Schizophyllan hydro-optical needle solution and preparation method thereof

Abstract

The invention relates to a schizophyllan hydro-optical needle solution and a preparation method thereof, and belongs to the technical field of medical cosmetology. The invention relates to a schizophyllan hydro-globular needle solution, which comprises schizophyllan, and the concentration of the schizophyllan is 5-10 mg / mL. The moisturizing cream not only has a good moisturizing effect, but also can maintain the effect for a long time.

Description

Technical Field

[0001] This application relates to the field of medical aesthetics technology, and in particular to a slit polysaccharide water light injection solution and its preparation method. Background Technology

[0002] Most mesotherapy solutions use non-crosslinked hyaluronic acid, whose molecular chains are loose, making it easy to diffuse and absorb. It is suitable for rapid superficial hydration and moisturization, providing immediate hydration to the skin. However, hyaluronic acid is broken down and metabolized by enzymes in the body, and its existence time in the human body is relatively short. In order to maintain its effect, some mesotherapy solutions will add a small amount of crosslinking agent (such as BDDE) for micro-crosslinking treatment. The purpose is to prolong the time it stays in the skin, but its degree of crosslinking is far lower than that of hyaluronic acid used for filling.

[0003] Schizophyllum polysaccharide is a neutral polysaccharide produced by extraction or submerged fermentation of Schizophyllum commune fruiting bodies. It possesses excellent moisturizing and hygroscopic properties and good biocompatibility, making it increasingly popular in cosmetic applications. Furthermore, the schizophyllum polysaccharide molecule has numerous hydroxyl groups, which can form dynamic cross-linking points through hydrogen bonding, exhibiting gel properties. Currently, it is used as a filler material in the form of hydrogels. Summary of the Invention

[0004] To address the shortcomings of existing technologies, the objective of this application is to provide a schizosaccharide-based mesotherapy solution and its preparation method. This solution offers both good moisturizing effects and can maintain its effectiveness for a longer period.

[0005] In one aspect, embodiments of this application provide a schizophyllum polysaccharide hyaluronic acid injection solution, which includes schizophyllum polysaccharide at a concentration of 5-10 mg / mL.

[0006] The schizopolysaccharide mesotherapy solution provided in this application includes a suitable concentration of schizopolysaccharide. Schizopolysaccharide has a good moisturizing effect and is not degraded by enzymes in the body, thus maintaining its effect for a long time. At the same time, the viscosity and storage modulus of the suitable concentration of schizopolysaccharide solution decrease sharply during injection, showing significant shear-thinning behavior. After injection, its viscosity and storage modulus quickly recover to the initial level, showing excellent thixotropic recovery and structural reversibility. Thus, it can be smoothly injected during the injection process and quickly recover its original viscoelastic state after injection into the tissue to maintain morphological stability and mechanical support, making it very suitable for use in mesotherapy solutions.

[0007] In some embodiments of this application, the concentration of schizopolysaccharide is 6-9 mg / mL.

[0008] This application further employs a more suitable concentration of schizopolysaccharide, which maintains a lower viscosity during injection and has a higher storage modulus after injection, thus exhibiting better morphological stability and mechanical support.

[0009] In some embodiments of this application, the active ingredient of the slit polysaccharide water light injection solution is slit polysaccharide.

[0010] The active ingredient of the schizophyllum polysaccharide mesotherapy solution provided in this application is schizophyllum polysaccharide, which has good moisturizing effect and can maintain the effect for a long time. At the same time, it can be smoothly injected during the injection process and can play a role in maintaining morphological stability and mechanical support after being injected into the tissue.

[0011] In some embodiments of this application, the storage modulus of the schizopolysaccharide hyaluronic acid injection solution at a shear frequency of 1 Hz is 10~40 Pa.

[0012] The slit polysaccharide mesotherapy solution provided in this application contains a suitable concentration of slit polysaccharide. The slit polysaccharide mesotherapy solution has a suitable storage modulus at a shear frequency of 1 Hz, which is beneficial for maintaining morphological stability and mechanical support after injection into the tissue.

[0013] In some embodiments of this application, the storage modulus of the schizopolysaccharide hyaluronic acid injection solution at a shear frequency of 1 Hz is 15~40 Pa.

[0014] The slit polysaccharide mesotherapy solution provided in this application further includes a more suitable concentration of slit polysaccharide, so that the slit polysaccharide mesotherapy solution has a more suitable storage modulus at a shear frequency of 1 Hz, which is beneficial to further play the role of maintaining morphological stability and mechanical support after injection into the tissue.

[0015] In some embodiments of this application, the schizopolysaccharide hyaluronic acid injection solution was used in 2.5 s -1 The viscosity at the shear rate is 1~4 Pa·s; preferably 1.5~4 Pa·s.

[0016] The schizophyllum polysaccharide mesotherapy solution provided in this application contains a suitable concentration of schizophyllum polysaccharide, and the schizophyllum polysaccharide mesotherapy solution is effective in 2.5 seconds. -1 It has a suitable viscosity at a shear rate to facilitate smooth injection of the slit polysaccharide water-light injection solution.

[0017] In some embodiments of this application, the pushing force of the slit polysaccharide water light injection solution is less than 13 N.

[0018] The slit polysaccharide mesotherapy solution provided in this application contains a suitable concentration of slit polysaccharide. Therefore, the slit polysaccharide mesotherapy solution has a small pushing force, which is conducive to smooth operation during injection and can reduce pain at the injection site. It is very beneficial for multi-point and large-area injection operations of mesotherapy.

[0019] Secondly, embodiments of this application provide a method for preparing the slit-fold polysaccharide hyaluronic acid injection solution as provided in the first aspect, comprising: S1, Dissolve the lyophilized powder of schizophyllum polysaccharide in physiological saline to obtain a solution with a schizophyllum polysaccharide concentration of 5~10 mg / mL; S2, centrifuge and sterilize the solution in S1 to obtain the schizopolysaccharide water light injection solution.

[0020] This application uses lyophilized schizophyllum polysaccharide powder dissolved in physiological saline to prepare a schizophyllum polysaccharide solution of appropriate concentration. After centrifugation and sterilization, the schizophyllum polysaccharide mesotherapy solution can be obtained. It has good moisturizing effect and can maintain the effect for a long time. At the same time, it can be smoothly injected during the injection process, and can play a role in maintaining morphological stability and mechanical support after being injected into the tissue.

[0021] In some embodiments of this application, in S1, the lyophilized powder of schizopolysaccharide is dissolved in physiological saline at 70-80 °C.

[0022] This application uses the method of dissolving the lyophilized schizophyllum polysaccharide powder in physiological saline at a suitable temperature, which facilitates a more complete dissolution of the lyophilized schizophyllum polysaccharide powder.

[0023] In some embodiments of this application, in step S2, the solution is centrifuged at 1200-2000 rpm for 3-5 min and sterilized at 125-130℃ for 5-8 min.

[0024] This application employs suitable conditions to centrifuge and sterilize the slit polysaccharide solution, in order to form a slit polysaccharide mesotherapy solution suitable for skin use. Attached Figure Description

[0025] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of this application and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0026] Figure 1 The infrared spectrum of the schizopolysaccharide provided in Experimental Example 1 of this application.

[0027] Figure 2 Image of the schizopolysaccharide hyaluronic acid injection solution provided in Experimental Example 2 of this application.

[0028] Figure 3 This is an image of the injection site in the mouse provided in Experiment Example 3 of this application.

[0029] Figure 4 HE staining image of mouse injection area tissue provided in Experiment Example 3 of this application.

[0030] Figure 5 The storage modulus curve of the schizopolysaccharide hyaluronic acid injection solution provided in Experimental Example 4 of this application is shown; where (A) is the schizopolysaccharide solution before sterilization; and (B) is the schizopolysaccharide hyaluronic acid injection solution after sterilization.

[0031] Figure 6 The storage modulus curve of the slit polysaccharide hydrogel solution provided in Experimental Example 4 of this application at a shear frequency of 1 Hz.

[0032] Figure 7 Viscosity diagram of the schizopolysaccharide hyaluronic acid injection solution provided in Test Example 5 of this application; wherein, (A) - schizopolysaccharide solution before sterilization; (B) - schizopolysaccharide hyaluronic acid injection solution after sterilization.

[0033] Figure 8 Viscosity diagram of the schizopolysaccharide hydrogel solution provided in Test Example 5 of this application at a shear frequency of 1 Hz.

[0034] Figure 9 The graph shows the storage modulus results of the slit polysaccharide hydrogel solution provided in Experimental Example 6 of this application.

[0035] Figure 10 The viscosity result of the schizopolysaccharide hydrogel solution provided in Test Example 6 of this application is shown in the figure.

[0036] Figure 11 The diagram shows the pushing force results of the slit polysaccharide water-light injection solution provided in Test Example 7 of this application.

[0037] Figure 12 The graph shows the results of the moisturizing performance provided in Test Example 8 of this application. Detailed Implementation

[0038] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions in the embodiments of this application will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0039] Currently, all existing mesotherapy solutions use non-crosslinked hyaluronic acid, which is broken down and metabolized by enzymes in the body and has a relatively short lifespan in the human body. In order to maintain its effect, some mesotherapy solutions will add a small amount of crosslinking agent for micro-crosslinking treatment, the purpose of which is to prolong the time it stays in the skin, but its degree of crosslinking is far lower than that of hyaluronic acid used for filling.

[0040] Therefore, this application provides a schizopolysaccharide hyaluronic acid injection solution, which includes schizopolysaccharide (SPG) at a concentration of 5-10 mg / mL.

[0041] The schizopolysaccharide mesotherapy solution provided in this application includes a suitable concentration of schizopolysaccharide. Schizopolysaccharide has a good moisturizing effect and is not degraded by enzymes in the body, thus maintaining its effect for a long time. At the same time, the viscosity and storage modulus of the suitable concentration of schizopolysaccharide solution decrease sharply during injection, showing significant shear-thinning behavior. After injection, its viscosity and storage modulus quickly recover to the initial level, showing excellent thixotropic recovery and structural reversibility. Thus, it can be smoothly injected during the injection process and quickly recover its original viscoelastic state after injection into the tissue to maintain morphological stability and mechanical support, making it very suitable for use in mesotherapy solutions.

[0042] In some embodiments of this application, the concentration of schizophyllum polysaccharide is 6-9 mg / mL. As examples, the concentration of schizophyllum polysaccharide may be, but is not limited to, 6 mg / mL, 6.5 mg / mL, 7 mg / mL, 7.5 mg / mL, 8 mg / mL, 8.5 mg / mL, and 9 mg / mL. Further, using a more suitable concentration of schizophyllum polysaccharide results in a lower viscosity during injection and a higher storage modulus after injection, thus providing better morphological stability and mechanical support.

[0043] It is understandable that other active ingredients can be added to the schizopolysaccharide mesotherapy solution as needed, as long as the added active ingredients do not affect the properties and efficacy of the schizopolysaccharide.

[0044] In some embodiments of this application, the active ingredient of the schizophyllum polysaccharide mesotherapy solution is schizophyllum polysaccharide. The schizophyllum polysaccharide mesotherapy solution uses only schizophyllum polysaccharide as its active ingredient, which provides both good moisturizing effects and a longer-lasting effect. Furthermore, it allows for smooth injection during the injection process and provides morphological stability and mechanical support after being injected into the tissue. It is understood that using only schizophyllum polysaccharide as the active ingredient in the schizophyllum polysaccharide mesotherapy solution can achieve good mesotherapy efficacy, improving biocompatibility while simplifying the process and reducing costs.

[0045] In some embodiments of this application, the storage modulus of the schizopolysaccharide mesotherapy solution at a shear frequency of 1 Hz is 10~40 Pa. Since the schizopolysaccharide mesotherapy solution contains a suitable concentration of schizopolysaccharide, it possesses a suitable storage modulus at a shear frequency of 1 Hz, which is beneficial for maintaining morphological stability and providing mechanical support after injection into tissue.

[0046] Furthermore, the storage modulus of the schizophyllum polysaccharide mesotherapy solution at a shear frequency of 1 Hz is 15~40 Pa. If the schizophyllum polysaccharide mesotherapy solution further includes a more suitable concentration of schizophyllum polysaccharide, then the schizophyllum polysaccharide mesotherapy solution will have an even more suitable storage modulus at a shear frequency of 1 Hz, which is beneficial for maintaining morphological stability and providing mechanical support after injection into tissues.

[0047] In some embodiments of this application, the schizopolysaccharide hyaluronic acid injection solution was used in 2.5 s -1 The viscosity at the shear rate is 1~4 Pa·s; preferably 1.5~4 Pa·s. The schizophyllum polysaccharide mesotherapy solution contains a suitable concentration of schizophyllum polysaccharide, and the schizophyllum polysaccharide mesotherapy solution has a viscosity of 2.5 s. -1 It has a suitable viscosity at a shear rate to facilitate smooth injection of the slit polysaccharide water-light injection solution.

[0048] In some embodiments of this application, the pushing force of the schizophyllum polysaccharide mesotherapy solution is less than 13 N. Since the schizophyllum polysaccharide mesotherapy solution contains a suitable concentration of schizophyllum polysaccharide, it has a low pushing force, which facilitates smooth injection and reduces pain at the injection site, making it highly suitable for multi-point, large-area injections of mesotherapy.

[0049] Secondly, embodiments of this application provide a method for preparing the above-mentioned slit-fold polysaccharide hyaluronic acid injection solution, comprising: S1, Dissolve the lyophilized powder of schizophyllum polysaccharide in physiological saline to obtain a solution with a schizophyllum polysaccharide concentration of 5~10 mg / mL; S2, centrifuge and sterilize the solution in S1 to obtain the schizopolysaccharide water light injection solution.

[0050] This application uses lyophilized schizophyllum polysaccharide powder dissolved in physiological saline to prepare a schizophyllum polysaccharide solution of appropriate concentration. After centrifugation and sterilization, the schizophyllum polysaccharide mesotherapy solution can be obtained. It has good moisturizing effect and can maintain the effect for a long time. At the same time, it can be smoothly injected during the injection process, and can play a role in maintaining morphological stability and mechanical support after being injected into the tissue.

[0051] In some embodiments of this application, in step S1, the lyophilized schizophyllum polysaccharide powder is dissolved in physiological saline at 70-80 °C. Dissolving the lyophilized schizophyllum polysaccharide powder in physiological saline at a suitable temperature facilitates more complete dissolution of the lyophilized schizophyllum polysaccharide powder.

[0052] In some embodiments of this application, in step S2, the solution is centrifuged at 1200-2000 rpm for 3-5 min and sterilized at 125-130℃ for 5-8 min. Centrifugation and sterilization of the schizopolysaccharide solution under suitable conditions facilitate the formation of a schizopolysaccharide mesotherapy solution suitable for skin use.

[0053] In an optional embodiment of this application, the preparation method of the lyophilized schizophyllum polysaccharide powder can be carried out according to the following process: the refined filtrate of schizophyllum polysaccharide (provided by Guangdong Marubi Biotechnology Co., Ltd.) is sequentially subjected to precipitation, centrifugation, reconstitution and concentration to obtain a sample solution; finally, the sample solution is freeze-dried using a vacuum freeze dryer to obtain the lyophilized schizophyllum polysaccharide powder. It is understood that commercially available lyophilized schizophyllum polysaccharide powder can also be used directly.

[0054] It should be noted that the refined filtrate of Schizophyllum polysaccharide refers to the refined filtrate obtained by filtering out the Schizophyllum mycelium after fermentation culture.

[0055] As an example, the filtrate of the polysaccharide was added with 3-4 times its volume of anhydrous ethanol and allowed to stand at 4 °C for 12-36 h to precipitate. The precipitate was then collected by centrifugation at 4000-6000 rpm for 15-20 min. The precipitate was reconstituted with distilled water at 50-55 °C and concentrated at below 75 °C using a rotary evaporator to obtain the sample solution.

[0056] As an example, the sample solution was pre-frozen at -40 ℃ and then transferred to a freeze dryer. It was freeze-dried for 24 to 36 hours under conditions where the cold trap temperature was below -50 ℃ and the vacuum degree was below 10 Pa to obtain the freeze-dried powder of schizopolysaccharide.

[0057] The features and performance of this application will be further described in detail below with reference to the embodiments.

[0058] Example 1 This embodiment provides a schizophyllum polysaccharide hyaluronic acid injection solution. The active ingredient of the schizophyllum polysaccharide hyaluronic acid injection solution is schizophyllum polysaccharide, and the concentration of schizophyllum polysaccharide is 5 mg / mL.

[0059] This embodiment provides a method for preparing a slit-fold polysaccharide hyaluronic acid injection solution, including: S1, the filtrate of the schizophyllum polysaccharide was added to 3 times the volume of anhydrous ethanol and allowed to stand at 4 ℃ overnight to precipitate. Then, the precipitate was collected by centrifugation at 4000 rpm for 15 min. The precipitate was redissolved in distilled water at 50 ℃ and concentrated by rotary evaporator at 70 ℃ to obtain the sample solution. The final sample solution was pre-frozen at -45 ℃ and then transferred to a freeze dryer. It was freeze-dried for 24 h under the conditions of cold trap temperature of -60 ℃ and vacuum degree of 8 Pa to obtain the lyophilized powder of schizophyllum polysaccharide. S2, the lyophilized powder of schizophyllum polysaccharide was dissolved in physiological saline in a 75 ℃ water bath with stirring to obtain a solution with a schizophyllum polysaccharide concentration of 5 mg / mL; S3. Centrifuge the solution in S2 at 1500 rpm for 5 min, take the supernatant and sterilize it at 125 ℃ for 6 min to obtain the schizopolysaccharide water light injection solution.

[0060] Example 2 This embodiment provides a schizophyllum polysaccharide hyaluronic acid injection solution. The active ingredient of the schizophyllum polysaccharide hyaluronic acid injection solution is schizophyllum polysaccharide, and the concentration of schizophyllum polysaccharide is 6 mg / mL.

[0061] The preparation method of the slit polysaccharide water light injection solution provided in this embodiment is the same as that in Example 1.

[0062] Example 3 This embodiment provides a schizophyllum polysaccharide hyaluronic acid injection solution. The active ingredient of the schizophyllum polysaccharide hyaluronic acid injection solution is schizophyllum polysaccharide, and the concentration of schizophyllum polysaccharide is 7 mg / mL.

[0063] The preparation method of the slit polysaccharide water light injection solution provided in this embodiment is the same as that in Example 1.

[0064] Example 4 This embodiment provides a schizophyllum polysaccharide hyaluronic acid injection solution. The active ingredient of the schizophyllum polysaccharide hyaluronic acid injection solution is schizophyllum polysaccharide, and the concentration of schizophyllum polysaccharide is 8 mg / mL.

[0065] The preparation method of the slit polysaccharide water light injection solution provided in this embodiment is the same as that in Example 1.

[0066] Example 5 This embodiment provides a schizophyllum polysaccharide hyaluronic acid injection solution. The active ingredient of the schizophyllum polysaccharide hyaluronic acid injection solution is schizophyllum polysaccharide, and the concentration of schizophyllum polysaccharide is 9 mg / mL.

[0067] The preparation method of the slit polysaccharide water light injection solution provided in this embodiment is the same as that in Example 1.

[0068] Example 6 This embodiment provides a schizophyllum polysaccharide hyaluronic acid injection solution. The active ingredient of the schizophyllum polysaccharide hyaluronic acid injection solution is schizophyllum polysaccharide, and the concentration of schizophyllum polysaccharide is 10 mg / mL.

[0069] The preparation method of the slit polysaccharide water light injection solution provided in this embodiment is the same as that in Example 1.

[0070] Comparative Example 1 This comparative example provides a schizophyllum polysaccharide hyaluronic acid injection solution. The active ingredient of the schizophyllum polysaccharide hyaluronic acid injection solution is schizophyllum polysaccharide, and the concentration of schizophyllum polysaccharide is 1 mg / mL.

[0071] The preparation method of the slit polysaccharide water-light injection solution provided in this comparative example is the same as that in Example 1.

[0072] Experimental Example 1 This experimental example performs qualitative tests on the lyophilized polysaccharide powder provided in Example 1. The specific methods are as follows: Before testing, the lyophilized polysaccharide powder was thoroughly dried in a desiccator to ensure the removal of moisture interference. Using the ATR-FTIR reflectance mode, a small amount of the lyophilized polysaccharide powder sample was placed on the clean, dry surface of an ATR crystal. The pressure device provided with the instrument was used to press the lyophilized polysaccharide powder sample evenly and firmly into contact with the ATR crystal. The wavenumber range was set to 4000-500 cm⁻¹. - ¹, resolution 4 cm - ¹, 32 scans were performed to collect the chromatogram of the lyophilized polysaccharide sample. Results are as follows: Figure 1 As shown.

[0073] Depend on Figure 1 The results show that 890-900 cm - ¹This is due to the bending vibration of the C1-H plane and is a characteristic absorption peak of the β-glycosidic bond; 1070 cm⁻¹ - ¹, and 1040 cm - ¹ is the COC / COH stretching vibration, often associated with the β-(1→3) glycosidic bond in the main chain; 1150-1000 cm⁻¹ - ¹ represents the stretching vibrations of COC and COH, with complex peak shapes, representing the "fingerprint region" of the pyranose ring; 1640 cm⁻¹ - ¹A sharp peak exists nearby, generated by the HOH bending vibration of bound water; 3400cm - The broad peak near ¹ is a characteristic peak of the O-H stretching vibration, which is a characteristic peak of hydrogen bonding. Thus, the lyophilized powder of the schizopolysaccharide was prepared.

[0074] Experimental Example 2 This experiment observed the pre-sterilization and post-sterilization schizophyllum polysaccharide solutions provided in Examples 1, 6, and Comparative Example 1. The results are as follows: Figure 2 As shown.

[0075] Depend on Figure 2The results show that as the concentration increases, the solution gradually exhibits gelation characteristics. Natural schizopolysaccharides exist stably in a triple helix structure. At low concentrations, the triple helix structures are far apart from each other and separated by water molecules, allowing them to flow freely. When the concentration increases, the distance between molecules shortens, and they come into contact and collide with each other. Because schizopolysaccharides contain a large number of hydroxyl groups, temporary cross-linking points are formed through hydrogen bonding, thereby maintaining the three-dimensional network structure of the gel.

[0076] Experimental Example 3 This experimental example assesses the biocompatibility of the schizopolysaccharide hyaluronic acid injection solutions provided in Examples 1 and 6. The specific methods are as follows: Four 8-week-old male BALB / c mice (purchased from Zhuhai Beston Biotechnology Co., Ltd.) were used for acclimatization for 3 days, followed by isoflurane inhalation anesthesia. After anesthesia, the fur on the backs of the mice was shaved, and the skin was disinfected with 75% ethanol. Subsequently, the sterilized schizosaccharide solution provided in Examples 1 and 6 was injected intradermally at two sites on each side of the back using a 30G needle, with an injection volume of 0.05 mL at each site. Physiological saline was used as a blank control group. Postoperatively, the injection areas were photographed at the same time each day. Figure 3 As shown; mice were sacrificed on day 9, and full-thickness skin tissue was harvested from each injection site. After fixation in 4% paraformaldehyde for 24 h, the tissue was embedded in paraffin and stained with hematoxylin-eosin (HE). The results are as follows. Figure 4 As shown, observe and assess the local inflammatory response. On the ninth day of dissection, residual polysaccharide solution was still visible to the naked eye, a phenomenon further confirmed in tissue sections. Figure 4 The results showed that the epidermal structure remained intact, and the dermis contained a large number of normally distributed sweat glands, hair follicles, and capillaries, with only a small amount of inflammatory cell infiltration. Small patches of schizopolysaccharide solution residue were observed below the fascia layer, surrounded by a large number of cell aggregates and mild inflammatory cell infiltration. Overall, the reaction was mild and within an acceptable safety range. This indicates that the sterilized schizopolysaccharide mesotherapy solution provided in Examples 1 and 6 exhibited good biocompatibility in the short term, without causing significant tissue damage or severe inflammatory reactions, preliminarily demonstrating its good biocompatibility.

[0077] Test Example 4 This experimental example performs rheological tests on the schizopolysaccharide solutions provided in Examples 1-6 before and after sterilization. The specific methods are as follows: A plate rotational rheometer (MCR 302e) from Anton Paar, Austria, was used. Before frequency scanning, the linear viscoelastic region of the schizopolysaccharide solution material needed to be determined. A PP50 parallel plate fixture was selected, and the test temperature was set to 25°C, the shear rate range was 0.01-100Hz, and the shear strain was fixed at 0.1%. Subsequently, dynamic frequency scanning was performed on the schizopolysaccharide solutions provided in Examples 1-6 before sterilization and after sterilization, with a fixed pressure of 0N. The storage modulus G' and loss modulus G'' at different frequencies f were recorded, and curves (double logarithmic coordinates) of G' and G'' as a function of frequency were plotted. The results are as follows: Figure 5 As shown.

[0078] Depend on Figure 5 The results showed that with the increase of schizophyllum polysaccharide concentration, the storage modulus (G′) of the sample significantly increased from 15 Pa to 40 Pa, and was higher than the loss modulus (G″) at all frequencies. This indicates that the schizophyllum polysaccharide solution transforms from a viscous liquid to an elastically dominated gel state with increasing concentration, forming a stable three-dimensional network structure inside. Furthermore, the modulus of the schizophyllum polysaccharide solution did not change significantly before and after moist heat sterilization, proving that the sterilization process is suitable for this material and does not affect its rheological properties and gel stability. This demonstrates that the appropriate concentration of schizophyllum polysaccharide solution provided in this application can still maintain excellent support properties after injection, and can be injected into the dermis as a mesotherapy formulation to achieve tightening, lifting, and slight depression filling effects from the inside. To standardize representation criteria and ensure data comparability, the following were selected: Figure 5 The energy storage modulus (G′) at 1 Hz in the frequency scan is used as the core evaluation index, such as... Figure 6 As shown; and commercially available products 1 (main ingredient: sodium hyaluronate, concentration 5 mg / mL), 2 (main ingredient: sodium hyaluronate, concentration 12 mg / mL), and 3 (main ingredient: sodium hyaluronate, concentration 12 mg / mL) were used as controls. By adjusting the concentration of the schizopolysaccharide solution within the range of 5–10 mg / mL, its storage modulus (G′) could be precisely adjusted between 10 and 40 Pa. This storage modulus range successfully covers the mechanical performance range corresponding to mainstream high-end mesotherapy products on the market.

[0079] Experimental Example 5 This experiment tested the steady-state shear viscosity of the schizopolysaccharide solutions provided in Examples 1-6 before and after sterilization. The specific method is as follows: An Austrian Anton Paar MCR 302e rotational rheometer was used with PP50 parallel plate clamps. The temperature was kept constant at 25°C, and the shear rate range was 0.01 - 100 s⁻¹. -¹; Steady-state flow scanning was performed on the schizopolysaccharide solutions provided in Examples 1-6 before and after sterilization, and the apparent viscosity as a function of shear rate was recorded. The results are as follows: Figure 7 As shown.

[0080] Depend on Figure 7 The results showed that the apparent viscosity of all schizopolysaccharide solutions decreased significantly with increasing shear rate. Simultaneously, at any given shear rate, the viscosity of the solution systematically increased with increasing schizopolysaccharide concentration. This rheological behavior primarily stems from the three-dimensional network structure constructed by the molecular chains of the schizopolysaccharide solution. Under static or low-shear conditions, the molecular chains form a stable network through hydrogen bonds and entanglement, encapsulating water molecules and thus exhibiting high viscosity. However, this dynamic network is disrupted under external shear stress. This significant shear-thinning characteristic directly determines the excellent injectability of the schizopolysaccharide solution. During syringe storage and transportation (high shear rate), the material viscosity is extremely low, easily passing through narrow needles; once injection is complete, the shear stress disappears, its three-dimensional network structure rapidly recovers, and the viscosity rebounds, thus maintaining its shape under the skin and providing the expected support, effectively avoiding the risk of clogging during injection.

[0081] To standardize representation criteria and ensure data comparability, the following were selected: Figure 7 During the frequency scan, 2.5s -1 The viscosity at that point is used as the core evaluation indicator, such as... Figure 8 As shown; and commercially available products 1 (main ingredient: sodium hyaluronate, concentration 5 mg / mL), 2 (main ingredient: sodium hyaluronate, concentration 12 mg / mL), and 3 (main ingredient: sodium hyaluronate, concentration 12 mg / mL) were used as controls. The viscosity of the schizopolysaccharide solution systematically increases with increasing concentration, and can be effectively controlled within the range of 1 Pa·s to 4 Pa·s. It is worth noting that this viscosity range is relatively low compared to some commercially available products characterized by high viscoelasticity, which brings unique application advantages to the schizopolysaccharide solution. Viscosity, as a key rheological parameter of mesotherapy preparations, directly affects its injectability and injection experience. Lower viscosity means that less pushing force is required during injection. This greatly reduces the difficulty of injection operation, allowing doctors to perform single-handed injection more easily and accurately, especially suitable for mesotherapy treatments that require multiple points and large areas of injection. Moreover, lower viscosity can effectively reduce pain during injection, thereby improving patient compliance and treatment satisfaction. In addition, the lower viscosity results in less flow resistance when passing through injection needles with small apertures, significantly reducing the probability of needle blockage and ensuring a smooth and safe treatment process.

[0082] Experimental Example 6 This experiment tested the rheological thixotropy of the polysaccharide solutions provided in Examples 1-6. The specific methods are as follows: The structural reversibility and recovery kinetics of the sterilized schizopolysaccharide mesotherapy solutions provided in Examples 1-6 were systematically characterized by a three-stage step shear experiment. First, alternating strain step tests were performed in oscillation mode: small strain (1%, simulating static structural response) and large strain (100%, simulating structural failure) were applied sequentially at a fixed oscillation frequency (10 Hz), followed by recovery to the small strain. This cycle was repeated three times, and the structural reversibility of the schizopolysaccharide solution during cyclic deformation was evaluated by modulus change. Figure 9 As shown; then, a three-step shear rate step test was performed in steady-state shear mode, first at a low shear rate ( = 1 s - ¹) Initial viscosity was measured to establish a rheological baseline before injection, followed by instantaneous application of a high shear rate ( = 100 s - ¹) Simulate the needle injection process, record the sudden drop in viscosity caused by shear thinning to reflect the degree of instantaneous structural damage, and then immediately restore to a low shear rate ( = 1 s - ¹) and continuously monitor the viscosity change over time to obtain kinetic parameters such as structural recovery rate and characteristic recovery time, systematically evaluating the material's performance recovery capability after injection, such as... Figure 10 As shown.

[0083] Depend on Figure 9 The results show that, at a constant oscillation frequency, the storage modulus (G′) of the schizopolysaccharide hyaluronic acid injection solution exhibits a systematic upward trend with increasing concentration, consistent with the rheological characterization results in Example 4, further confirming the structure-property relationship of its internal physical cross-linked network increasing with concentration. In the alternating strain step test, the schizopolysaccharide hyaluronic acid injection solution exhibits a stable linear viscoelastic response at a small strain of 1%, with the modulus remaining constant. When the strain instantaneously steps to 100%, the modulus drops sharply and remains at a low plateau in the high strain stage, indicating that the internal structure of the schizopolysaccharide hyaluronic acid injection solution is significantly damaged under larger strains. Subsequently, when the strain recovers to 1%, the modulus can quickly recover to the initial level, and no significant modulus decay is observed after three complete cycles. This demonstrates that the schizopolysaccharide hyaluronic acid injection solution provided in this application possesses rapid and almost complete structural self-recovery ability after experiencing high-intensity shear failure, with negligible modulus loss, exhibiting excellent thixotropic recovery and structural reversibility.

[0084] Depend on Figure 10The results show that the steady-state viscosity of the schizopolysaccharide hydrogel solution systematically increases with increasing concentration, consistent with the rheological characterization in Example 5, further confirming the structure-activity relationship of its internal physical network enhancing with concentration. At low shear rates ( = 1 s - Under the conditions described in ¹), the schizopolysaccharide solution exhibited a high initial viscosity, reflecting its structured state under static or low-speed shear; when the shear rate instantaneously stepped to 100 s⁻¹, the viscosity remained relatively high. - At ¹, its viscosity decreased sharply, exhibiting significant shear-thinning behavior, indicating that the internal structure of the schizopolysaccharide mesotherapy solution underwent rapid and reversible dissociation under high shear, corresponding to the fluidity required when passing through a narrow needle during injection. Subsequently, when the shear rate recovered to 1 s⁻¹, the viscosity decreased. - ¹ At this point, the viscosity rapidly recovers to its initial level, and in three consecutive complete "low-high-low" shear rate cycles, the schizopolysaccharide mesotherapy solution exhibits almost complete reversible recovery behavior, without significant viscosity decay or thixotropic hysteresis. This indicates that the schizopolysaccharide mesotherapy solution provided in this application not only possesses excellent shear-thinning properties to adapt to the injection process, but also has good structural reconstruction ability and reversibility, thereby ensuring that it can quickly recover its original viscoelastic properties after injection into the skin, maintaining stable filling and support functions.

[0085] In practical applications, this characteristic indicates that the slit polysaccharide mesotherapy solution provided in this application can achieve smooth injection through shear thinning during the injection process, and quickly recover its original viscoelastic state after being injected into the skin tissue, thereby maintaining morphological stability and mechanical support function. Therefore, it is indicated that the slit polysaccharide mesotherapy solution provided in this application can be considered as a type of mesotherapy preparation with good injection and filling performance, suitable for medical aesthetic and biomaterial applications that require both injection fluidity and in vivo structural stability.

[0086] Experimental Example 7 This experiment tested the extrusion force of the schizopolysaccharide solutions provided in Examples 1-6 before and after sterilization. The specific method is as follows: Each schizopolysaccharide solution, before and after sterilization, was loaded into a pre-filled syringe. The matching core rod and injection needle (30G) were installed, and a small amount of air was expelled from the syringe tip. The compression clamp was installed on a universal testing machine. The test temperature was room temperature, and the room temperature equilibration time (for products stored under refrigeration) was 1 hour after the product was removed from the machine. The test distance was set to full scale, and the pushing speed was set to 30 mm / min. The pushing force was then tested according to the prescribed method. Commercially available products 1 (main ingredient: sodium hyaluronate, concentration 5 mg / mL), 2 (main ingredient: sodium hyaluronate, concentration 12 mg / mL), and 3 (main ingredient: sodium hyaluronate, concentration 12 mg / mL) were used as controls. The maximum, minimum, and average pushing forces in the plateau region of the pushing force curve were recorded. The results are as follows: Figure 11 As shown.

[0087] Depend on Figure 11 The results show that as the concentration of schizopolysaccharide increases, the number of molecular chains per unit volume increases, the density of intermolecular hydrogen bonds and chain entanglement increases, and the hydroxyl groups in the molecules form hydrogen bonds, resulting in a denser three-dimensional network structure. Fluid flow requires breaking more energy, which macroscopically manifests as an increase in the pushing force of the solution. The schizopolysaccharide aqueous solution provided in this application has a pushing force of less than 20N. This value range is considered to ensure smooth one-handed injection by doctors while effectively reducing patient pain caused by excessive injection resistance.

[0088] Experimental Example 8 This experiment tested the moisturizing properties of the schizopolysaccharide solution, and the specific method is as follows: Using a simulated human body temperature (37℃), the in vitro moisturizing performance of schizopolysaccharide (SPG) solution was quantitatively evaluated and compared with that of commonly used moisturizers (glycerin, hyaluronic acid / HA) and occlusive agents (olive oil) in an open environment was assessed. 0.1 mg / mL and 1 mg / mL SPG solutions were placed in a constant temperature (37℃) dry environment for 50 h to allow surface moisture to evaporate freely. The rate of change in sample mass was calculated by periodically and accurately weighing the samples to obtain the residual moisture rate. Glycerin was used as a positive control, olive oil as a negative control, 0.1 mg / mL and 1 mg / mL hyaluronic acid (HA) solutions as references for mainstream products on the market, and deionized water as a blank control. Results are as follows: Figure 12 As shown.

[0089] Depend on Figure 12The results show that glycerin, as a representative of hygroscopic moisturizers, exhibits excellent hygroscopic capacity; olive oil, as a representative of occlusive agents, maintained a relatively constant mass throughout the experiment, confirming its physical barrier function by forming a hydrophobic oil film on the liquid surface to effectively prevent water evaporation. At the same concentration, the residual water content of the schizopolysaccharide solution was consistently slightly higher than that of the hyaluronic acid (HA) solution, indicating that SPG possesses a slightly superior long-lasting moisturizing ability. Although both HA and SPG are hydrophilic polysaccharides that can bind water by forming hydrogen bond networks with water through hydroxyl groups on their molecular chains, SPG's unique β-(1,3)- and β-(1,6)-glycosidic bond structure endows its molecular chains with higher rigidity and a stronger ability to form a stable three-dimensional network. This denser and more stable network structure can more effectively "capture" and fix free water within its mesh, thus more effectively delaying water loss under dynamic evaporation conditions and exhibiting superior long-lasting moisturizing performance.

[0090] The embodiments described above are some, but not all, of the embodiments of this application. The detailed description of the embodiments of this application is not intended to limit the scope of the claimed application, but merely to illustrate selected embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of this application without inventive effort are within the scope of protection of this application.

Claims

1. A riparian polysaccharide hydrogel solution, characterized in that, The schizophyllan hydro-optical needle solution comprises schizophyllan, and the concentration of the schizophyllan is 5-10 mg / mL.

2. The riparian polysaccharide hydro-optical needle solution according to claim 1, characterized in that, The concentration of the schizophyllan is 6-9 mg / mL.

3. The fissured polysaccharide hydrophilic needle solution according to claim 1, wherein, The active ingredient of the schizophyllan hydro-optical needle solution is schizophyllan.

4. The sclerosing polysaccharide light solution according to any one of claims 1 to 3, characterized in that, The storage modulus of the schizophyllan hydro-optical needle solution under a shear frequency of 1 Hz is 10-40 Pa.

5. The riparian polysaccharide hydrogel solution of claim 4, wherein, The storage modulus of the schizophyllan hydro-optical needle solution under a shear frequency of 1 Hz is 15-40 Pa.

6. The sclerosing polysaccharide light needle solution according to any one of claims 1 to 3, characterized in that, The celloidal polysaccharide hydrogel solution has a viscosity of 1-4 Pa-s at a shear rate of 2.5 s -1 The viscosity at a shear rate of 1-4 Pa-s; preferably 1.5-4 Pa-s.

7. The sclerosing polysaccharide light needle solution according to any one of claims 1 to 3, characterized in that, The push force of the schizophyllan hydro-optical needle solution is less than 13 N.

8. A method of preparing a Schizophyllan aqueous light needle solution according to any one of claims 1 to 7, characterized by, Comprise: S1, dissolving schizophyllan freeze-dried powder in physiological saline to obtain a solution with a schizophyllan concentration of 5-10 mg / mL; S2, centrifuging and sterilizing the solution in S1 to obtain a schizophyllan hydro-optical needle solution.

9. The production method according to claim 8, characterized by, In S1, the schizophyllan freeze-dried powder is dissolved in physiological saline at 70-80 ℃.

10. The production method according to claim 8, characterized by, In S2, the solution is centrifuged at 1200-2000 rpm for 3-5 min, and sterilized at 125-130 ℃ for 5-8 min.

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