Composition containing PDRN microspheres as well as preparation method and application of composition

Abstract

The invention discloses a composition containing PDRN microspheres as well as a preparation method and application thereof, and relates to the technical field of repair. The composition containing the PDRN microspheres is prepared from the following components in parts by weight: 1 to 10 parts of the PDRN microspheres, 30 to 60 parts of sodium hyaluronate, 0.05 to 1 part of a cross-linking agent, 0.5 to 5 parts of bisabolol and 1 to 8 parts of trehalose, the molecular weight of the sodium hyaluronate is 300 kDa to 800 kDa. By adding the PDRN microspheres and selecting reasonable component compatibility, the stability is remarkably improved, and a synergistic effect is reflected in the aspect of repairing.

Description

Technical Field

[0001] The present invention relates to the field of repair technology, and in particular to a composition comprising PDRN microspheres, and a preparation method and application thereof. Background Art

[0002] Polydeoxyribonucleotide (PDRN) is a low molecular weight DNA derivative of natural origin, with a molecular weight between 50kDa and 1500kDa. The most representative molecular weight of PDRN is 80kDa-200kDa, and the peak of Gaussian distribution is about 132kDa. The base length of PDRN is 50bp-2000bp. It is a linear polymer of deoxyribonucleotides with phosphodiester bonds. PDRN contains 50% double-stranded deoxyribonucleotides. Two polydeoxyribonucleotides are connected by hydrogen bonds between base pairs, eventually forming a double helix structure. PDRN is mainly extracted and purified from sperm cells of salmon or rainbow trout.

[0003] The base composition of PDRN is 98% similar to human DNA. Based on this biological effect, its clinical application scope has gradually expanded, showing significant effects in tissue repair, wound healing, anti-ischemia, anti-inflammation, etc. In vitro and in vivo experiments have shown that the most relevant mechanism of action of PDRN is as an agonist of adenosine A2A receptors, activating A2A receptors and producing a variety of physiological effects.

[0004] Chinese patent CN114642606A discloses a composition with skin barrier repair function, its preparation method and application, which includes the following ingredients: high molecular weight hyaluronic acid or its salt 0.01%-0.5%, hydrolyzed hyaluronic acid or its salt 0.1%-1.0%, silk protein 0.01%-10%, polydeoxyribonucleic acid 0.05%-1%, and bosine 0.1%-10%. The high molecular weight hyaluronic acid or its salt of the invention can target the stratum corneum of the skin and form a moist and breathable protective film on the surface of the skin. The extremely low molecular weight hydrolyzed hyaluronic acid or its salt can be absorbed through the skin into the dermis layer of the skin to stimulate angiogenesis. PDRN, silk protein and bosine can promote the synthesis of collagen and promote cell migration. These ingredients work together to have outstanding wound healing effects, can repair damaged skin barriers, and achieve the effect of quickly repairing skin wounds.

[0005] Chinese patent CN112932988A discloses an anti-aging composition containing polydeoxyribonucleic acid, including honey and polydeoxyribonucleic acid; the invention also discloses a skin care product, including the anti-aging composition containing polydeoxyribonucleic acid and an adjuvant acceptable in cosmetics. The anti-aging composition disclosed in the invention and the skin care product including the anti-aging composition enhance the effects of promoting skin cell regeneration and anti-aging through the synergistic effect of honey and polydeoxyribonucleic acid.

[0006] At present, PDRN has the problem of poor stability and still has certain limitations when used in repair products. Summary of the invention

[0007] In view of this, the purpose of the present invention is to provide a composition comprising PDRN microspheres, and a preparation method and application thereof. By adding PDRN microspheres and selecting a reasonable composition combination, the stability is significantly improved, and a synergistic effect is shown in the repair aspect.

[0008] To achieve the above-mentioned object of the invention, the technical solution of the present invention is as follows: On the one hand, the present invention provides a composition comprising PDRN microspheres, which is composed of the following ingredients in parts by weight: 1-10 parts of PDRN microspheres, 30-60 parts of sodium hyaluronate, 0.05-1 part of a cross-linking agent, 0.5-5 parts of bisabolol and 1-8 parts of trehalose.

[0009] Preferably, the composition containing PDRN microspheres consists of the following ingredients, in parts by weight: 3-8 parts of PDRN microspheres, 35-55 parts of sodium hyaluronate, 0.1-0.6 parts of a cross-linking agent, 1-4 parts of bisabolol, and 2-6 parts of trehalose.

[0010] Further preferably, the composition containing PDRN microspheres consists of the following ingredients, in parts by weight: 6 parts of PDRN microspheres, 50 parts of sodium hyaluronate, 0.5 parts of a cross-linking agent, 3 parts of bisabolol and 5 parts of trehalose.

[0011] Preferably, the molecular weight of the sodium hyaluronate is 300 kDa-800 kDa.

[0012] More preferably, the molecular weight of the sodium hyaluronate is 400 kDa-600 kDa.

[0013] If the molecular weight of sodium hyaluronate is too large, the strength of the cross-linking system will be too high, the feel of use will be poor, and it will be unfavorable for the release of active ingredients; if the molecular weight of sodium hyaluronate is too small, the stability will be reduced. Selecting the appropriate molecular weight range can ensure the comprehensive performance of the composition.

[0014] Preferably, the crosslinking agent is selected from at least one of divinyl sulfone, 1,4-butanediol diglycidyl ether, carbodiimide, and N-hydroxysuccinimide. More preferably, it is a combination of carbodiimide and N-hydroxysuccinimide. Furthermore, the molar ratio of carbodiimide to N-hydroxysuccinimide is 1-4:1.

[0015] Preferably, the particle size of the PDRN microspheres is less than 500 μm. In the present invention, there is no need to impose too many restrictions on the particle size of the PDRN microspheres, and the particle size is not a key influencing factor. The current forms of PDRN microspheres can achieve synergistic effects after the above compatibility, thereby improving the repair activity of the product. Considering the particle size distribution of the current microsphere products and reducing the impact on the preparation process, it is more preferred to select PDRN microspheres with a particle size of 10 μm-200 μm.

[0016] In another aspect, the present invention provides a method for preparing the above-mentioned composition comprising PDRN microspheres, comprising the following steps: (1) dissolving the formulated amount of trehalose and bisabolol in a solvent, and then adding the formulated amount of PDRN microspheres and sodium hyaluronate to obtain a mixed solution; (2) dissolving the cross-linking agent in a solvent, adding dropwise to the mixed solution obtained in step (1), treating in an ice bath, adjusting the pH, and heating the solution to obtain a reactant; (3) Concentrating the reactant obtained in step (2) to obtain a composition containing PDRN microspheres.

[0017] Preferably, in step (1), the solvent is a combination of phosphate buffered saline (PBS) and propylene glycol. More preferably, the pH of the phosphate buffered saline is 5.0-6.0, and more preferably 5.5. More preferably, the volume ratio of the phosphate buffered saline (PBS) to propylene glycol is 1-10:1, and most preferably 5:1.

[0018] Preferably, in step (1), the mass content of sodium hyaluronate in the mixed solution is 1%-5%, more preferably 2%.

[0019] Preferably, in step (2), the solvent is propylene glycol.

[0020] Preferably, in step (2), the ice bath treatment time is 5 min-30 min, more preferably 15 min.

[0021] Preferably, in step (2), the pH is adjusted to 5.0-6.5, more preferably 5.5-6.0.

[0022] Ice bath treatment for an appropriate time and at an appropriate pH can activate the carboxyl groups to the greatest extent, which is conducive to the cross-linking reaction.

[0023] Preferably, in step (2), the temperature-raising reaction condition is: heating to 20°C-35°C for 2h-5h, more preferably heating to 30°C for 2.5h.

[0024] Preferably, in step (3), the concentration is ultrafiltration concentration.

[0025] As a specific example of the present invention, the preparation method of the composition containing PDRN microspheres comprises the following steps: (1) dissolving the formulated amount of trehalose and bisabolol in a solvent, and then adding the formulated amount of PDRN microspheres and sodium hyaluronate to obtain a mixed solution, wherein the mass content of sodium hyaluronate in the mixed solution is 1%-5%; (2) dissolving the cross-linking agent in a small amount of solvent, adding dropwise to the mixed solution obtained in step (1), stirring in an ice bath for 5 min-30 min, adjusting the pH to maintain at 5.5-6.0, heating to 20°C-35°C for reaction for 2 h-5 h, adding ethanolamine to the obtained reactant, stirring to quench the reaction, and obtaining a gel-like reactant; (3) The gel-like reaction product obtained in step (2) is concentrated using a ceramic ultrafiltration membrane to obtain a composition containing PDRN microspheres.

[0026] Finally, the present invention provides the use of the above-mentioned composition containing PDRN microspheres in the preparation of repair products.

[0027] Preferably, the repair products include cosmetics and medicines.

[0028] Further preferably, the repair is to promote wound healing.

[0029] The beneficial effects of the present invention are: (1) The present invention selects PDRN microspheres to replace conventional PDRN components, which can significantly improve the repair activity of the product and improve the product stability; (2) In the present invention, PDRN microspheres are used in combination with sodium hyaluronate, bisabolol, and trehalose, and are prepared into a composite gel at a suitable ratio, which can exert a synergistic effect and achieve a very significant improvement in stability and repair activity.

[0030] (3) The present invention selects a suitable preparation method and sodium hyaluronate with a specific molecular weight, and encapsulates PDRN microspheres and bisabolol in the prepared network cross-linked structure, thereby achieving a long-term effect of PDRN and bisabolol on the epidermis, which can exert a repair function for a long period of time after use, significantly improving the actual repair effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1It is a graph showing the PDRN retention rate test results of the composition of the present invention. DETAILED DESCRIPTION

[0032] In order to make the technical means, creative features, purpose and efficacy of the present invention easy to understand, the present invention is further explained in conjunction with specific embodiments below, but the following embodiments are only preferred embodiments of the present invention, not all. Based on the embodiments in the implementation mode, other embodiments obtained by those skilled in the art without creative work all belong to the protection scope of the present invention. In the following embodiments, unless otherwise specified, the operating methods used are conventional operating methods, the equipment used are conventional equipment, and the equipment materials used in each embodiment are the same.

[0033] In the following examples, the sources of raw materials are as follows: Carbodiimide (dicyclohexylcarbodiimide is specifically used in the embodiment) and N-hydroxysuccinimide were purchased from Guangdong Yuanfeng; bisabolol was purchased from Xi'an Xirun, item number XRHZ-207, purity 99%; trehalose was Linyuan brand, particle size 100 mesh; Centella asiatica extract was purchased from Shaanxi Zelang, item number HJDT0020, purity >90%; sodium alginate was purchased from Jiangsu Nuojia, particle size 40 mesh-60 mesh.

[0034] PDRN microspheres are prepared in the following manner, which does not limit the present invention: (1) Material preparation: Prepare a 20 mg / mL PDRN aqueous solution; Prepare a poly-L-lactic acid organic phase solution (the organic phase is dichloromethane) with a mass volume concentration of 5%; Prepare an organic phase solution of zinc sulfate with a mass volume concentration of 5% (the organic phase is dichloromethane); A polyvinyl alcohol aqueous solution with a mass volume concentration of 60% was prepared.

[0035] “Mass volume concentration” means: the ratio of the mass of the solute in the solution (g) to the volume of the solution (mL).

[0036] (2) Microsphere preparation: The zinc sulfate organic phase solution and the poly-L-lactic acid organic phase solution were mixed in a volume ratio of 1:1 to obtain a mixed solution 1; under high-speed shearing conditions, the PDRN aqueous phase solution was injected, and the volume ratio of the PDRN aqueous phase solution to the mixed solution 1 was 1:1 to obtain an oil-in-water emulsion; under high-speed stirring conditions, the oil-in-water emulsion was slowly injected into the polyvinyl alcohol aqueous solution, and the volume ratio of the oil-in-water emulsion to the polyvinyl alcohol aqueous solution was 1:1 to obtain a water-in-oil-in-water emulsion; stirred at 1000 rpm until microspheres were formed, the solvent was removed, and washed to obtain PDRN microspheres. It was determined that the particle size distribution was 40μm-90μm.

[0037] Example 1 Formula: 6 parts of PDRN microspheres, 50 parts of sodium hyaluronate (400kDa-600kDa, purity >95%), 0.5 parts of cross-linking agent (carbodiimide and N-hydroxysuccinimide mixed in a molar ratio of 2:1), 3 parts of bisabolol and 5 parts of trehalose.

[0038] Preparation process: (1) Dissolve the formulated amount of trehalose in a solvent, wherein the solvent is PBS (pH 5.5) and propylene glycol in a volume ratio of 5:1 to obtain a trehalose solution, and then add the formulated amount of PDRN microspheres and sodium hyaluronate, mix thoroughly, and obtain a mixed solution; in the mixed solution, the mass content of sodium hyaluronate is 2%; (2) Dissolve the cross-linking agent in a small amount of propylene glycol, add dropwise to the mixed solution obtained in step (1), stir in an ice bath for 15 min, adjust the pH to maintain at 5.5, heat to 30°C and react for 2.5 h, add ethanolamine to the obtained reactant and stir at room temperature for 1 hour to quench the reaction, and obtain a gel-like reactant; (3) The gel-like reaction product obtained in step (2) is concentrated using a ceramic ultrafiltration membrane (100 kDa) to obtain a composition containing PDRN microspheres.

[0039] Example 2 Formula: 10 parts of PDRN microspheres, 60 parts of sodium hyaluronate (300kDa-600kDa, purity >95%), 1 part of cross-linking agent (carbodiimide), 0.5 parts of bisabolol and 1 part of trehalose.

[0040] The preparation process is the same as that in Example 1.

[0041] Example 3 Formula: 1 part of PDRN microspheres, 30 parts of sodium hyaluronate (500kDa-800kDa, purity >95%), 0.05 parts of cross-linking agent (N-hydroxysuccinimide), 5 parts of bisabolol and 8 parts of trehalose.

[0042] The preparation process is the same as that in Example 1.

[0043] Example 4 Formula: 3 parts of PDRN microspheres, 35 parts of sodium hyaluronate (400kDa-600kDa, purity >95%), 0.1 parts of cross-linking agent (carbodiimide and N-hydroxysuccinimide mixed in a molar ratio of 2:1), 4 parts of bisabolol and 2 parts of trehalose.

[0044] The preparation process is the same as that in Example 1.

[0045] Example 5 Formula: 8 parts of PDRN microspheres, 55 parts of sodium hyaluronate (400kDa-600kDa, purity >95%), 0.6 parts of cross-linking agent (carbodiimide and N-hydroxysuccinimide mixed in a molar ratio of 2:1), 1 part of bisabolol and 6 parts of trehalose.

[0046] The preparation process is the same as that in Example 1.

[0047] Example 6 The recipe is the same as Example 1.

[0048] Preparation process: (1) Dissolve the formulated amount of trehalose in a solvent, wherein the solvent is PBS (pH 5.5) and propylene glycol in a volume ratio of 5:1 to obtain a trehalose solution, and then add the formulated amount of PDRN microspheres and sodium hyaluronate, mix thoroughly, and obtain a mixed solution; in the mixed solution, the mass content of sodium hyaluronate is 5%; (2) dissolving the cross-linking agent in the formula amount with a small amount of propylene glycol, adding dropwise to the mixed solution obtained in step (1), stirring in an ice bath for 30 minutes, adjusting the pH to maintain at 6.0, heating to 35° C. for reaction for 2 hours, adding ethanolamine to the obtained reactant, stirring at room temperature for 1 hour, quenching the reaction, and obtaining a gel-like reactant; (3) The gel-like reaction product obtained in step (2) is concentrated using a ceramic ultrafiltration membrane (100 kDa) to obtain a composition containing PDRN microspheres.

[0049] Example 7 The recipe is the same as Example 1.

[0050] Preparation process: (1) Dissolve the formulated amount of trehalose in a solvent, wherein the solvent is PBS (pH 5.5) and propylene glycol in a volume ratio of 5:1 to obtain a trehalose solution, and then add the formulated amount of PDRN microspheres and sodium hyaluronate, mix thoroughly, and obtain a mixed solution; in the mixed solution, the mass content of sodium hyaluronate is 1%; (2) Dissolve the cross-linking agent in a small amount of propylene glycol, add dropwise to the mixed solution obtained in step (1), stir in an ice bath for 5 min, adjust the pH to maintain at 5.5, heat to 20° C. and react for 5 h, add ethanolamine to the obtained reactant and stir at room temperature for 1 hour to quench the reaction, and obtain a gel-like reactant; (3) The gel-like reaction product obtained in step (2) is concentrated using a ceramic ultrafiltration membrane (100 kDa) to obtain a composition containing PDRN microspheres.

[0051] Comparative Example 1 Different from Example 1, the formula selection is: 6 parts of PDRN microspheres, 50 parts of sodium hyaluronate (400kDa-600kDa, purity>95%), 0.5 parts of a cross-linking agent (carbodiimide and N-hydroxysuccinimide mixed at a molar ratio of 2:1), 3 parts of Centella asiatica extract and 5 parts of trehalose. The rest are the same.

[0052] In Comparative Example 1, the Centella asiatica extract was used without using bisabolol.

[0053] Comparative Example 2 Different from Example 1, the formula selection is: 15 parts of PDRN microspheres, 50 parts of sodium hyaluronate (400kDa-600kDa, purity>95%), 0.5 parts of a cross-linking agent (carbodiimide and N-hydroxysuccinimide mixed at a molar ratio of 2:1), 0.2 parts of bisabolol and 7 parts of trehalose. The rest are the same.

[0054] The composition ratio of Comparative Example 2 is different from that of Example 1.

[0055] Comparative Example 3 The difference from Example 1 is that the molecular weight of sodium hyaluronate is 1000 kDa-1200 kDa and the purity is >95%. The rest are the same.

[0056] Comparative Example 4 The difference from Example 1 is that the molecular weight of sodium hyaluronate is 100 kDa-200 kDa and the purity is greater than 95%. The rest are the same.

[0057] Comparative Example 5 The difference from Example 1 is that the formula selection is: 6 parts of PDRN microspheres, 55 parts of sodium hyaluronate (400kDa-600kDa, purity>95%), 0.5 parts of a cross-linking agent (carbodiimide and N-hydroxysuccinimide mixed at a molar ratio of 2:1) and 3 parts of bisabolol. The rest are the same.

[0058] In Comparative Example 5, trehalose was not used.

[0059] Comparative Example 6 Different from Example 1, the formula selection is: 6 parts of PDRN microspheres, 50 parts of sodium hyaluronate (400kDa-600kDa), 0.5 parts of a cross-linking agent (carbodiimide and N-hydroxysuccinimide mixed at a molar ratio of 2:1) and 5 parts of trehalose. The rest are the same.

[0060] In Comparative Example 6, no bisabolol was used.

[0061] Comparative Example 7 The difference from Example 1 is that in step (2), no ice bath treatment is performed. The rest is the same.

[0062] Comparative Example 8 The difference from Example 1 is that the same amount of PDRN is used as the raw material, and PDRN microspheres are not used. The rest are the same.

[0063] Comparative Example 9 Different from Example 1, the formula selection is: 6 parts of PDRN microspheres, 50 parts of sodium hyaluronate (400kDa-600kDa, purity>95%), 0.5 parts of a cross-linking agent (carbodiimide and N-hydroxysuccinimide mixed at a molar ratio of 2:1), 3 parts of bisabolol and 5 parts of sodium alginate. The rest are the same.

[0064] In Comparative Example 9, sodium alginate was used instead of trehalose.

[0065] Results 1. Stability test In the wound inflammation stage, as well as in the proliferation stage of epidermal cells and fibroblasts, DNA enzymes such as DNase I are released to remove free DNA, and their enzymatic ability also acts on PDRN, causing PDRN to decompose and thus affect the product effect. Therefore, the present invention simulates the enzymatic hydrolysis process of PDRN by DNase I to verify product stability.

[0066] Take 0.5g of the composition prepared by Example 1-Example 7 and Comparative Example 1-Comparative Example 9 respectively, add 50mL of water, then add DNase I (the amount added is 100u / mL according to the enzyme activity), seal, and place in a constant temperature water bath shaker for enzymolysis (enzymolysis temperature 37°C, enzymolysis time 2h, stirring speed 100rpm), after the enzymolysis is completed, add 20mL EDTA standard solution (concentration 0.01mol / L) and 1mL sodium dodecyl sulfate aqueous solution (mass fraction 10%), stir at 1500rpm for 20min, obtain a mixed solution, place the mixed solution in a centrifuge, and centrifuge at 10000rpm for 20min. After the centrifugation is completed, take the supernatant, test the PDRN content by the diphenylamine method, and record it as M1. Each group is set up with a blank treatment group without enzymolysis, and the PDRN content of the blank treatment group is tested by the diphenylamine method, which is recorded as M0.

[0067] The retention rate is calculated as follows: Retention rate = M1 / M0×100%.

[0068] The results are as follows Figure 1 As shown, Figure 1 The results of Examples 1 to 7 show that the hydrogel-encapsulated PDRN microspheres prepared by the present invention can isolate the contact with DNase I, delay the enzymatic hydrolysis of DNase I, and ensure its smooth release. In our relevant preliminary tests, the retention rate of the prepared pure PDRN microspheres can only reach about 58%. Figure 1 The experimental results show that the retention rates of Examples 1 to 7 and Comparative Example 3 are greater than 88%, which shows that the stability is significantly improved. At the same time, the results of Comparative Examples 1 to 9 show that although hydrogel encapsulation can improve the retention rate of PDRN microspheres, the molecular weight and preparation process of sodium hyaluronate will also affect the cross-linking process and thus affect the retention rate. At the same time, the addition of bisabolol and trehalose will also significantly improve the stability of PDRN microspheres. ‌

[0069] 2. Repair effect detection The embodiments and comparative examples with a retention rate of 70% were selected to further conduct repair effect detection experiments.

[0070] Male SD rats (weight 220g-230g) were selected as experimental animals. After one week of adaptive feeding, they were randomly divided into groups of 8 rats in each group. All rats were anesthetized with 10 mg / kg xylazine and 80 mg / kg ketamine, the hair on the back was removed, and a 1.5×1.5 cm 2 The full-thickness skin wound can inhibit the self-healing ability of rats. The control group used commercially available sodium hyaluronate gel (Tonon Pharmaceutical, the main component is sodium hyaluronate), and the experimental groups used the products of Example 1-Example 7, Comparative Example 1-Comparative Example 3, Comparative Example 6, and Comparative Example 9, which were fully applied to the wound, with a thickness of 1mm-2mm above the epidermis, and bandaged with medical transparent tape.

[0071] The wound surface areas at 0 days, 5 days, and 9 days were counted and the percentage of the original wound surface area was calculated. The results are shown in Table 1: Table 1

[0072] Note: In the table, Compared with the control group, p < 0.05, Represents p < 0.01 compared with the control group.

[0073] On the 9th day after wound treatment, the wound area was counted, the rats were anesthetized and killed, the wounds were excised, and the rats were divided into two groups. One group used CD68 to mark macrophages for inflammation diagnosis, and the other group used KI-67 fluorescent staining to observe the cell growth at the wound. The results are shown in Table 2: Table 2

[0074] Note: In the table, Compared with the control group, p < 0.05, Represents p < 0.01 compared with the control group.

[0075] The data in Table 1 show that self-healing can be achieved by using only commercially available sodium hyaluronate gel. When the products of Examples 1 to 7 of the present invention are used, a more obvious repair effect of promoting healing is shown, with the repair effect of Example 1 being the best. At the same time, the composition of the present invention has a long-lasting effect, so after one application, a continuous repair effect is shown within a 9-day experimental period.

[0076] When trauma occurs, the body triggers a self-defense mechanism, which leads to inflammation. Early macrophages can remove pollutants and inhibit inflammatory necrotic tissue, but excessive inflammatory responses can delay wound healing and cause scars. Therefore, it is critical to control inflammation during the repair phase. By observing the proportion of KI-67 positive cells, the cell proliferation during wound healing can be known to a certain extent. The results in Table 2 show that the product prepared by the embodiment of the present invention can significantly slow down the early inflammatory response and promote cell proliferation, thereby achieving the purpose of promoting repair.

[0077] Although the results of the comparative examples also show significant differences compared with the control group, there is still a significant gap compared with the examples. The results of comparative examples 1, 6 and 9 show that bisabolol and trehalose are important ingredients, which can synergistically improve the repair effect with PDRN microspheres, and all three are indispensable; the results of comparative example 2 show that the selection of raw materials in a suitable proportion is more conducive to wound repair; comparative example 3 uses sodium hyaluronate with a higher molecular weight, which shows a high stability in the stability experiment, but is not conducive to the release of PDRN to a certain extent, resulting in a decrease in the repair effect.

[0078] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A composition comprising PDRN microspheres, characterized in that The invention is composed of the following ingredients in parts by weight: 1-10 parts of PDRN microspheres, 30-60 parts of sodium hyaluronate, 0.05-1 part of a cross-linking agent, 0.5-5 parts of bisabolol and 1-8 parts of trehalose; the molecular weight of the sodium hyaluronate is 300kDa-800kDa.

2. The composition comprising PDRN microspheres according to claim 1, characterized in that The composition is composed of the following ingredients in parts by weight: 3-8 parts of PDRN microspheres, 35-55 parts of sodium hyaluronate, 0.1-0.6 parts of cross-linking agent, 1-4 parts of bisabolol and 2-6 parts of trehalose.

3. The composition comprising PDRN microspheres according to claim 2, characterized in that The composition is composed of the following ingredients in parts by weight: 6 parts of PDRN microspheres, 50 parts of sodium hyaluronate, 0.5 parts of a cross-linking agent, 3 parts of bisabolol and 5 parts of trehalose.

4. The composition comprising PDRN microspheres according to claim 1, characterized in that The molecular weight of the sodium hyaluronate is 400kDa-600kDa.

5. The composition comprising PDRN microspheres according to claim 1, characterized in that The cross-linking agent is selected from at least one of divinyl sulfone, 1,4-butanediol diglycidyl ether, carbodiimide, and N-hydroxysuccinimide.

6. The composition comprising PDRN microspheres according to claim 5, characterized in that The cross-linking agent is a combination of carbodiimide and N-hydroxysuccinimide.

7. The composition comprising PDRN microspheres according to claim 6, characterized in that The molar ratio of the carbodiimide to N-hydroxysuccinimide is 1-4:

1.

8. The method for preparing a composition comprising PDRN microspheres according to any one of claims 1 to 7, characterized in that The following steps are involved: (1) dissolving the formulated amount of trehalose and bisabolol in a solvent, and then adding the formulated amount of PDRN microspheres and sodium hyaluronate to obtain a mixed solution; (2) dissolving the cross-linking agent in a solvent, adding dropwise to the mixed solution obtained in step (1), treating in an ice bath, adjusting the pH, and heating the solution to obtain a reactant; (3) Concentrating the reactant obtained in step (2) to obtain a composition containing PDRN microspheres.

9. The preparation method according to claim 8, characterized in that: In step (1), the mass content of sodium hyaluronate in the mixed solution is 1%-5%.

10. The preparation method according to claim 9, characterized in that: In step (1), the mass content of sodium hyaluronate in the mixed solution is 2%.

11. The preparation method according to claim 8, characterized in that: In step (2), the ice bath treatment time is 5 min-30 min.

12. The preparation method according to claim 11, characterized in that: In step (2), the ice bath treatment time is 15 minutes.

13. The preparation method according to claim 8, characterized in that: In step (2), the pH is adjusted to 5.0-6.

5.

14. The preparation method according to claim 13, characterized in that: In step (2), the pH is adjusted to 5.5-6.

0.

15. The preparation method according to claim 8, characterized in that: In step (2), the temperature-raising reaction conditions are: raising the temperature to 20°C-35°C and reacting for 2h-5h.

16. The preparation method according to claim 8, characterized in that: The following steps are involved: (1) dissolving the formulated amount of trehalose and bisabolol in a solvent, and then adding the formulated amount of PDRN microspheres and sodium hyaluronate to obtain a mixed solution; in the mixed solution, the mass content of sodium hyaluronate is 1%-5%; (2) dissolving the cross-linking agent in a small amount of solvent, adding dropwise to the mixed solution obtained in step (1), stirring in an ice bath for 5 min-30 min, adjusting the pH to maintain at 5.5-6.0, heating to 20°C-35°C for reaction for 2 h-5 h, adding ethanolamine to the obtained reactant, stirring to quench the reaction, and obtaining a gel-like reactant; (3) The gel-like reaction product obtained in step (2) is concentrated using a ceramic ultrafiltration membrane to obtain a composition containing PDRN microspheres.

17. Use of a composition comprising PDRN microspheres as described in any one of claims 1 to 7 in the preparation of a repair product.

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