Physiological body fluid environmental response type medical beauty subcutaneous tissue injection moisturizing filler and application thereof

Abstract

The invention relates to the technical field of biological medicine, in particular to a physiological body fluid environment response type medical beauty subcutaneous tissue injection moisturizing filler and application thereof. The composition specifically comprises the following components in parts by weight: 0.2-15 parts of a body fluid response material, 0.2-50 parts of a whitening and anti-wrinkle functional component, 0.2-15 parts of a structural component, 50-100 parts of an osmotic pressure regulator and 0.3-5 parts of a local pain relieving agent. According to the body fluid environment response type medical beauty injection filling agent provided by the invention, a three-dimensional gel network is rapidly formed by the ion sensitive material and tissue fluid cations subcutaneously, so that long-acting water locking can be realized, the release of functional components can be delayed, and the water replenishing and filling durability can be improved. The self-healing property of the material can be reshaped, liquid leakage and diffusion after injection are avoided, meanwhile, enzymolysis of structural components such as collagen is remarkably slowed down, and the maintaining effect is prolonged. The product has good biocompatibility and supporting force, and an innovative solution is provided for medical beauty moisturizing and tissue filling.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to a physiological fluid environment-responsive subcutaneous tissue injection hydrating filler and its application. Background Technology

[0002] Subcutaneous injection techniques in aesthetic medicine, especially hydration and filling solutions centered around mesotherapy, have become one of the most watched core technologies in the current anti-aging and beauty field due to their significant minimally invasive characteristics. Currently, various materials have been applied in this field and commercialized, including sodium hyaluronate, bio-derived and recombinant collagen, polynucleotides, polycaprolactone, polylactic acid, carboxymethyl cellulose, hydroxyapatite, polymethyl methacrylate, and agarose. These materials have been proven effective in long-term clinical applications; however, due to increasingly severe product homogenization, the market has entered a period of fierce competition. More importantly, existing materials generally lack the ability to intelligently respond to the physiological environment within the human body (such as ion concentration and pH value), and their function largely depends on physical filling or passive degradation. Therefore, developing innovative materials with unique properties that can respond to the physiological environment is particularly urgent and necessary.

[0003] From the perspective of product classification and technological approach, current filler products are mainly based on hyaluronic acid (HA), adjusting its mechanical properties and in vivo degradation rate through different cross-linking technologies to adapt to different clinical scenarios such as contouring (high cross-linking degree) and soft tissue filling (medium to low cross-linking degree). Meanwhile, the market share of "regenerative materials" such as collagen and poly-L-lactic acid (PLLA), which can stimulate autologous tissue regeneration, is gradually increasing, initially forming a dual technological approach of "physical filling" and "collagen regeneration." In hydrating products, small-molecule sodium hyaluronate is commonly used as a carrier, combined with functional ingredients such as vitamins, amino acids, and peptides, aiming to achieve a combination of immediate hydration and medium- to long-term skin improvement through multi-component synergy.

[0004] Although existing material systems are relatively mature, their performance still has inherent limitations. Traditional cross-linked hyaluronic acid fillers typically only last 6 to 12 months, requiring regular, repeated injections to maintain the effect. Small-molecule hyaluronic acid and its compound components used in mesotherapy have an even shorter residence time in the dermis, with a single treatment typically lasting 1 to 3 months, often requiring multiple treatments to consolidate the effect. While regenerative materials can extend the duration of action, the non-absorbable microspheres pose a risk of overstimulation or even granulomas, and removal is difficult once problems occur. Fully absorbable materials (such as PLLA) face challenges due to significant individual differences in collagen regeneration stimulation efficiency and the difficulty in accurately predicting the results. Furthermore, the injection procedure itself may carry risks such as infection and vascular embolism. Another common and impactful issue affecting efficacy is that the injected fluid often leaks from the injection site. Since the dosage per injection is limited, leakage directly leads to the loss of effective ingredients, significantly weakening the expected hydration and filling effect.

[0005] To address these challenges, innovation in medical aesthetic fillers and hydrating materials is evolving towards longer-lasting and more functional applications. In the field of fillers, research focus is shifting from simple physical placement to functional regenerative materials. The aim is to construct biocompatible porous scaffold structures to guide autologous cell migration and tissue regeneration, thereby reducing the risk of foreign body reactions and achieving more natural, long-lasting filling effects. In the field of hydrating and functional products, efforts are being made to develop novel sustained-release technologies, such as microencapsulation and in-situ gelation, to allow active ingredients to be released slowly and continuously at the injection site, thus extending the duration of action and improving the efficacy of a single treatment. However, currently widely used materials such as sodium hyaluronate and collagen typically involve complex chemical cross-linking and physical pulverization processes to form injectable gel particles. These processes are cumbersome and still cannot completely solve the problem of rapid degradation in vivo. Fillers made from materials such as polylactic acid and polycaprolactone often require a long reconstitution process before use, resulting in inconvenience. Traditional mesotherapy formulations are mostly liquid mixtures. After injection into tissues, their small molecule components diffuse and are absorbed rapidly, resulting in a very short residence time at the target site, which limits their full functionality. Therefore, developing an injectable material that can intelligently respond to the physiological environment, quickly form a stable presence, and achieve sustained release of functional components has significant clinical and market value. Summary of the Invention

[0006] The purpose of this invention is to provide a physiological fluid-responsive subcutaneous injectable hydrating filler for aesthetic purposes. Its core mechanism lies in the following: when the filler is injected into the subcutaneous tissue, the cations such as sodium, potassium, calcium, and magnesium in the tissue fluid can undergo a specific ionic response with the fluid-responsive material (such as deacetylated gellan gum) in the filler, rapidly forming a non-flowing, solid, three-dimensional network gel scaffold. This gel structure effectively locks in moisture and stabilizes functional ingredients such as whitening and anti-wrinkle agents, as well as macromolecules such as collagen, within the network, thereby significantly prolonging the retention time of active ingredients at the injection site. Simultaneously, the gel scaffold itself can act as a sustained-release matrix, allowing functional ingredients to be slowly released into the surrounding tissues for a long-lasting effect. Furthermore, the gel network can protect structural components such as collagen from enzymatic degradation, delaying their degradation, and providing three-dimensional space for cell migration and collagen regeneration, thus achieving both immediate filling and tissue regeneration promotion effects.

[0007] The product of this invention exhibits excellent performance in in vitro simulation tests: upon contact with ion-containing simulated tissue fluid, its viscosity rapidly increases and solidifies into a supportive gel, effectively filling tissue gaps. Further research confirms that after being damaged by external forces (such as vibration), the gel can spontaneously recover to a solid state upon standing, demonstrating good self-healing and plasticity, making it suitable for shaping and filling different areas of the face. Most importantly, because this material solidifies in situ upon contact with tissue fluid, it can seal the needle hole instantly upon needle removal, significantly reducing or even avoiding common problems in traditional mesotherapy such as leakage of medication, bleeding from the needle hole, or tissue fluid seepage, thus improving treatment efficiency and patient experience.

[0008] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a physiological fluid-responsive subcutaneous tissue injection hydrating filler, comprising, by weight: 0.2-15 parts of fluid-responsive material, 0.2-50 parts of whitening and anti-wrinkle functional ingredients, 0.2-15 parts of structural ingredients, 50-100 parts of osmotic pressure regulator, and 0.3-5 parts of local analgesic.

[0009] Preferably, the body fluid responsive material is one or both of deacetylated low-acyl gellan gum or non-acyl gellan gum.

[0010] Preferably, the whitening and anti-wrinkle functional ingredients are selected from one or more of the following: vitamin C and its derivatives, B vitamins, glutathione, tranexamic acid, amino acids, lipoic acid, hexapeptide, copper peptide, anti-aging / repair complex peptide, dipotassium glycyrrhizate, allantoin, nicotinamide, retinol and its derivatives, pro-xylene, asiaticoside, polydeoxynucleotides, oligodeoxynucleotides, fibronectin, elastin, silk fibroin, mucin, chondroitin sulfate, exosomes, ergothioneine, L-carnosine, deoxycholic acid, and botulinum toxin.

[0011] Preferably, the structural component is selected from one or more of alginate, animal-derived collagen, allogeneic collagen, recombinant human collagen, recombinant humanized collagen, recombinant humanoid collagen, gelatin, carboxymethyl chitosan, sodium hyaluronate, hydroxypropyl methylcellulose, hydroxyethylcellulose, polylactic acid microspheres, polycaprolactone microspheres, hydroxyapatite microspheres, and fibroblasts.

[0012] Preferably, the osmotic pressure regulator is selected from one or more of glycerol, glucose, mannitol, sorbitol, propylene glycol, and polyethylene glycol.

[0013] Preferably, the local analgesic is selected from one or more of lidocaine, prilocaine, and tetracaine.

[0014] Preferably, it also includes a solvent, wherein the solvent is 400-1200 parts of water for injection.

[0015] Preferably, the water for injection is deionized water.

[0016] This invention provides the application of the physiological fluid environment-responsive subcutaneous tissue injection hydrating filler in medical aesthetic hydration and / or tissue filling.

[0017] Compared with the prior art, the present invention has the following beneficial effects: The fluid-responsive material used in this invention is the only biomaterial capable of directly reacting with cations in body fluids to form a gel under physiological saline conditions. Currently used clinical fillers, such as sodium alginate, collagen, sodium hyaluronate, chitosan, and cellulose, do not possess the ability to specifically bind to cations in body fluids and rapidly form a stable gel. Human tissue fluid contains various cations, including sodium, potassium, calcium, and magnesium ions. This invention utilizes the characteristic of fluid-responsive materials to bind with these cations and form a gel to design and develop a novel tissue filler material. After injection into human tissue, this material rapidly reacts with cations in the tissue fluid, forming a gel in situ, thereby achieving long-lasting hydration and effective filling.

[0018] Under physiological conditions, the salt ion concentration in human tissue fluid is approximately 145.1 mEq / L. The fluid-responsive material used in this invention exhibits significant salt ion sensitivity, enabling it to bind with cations at this concentration to form a stable gel. Specifically, cations in the tissue fluid can complex with carbonyl groups on the gellan gum polymer chains, promoting the formation of a stable double-helix structure between molecular chains through hydrogen bonding. Further, through the reverse aggregation of the double helix, a three-dimensional gel network is constructed. This network structure significantly reduces the material's fluidity, thereby prolonging its retention time at the injection site.

[0019] Given that the injectable filler of this invention is sensitive to salt ions, to avoid ion interference with its gel formation process, the selected osmotic pressure regulators are all non-ionic substances, such as glycerol, glucose, mannitol, sorbitol, propylene glycol, and polyethylene glycol. By adjusting the formulation to an isotonic state, tissue and cell damage caused by improper osmotic pressure can be effectively avoided.

[0020] To prevent the denaturation and inactivation of heat-sensitive ingredients such as collagen and peptides during high-temperature processing, this invention employs a two-component formulation. Specifically, the heat-sensitive ingredients are freeze-dried into a solid powder and then sterilized by irradiation; while the fluid-responsive materials and other heat-resistant ingredients are prepared as a liquid and sterilized at high temperature. The two components are mixed before use, ensuring both sterility and the stability of the active ingredients.

[0021] The body fluid responsive material used in this invention, after dissolving in conventional hot water (below 100°C), undergoes thermo-coagulation upon cooling to room temperature, making it unsuitable for direct preparation into an injectable liquid formulation. Therefore, the liquid formulation containing this material requires high-temperature treatment (thermal sterilization) at 121°C–132°C for 15–60 minutes. After this treatment, the material no longer undergoes thermo-coagulation at room temperature, retaining only its ion-responsive gel properties in physiological body fluid environments, thus meeting the stability requirements for clinical injection.

[0022] Furthermore, the solid gel formed by the injectable filler of this invention possesses excellent self-healing properties. When the gel structure is disrupted by external forces (such as rapid vibration) and transforms into a fluid state, it can revert to a solid gel after being left to stand for a period of time. This characteristic allows the material to retain a certain degree of plasticity after injection, making it suitable for hydration and tissue filling needs in different areas and shapes within the medical aesthetics field. Attached Figure Description

[0023] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0024] Figure 1 The images show the products prepared in Examples 1 to 5. Detailed Implementation

[0025] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0026] Example 1

[0027] Unless otherwise specified, all chemicals used in the embodiments of this invention are common chemicals that can be purchased commercially. Gellan gum is produced by Zhengzhou Hongxiang Chemical Co., Ltd.

[0028] Example 1

[0029] A physiological fluid-responsive subcutaneous tissue injection hydrating filler for cosmetic purposes, comprising the following components by weight: 5 grams of low-acyl gellan gum 2 grams of sodium alginate Cysteine-type Vitamin C 5 grams Mannitol 50.7g 3 grams of lidocaine The specific preparation method is as follows: First, 5 grams of low-acyl gellan gum are evenly dispersed in 800 ml of deionized water. Then, the mixture is heated to 80°C and stirred until completely dissolved. 2 grams of sodium alginate, 5 grams of cysteine-type vitamin C, 50.7 grams of mannitol, and 3 grams of lidocaine are added and stirred until completely dissolved. Then, the volume is adjusted to 1000 ml by adding water to make up the evaporated volume. Each bottle contains 10 ml and is sterilized at 121°C for 30 minutes to obtain the injectable filler.

[0030] Example 2

[0031] A physiological fluid-responsive subcutaneous tissue injection hydrating filler for cosmetic purposes, comprising the following components by weight: 5 grams of low-acyl gellan gum 2 grams of sodium hyaluronate 30 grams of glutathione Mannitol 50.7g 3 grams of lidocaine The specific preparation method is as follows: First, 5 grams of low-acyl gellan gum are evenly dispersed in 800 ml of deionized water. Then, the mixture is heated to 90°C and stirred until completely dissolved. 2 grams of sodium hyaluronate, 30 grams of glutathione, 50.7 grams of mannitol, and 3 grams of lidocaine are added and stirred until completely dissolved. Then, the volume is adjusted to 1000 ml by adding water to make up the evaporated volume. Each bottle contains 10 ml and is sterilized at 121°C for 30 minutes to obtain the injectable filler.

[0032] Example 3

[0033] A physiological fluid-responsive subcutaneous tissue injection hydrating filler for cosmetic purposes, comprising the following components by weight: 10g of low-acyl gellan gum 2 grams of hydroxypropyl methylcellulose Cysteine-type Vitamin C 5 grams Mannitol 50.7g 3 grams of lidocaine The specific preparation method is as follows: First, 10 grams of low-acyl gellan gum is evenly dispersed in 800 ml of deionized water. Then, it is heated to 70°C and stirred until completely dissolved. 2 grams of hydroxypropyl methylcellulose, 5 grams of cysteine-type vitamin C, 50.7 grams of mannitol, and 3 grams of lidocaine are added and stirred until completely dissolved. Then, the volume is adjusted to 1000 ml by adding water to make up the evaporated volume. Each bottle contains 10 ml and is sterilized at 121°C for 30 minutes to obtain the injectable filler.

[0034] Example 4

[0035] A physiological fluid-responsive subcutaneous tissue injection hydrating filler for cosmetic purposes, comprising the following components by weight: 5 grams of low-acyl gellan gum 10 grams of recombinant collagen Cysteine-type Vitamin C 5 grams Mannitol 50.7g 3 grams of lidocaine The specific preparation method is as follows: First, 5 grams of low-acyl gellan gum are evenly dispersed in 800 ml of deionized water. Then, the mixture is heated to 80°C and stirred until completely dissolved. Then, the evaporated water is added to make up the volume to 1000 ml. Each bottle contains 10 ml and is sterilized at 121°C for 30 minutes to obtain the liquid component of the injection filler.

[0036] 10g of recombinant collagen, 5g of cysteine-type vitamin C, 50.7g of mannitol, and 3g of lidocaine were stirred until completely dissolved. Then, water was added to make up the volume to 1000ml. The solution was packaged in 10ml vials, freeze-dried into a powder, and then sterilized by irradiation to obtain the solid component of the injectable filler. When using, the liquid component is injected into the solid component using a syringe until completely dissolved before use.

[0037] Example 5

[0038] A physiological fluid-responsive subcutaneous tissue injection hydrating filler for cosmetic purposes, comprising the following components by weight: 5 grams of low-acyl gellan gum 10 grams of recombinant collagen 5 grams of hexapeptide Mannitol 50.7g 3 grams of lidocaine The specific preparation method is as follows: First, 5 grams of low-acyl gellan gum are evenly dispersed in 800 ml of deionized water. Then, the mixture is heated to 80°C and stirred until completely dissolved. Then, the evaporated water is added to make up the volume to 1000 ml. Each bottle contains 10 ml and is sterilized at 121°C for 30 minutes to obtain the liquid component of the injection filler.

[0039] 10g of recombinant collagen, 5g of hexapeptide, 50.7g of mannitol, and 3g of lidocaine were stirred until completely dissolved. Then, water was added to make up the volume to 1000ml. The mixture was packaged in 10ml vials, freeze-dried into a powder, and sterilized by irradiation to obtain the solid component of the injectable filler. When using, the liquid component is injected into the solid component using a syringe until completely dissolved before use.

[0040] Experimental Example 1

[0041] The products from Examples 1 to 5 above are respectively used as Sample 1 to Sample 5 (e.g. Figure 1 (As shown), used for relevant measurement tests.

[0042] 1. Dialysis membrane diffusion gel method

[0043] 1) Experimental Principle

[0044] This experiment simulates the process by which tissue fluid diffuses into the filler after it is injected into subcutaneous tissue, causing the filler to gradually solidify. The filler was placed in a dialysis bag, which only allows small molecules to enter and exit, while large molecules are prevented from doing so. This allows salt ions from the saline solution to enter the dialysis bag, gradually increasing the viscosity of the filler until it eventually solidifies.

[0045] 2) Test methods

[0046] In the experiment, 5 ml of sample was placed in a 16 mm diameter dialysis bag, sealed at both ends, and completely immersed in physiological saline for 0, 5, 15, 30, 60, and 120 minutes. The sample was then removed, poured out, and its viscosity was measured. The viscosity of a sample that had not undergone dialysis before the experiment was taken as the viscosity at 0.

[0047] 3) Test Results

[0048] Table 1. Viscosity Measurement Results (mPa·s)

[0049] The experiment showed that the viscosity of the filler began to increase after 5 minutes of contact with body fluids, and it became jelly-like after 15 minutes. Furthermore, the viscosity gradually increased and solidified over time. This experiment demonstrates that the filler can interact with body fluids in the in vivo environment to form a solid gel, filling interstitial spaces and providing support.

[0050] 2. Drug sustained-release test

[0051] This experiment used sodium fluorescein ultraviolet spectrophotometry to determine the sustained-release effect of implanted drugs. The principle is to quantify the effect by utilizing the characteristic that the absorbance of sodium fluorescein at a specific wavelength (λmax=330nm) is proportional to its concentration.

[0052] 1) Plotting a standard curve: Prepare sodium fluorescein solutions of different concentrations (0~100µg / mL) and measure the absorbance at 330nm; immerse the sodium fluorescein-loaded hydrogel in PBS, take samples at regular intervals and measure the absorbance, and calculate the concentration through the standard curve.

[0053] 2) Sample preparation: Mix sodium fluorescein with the filler solution (add 0.1 mL of sodium fluorescein solution to every 1 mL of colloidal solution, mix thoroughly, then pour into a culture dish and fix with physiological saline for 30 minutes) to form a drug-loaded hydrogel, achieving a final concentration of 0.5–5 mg / mL. PBS (pH 7.4) is used as the release medium to simulate the physiological environment, and its volume must cover the gel surface.

[0054] 3) Sampling and detection: Initial intensive (30min, 1h, 4h), later extended (12h, 1d, 2d, 3d); 1mL of release solution was taken each time, and fresh medium of equal volume and temperature was added. The cumulative release rate was calculated after measurement.

[0055] 4) Test Results

[0056] Table 2 Results of the Drug Sustained-Release Survey

[0057] The experimental results show that the filler, after coagulation with small molecule drugs, has the effect of slow drug release. It can fix functional small molecule substances in the gel and release them slowly, thus allowing them to exert a longer-lasting effect locally.

[0058] 3. Collagen Anti-degradation Test

[0059] 1) Experimental Objectives and Principles

[0060] This study simulates an in vivo environment to determine the degradation rate of collagen materials under the action of collagenase, and evaluates the anti-degradation effect of the implant's collagen by assessing the degradation status. Collagenase specifically hydrolyzes peptide bonds in the triple helix structure of collagen. A quantitative amount of collagen material was placed in a buffer solution containing a specific concentration of collagenase and incubated at a constant temperature. The degree of enzymatic resistance of the implant was quantified by periodically measuring the increase in hydroxyproline, a characteristic protein hydrolysis product of collagen. A high hydroxyproline content in the degradation solution indicates faster degradation, while a low hydroxyproline content indicates slower degradation.

[0061] 2) Experimental materials and reagents

[0062] (1) Samples: Samples 4 and 5. The low-acyl gellan gum in product 4 was replaced with an equal amount of sodium hyaluronate as a control.

[0063] (2) Enzyme solution: Tris-HCl buffer (50 mM, pH 7.4) containing CaCl2 (5~10 mM). Calcium ions are an essential cofactor for collagenase activity. The concentration of collagenase is 100 U / mL, depending on the experimental purpose and sample properties.

[0064] Equipment and apparatus: High-performance liquid chromatograph; constant temperature water bath shaker (set to 37℃); analytical balance (accuracy 0.1mg)

[0065] 3) Test Procedure

[0066] (1) Sample preparation and weighing: Accurately weigh 2 grams of sample and 2 grams of control.

[0067] (2) Preparation of enzyme hydrolysate: Prepare the collagenase working solution of the required concentration using preheated buffer (containing CaCl2). Ensure complete dissolution.

[0068] (3) Incubation reaction: Place each weighed sample into a separate test tube. Add sufficient collagenase working solution to the test tubes of the experimental group (ensure that the sample is completely submerged). Place all test tubes in a constant temperature water bath shaker at 37°C and shake at a low speed (e.g., 60 rpm) to ensure thorough mixing and reaction.

[0069] (4) Termination of reaction and sampling: Parallel samples from the experimental and control groups were collected at predetermined time points (6h, 12h, 24h, 48h, 72h). Immediately after sample collection, EDTA (ethylenediaminetetraacetic acid, final concentration 20 mM) was added to chelate Ca²⁺. + This terminates enzyme activity.

[0070] (5) Hydroxyproline determination method: Hydroxyproline is an amino acid unique to collagen. The hydroxyproline content in the enzymatic hydrolysate was determined by high performance liquid chromatography. A standard curve was plotted using hydroxyproline standards, and the hydroxyproline content in the degradation solution was calculated.

[0071] Table 3. Results of the Collagen Anti-Degradation Test

[0072] The experimental results showed that samples 4 and 5, when containing salt ions and collagenase, formed a gel, encapsulating the collagen within the gel structure. This prevented collagenase from easily diffusing into the gel structure, thus inhibiting its degradation of collagen. However, when containing sodium hyaluronate with the same amount of collagen, sodium hyaluronate could not form a gel structure, allowing collagenase to easily contact and degrade the collagen into peptides and amino acids. Therefore, the filler of this invention, when containing collagen, has an anti-collagen degradation effect, allowing collagen to remain at the injection site for a longer time and maintaining a longer-lasting cosmetic effect.

[0073] 4. Plasticity Test

[0074] In the experiment, 10 ml of sample was placed in a 16 mm diameter dialysis bag, sealed at both ends, and completely immersed in physiological saline for 120 minutes. Afterward, the solidified colloid was removed, poured out, and injected into a 10 ml vial using a syringe. The vial was then sealed, and vigorous shaking was applied to allow the gel to flow. The gel was then allowed to stand for 10 minutes to observe its solidification. The results showed that all five samples solidified again, indicating that the filler has plasticity.

[0075] 5. Safety Research and Testing

[0076] The injectable fillers prepared in Examples 1-5 were subjected to the following biological tests.

[0077] 1) Intradermal irritation: The test solution was prepared by adding physiological saline at a ratio of 0.2 g / ml. The intradermal irritation test was conducted according to the animal intradermal irritation test method specified in the national standard GB / T16886.23, Biological Evaluation of Medical Devices. The results showed that the irritation index was 0 for all samples, indicating no intradermal irritation.

[0078] 2) Cytotoxicity: The test solution was prepared by adding cell culture medium at a ratio of 0.2 g / ml. Then, the cytotoxicity test was performed using the MTT assay as specified in the national standard GB / T16886.5 Biological Evaluation of Medical Devices. The results of the cytotoxicity test showed that the cell proliferation rate was in the range of 85%~105%, and there was no cytotoxicity.

[0079] 3) Sensitization test: The test solution was prepared by adding physiological saline at a ratio of 0.2 g / ml. Then, skin sensitization tests were performed according to the methods specified in the national standard GB / T16886.10 Biological Evaluation of Medical Devices, and no sensitization effect was observed.

[0080] 4) Subcutaneous implantation test: The subcutaneous implantation test was conducted according to the methods specified in the national standard GB / T16886.6 Biological Evaluation of Medical Devices, with 0.5 ml of sample injected subcutaneously at the implantation site. A control group was injected subcutaneously with 0.5 ml of a 20 mg / ml sodium hyaluronate solution. Histopathological reactions were observed at 2, 4, and 8 weeks post-implantation. The results showed no local irritation from the implant.

[0081] 6. Skin Filling Support Effect Test

[0082] In clinical practice, the dosage of cosmetic fillers at each injection point is generally 0.05ml to 0.2ml. This experiment evaluates the post-injection support strength by observing the collapse over time after subcutaneous injection of an excessive amount of filler. A sustained height of the injection site indicates good support, while gradual collapse over time indicates weak support.

[0083] 1) Test materials: (1) Three fresh detached pig skins were purchased from the local market and stored in a refrigerator at 4°C when needed. The experimental time was no more than 24 hours after the pig slaughter time.

[0084] (2) Experimental table, syringe, timer, pins, thread, camera, ruler, white foam board. 23G injection needle.

[0085] 2) Test methods

[0086] Pigskin was fixed to a square foam board using pins. One ml of each of the six samples (samples 1-5) of filler and a 20 mg / ml sodium hyaluronate control solution was injected subcutaneously. Immediately, a bulge formed under the pig skin. The height of each bulge (0 min) was measured using a ruler and silk thread, and photographs were taken perpendicular to the plane of the skin using a digital camera to accurately record the height (mm) of each bulge. The pigskin was then immersed in physiological saline, and the bulge height was measured again at the same location after 15, 30, 60, and 120 minutes, with photographs taken simultaneously. Each subcutaneous injection experiment was repeated independently three times. At the end of the experiment, the injection site was cut open with scissors to observe gel formation. The results indicate that the samples have a good skin-filling effect.

[0087] 3) Test Results

[0088] Table 4. Results of the Skin Filling Support Effect Test (Unit: mm)

[0089] The experimental results show that after the sample of the present invention is injected subcutaneously, it forms a solid gel due to rapid interaction with the tissue fluid and does not diffuse rapidly, thus providing a good filling and support effect.

[0090] In summary, it is evident that this invention systematically verifies the performance of the invented physiological fluid-responsive injectable hydrating filler. In experiments simulating in vivo gel formation and support effects, the sample rapidly thickens and solidifies upon contact with physiological saline, exhibiting excellent interstitial filling and support capabilities, while also possessing self-healing properties and the ability to be repeatedly reshaped. In drug sustained-release and anti-degradation tests, the gel structure effectively encapsulates functional small molecules and collagen, achieving sustained-release function and significantly resisting collagenase degradation, thereby prolonging the duration of action of the active ingredients.

[0091] Furthermore, biological safety assessments showed that the material was non-irritating, non-cytotoxic, and non-sensitizing, meeting medical device safety standards. Ex vivo porcine skin filling experiments further confirmed that the gel formed after injection stably maintained the subcutaneous elevation, demonstrating excellent filling effects. In summary, this filler exhibits significant advantages in responsive gel formation, functional sustained release, long-lasting support, and biological safety, making it suitable for cosmetic hydration and tissue filling applications.

[0092] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A physiological fluid-responsive subcutaneous tissue injection hydrating filler, characterized in that, By weight, it includes: 0.2-15 parts of body fluid responsive material, 0.2-50 parts of whitening and anti-wrinkle functional ingredients, 0.2-15 parts of structural ingredients, 50-100 parts of osmotic pressure regulator, and 0.3-5 parts of local analgesic.

2. The water-replenishing filler according to claim 1, characterized in that, The fluid-responsive material is one or both of deacetylated low-acyl gellan gum or non-acyl gellan gum.

3. The water-replenishing filler according to claim 1, characterized in that, The whitening and anti-wrinkle functional ingredients are selected from one or more of the following: Vitamin C and its derivatives, B vitamins, glutathione, tranexamic acid, amino acids, lipoic acid, hexapeptide, copper peptide, anti-aging / repair complex peptide, dipotassium glycyrrhizate, allantoin, nicotinamide, retinol and its derivatives, pro-xylene, asiaticoside, polydeoxynucleotides, oligodeoxynucleotides, fibronectin, elastin, silk fibroin, mucin, chondroitin sulfate, exosomes, ergothioneine, L-carnosine, deoxycholic acid, and botulinum toxin.

4. The water-replenishing filler according to claim 1, characterized in that, The structural components are selected from one or more of the following: alginate, animal-derived collagen, allogeneic collagen, recombinant human collagen, recombinant humanized collagen, recombinant humanoid collagen, gelatin, carboxymethyl chitosan, sodium hyaluronate, hydroxypropyl methylcellulose, hydroxyethylcellulose, polylactic acid microspheres, polycaprolactone microspheres, hydroxyapatite microspheres, and fibroblasts.

5. The water-replenishing filler according to claim 1, characterized in that, The osmotic pressure regulator is selected from one or more of glycerol, glucose, mannitol, sorbitol, propylene glycol, and polyethylene glycol.

6. The water-replenishing filler according to claim 1, characterized in that, The local analgesic is selected from one or more of lidocaine, prilocaine, and tetracaine.

7. The water-replenishing filler according to claim 1, characterized in that, It also includes a solvent, wherein the solvent is 400-1200 parts of water for injection.

8. The application of a physiological fluid environment-responsive subcutaneous tissue injection hydrating filler as described in any one of claims 1 to 7 in medical aesthetic hydration and / or tissue filling.

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