A PDRN non-crosslinked sodium hyaluronate gel composition, preparation method, related products and applications

Abstract

The present invention provides a PDRN non-crosslinked sodium hyaluronate gel composition, a preparation method, related products and applications, belonging to the technical fields of medicine, medical devices and cosmetics. The PDRN gel composition of the present invention is composed of the following components by weight: 5-15 parts of sodium hyaluronate 1, 1-3 parts of sodium hyaluronate 2, 1-2.7 parts of PDRN, 0.05-0.15 parts of phytic acid, 0.025-0.075 parts of zinc chloride and 200-300 parts of water; wherein, the molecular weight of sodium hyaluronate 1 is 1000 kDa-2000 kDa; the molecular weight of sodium hyaluronate 2 is less than 10 kDa. The composition provided by the present invention can prepare a PDRN sodium hyaluronate gel without chemical crosslinking agents, and can achieve the controlled release of PDRN and better storage stability.

Description

Technical Field

[0001] The present invention belongs to the technical fields of medicine, medical devices and cosmetics, and relates to a PDRN non-crosslinked sodium hyaluronate gel composition, a preparation method, related products and applications. Background Art

[0002] Sodium hyaluronate is a sodium salt of glucuronic acid and exists naturally in mammalian connective tissues, skin, eyes and synovial fluid. It is an important biopolymer. Its corresponding acid is hyaluronic acid, also known as hyaluronic acid, which is one of the main components of the extracellular matrix. Hyaluronic acid and sodium hyaluronate have good biocompatibility and have been widely used in the fields of beauty, cosmetics, daily chemicals, etc.

[0003] In order to improve the physical and chemical properties of sodium hyaluronate to adapt to the characteristics of products containing sodium hyaluronate components in different scenarios, the chemical cross-linking method is often used to transform the microstructure of sodium hyaluronate. Since sodium hyaluronate contains active groups such as carboxyl groups (sodium salts) and hydroxyl groups, chemical reagents that can react with it are often used for cross-linking. For example, Mi Pengcheng et al. provided a technique for preparing cross-linked sodium hyaluronate by using divinyl sulfone as a cross-linking agent in the article "Preparation and Biocompatibility of Divinyl Sulfone Cross-linked Sodium Hyaluronate Gel" (Chinese Journal of Tissue Engineering Research and Clinical Rehabilitation, 2008, 12(14): 4). The prepared sodium hyaluronate has good biocompatibility and its stability has been improved to a certain extent. However, covalent cross-linking of sodium hyaluronate with a chemical cross-linking agent introduces small molecule substances of the cross-linking agent that do not exist in the biological system. Although the sodium hyaluronate cross-linked material can be degraded macroscopically (visible to the naked eye), it does not mean that the molecular fragments containing the cross-linking agent after degradation will not pose a biocompatibility risk. In addition, the degree of the cross-linking reaction is limited, so chemical cross-linking agents may remain. These all affect the biocompatibility of chemically cross-linked sodium hyaluronate materials.

[0004] Phytic acid, also known as inositol hexaphosphate and cyclohexanehexol hexaphosphate, is an organic phosphate compound extracted from plant seeds. Phytic acid is a natural small molecule with a chemical structure having multiple phosphate groups, which can produce intermolecular interactions with various biomolecules such as proteins, polysaccharides, polypeptides, nucleic acids, etc. At the same time, it is a humectant and antioxidant and is widely used in the cosmetics industry. The interaction force between phytic acid and biomacromolecules belongs to intermolecular interaction and does not affect the covalent structure of the biomacromolecules themselves. However, due to the relatively weak binding force of intermolecular interaction compared with covalent bonds, the role of phytic acid itself in the stability of the complex gel system containing various biomacromolecules is relatively complex and difficult to predict.

[0005] PDRN (polydeoxyribonucleotide), also known as "polydeoxyribonucleotide", is a class of DNA derivatives of natural origin, consisting of a mixture of DNA fragments with a chain length of 50 - 2000 base pairs. Currently, PDRN is mainly extracted and purified from the sperm cells of salmon. PDRN has shown definite effects in aspects such as anti - inflammation, wound healing, tissue restoration, and anti - ischemia.

[0006] Chinese invention patent CN118873426A provides a PDRN - crosslinked sodium hyaluronate composition and its application. This composition includes PDRN, oxidized sodium hyaluronate, polyamino acid, and deionized water. This technology uses sodium periodate to oxidize sodium hyaluronate, and the oxidized sodium hyaluronate contains aldehyde groups; then PDRN is cross - linked with sodium hyaluronate containing aldehyde groups to form a PDRN - crosslinked sodium hyaluronate composition. Although this method does not add chemical cross - linkers, it changes the chemical structure of sodium hyaluronate, and the cross - linking position of the generated PDRN - crosslinked sodium hyaluronate is uncertain, which has potential impacts on the anti - inflammatory and tissue restoration properties of PDRN itself. Finally, the oxidative modification of the gel itself improves the oxidation value of the gel as a whole, which is not conducive to the manifestation of the anti - inflammatory and antioxidant effects of the gel.

[0007] In summary, the prior art has not provided a PDRN - sodium hyaluronate gel system that does not undergo covalent cross - linking, does not change the covalent structure of each raw material, and can exist stably at the same time. Summary of the Invention

[0008] In view of this, aiming at the problems existing in the prior art, the purpose of the present invention is to provide a PDRN non - crosslinked sodium hyaluronate gel composition, related products, preparation methods, and applications.

[0009] To achieve the above - mentioned invention purpose, on the one hand, the present invention provides a PDRN non - crosslinked sodium hyaluronate gel composition, which is composed of the following components by weight:

[0010] 5 - 15 parts of sodium hyaluronate 1, 1 - 3 parts of sodium hyaluronate 2, 1 - 2.7 parts of PDRN, 0.05 - 0.15 parts of phytic acid, 0.025 - 0.075 parts of zinc chloride, and 200 - 300 parts of water;

[0011] Among them, the molecular weight of sodium hyaluronate 1 is 1000 kDa - 2000 kDa; the molecular weight of sodium hyaluronate 2 is less than 10 kDa.

[0012] Preferably, the PDRN non - crosslinked sodium hyaluronate gel composition is composed of the following components by weight:

[0013] 8 - 12 parts of sodium hyaluronate 1, 1.5 - 2.5 parts of sodium hyaluronate 2, 1.4 - 2.3 parts of PDRN, 0.08 - 0.12 parts of phytic acid, 0.04 - 0.06 parts of zinc chloride and 200 - 250 parts of water.

[0014] More preferably, the PDRN non - crosslinked sodium hyaluronate gel composition is composed of the following components by weight:

[0015] 10 parts of sodium hyaluronate 1, 2 parts of sodium hyaluronate 2, 1.85 parts of PDRN, 0.1 part of phytic acid, 0.05 part of zinc chloride and 236 parts of water.

[0016] Preferably, and as a specific example of the present invention, the molecular weight of the sodium hyaluronate 1 is 1500 kDa ± 1 kDa.

[0017] On the other hand, the present invention provides a method for preparing PDRN non - crosslinked sodium hyaluronate gel using the above - mentioned PDRN non - crosslinked sodium hyaluronate gel composition, including the following steps:

[0018] S1. Mix the formulated amounts of sodium hyaluronate 2, PDRN, zinc chloride and water, and stir to obtain solution 1;

[0019] S2. Mix the formulated amount of phytic acid with water to obtain solution 2;

[0020] S3. Dropwise add solution 2 into the stirring solution 1 to obtain solution 3;

[0021] S4. Mix the formulated amount of sodium hyaluronate 1 with the remaining amount of water in the formulated amount, and then add solution 3 and mix evenly to obtain PDRN non - crosslinked sodium hyaluronate gel.

[0022] Preferably, in step S1, the weight of the water is 16 - 24 times, more preferably 20 times, the sum of the weights of sodium hyaluronate 2, PDRN and zinc chloride.

[0023] Preferably, in step S2, the weight of the water is 20 - 40 times, more preferably 30 times, the weight of phytic acid.

[0024] Preferably, in step S1, the stirring speed is 800 - 1200 rpm, more preferably 1000 rpm.

[0025] Preferably, in step S1, the stirring time is 8 - 18 h, more preferably 9 h.

[0026] Preferably, in step S3, the dropping speed is 0.01 - 0.05 mL / min, more preferably 0.03 mL / min.

[0027] Preferably, in step S3, the stirring speed is 800 - 1200 rpm, more preferably 1000 rpm.

[0028] Preferably, in step S4, the mixing specifically means stirring for 24 - 48 h, and more preferably, the stirring speed is 500 - 800 rpm.

[0029] In the above method, a sub - micron / micron - sized small cross - linker can be formed between sodium hyaluronate with small molecular weight, zinc chloride, PDRN, and phytic acid. On the one hand, it can play a certain role in stabilizing and encapsulating PDRN. On the other hand, after mixing with sodium hyaluronate with large molecular weight, the above - mentioned micron - sized small cross - linker can serve as a node for the non - covalent connection of sodium hyaluronate with large molecular weight, that is, through the intermolecular interaction between the groups on the surface of the small cross - linker and sodium hyaluronate with large molecular weight, the gel - forming performance of the gel is improved. And in this process, the covalent structures of PDRN and sodium hyaluronate remain unchanged.

[0030] On the other hand, the present invention provides a PDRN non - crosslinked sodium hyaluronate gel prepared by the above method.

[0031] On the other hand, the present invention provides the use of the above - mentioned PDRN non - crosslinked sodium hyaluronate gel composition or the PDRN non - crosslinked sodium hyaluronate gel prepared by the above method in the preparation of drugs, medical devices, or cosmetics.

[0032] On the other hand, the present invention provides a drug, and the product contains the above - mentioned PDRN non - crosslinked sodium hyaluronate gel composition or the PDRN non - crosslinked sodium hyaluronate gel prepared by the above method.

[0033] On the other hand, the present invention provides a medical device, and the product contains the above - mentioned PDRN non - crosslinked sodium hyaluronate gel composition or the PDRN non - crosslinked sodium hyaluronate gel prepared by the above method.

[0034] Finally, the present invention provides a cosmetic, and the product contains the above - mentioned PDRN non - crosslinked sodium hyaluronate gel composition or the PDRN non - crosslinked sodium hyaluronate gel prepared by the above method.

[0035] The drugs, medical devices, and cosmetics can be external application products acting on the skin, or products suitable for intradermal introduction or implantation, preferably in dosage forms such as dressings, emulsions, ointments, sprays, injections, and microneedles.

[0036] Compared with the prior art, the present invention has the following beneficial effects:

[0037] (1)The present invention provides a PDRN non-crosslinked sodium hyaluronate gel composition. By compounding PDRN with sodium hyaluronate of different molecular weights, zinc chloride and phytic acid, a PDRN non-crosslinked sodium hyaluronate gel with PDRN sustained-release performance, good storage stability and good biosafety can be obtained.

[0038] (2)The present invention provides a preparation method of a PDRN non-crosslinked sodium hyaluronate gel. The preparation method is simple and easy to implement, and is conducive to industrial production. Specific Embodiments

[0039] Terms and Statements of the Present Invention:

[0040] 1. Articles "a", "an" and "the": Unless otherwise explicitly limited to one (kind) of object, they include plural objects.

[0041] 2. Numerical ranges: Unless otherwise explicitly indicated, all ranges or ratios disclosed herein will be understood to include any and all sub-ranges or sub-ratios contained therein. For example, the stated range or ratio of 1 to 30 should be considered to be included between the minimum value of 1 and the maximum value of 30, and any sub-range or sub-ratio, integer, decimal, or sub-range or sub-ratio composed of integers or decimals including the endpoints.

[0042] The following non-limiting examples can enable those of ordinary skill in the art to more comprehensively understand the present invention, but do not limit the present invention in any way. The following content is merely an exemplary illustration of the scope claimed by the present invention. Those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes and modifications should also fall within the scope claimed by the present invention.

[0043] The present invention will be further described below in the form of specific examples. All kinds of chemical reagents used in the examples of the present invention are obtained through conventional commercial channels unless otherwise specified. Unless otherwise specified, the contents described below are mass contents. Unless otherwise specified, it is understood to be carried out at room temperature.

[0044] In the following examples, the sources of some reagents are as shown in Table 1 below:

[0045] Table 1

[0046]

[0047] In the following examples, the instrument and equipment used are as shown in Table 2 below:

[0048] Table 2

[0049]

[0050] Example 1

[0051] Preparation of PDRN non-crosslinked sodium hyaluronate gel is as follows.

[0052] S1. Add 200 mg of sodium hyaluronate (molecular weight < 10 kDa), 185 mg of PDRN, and 5 mg of zinc chloride to 8 mL (8 g) of deionized water successively under stirring at 1000 rpm. Under airtight conditions, stir for 9 h to disperse evenly to obtain Solution 1.

[0053] S2. Take 20 mg of phytic acid aqueous solution (weight fraction 50%, containing 10 mg of phytic acid and 10 mg of water) and mix it with 290 mg of water for dilution to obtain Solution 2.

[0054] S3. Suck Solution 2 into a syringe, and use a micro-injection pump to drop Solution 2 in the syringe into Solution 1 under stirring at 1000 rpm at a constant speed of 0.03 mL / min to obtain Solution 3.

[0055] S4. Mix 1000 mg of sodium hyaluronate (1500 kDa ± 1 kDa) with 15.3 g of water, stir overnight under the condition of 1000 rpm to mix evenly, and then pour Solution 3 into this mixed solution and mix well (stir at 600 rpm for 36 h) to obtain PDRN non-crosslinked sodium hyaluronate gel.

[0056] Example 2

[0057] Preparation of PDRN non-crosslinked sodium hyaluronate gel is as follows.

[0058] S1. Add 250 mg of sodium hyaluronate (molecular weight < 10 kDa), 230 mg of PDRN, and 4 mg of zinc chloride to 9.68 mL (9.68 g) of deionized water successively under stirring at 1000 rpm. Under airtight conditions, stir for 9 h to disperse evenly to obtain Solution 1.

[0059] S2. Take 24 mg of phytic acid aqueous solution (weight fraction 50%, containing 12 mg of phytic acid and 12 mg of water) and mix it with 348 mg of water for dilution to obtain Solution 2.

[0060] S3. Suck Solution 2 into a syringe, and use a micro-injection pump to drop Solution 2 in the syringe into Solution 1 under stirring at 1000 rpm at a constant speed of 0.03 mL / min to obtain Solution 3.

[0061] S4. Mix 800 mg of sodium hyaluronate (1500 kDa ± 1 kDa) with 9.96 g of water, stir overnight under the condition of 1000 rpm to mix evenly, and then pour Solution 3 into this mixed solution and mix well (stir at 600 rpm for 36 h) to obtain PDRN non-crosslinked sodium hyaluronate gel.

[0062] Example 3

[0063] The preparation of PDRN non-crosslinked sodium hyaluronate gel is as follows.

[0064] S1. Add 150 mg of sodium hyaluronate (molecular weight < 10 kDa), 140 mg of PDRN, and 6 mg of zinc chloride to 5.92 mL (5.92 g) of deionized water successively under stirring at 1000 rpm. Under airtight conditions, stir for 9 h to disperse evenly to obtain Solution 1.

[0065] S2. Take 16 mg of phytic acid aqueous solution (weight fraction 50%, containing 8 mg of phytic acid and 8 mg of water) and mix and dilute it with 232 mg of water to obtain Solution 2.

[0066] S3. Draw Solution 2 into a syringe, and use a micro-injection pump to slowly drip Solution 2 in the syringe into Solution 1 under stirring at 1000 rpm at a constant speed of 0.03 mL / min to obtain Solution 3.

[0067] S4. Mix 1200 mg of sodium hyaluronate (1500 kDa ± 1 kDa) with 18.84 g of water, stir overnight under the condition of 1000 rpm to mix evenly, and then pour Solution 3 into the mixed solution and mix well (stir at 600 rpm for 36 h) to obtain PDRN non-crosslinked sodium hyaluronate gel.

[0068] Example 4

[0069] The preparation of PDRN non-crosslinked sodium hyaluronate gel is as follows.

[0070] S1. Add 300 mg of sodium hyaluronate (molecular weight < 10 kDa), 270 mg of PDRN, and 2.5 mg of zinc chloride to 9.16 mL (9.16 g) of deionized water successively under stirring at 1200 rpm. Under airtight conditions, stir for 8 h to disperse evenly to obtain Solution 1.

[0071] S2. Take 30 mg of phytic acid aqueous solution (weight fraction 50%, containing 15 mg of phytic acid and 15 mg of water) and mix and dilute it with 585 mg of water to obtain Solution 2.

[0072] S3. Draw Solution 2 into a syringe, and use a micro-injection pump to slowly drip Solution 2 in the syringe into Solution 1 under stirring at 800 rpm at a constant speed of 0.05 mL / min to obtain Solution 3.

[0073] S4. Mix 500 mg of sodium hyaluronate (1500 kDa ± 1 kDa) with 10.24 g of water, stir overnight at 800 rpm until well mixed, then pour solution 3 into this mixed solution and mix well (stir at 600 rpm for 36 h) to obtain PDRN non-crosslinked sodium hyaluronate gel.

[0074] Example 5

[0075] The preparation of PDRN non-crosslinked sodium hyaluronate gel is as follows.

[0076] S1. Add 100 mg of sodium hyaluronate (molecular weight < 10 kDa), 100 mg of PDRN, and 7.5 mg of zinc chloride to 4.98 mL (4.98 g) of deionized water successively under stirring at 800 rpm. Under sealed conditions, stir for 18 h until fully dispersed and well mixed to obtain solution 1.

[0077] S2. Take 10 mg of phytic acid aqueous solution (weight fraction 50%, containing 5 mg of phytic acid and 5 mg of water) and mix and dilute it with 95 mg of water to obtain solution 2.

[0078] S3. Aspirate solution 2 into a syringe, and use a micro-injection pump to slowly drip the solution 2 in the syringe into solution 1 under stirring at 1200 rpm at a speed of 0.01 mL / min to obtain solution 3.

[0079] S4. Mix 1500 mg of sodium hyaluronate (1500 kDa ± 1 kDa) with 24.92 g of water, stir overnight at 1200 rpm until well mixed, then pour solution 3 into this mixed solution and mix well (stir at 600 rpm for 36 h) to obtain PDRN non-crosslinked sodium hyaluronate gel.

[0080] The PDRN non-crosslinked sodium hyaluronate gels obtained in Examples 1 - 5 are respectively denoted as Gel 1 - Gel 5 in sequence. The formulation summaries of the PDRN non-crosslinked sodium hyaluronate gel compositions corresponding to Gel 1 - Gel 5 are summarized in Table 3:

[0081] Table 3

[0082]

[0083] Comparative Example 1

[0084] Compared with Example 1, in step S1, the dosage of sodium hyaluronate (molecular weight < 10 kDa) is changed to 600 mg; in step S4, the dosage of sodium hyaluronate (1500 kDa ± 1 kDa) is changed to 600 mg, and the other conditions are the same.

[0085] Comparative Example 2

[0086] Compared with Example 1, in step S1, zinc chloride was replaced with calcium chloride of the same mass, and the rest were the same.

[0087] Comparative Example 3

[0088] Compared with Example 1, in step S1, zinc chloride was replaced with magnesium chloride of the same mass, and the rest were the same.

[0089] Comparative Example 4

[0090] Compared with Example 1, in step S1, the use of zinc chloride was omitted, and the dosage of PDRN was changed to 190 mg, and the rest were the same.

[0091] Comparative Example 5

[0092] Compared with Example 1, in step S2, solution 2 was 310 mg of deionized water, and the rest were the same.

[0093] Comparative Example 6

[0094] Compared with Example 1, in step S3, instead of using the dropping method, solution 2 and solution 1 were mixed by direct mixing, and the other conditions were the same.

[0095] Comparative Example 7

[0096] Compared with Example 1, in step S1, sodium hyaluronate (molecular weight < 10 kDa) was replaced with sodium hyaluronate of the same weight (Mw 100 kDa - 200 kDa), and the rest were the same.

[0097] Comparative Example 8

[0098] Compared with Example 1, in step S4, sodium hyaluronate (1500 kDa ± 1 kDa) was replaced with sodium hyaluronate of the same weight (Mw 100 kDa - 200 kDa), and the rest were the same.

[0099] The PDRN non-crosslinked sodium hyaluronate gels obtained from Comparative Example 1 to Comparative Example 8 were respectively denoted as gel d1 - gel d8 in sequence.

[0100] Effect evaluation

[0101] 1. Characterization of the formation of small cross-linked bodies by sodium hyaluronate, zinc chloride, phytic acid, and PDRN.

[0102] Take the solution 3 obtained in step S3 of Examples 1 - 5 and Comparative Examples 1 - 7, dilute it with water under stirring with a glass rod, and gradually dilute it to 100 times the original volume. Take an appropriate amount of the diluted solution and use a DLS particle size analyzer to detect its average particle size. The results are shown in Table 4:

[0103] Table 4

[0104]

[0105] It can be seen that in Examples 1 - 5 of the present invention, sodium hyaluronate with a molecular weight < 10 kDa can form a submicron - sized particle solution with PDRN, phytic acid, and zinc chloride. In Comparative Example 1, the excessive amount of sodium hyaluronate interfered with the formation of stable small cross - linked bodies. In Comparative Examples 2 and 3, calcium chloride and magnesium chloride were used to replace an equal amount of zinc chloride respectively. Although calcium ions and magnesium ions are both divalent metal cations like zinc ions, Comparative Examples 2 and 3 showed that the use of calcium chloride and magnesium chloride could not achieve the formation of a stable submicron - sized particle solution. In Comparative Example 4, zinc chloride was omitted; in Comparative Example 5, phytic acid was omitted. As a result, no submicron - sized particle solution was seen in the solution without the use of zinc chloride or phytic acid, which indicates that zinc chloride and phytic acid are essential components for the formation of the sodium hyaluronate - PDRN small cross - linked body. In Comparative Example 6, Solution 2 and Solution 1 were mixed by direct mixing instead of dropping. Due to the uneven mixing process, the formed sodium hyaluronate - PDRN cross - linked body particles were significantly larger. In Comparative Example 7, sodium hyaluronate with a larger weight - average molecular weight was cross - linked with PDRN, phytic acid, and zinc chloride. Since the molecular weight of sodium hyaluronate was larger, multiple sites for small cross - linked bodies might be generated on a single sodium hyaluronate molecule, thereby forming a network - like structure, which further led to the sedimentation of the cross - linked body.

[0106] 2. Sustained - release effect of Gel 1 - Gel 5 and Gel d1 - Gel d8

[0107] First, the nucleic acid content in Gel 1 - Gel 5 and Gel d1 - Gel d8 with unit concentration was measured by determining the absorbance at 260 nm. Then, 1 g of each of Gel 1 - Gel 5 and Gel d1 - Gel d8 was placed in a dialysis bag with a molecular weight cut - off of 5000 Da. The dialysis bag was placed in 100 mL of PBS aqueous solution with a pH of 7.4, and the stirring speed was 350 rpm. 1 mL of the buffer solution outside the dialysis bag was taken at different times to measure the absorbance at 260 nm to determine the nucleic acid concentration and total nucleic acid amount outside the dialysis bag. After each test, the solution was poured back into the buffer solution beaker. The nucleic acid release percentage was calculated.

[0108] Nucleic acid concentration (μg / mL)=A÷0.020.

[0109] Wherein, A is the light absorption peak value at 260 nm in a 1 - cm optical path length cuvette of the solution, and 0.020 is the light absorption concentration coefficient of DNA.

[0110] The nucleic acid release percentages of each group of gels at 24 h, 48 h, and 72 h measured by the above - mentioned method are shown in Table 5 (“ - ” indicates that it is close to complete release and not detected):

[0111] Table 5

[0112]

[0113] It can be seen that the gels 1 - 5 provided by the present invention can achieve a better nucleic acid sustained - release effect, and the nucleic acid release percentage within 24 h is only 46.90% - 58.10%, and the nucleic acid release percentage within 48 h is only 62.84% - 78.81%. However, the gels d1 - d8 provided in each comparative example do not have the sustained - release effect achieved by each example.

[0114] 3. Storage stability evaluation

[0115] Take 300 μL of sterile gel and inject it into a 1 mL glass transparent vial. Store it at 20 °C and 40 °C for 1 day, 15 days, 30 days and 60 days respectively, and visually observe its color, transparency and gel state. The results of the storage stability experiment at 20 °C are summarized in Table 6:

[0116] Table 6

[0117]

[0118] The results of the storage stability experiment at 40 °C are summarized in Table 7:

[0119] Table 7

[0120]

[0121] It can be seen that the sodium hyaluronate - PDRN gels provided in Examples 1 - 5 of the present invention have better storage stability at 20 °C and 40 °C.

[0122] 4. Biosafety evaluation

[0123] Cytotoxicity evaluation of human immortalized keratinocyte HaCaT cells.

[0124] Culture medium: DMEM medium + 10% fetal bovine serum (FBS) + 1% penicillin - streptomycin.

[0125] Culture conditions: 37 °C, 5% CO2, humidity 95%.

[0126] Experimental method:

[0127] The resuscitated HaCaT cells were subcultured until the logarithmic growth phase, digested, resuspended with the culture medium, and counted. After adjusting the cell concentration with the culture medium, 10,000 cells / well (containing 0.1 mL of the culture medium) were inoculated in a 96-well plate. After 24 h of colonization, the culture medium was aspirated, and 0.1 mL of the culture medium mixture containing gels with different concentrations was added to each well and cultured for 24 h. The culture medium was aspirated, and the MTT cell viability kit was used to detect the activity of HaCaT cells. The group with a gel concentration of 0 μg / mL was used as the benchmark for relative viability to calculate the relative viability of HaCaT cells at other gel concentrations. The experimental results are shown in Table 8 below (average relative viability, dimensionless quantity, with the average relative viability of cells when the gel concentration is 0 μg / mL as 1):

[0128] Table 8

[0129]

[0130] It can be seen that the gels provided in each example and comparative example did not show significant toxicity to HaCaT cells.

[0131] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than limiting the protection scope of the present invention. Any simple modification or equivalent replacement of the technical solution of the present invention by those of ordinary skill in the art shall not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A PDRN non-cross-linked sodium hyaluronate gel composition, characterized in that: The composition is as follows by weight: 5-15 parts of sodium hyaluronate 1, 1-3 parts of sodium hyaluronate 2, 1-2.7 parts of PDRN, 0.05-0.15 parts of phytic acid, 0.025-0.075 parts of zinc chloride and 200-300 parts of water; Wherein, the molecular weight of sodium hyaluronate 1 is 1000kDa-2000kDa; the molecular weight of sodium hyaluronate 2 is less than 10kDa; The method for preparing PDRN non-cross-linked sodium hyaluronate gel using the PDRN non-cross-linked sodium hyaluronate gel composition comprises the following steps: S1. Mix the formulated amount of sodium hyaluronate 2, PDRN, zinc chloride and water, and stir to obtain solution 1; S2, mixing the formulated amount of phytic acid with water to obtain solution 2; S3, adding solution 2 dropwise to the stirring solution 1 to obtain solution 3; S4. Mix the formulated amount of sodium hyaluronate 1 with the remaining amount of water, then add solution 3 and mix well to obtain PDRN non-cross-linked sodium hyaluronate gel.

2. The PDRN non-crosslinked sodium hyaluronate gel composition according to claim 1, characterized in that The composition is as follows by weight: 8-12 parts of sodium hyaluronate 1, 1.5-2.5 parts of sodium hyaluronate 2, 1.4-2.3 parts of PDRN, 0.08-0.12 parts of phytic acid, 0.04-0.06 parts of zinc chloride and 200-250 parts of water.

3. The PDRN non-crosslinked sodium hyaluronate gel composition according to claim 2, characterized in that The composition is as follows by weight: 10 parts of sodium hyaluronate 1, 2 parts of sodium hyaluronate 2, 1.85 parts of PDRN, 0.1 parts of phytic acid, 0.05 parts of zinc chloride and 236 parts of water.

4. The PDRN non-crosslinked sodium hyaluronate gel composition according to claim 1, characterized in that The molecular weight of the sodium hyaluronate 1 is 1500 kDa±1 kDa.

5. A method for preparing a PDRN non-crosslinked sodium hyaluronate gel using the PDRN non-crosslinked sodium hyaluronate gel composition according to any one of claims 1 to 4, characterized in that: The following steps are involved: S1. Mix the formulated amount of sodium hyaluronate 2, PDRN, zinc chloride and water, and stir to obtain solution 1; S2, mixing the formulated amount of phytic acid with water to obtain solution 2; S3, adding solution 2 dropwise to the stirring solution 1 to obtain solution 3; S4. Mix the formulated amount of sodium hyaluronate 1 with the remaining amount of water, then add solution 3 and mix well to obtain PDRN non-cross-linked sodium hyaluronate gel.

6. The method according to claim 5, characterized in that In step S1, the weight of the water is 16-24 times the sum of the weights of sodium hyaluronate 2, PDRN and zinc chloride; in step S2, the weight of the water is 20-40 times the weight of phytic acid.

7. The method according to claim 5, characterized in that In step S1, the stirring speed is 800-1200rpm, and the stirring time is 8-18h; in step S3, the dropping speed is 0.01-0.05mL / min; the stirring speed is 800-1200rpm; in step S4, the mixing is specifically stirred for 24-48h.

8. A PDRN non-cross-linked sodium hyaluronate gel, characterized in that: Made by the method described in any one of claims 5-7.

9. Use of the PDRN non-crosslinked sodium hyaluronate gel composition described in any one of claims 1-4 or the PDRN non-crosslinked sodium hyaluronate gel prepared by the method described in any one of claims 5-7 in the preparation of medicines, medical devices or cosmetics.

10. A medicine, characterized in that: A PDRN non-cross-linked sodium hyaluronate gel composition containing any one of claims 1 to 4 or a PDRN non-cross-linked sodium hyaluronate gel prepared by the method described in any one of claims 5 to 7.

11. A medical device, characterized in that: A PDRN non-cross-linked sodium hyaluronate gel composition containing any one of claims 1 to 4 or a PDRN non-cross-linked sodium hyaluronate gel prepared by the method described in any one of claims 5 to 7.

12. A cosmetic, characterized in that: A PDRN non-cross-linked sodium hyaluronate gel composition containing any one of claims 1 to 4 or a PDRN non-cross-linked sodium hyaluronate gel prepared by the method described in any one of claims 5 to 7.

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