 1:1;

[0037] In some embodiments, the step (S2) further includes adjusting the pH of the cellulose solution before mixing, wherein the pH is adjusted within a range of 3-11, and the reagents used to adjust the pH include acid solutions, alkaline solutions or buffer solutions, commonly used acid solutions include hydrochloric acid, sulfuric acid, acetic acid, etc., commonly used alkaline solutions include sodium hydroxide, sodium bicarbonate, concentrated ammonia solution, etc., and commonly used buffer solutions include phosphate buffer, borate buffer, boric acid buffer, etc.

[0038] In some embodiments, step (S2) further comprises adjusting the pH of the formed complex or compound system after mixing, wherein the pH is adjusted to be within the range of 5-11, for example, to 7.0 or 8.0;

[0039] In some embodiments, the rotation speed of the high-speed vortex oscillation in step (S4) is 2000-3000 rpm, and the oscillation time is 5-10 minutes.

[0040] The present invention also provides use of the aforementioned composition in the preparation of medicines, for example, use in the preparation of medicines for repairing damaged skin, healing wounds or treating skin diseases.

[0041] The present invention also provides a method for repairing damaged skin, healing wounds or treating skin diseases, which comprises administering the above composition to a patient in need.

[0042] According to an embodiment of the present invention, the patient in need thereof may be a mammal, such as a human, livestock or pet.

[0043] The present invention also provides a medicine for repairing damaged skin, healing wounds or treating skin diseases, which comprises the above composition.

[0044] In some embodiments, the drug product further contains one or more pharmaceutically acceptable excipients.

[0045] In some embodiments, the drug product is applied to the skin and can be formulated as a topical preparation; suitable topical preparations include, but are not limited to, ointments, creams, lotions, pastes, aerosol sprays, roll-ons, or aerosol foam (mousse) compositions.

[0046] In some embodiments, the drug is applied to 3D-printed artificial skin to promote the repair or healing of defect wounds.

[0047] The present invention also provides the use of the aforementioned composition in daily chemical products, foods, beverages, special medical purpose formula foods or health products, such as cosmetics for hair care and skin care; and provides a method for skin care and hair care, which comprises administering the aforementioned composition to an individual in need.

[0048] The present invention also provides a cosmetic comprising the above composition.

[0049] In some embodiments, the cosmetic further contains cosmetic ingredients including fragrance, water, oil, surfactant, moisturizer, polyol, polymer, preservative, colorant, emollient, anti-wrinkle agent, antioxidant, or whitening agent.

[0050] Beneficial effects

[0051] In the composition provided by the present invention, the active substance matches the cellulose Zeta potential, and the two are more tightly bound by strong electrostatic adsorption, hydrogen bonding, and physical embedding, and can adapt to active substances of different structures. The composition forms a barrier between the active substance and the protease to protect it from protease cleavage, thereby playing a role in isolating external interference, and provides a new idea and method for protecting polypeptide active substances that are easily attacked by proteases. Furthermore, by adjusting the pH value of the cellulose and the active substance before mixing, the Zeta potential of the cellulose and the active substance is controlled, and then the two are mixed to obtain a composition, so that the cellulose and the active substance can be tightly bound to achieve protection of the active substance, thereby avoiding the destruction of the structure and activity of the active substance itself by changes in pH value of subsequent complexes or composites in different application systems.

[0052] In the composition provided by the present invention, cellulose can also be degraded by adding cellulase, causing the cellulose network structure to collapse, thereby releasing active substances adsorbed on the cellulose surface or embedded in its network structure.

[0053] Compared with existing recombinant collagen technology, the composition provided by the present invention is safer and more effective, has a simple preparation process, low cost, and is easy to produce on a large scale, which is of great significance for promoting the industrialization of pharmaceutical and other technologies. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1 is the relationship between the adsorption rate and adsorption time of the nanocellulose surface in Example 1;

[0055] Figure 2 The relationship between keratin stability and adsorption time after adding nanocellulose in Example 1;

[0056] Figure 3 This is a scanning microscope image of the hair scales repaired with nanocellulose in Example 7. DETAILED DESCRIPTION

[0057] [Cellulose]

[0058] The cellulose involved in the present invention is a polysaccharide structure composed of hundreds to thousands of glucose units, and is a linear high molecular polymer formed by glucose molecules connected by β-1,4-glycosidic bonds.

[0059] In some embodiments, the absolute value of the cellulose Zeta potential is 15-100 mV, preferably 20-60 mV, and further preferably 30-50 mV. For example, under pH 7.0, the absolute value of the cellulose Zeta potential is 30 mV, 32 mV, 34 mV, 36 mV, 38 mV, 39 mV, 40 mV, 41 mV, 42 mV, 43 mV, 44 mV, 45 mV, 46 mV, 47 mV, 48 mV, 49 mV, or 50 mV.

[0060] In some embodiments, the Zeta potential of cellulose is controlled by adjusting the pH value of the system, and the pH is adjusted within the range of 3-11; for example, at pH 7.0, the Zeta potential of the modified cellulose with a negative charge obtained by replacing the hydroxyl groups in the cellulose with carboxymethyl or carboxyl groups is -60 mV to -20 mV, and when the pH is adjusted within the range of 3.0-5.0, the Zeta potential of the cellulose is -50 mV to -10 mV; at pH 7.0, the Zeta potential of the modified cellulose with a positive charge obtained by replacing the hydroxyl groups in the cellulose with quaternary ammonium groups is +20 mV to +60 mV, and when the pH is adjusted within the range of 8.0-11.0, the Zeta potential of the cellulose is +10 mV to +50 mV.

[0061] [Nanocellulose]

[0062] The nanocellulose involved in the present invention is a nanoscale fiber made by treating cellulose chemically, physically, or by a combination of methods. Its diameter is usually between a few nanometers and a few hundred nanometers, and its length can reach hundreds of nanometers to a few microns. It has a high specific surface area and good biocompatibility.

[0063] In some embodiments, the diameter of the nanocellulose is less than 100 nanometers, preferably less than 50 nanometers, and more preferably 5-20 nanometers.

[0064] In some embodiments, the specific surface area of the nanocellulose is greater than 100 m 2 / g, preferably greater than 200 m 2 / g, more preferably greater than 300m 2 / g.

[0065] [Modified cellulose]

[0066] The modified cellulose disclosed herein is modified by converting hydroxyl groups in the cellulose into groups with positive or negative charges. For example, the modified cellulose may be obtained by replacing hydroxyl groups in the cellulose with carboxymethyl, carboxyl, phosphoric acid, or sulfonic acid groups to obtain a negatively charged modified cellulose; or by replacing hydroxyl groups in the cellulose with quaternary ammonium groups to obtain a positively charged modified cellulose.

[0067] In some embodiments, 1% to 80% of the hydroxyl groups in the modified cellulose are substituted with carboxymethyl, carboxyl, phosphoric acid, sulfonic acid, or quaternary ammonium groups.

[0068] In some embodiments, the crystallinity of the modified cellulose is less than 90%, preferably less than 85%, and more preferably the crystallinity of the modified cellulose is 60-85%.

[0069] In some embodiments, the surface charge of the modified cellulose is greater than 0.5 mmol / g, such as greater than 1 mmol / g, greater than 1.2 mmol / g, or greater than 1.5 mmol / g.

[0070] [Cellulose with positive or negative charge]

[0071] The cellulose with positive or negative charge involved in the present invention is cellulose with controllable surface charge characteristics achieved by introducing carboxyl, carboxymethyl, phosphoric acid, sulfonic acid or quaternary ammonium groups through surface modification.

[0072] By substituting the hydroxyl groups in cellulose with carboxymethyl, carboxyl, phosphoric acid or sulfonic acid groups, cellulose with a negative charge is obtained; by substituting the hydroxyl groups in cellulose with quaternary ammonium groups, cellulose with a positive charge is obtained.

[0073] [Source of cellulose raw materials]

[0074] The raw material source of the cellulose involved in the present invention includes at least one of natural cellulose, regenerated cellulose, and bacterial cellulose, such as cellulose extracted from cotton, seaweed, hemp, bamboo, trees, wood, grains, beans, plant straw, and cellulose derived from bacterial fermentation. The raw material source of the cellulose involved in the present invention also includes microcrystalline cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, microcrystalline cellulose, and methylcellulose. Preferably, the raw material source of the cellulose is cellulose fiber with a degree of polymerization of 700-1200, more preferably cellulose fiber with a degree of polymerization of 800-1000.

[0075] [Active substance]

[0076] The active substance involved in the present invention is selected from biologically active proteins, such as keratin (for example, α-keratin, β-keratin), elastin (for example, tropoelastin, etc.), fibrillin (for example, fibrillin-1 (the main component of microfibrils), fibrillin-2 (component for elasticity generation), fibrillin-3 (present in the brain), fibrillin-4 (component for elasticity generation)), etc.), fibrinogen / fibrin / thrombin (for example, fibrinogen is converted into fibrin by thrombin during wound healing), fibronectin, laminin, silk protein, collagen (for example, collagen I present in skin, tendon and bone, collagen II present in cartilage or collagen III present in connective tissue, collagen IV present in extracellular matrix protein, collagen V present in hair, etc.), from a wide range of sources. Collagen extracted from various sources (e.g., porcine, bovine, human, fish, or rat tail), such as human recombinant collagen (recombinant collagen type III or XVII), transgenic human tissue collagen, porcine collagen, human placental collagen, bovine collagen, autologous collagen, collagen fibers and human tissue collagen matrix, cell growth factors (fibroblast growth factor, endothelial growth factor, or insulin-like growth factor); bioactive peptides, including polypeptides, oligopeptides, and cyclic peptides based on molecular weight and structure, and signal peptides, neurotransmitter inhibitory peptides, carrier peptides, or enzyme inhibitory peptides based on function (oligopeptide-1, tripeptide-1, hexapeptide-1, hexapeptide-3, acetyl oligopeptide, palmitoyl oligopeptide, carnosine, glutathione, or blue copper peptide); non-colloidal active small molecule compounds, such as dasatinib, quercetin, fisetin, epigallocatechin gallate, proanthocyanidin C1, hydroquinone, arbutin, kojic acid, phenylethylresorcinol, or niacinamide.

[0077] In some embodiments, when the active substance has a Zeta potential, the absolute value of the Zeta potential of the active substance is 10-100 mV, preferably 20-50 mV, and further preferably 30-40 mV, for example, 26 mV, 29 mV, 30 mV, 31 mV, 32 mV, 33 mV, 34 mV, 35 mV, 36 mV, 37 mV, 38 mV, 39 mV, and 40 mV.

[0078] In some embodiments, the Zeta potential of the active substance is controlled by adjusting the pH value of the system, and the pH is adjusted in the range of 3-11; for example, the isoelectric point of collagen is 7.5-7.8, and the pH is adjusted in the range of 5.0-11.0, and the Zeta potential of collagen is in the range of +45 mV to -30 mV; the isoelectric point of blue copper peptide is 10.0, and the pH is adjusted in the range of 5.0-11.0, and the Zeta potential of blue copper peptide is in the range of +35 mV to -15 mV; the isoelectric point of fibroblast growth factor is 9.6, and the pH is adjusted in the range of 5.0-11.0, and the Zeta potential of fibroblast growth is in the range of +37 mV to -10 mV.

[0079] In some embodiments, the active substance is selected from a non-colloidal active small molecule compound, and the ionization or hydrolysis equilibrium of the non-colloidal active small molecule compound is controlled by adjusting the pH value of the system; for example, the niacinamide solution itself is relatively stable between pH 5-7, and the pH is adjusted within the range of 5.0-7.0 to control the ionization or hydrolysis of niacinamide and prevent excessive hydrolysis to niacin and skin irritation; quercetin is relatively stable between pH 5-8, and the pH is adjusted within the range of 5.0-8.0 to control quercetin autooxidation and prevent the loss of quercetin's free radical scavenging activity.

[0080] [Introduction of carboxyl groups into cellulose surface hydroxyl groups]

[0081] The present invention relates to the introduction of carboxyl groups into cellulose surface hydroxyl groups, including but not limited to one or more of TEMPO (tetramethylpiperidinium oxide) oxidation, H2O2 oxidation, HIO4 oxidation, and NaClO2 oxidation. More preferably, the surface-carboxylated nanocellulose is prepared by TEMPO oxidation. Further preferably, the TEMPO oxidation method comprises dispersing cellulose fibers in an aqueous solution of TEMPO and NaBr, adding NaClO, and conducting an oxidation reaction for 6-12 hours. During the reaction, the system is maintained alkaline by dropwise addition of an alkaline solution, such as potassium hydroxide, sodium hydroxide, potassium carbonate, or sodium carbonate solution, to maintain a pH of 10.0-10.5.

[0082] [Introduction of carboxymethyl groups into hydroxyl groups on the cellulose surface]

[0083] The present invention relates to the introduction of carboxymethyl groups into hydroxyl groups on the surface of cellulose, including but not limited to the alkalization-etherification method. More preferably, the alkalization is carried out using an aqueous solution of an organic base or an inorganic base. The organic base or inorganic base is, for example, sodium hydroxide, potassium hydroxide, calcium hydroxide, carbonic acid, potassium carbonate, triethylamine, ammonia water, etc. The etherification is carried out using sodium chloroacetate or chloroacetic acid, for example, the etherification is carried out using a methanol or ethanol solution of sodium chloroacetate. Wherein, the alkalization is carried out at 10-50°C for 10-60 minutes, preferably at 25°C for 30 minutes; the etherification is reacted at 50-100°C for 30-150 minutes, preferably at 60-80°C for 60-120 minutes.

[0084] [Introduction of quaternary ammonium groups into hydroxyl groups on the cellulose surface]

[0085] The present invention relates to the introduction of quaternary ammonium groups onto hydroxyl groups on the surface of cellulose, including but not limited to a substitution reaction between a quaternizing agent and cellulose under alkaline conditions. More preferably, the quaternizing agent is selected from one or more of 3-chloro-2-hydroxypropyltrimethylammonium chloride, 2,3-epoxypropyltrimethylammonium chloride, 3-chloro-2-hydroxypropyltrimethylammonium bromide, and 2,3-epoxypropyltrimethylammonium bromide. Further preferably, the reaction temperature for introducing the quaternary ammonium groups is 50-100°C, such as 65°C.

[0086] [Introduction of phosphoric acid groups into hydroxyl groups on the cellulose surface]

[0087] The present invention relates to introducing phosphate groups into cellulose surface hydroxyl groups, including but not limited to reacting a phosphating agent with the hydroxyl groups on the cellulose surface. More preferably, the phosphating agent comprises diammonium hydrogen phosphate or a mixture of diammonium hydrogen phosphate and urea. The method comprises, for example, impregnating the cellulose with the phosphating agent (at a mass ratio of cellulose: diammonium hydrogen phosphate: urea of 1:0.8:3.7) for 1 hour, followed by placing the cellulose in a 170°C oven and continuing the reaction for 30 minutes.

[0088] [Introduction of sulfonic acid groups into hydroxyl groups on the cellulose surface]

[0089] The present invention involves introducing sulfonic acid groups into hydroxyl groups on the surface of cellulose, including but not limited to reacting a sulfonating agent with the hydroxyl groups on the surface of the cellulose. More preferably, the reaction comprises hydrolyzing the amorphous region of the cellulose fiber using 64% sulfuric acid at 45°C, while simultaneously introducing sulfonic acid groups into the surface of the crystalline region of the cellulose via an esterification reaction for 60 minutes.

[0090] [Cellulose pretreatment and post-treatment]

[0091] Prior to introducing carboxyl, carboxymethyl, phosphoric acid, sulfonic acid, or quaternary ammonium groups, the method of the present invention may further optionally include a step of pretreating the cellulose. Preferably, the pretreatment comprises pretreating the cellulose with an alkali and an acid. Further preferably, the cellulose is dispersed with an alkali solution, heated, and washed with water until neutral, and then dispersed with an acid solution, heated, and washed with water until neutral.

[0092] Preferably, in the step of pretreating the cellulose, the alkali includes organic alkalis and inorganic alkalis, such as sodium hydroxide, potassium hydroxide, calcium hydroxide, carbonic acid, potassium carbonate, triethylamine, ammonia water, etc.; the acid includes organic acids and inorganic acids, such as hydrochloric acid, sulfuric acid, phosphoric acid, formic acid, acetic acid, citric acid, etc. When pretreating the cellulose with alkali and acid, an alkali solution or an acid solution is added to the cellulose and the slurry concentration is adjusted to 0.1-10 wt%, preferably 2.5 wt%.

[0093] Preferably, in the step of pretreating the cellulose, the heating treatment temperature is 100-150° C., and the time is 10-60 minutes; preferably, the heating treatment temperature is 120-130° C., and the time is 30 minutes.

[0094] After the reaction of introducing carboxyl, carboxymethyl, phosphoric acid, sulfonic acid, or quaternary ammonium groups is completed, the method of the present invention may optionally include a post-treatment step. Preferably, the post-treatment step includes adjusting the pH to neutral, washing the reaction product with water, and filtering the reaction product until the conductivity of the filtrate is less than 50 μS / cm, preferably until the conductivity of the filtrate is less than 5 μS / cm.

[0095] [High-pressure homogenization]

[0096] The high-pressure homogenization process described herein refers to the process of uniformly mixing the components of a substance to form a homogeneous system by applying high pressure. The high-pressure homogenization process described herein may include adjusting the cellulose fibers obtained in step (S1) to a concentration of 1 wt% with water, pre-shearing and dispersing them using a disperser at 2000 rpm for 5 minutes, and then subjecting them to high-pressure homogenization at a pressure of 200-1200 bar.

[0097] [Sum of the absolute values of the zeta potential of cellulose and active substances]

[0098] The sum of the absolute values of the Zeta potential of the cellulose and the active substance involved in the present invention refers to the sum of the absolute values of the Zeta potential of the cellulose and the active substance when the cellulose and the active substance have opposite charges.

[0099] [Cellulase]

[0100] The cellulase involved in the present invention refers to an enzyme that can degrade cellulose to produce glucose, including at least one of endo-enzymes, exo-enzymes, and glucosidases, such as the composite cellulase derived from Trichoderma reesei.

[0101] [Flexibility / rigidity of cellulose]

[0102] The cellulose flexibility involved in the present invention refers to cellulose with a crystallinity of 60-85%, or an aspect ratio higher than 250, or an absolute value of Zeta potential higher than 40 mV, which has relatively strong flexibility.

[0103] The cellulose rigidity involved in the present invention refers to cellulose with a crystallinity of 85% or above, or an aspect ratio of less than 150, or an absolute value of Zeta potential of less than 30 mV, which has relatively strong rigidity.

[0104] [Daily Chemicals]

[0105] The daily chemical products covered by this invention are chemical products manufactured from certain chemicals or natural products, related to daily life, and intended to cleanse and beautify people's homes and meet the health, cleanliness, beauty, and comfort needs of animals and pets. Based on their intended use, these products can be categorized as toiletries, household products, kitchen and bathroom products, decorative items, cosmetics, and animal or pet care products. Based on industry practices, these products can be categorized as cosmetics (such as skincare, beauty, cleaning, and hair products), cleaning products (such as soaps, laundry detergents, and detergents), oral products (such as toothpaste and mouthwash), fragrances, deodorants, insect repellents, and animal or pet care products (such as pet-specific bathing and cleaning products, oral care products, insect repellents and flea control products, and skin care products).

[0106] The technical solutions of the present invention will be described in further detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are encompassed within the scope of protection that the present invention is intended to protect.

[0107] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0108] Unless otherwise specified, % in the following preparation examples and embodiments refers to mass percentage.

[0109] Zeta potential test method of modified nanocellulose: Take the prepared modified nanocellulose, dilute it with ultrapure water to 0.05% solid content, and use a Zeta potential analyzer to detect the Zeta potential of the modified nanocellulose.

[0110] Zeta potential test method for active substances: prepare an active substance solution with a solid content of 0.01% with ultrapure water, and use a Zeta potential analyzer to detect its Zeta potential.

[0111] Crystallinity test method: The prepared modified nanocellulose was freeze-dried at -50°C and a vacuum of 0.01 mbar for 72 hours. An X-ray diffractometer was used to scan the sample at 5-40 degrees, and the X-ray diffraction pattern of the sample was analyzed to calculate the crystallinity of the sample.

[0112] Diameter and aspect ratio testing: Prepared modified nanocellulose was diluted with ultrapure water to 0.01% solids, ultrasonically dispersed for 10 minutes, and then dripped onto a copper mesh. After air-drying, the modified nanocellulose morphology was imaged using a transmission electron microscope at a magnification of 10,000-20,000. ImageJ software was used to measure and calculate the length and diameter of the modified nanofibers and calculate the aspect ratio.

[0113] Specific surface area test method: Take 1% modified nanocellulose sample and place it in a centrifuge tube, immerse it in liquid nitrogen for rapid freezing, transfer it to -50 degrees Celsius, freeze-dry the sample under vacuum conditions of 0.05 mbar for 72 hours, and use a specific surface area analyzer to detect the specific surface area of the sample.

[0114] Test method for surface charge of modified cellulose:

[0115] 5 g of 1% modified nanocellulose was diluted to 0.1% with deionized water. 2 ml of 0.1 M HCl and 1 ml of 50 mM sodium chloride solution were added dropwise, and the suspension was stirred at room temperature for 30 minutes. The suspension was titrated with 0.1 M NaOH, monitoring the conductivity and pH of the suspension until pH 11. The charge of the anionic groups on the cellulose surface (including introduced carboxyl, carboxymethyl, and sulfonic acid groups) was calculated from the titration curve.

[0116] Take 5 g of 1% modified nanocellulose, dilute it to 0.1% with deionized water, use 0.01 M AgNO3 to titrate and monitor the conductivity of the suspension. Titrate until the conductivity value continues to increase steadily. Calculate the charge of the cationic quaternary ammonium salt group on the cellulose surface through the titration curve.

[0117] Adsorption rate test method:

[0118] The active substance to be adsorbed is weighed and dissolved or dispersed in deionized water. For active substances with characteristic ultraviolet absorption wavelengths, a working curve between the active substance concentration and the absorbance value is established using an ultraviolet spectrophotometer. For active substances without characteristic ultraviolet absorption, a working curve between the active substance concentration and the detection response value is established using high performance liquid chromatography or a corresponding enzyme-linked immunosorbent assay kit.

[0119] Weigh cellulose and adjust the solids content to 0.1% with deionized water. Add the active ingredient and mix thoroughly with vortexing. Incubate for a specified period of time, centrifuge at 10,000 rpm for 15 minutes, and filter through a 0.2 μm filter. Determine the concentration of free active ingredient using a UV spectrophotometer, high-performance liquid chromatography, or an appropriate enzyme-linked immunosorbent assay kit, and calculate the amount of active ingredient adsorbed on the cellulose surface.

[0120] Establishment of absorbance calibration curve:

[0121] (1) Keratin was dissolved in deionized water to form a 0.01% keratin solution, and the maximum absorption wavelength was determined by full wavelength scanning;

[0122] (2) Gradient dilution of the keratin solution was performed, and the absorbance was measured at the maximum absorption wavelength to establish a concentration-absorbance standard curve.

[0123] Preparation Example 1: Preparation of TJ-300:

[0124] Use 0.1M NaOH solution to disperse cellulose fibers with a degree of polymerization of 800-1000, a cellulose alpha content >90%, and a whiteness >85% ISO. Maintain a slurry concentration of 2.5%, heat at 121°C for 30 minutes, rinse with sterile water, and filter until the filtrate has a neutral pH. Subsequently, use 0.3M HCl solution to disperse the slurry, adjust the slurry concentration to 2.5%, heat at 121°C for 30 minutes, and rinse with sterile water and filter until the filtrate has a neutral pH.

[0125] Weigh (5.0 ± 0.1) g of cellulose fibers (absolute dry weight) after the pretreatment described above and knead and mix thoroughly with (15 ± 0.1) g of 30% NaOH solution at 25°C. Immerse for 30 min. After immersion, add (7 ± 0.1) g of sodium chloroacetate (preheated to 70°C and dispersed in 100 mL of 95% ethanol). Stir at 500 rpm in a water bath at 60-80°C for 30-120 min. After the reaction, adjust the pH to neutral with glacial acetic acid. Wash the reaction product repeatedly with ultrapure water and filter it using an ultrafiltration system until the filtrate has a conductivity of less than 5 μS / cm. Adjust the concentration of the cellulose fibers to 1% with sterile water and homogenize them in a sterile room using a high-pressure homogenizer at pressures of 200, 400, 600, 800, 1000, and 1200 bar, sequentially. Store in a refrigerator at 4°C until needed.

[0126] In this preparation example, by regulating the etherification reaction temperature and time (60°C, 30 minutes; 70°C, 100 minutes; 80°C, 120 minutes), the TJ-300 prepared has high flexibility. The characterization results obtained through testing and calculation are shown in Table 1:

[0127] Table 1: Characterization results of TJ-300 prepared at different etherification reaction temperatures and times

[0128]

[0129] Preparation Example 2: Preparation of TJ-321T

[0130] Use 0.1M NaOH solution to disperse cellulose fibers with a degree of polymerization of 800-1000, a cellulose alpha content of >90%, and a whiteness of >85% ISO. Maintain a slurry concentration of 2.5%, heat at 121°C for 30 minutes, rinse with sterile water, and filter until the filtrate has a neutral pH. Then, use 0.3M HCl solution to disperse the slurry, adjust the slurry concentration to 2.5%, heat at 121°C for 30 minutes, and rinse with sterile water and filter until the filtrate has a neutral pH.

[0131] Weigh (10.0 ± 0.1) g of cellulose fibers (absolute dry mass) after the above pretreatment and disperse them in a solution containing 1.6 ± 0.1 g of TEMPO and 1.0 ± 0.1 g of NaBr at 25°C. Stir continuously at 500 rpm and dropwise add 75 ml of 10% NaClO to initiate the oxidation reaction for 6–12 hours. During the reaction, monitor and maintain the pH of the reaction suspension at 10.0 with the addition of 0.5 M NaOH solution. After the reaction, adjust the pH to neutral with 1.0 M HCl. The reaction product is repeatedly washed with ultrapure water and filtered through an ultrafiltration system until the filtrate has a conductivity of less than 5 μS / cm. The oxidized cellulose fibers are adjusted to a 1% concentration with sterile water and homogenized in a sterile room using a high-pressure homogenizer at pressures of 200, 400, 600, 800, 1000, and 1200 bar, sequentially. Store in a refrigerator at 4°C until further use.

[0132] In this preparation example, TJ-321T obtained by regulating the TEMPO catalytic oxidation time (6h; 8h; 12h) has moderate flexibility. After testing and calculation, the characterization results are shown in Table 2.

[0133] Table 2: Characterization results of TJ-321T prepared under different TEMPO catalytic oxidation times

[0134]

[0135] Preparation Example 3: QCNF Preparation

[0136] Use 0.1M NaOH solution to disperse cellulose fibers with a degree of polymerization of 800-1000, a cellulose alpha content of >90%, and a whiteness of >85% ISO. Maintain a slurry concentration of 2.5%, heat at 121°C for 30 minutes, rinse with sterile water, and filter until the filtrate has a neutral pH. Then, use 0.3M HCl solution to disperse the slurry, adjust the slurry concentration to 2.5%, heat at 121°C for 30 minutes, and rinse with sterile water and filter until the filtrate has a neutral pH.

[0137] Weigh (15.0 ± 0.1) g of cellulose fibers (absolute dry mass) after the pretreatment described above and disperse them in 200 ml of 5% NaOH solution at room temperature. Stir and immerse for 30 minutes. After immersion, add 7.5 to 10 g of 2,3-epoxypropyltrimethylammonium chloride, heat to 65°C, and stir at 500 rpm for 1-8 hours. After the reaction, adjust the pH to neutral with 1.0 M HCl. The reaction product is repeatedly washed with ultrapure water and filtered using an ultrafiltration system until the filtrate has a conductivity of less than 5 μS / cm. The reacted cellulose fibers are adjusted to a 1% concentration with sterile water and homogenized in a sterile room using a high-pressure homogenizer at pressures of 200, 400, 600, 800, 1000, and 1200 bar, sequentially. Store in a refrigerator at 4°C until needed.

[0138] In this preparation example, by adjusting the amount of etherifying agent and reaction time (7.5 g, 1 h; 8.5 g, 6 h; 10 g, 8 h), the QCNF obtained has a certain rigidity (+15 / +46 mV) or flexibility (+79 mV). After testing and calculation, the characterization results are shown in Table 3.

[0139] Table 3: Characterization results of QCNF prepared at different etherifying agent dosages and reaction times

[0140]

[0141] Example 1: Adsorption and stability of keratin on nanocellulose surface

[0142] (1) Select TJ-300 (Zeta potential -49 mV at pH 7.0), TJ-321T (Zeta potential -47 mV at pH 7.0), and QCNF (Zeta potential +15 / +46 / +79 mV at pH 7.0) nanocellulose prepared in Preparation Example 1-3, adjust the solid content to 0.1%, add keratin solution (10 mg / mL, Zeta potential -26 mV at pH 7.0), wherein the volume ratio of keratin solution to nanocellulose prepared in Preparation Example 1-3 is 1:1, shake and mix, and incubate at 37°C for 0 to 24 hours.

[0143] (2) Samples at six time points (0 h, 2 h, 4 h, 6 h, 8 h, 12 h, and 24 h) were centrifuged (10,000 rpm, 15 min) and filtered through a 0.2 μm filter membrane.

[0144] (3) Set an equal volume of deionized water as a control, measure the absorbance of the filtrate, and calculate the adsorption rate of keratin on the nanocellulose surface based on the concentration-absorbance curve. The relationship between the adsorption rate of keratin on the nanocellulose surface of TJ-300, TJ-321T, and QCNF and the adsorption time is as follows: Figure 1 shown.

[0145] (4) Add keratinase (50 IU / g) to the nanocellulose-keratin complex after 24 h of adsorption stabilization in (1) and incubate at 37 °C for 0 to 24 h.

[0146] (5) Samples were taken at six time points: 0h, 2h, 4h, 6h, 8h, 12h, and 24h. The keratin in the samples was quantified using a keratin ELISA kit. The keratin content in the sample at the 0h time point was used as a control to calculate the keratin stability after adding nanocellulose. The results are as follows: Figure 2 shown.

[0147] Example 2: Preparation of collagen-nanocellulose composite and experiment on stabilization and protection of collagen by nanocellulose

[0148] (1) Prepare collagen at a concentration of 1.0 wt%, including recombinant collagen type III (Zeta potential -29 mV at pH 7.0) and type XVII (Zeta potential -31 mV at pH 7.0), and refrigerate at 4°C until use.

[0149] (2) Pure water and 1% QCNF (Zeta potential +46 mV at pH 7.0) nanocellulose obtained in Preparation Example 1 were used to dilute collagen to an effective concentration of 0.1% in the solution. The mixture was incubated at 25°C and 60°C for 1 hour, and then cooled to room temperature.

[0150] (3) Circular dichroism spectrometer was used to scan the wavelength range of 190-260 nm at a speed of 1 nm / s to record the ellipticity. The maximum positive absorption and negative absorption peaks of the ellipticity of each sample were recorded. The results are shown in Table 4 (Col-III represents type III recombinant collagen, Col-XVII represents type XVII recombinant collagen, Col-III-QCNF represents type III recombinant collagen and QCNF complex, and Col-XVII-QCNF represents type XVII recombinant collagen and QCNF complex).

[0151] Table 4: Maximum positive and negative absorption peaks of sample ellipticity

[0152]

[0153] As shown in Table 4, at 25°C, the circular dichroism ellipticity of type III and type XVII recombinant collagens (Col-III, 25°C and Col-XVII, 25°C) exhibited maximum positive absorption peaks at 224-225 nm and 220 nm, respectively, and maximum negative absorption peaks at 192-194 nm and 197-198 nm, respectively. These absorption peaks confirm the triple-helical structure of type III and type XVII recombinant collagens. After incubation at 60°C for 1 hour, the maximum positive absorption peaks of the circular dichroism ellipticity of type III and type XVII recombinant collagens (Col-III, 60°C and Col-XVII, 60°C) disappeared, indicating that the collagen triple-helical structure was disrupted. After incubation of type III and type XVII recombinant collagen and QCNF complexes (Col-III-QCNF and Col-XVII-QCNF) at 60°C for 1 hour, the circular dichroism ellipticity retained the maximum positive absorption peak at 220 nm and the maximum negative absorption peak at 197 nm, indicating that collagen did not lose its triple helical structure under the protection of QCNF.

[0154] Example 3: Wound healing experiment

[0155] 2 ml of the type III recombinant collagen and QCNF complex (Col-III-QCNF) prepared in Example 2 was plated in a 6-well plate. 20 mm x 20 mm medical gauze was cut and impregnated with Col-III-QCNF. The mixture was incubated at 4°C for 24 hours, removed, placed in a sterile Petri dish, and sterilized by UV irradiation for 15 minutes. This served as the experimental group. QCNF nanocellulose-impregnated gauze, prepared under the same conditions, served as a blank control, untreated medical gauze served as a negative control, and a commercial collagen wound dressing (3MPromogran Prisma) served as a positive control. Full-thickness skin wounds (8 mm in diameter) were applied to the backs of SPF-grade Sprague-Dawley rats (Purchased from the Animal Experimental Research Center of Jiangsu University, 20 males aged 5-6 weeks, weighing approximately 220-240 g, and divided equally into four groups) to evaluate the material's effectiveness in promoting skin wound healing. The dressings were changed and the wound diameters were measured on days 3, 7, 14, and 21 after surgery, and the wound healing rates were calculated. The results are shown in Table 5.

[0156] Table 5: Collagen-nanocellulose QCNF composite impregnated gauze promotes wound healing rate (%)

[0157]

[0158] Example 4: Composite cellulase sustained release experiment

[0159] (1) Prepare 1.0 wt% recombinant human collagen solution (Zeta potential of +35 mV at pH 7.0) and composite cellulase solution (the composite cellulase is a commercial composite cellulase purchased from Shanghai Yuanye Biotechnology Co., Ltd., containing endo-enzyme, exo-enzyme, glucosidase, and filter paper enzyme activity FPA of 45 U / mg protein), and refrigerate at 4°C for use.

[0160] (2) The collagen solution prepared in (1) was mixed with 1% TJ-300 (Zeta potential -49 mV at pH 7.0) nanocellulose obtained in Preparation Example 1 or 1% QCNF (Zeta potential +46 mV at pH 7.0) nanocellulose obtained in Preparation Example 3 at a volume ratio of 1:100 (pH 7.0). After mixing, the mixture was vortexed at high speed (2000-3000 rpm) for 5-10 minutes to obtain a collagen-nanocellulose composite, which was then refrigerated at 4°C for use.

[0161] (3) Take the composite cellulase solution prepared in (1) and add it to the collagen-nanocellulose complex prepared in (2) at an enzyme dosage of 100-500 U / g cellulose absolute dry mass. After mixing, vortex at high speed (2000-3000 rpm) for 1 minute to prepare the nanocellulose degradation complex loaded with collagen, and refrigerate at 4°C for use.

[0162] (4) The complex prepared in (3) was placed in a 96-well plate and placed in a 37°C constant temperature incubator for complex degradation and collagen release monitoring. Samples were taken at different degradation time points (0, 1, 2, 4, 8, 16, and 24 h), centrifuged at 5000 rpm, and the supernatant was collected. The released collagen was quantitatively detected using an enzyme-linked immunosorbent assay kit, and the amount of free glucose produced by cellulase degradation of TJ-300 and QCNF was detected using a DNS colorimetric assay.

[0163] (5) The complex prepared in (3) was placed in a 50 ml centrifuge tube and placed in a 37°C constant temperature incubator for complex degradation and complex viscosity monitoring. Samples were taken out at different degradation time points (0, 1, 2, 4, 8, 16, and 24 h) and cooled to room temperature. The viscosity of the complex was measured using a rotational viscometer with a 4# rotor at a speed of 12 rpm to characterize the enzymatic degradation of the nanocellulose gel network.

[0164] Among them, the results of the enzyme degradation test are shown in Tables 6 and 7.

[0165] Table 6 Enzyme degradation test results of TJ-300 nanocellulose degradation complex loaded with collagen

[0166]

[0167] The isoelectric point (PI) of recombinant human collagen is 8.0-9.0. Under neutral conditions, the Zeta potential of recombinant human collagen is +35 mV. It can be adsorbed on the negatively charged surface of TJ-300 (Zeta potential -49 mV). The sum of the absolute values of the Zeta potential of TJ-300 and recombinant human collagen is 84 mV.

[0168] As shown in Table 6, with increasing cellulase dosage and degradation time, the cumulative release of collagen and glucose increased, while the viscosity of the composite decreased. When the cellulase dosage reached 500 U / g, the cumulative release of collagen reached over 50% and the cumulative release of glucose reached over 10 mg after degradation for more than 4 hours, indicating that the nanocellulose network structure collapsed and collagen adsorbed on the surface or embedded in the network structure could be effectively released.

[0169] Table 7 Enzyme degradation test results of QCNF nanocellulose degradation complex loaded with collagen

[0170]

[0171] At pH 7.0, the zeta potentials of QCNF and recombinant human collagen were +46 mV and +35 mV, respectively. Because both have the same surface charge, electrostatic repulsion hinders the adsorption of collagen onto the QCNF surface. Some collagen (approximately 27%) remains free in the solution, while some is embedded within the QCNF network through hydrogen bonding and physical entrapment. Some collagen or QCNF forms aggregates with the negatively charged cellulase (zeta potential -28 mV), resulting in a low overall viscosity of the complex and impaired cellulose degradation by the cellulase.

[0172] Example 5: Single cellulase sustained release experiment

[0173] This example refers to Example 4, with the only difference being that: in step (1), the composite cellulase is replaced by single cellulases of endo-, exo-, and glucosidase, respectively; in step (2), 1% TJ-300 (Zeta potential -49 mV at pH 7.0) nanocellulose obtained in Preparation Example 1 is mixed with the collagen solution; in step (3), an enzyme dosage of 500 U / g cellulose absolute dry mass is added to the collagen-nanocellulose composite prepared in step (2); the cumulative collagen release amount, the cumulative glucose release amount, and the composite viscosity are detected for 24 hours; and the remaining steps are consistent with Example 4.

[0174] The test results showed that after adding endo-, exo- and glucosidase to hydrolyze the TJ-300 and collagen complex for 24 hours, the cumulative collagen release, cumulative glucose release and complex viscosity were: endo-, exo- and glucosidase: 53%, 3.5 mg, 9920 mP s; exo- and glucosidase: 16%, 0.52 mg, 13010 mP s; glucosidase: 8%, 0.22 mg, 14370 mP s.

[0175] Example 6: Preparation of fibronectin-nanocellulose composite and the effect of pH on the composite

[0176] (1) Prepare a 1 mg / ml fibronectin solution using 50 mM Tris buffer at pH 7.0 (the fibronectin solution was purchased from Sigma, with an isoelectric point (PI) of 5.5 and a Zeta potential of -30 mV at pH 7.0). Adjust the pH to 4.5 using 0.1 M HCl, at which the Zeta potential of fibronectin is +25 mV.

[0177] (2) Take 1% TJ-300 (Zeta potential -49 mV at pH 7.0) prepared in Preparation Example 1 and adjust the pH to 4.5 using 0.1 M HCl. At this pH, the Zeta potential of TJ-300 is -37 mV.

[0178] (3) The fibronectin solution was mixed with TJ-300 at a mass ratio of 1:100 and vortexed at high speed (2000-3000 rpm) for 5-10 minutes to obtain a solution containing the fibronectin-nanocellulose complex. The pH of the complex solution was adjusted to 7.0 using 0.1 M NaOH.

[0179] (4) Take the fibronectin-nanocellulose complex solution prepared in step (3), centrifuge it at 10,000 rpm for 10 minutes, take the supernatant, and detect the free fibronectin content.

[0180] (5) Take the unadjusted pH fibronectin solution (Zeta potential -30 mV at pH 7.0) and TJ-300 (Zeta potential -49 mV at pH 7.0) from steps (1) and (2), and follow the procedures of steps (3) and (4) to detect the free fibronectin content.

[0181] The test results showed that adjusting the pH of the system to below the isoelectric point of fibronectin before preparing the fibronectin-nanocellulose complex caused the fibronectin zeta potential to change from negative to positive, thereby enhancing its binding to the negatively charged nanocellulose. After the complex was formed, the pH of the system was adjusted back to neutral, and only a small amount of fibronectin (<5%) remained free in the supernatant. In contrast, when the pH of the system was not adjusted before preparing the fibronectin-nanocellulose complex, the nanocellulose in the complex only provided a physical coating and protection, resulting in a loose binding between the two, with over 35% of the fibronectin detected as free in the supernatant.

[0182] Example 7: Preparation of keratin-nanocellulose composite and experiment on repairing hair scales

[0183] (1) Collect adult female hair, wash it, and soak it in 5% ammonia water. Incubate it at 50℃ for 2 hours, take it out and wash it to prepare simulated damaged hair with split hair scales.

[0184] (2) Take 1% of TJ-300 (Zeta potential -49 mV at pH 7.0), TJ-321T (Zeta potential -47 mV at pH 7.0), and QCNF (Zeta potential +15 mV / +46 mV / +79 mV at pH 7.0) nanocellulose prepared in Preparation Examples 1-3, respectively, and mix them with 1% of keratin (Zeta potential -26 mV at pH 7.0) aqueous solution (modified nanocellulose and keratin are mixed in equal volumes), and treat with high-speed vortex oscillation (2000-3000 rpm) for 5-10 minutes to obtain a solution containing keratin-nanocellulose complex. Immerse damaged hair with split scales in the solution (immerse 10 hairs of 10 cm in 10 mL of the mixture), incubate at 37°C for 8 hours, remove and wash to obtain repaired hair.

[0185] (3) After drying, use a scanning electron microscope to take pictures and observe the results. Figure 3 shown.

[0186] The above describes exemplary embodiments of the present invention. However, the scope of protection of this application is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc. made by those skilled in the art within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A composition, characterized in that The composition comprises nano-modified cellulose and an active substance, wherein the Zeta potential or charging property of the active substance is opposite to that of the nano-modified cellulose, wherein the absolute value of the Zeta potential of the nano-modified cellulose is 30-50 mV, the absolute value of the Zeta potential of the active substance is 30-40 mV, the sum of the absolute values of the Zeta potential of the nano-modified cellulose and the active substance is 60-90 mV, the diameter of the nano-modified cellulose is 5-20 nanometers, and the nano-modified cellulose having a positive charge is obtained by replacing the hydroxyl groups in the cellulose with quaternary ammonium groups, and the active substance is keratin; the nano-modified cellulose and the active substance form a complex or a composite.

2. The composition according to claim 1, characterized in that The Zeta potential of the nano-modified cellulose and the active substance in the composition is controlled by adjusting the pH value of the system, and the pH is adjusted within the range of 7-11.

3. The composition according to claim 1, characterized in that The composition also includes cellulase.

4. The composition according to claim 3, characterized in that The cellulase comprises at least one of endo-enzyme, exo-enzyme and glucosidase.

5. The composition according to claim 4, characterized in that The cellulase has one or both of the following conditions: (1) The added amount of the cellulase is 100-500 U / g, based on the absolute dry weight of cellulose; (2) the filter paper enzyme activity of the cellulase is 30-60 U / mg.

6. The composition according to claim 1, characterized in that The surface adsorption rate of the nano-modified cellulose in the composition to the active substance is: an adsorption rate of 60% or more after 5 hours of adsorption; or an adsorption rate of 85% or more after 10 hours of adsorption.

7. A method for preparing the composition according to claim 1, comprising: (S1) preparing a solution of the active substance; (S2) adding the nano-modified cellulose solution to (S1) and mixing; And further comprising optional steps (S3) and (S4): (S3) preparing a cellulase solution and refrigerating it for later use; (S4) Mixing the cellulase solution in step (S3) and the solution of the composition obtained in step (S2).

8. The method according to claim 7, characterized in that The preparation method of the nano-modified cellulose comprises: (SS1) converts the hydroxyl groups on the cellulose surface into positively charged groups by replacing the hydroxyl groups in cellulose with quaternary ammonium groups; (SS2) High-pressure homogenization is performed after step (SS1).

9. The method according to claim 7 or 8, characterized in that The concentration of the active substance solution in step (S1) is 0.01-25 wt%; Alternatively, the step (S1) further comprises adjusting the pH of the solution of the active substance after preparing the solution, wherein the pH is adjusted to be within a range of 7-11; Alternatively, the step (S2) further comprises adjusting the pH of the nano-modified cellulose solution before mixing, wherein the pH is adjusted to be within a range of 7-11; Alternatively, step (S2) further comprises adjusting the pH of the formed complex or compound system after mixing, and adjusting the pH to be within the range of 5-11; Alternatively, step (S4) is mixed by high-speed vortex oscillation, the rotation speed of the high-speed vortex oscillation is 2000-3000 rpm, and the oscillation time is 5-10 minutes.

10. Use of the composition according to any one of claims 1 to 6 in the preparation of medicines for repairing damaged skin, healing wounds, and treating skin diseases.

11. A medicine for repairing damaged skin, healing wounds, and treating skin diseases, characterized in that: The medicine comprises the composition according to any one of claims 1 to 6.

12. Use of the composition according to any one of claims 1 to 6 in the preparation of daily chemical products.

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