Composite dissoluble microneedle gel patch mask and preparation method thereof

Abstract

The invention relates to a composite dissoluble microneedle gel patch mask and a preparation method thereof. Comprising the following steps: (1) preparing a lipid oil phase solution and a water phase solution; mixing the lipid oil phase solution with the water phase solution to obtain a liposome suspension; (2) preparing recombinant collagen hydrogel; (3) adding the tea polyphenol and the lipidosome suspension into the recombinant collagen hydrogel to obtain composite gel; and (4) injecting the composite gel into a mold, carrying out vacuum drying, and demolding to obtain the composite dissoluble microneedle gel patch mask. According to the composite dissoluble microneedle gel patch mask provided by the invention, the tea polyphenol, the plant exosome and the liposome carrier are integrated into the microneedle gel patch mask for the first time, the oxidation resistance is improved by utilizing the tea polyphenol, the inflammation resistance is improved by utilizing the plant exosome, the transdermal effect is enhanced by utilizing the liposome, the transdermal efficiency reaches 35.2-42.5 mu g / cm < 2 > / h, the medical beauty effect is obvious, and the application prospect is wide. The obvious advantages are realized in the aspects of safety, transdermal efficiency and repairing effect.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetics, and in particular relates to a composite soluble microneedle gel patch mask and a preparation method thereof. Background Art

[0002] Human skin is a typical "brick wall structure" with layers, forming a series of hard barriers to protect the skin from harmful external substances. However, it is also based on this feature that makes it difficult for medications or beauty products applied to the skin surface to enter the skin, enter the dermis or cells to take effect. Conventional medical masks or skin care products contain many proteins, lipids, and polysaccharide macromolecules that are extremely friendly to the skin. These substances may reduce fine lines on the skin, promote pigment metabolism, reduce the skin's antioxidant effect, etc. However, due to the special structure of the skin, these substances are difficult to be absorbed by the skin. There are many ways to promote the absorption of skin substances, such as water light injections and microneedle operations, but these operations are cumbersome and require personnel with corresponding expertise to operate. The addition of corresponding substances such as skin penetration promoters will cause safety hazards to a certain extent.

[0003] Collagen is the most abundant protein in mammals, comprising 25%-30% of the total protein content. This protein plays a key role in maintaining tissue shape and form through molecular and cellular interactions within the extracellular matrix. However, natural collagen has low bioactivity, carries the risk of contamination from poultry and animal diseases, and differs from human collagen fragments, which can easily lead to adverse reactions such as immune rejection. Recombinant collagen, with its low molecular weight, excellent water solubility, and high absorption rate, has become an ideal biofiller for promoting cell production and metabolism in the medical aesthetics field.

[0004] While collagen hydrogels and microneedle patches can improve skin problems, they pose challenges such as crosslinker residue, poor stability, and low transdermal efficiency. For one thing, the risk of crosslinker residue can lead to facial redness, swelling, allergies, stiffness, and unevenness, resulting in a poor user experience. Furthermore, poor product stability can lead to degradation during long-term storage, and the recombinant collagen in the mask cannot maintain a stable three-dimensional structure, further contributing to low transdermal efficiency and poor efficacy.

[0005] Therefore, how to prepare a microneedle skin care product that can not only improve the permeability of effective substances and promote skin self-repair, but also has no cross-linking agent residue and is harmless to the skin is a problem that needs to be overcome urgently and has great practical significance. Summary of the Invention

[0006] To address the shortcomings of existing technologies, the present invention provides a composite dissolvable microneedle gel patch mask and its preparation method. Compared to traditional recombinant collagen obtained through chemical crosslinking, this mask has no chemical residue, high stability, and strong resistance to degradation. It successfully transcends the limitations of traditional technologies, reduces the risks of cosmetic procedures for consumers, and improves their comfort after use.

[0007] The technical solutions of the present invention are as follows:

[0008] A method for preparing a composite soluble microneedle gel patch mask comprises the following steps:

[0009] (1) dissolving lecithin and cholesterol in ethanol to obtain a lipid oil phase solution; dissolving plant exosomes and sodium deoxycholate in PBS buffer to obtain an aqueous phase solution; and then mixing the lipid oil phase solution with the aqueous phase solution through a microfluidic device to obtain a liposome suspension;

[0010] (2) dissolving the recombinant collagen and the high molecular weight polymer in an alkaline solution, stirring and mixing uniformly, then adding tetrakis(hydroxymethyl)phosphonium chloride, and magnetically stirring the solution at 4-37°C for a cross-linking reaction for 0.5-4 hours to obtain a recombinant collagen hydrogel;

[0011] (3) adding the tea polyphenols and the liposome suspension obtained in step (1) to the recombinant collagen hydrogel obtained in step (2), and homogenizing to obtain a composite gel;

[0012] (4) The composite gel is injected into a mold, vacuum dried, and then demoulded to obtain a composite soluble microneedle gel patch mask.

[0013] Preferably, according to the present invention, in step (1), the mass volume ratio of lecithin, cholesterol and ethanol is (40-80): (15-25): (3-8), unit: mg / mg / mL.

[0014] According to the preferred embodiment of the present invention, in step (1), the plant exosomes are Prinus utilis exosomes or Centella asiatica exosomes.

[0015] Preferably, according to the present invention, in step (1), the mass volume ratio of the plant exosomes, sodium deoxycholate and PBS buffer is (0.5-1.5): (0.1-0.3): (8-12), unit: mg / mg / mL.

[0016] Preferably, according to the present invention, in step (1), the flow rate ratio of the lipid oil phase solution to the aqueous phase solution in the microfluidic device is (10-20):1.

[0017] Preferably, according to the present invention, in step (2), the high molecular weight polymer is hyaluronic acid, hydroxypropyl methylcellulose (HPMC) or polyethylene glycol (PEG).

[0018] Preferably, according to the present invention, in step (2), the alkaline solution is a 0.0.5-0.15M NaOH solution.

[0019] Preferably, according to the present invention, in step (2), the mass volume ratio of the recombinant collagen, high molecular polymer and alkaline solution is (100-200): (50-100): (500-1000), unit: mg / mg / mL.

[0020] According to the preferred embodiment of the present invention, in step (2), the amount of tetrakis(hydroxymethyl)phosphonium chloride added is 0.02-1% of the mass of the recombinant collagen.

[0021] Preferably, according to the present invention, in step (3), the mass volume ratio of the tea polyphenols, liposome suspension and recombinant collagen hydrogel is (8-12): (3-8): (40-60), unit: mg / mL / mg.

[0022] According to a preferred embodiment of the present invention, in step (3), the homogenization parameters are: homogenization at 8000-12000 rpm for 3-8 minutes.

[0023] The present invention also provides a composite soluble microneedle gel patch mask prepared by the above method.

[0024] The technical features and beneficial effects of the present invention are as follows:

[0025] 1. The composite soluble microneedle gel patch mask provided by the present invention integrates tea polyphenols, plant exosomes and liposome carriers into the microneedle gel patch mask for the first time, using tea polyphenols to improve antioxidant properties, using plant exosomes to improve anti-inflammatory properties, and using liposomes to enhance transdermal effects, with a transdermal efficiency of 35.2-42.5 μg / cm 2 / h. It forms a multi-dimensional synergistic effect with the repair function of recombinant collagen, resulting in significant aesthetic results. It also offers significant advantages in safety, transdermal efficiency, and repair effectiveness, and can be used in the development of products for wrinkle improvement, photoaging, and post-operative repair.

[0026] 2. The present invention first encapsulates plant exosomes using lecithin, cholesterol, and sodium deoxycholate under specific parameters, achieving a high liposome encapsulation efficiency (≥85%) and activity retention rate (≥93%), increasing the exosome activity retention rate to 93%-97%, breaking through the bottleneck of insufficient stability of existing liposome technology. THPC is then used as a crosslinker for recombinant collagen, significantly improving the mechanical properties of the hydrogel (elastic modulus ≥400Pa), while avoiding the toxic residues of traditional crosslinkers, eliminating the risk of chemical reagent residues, and greatly reducing the occurrence of allergic reactions such as redness, swelling, and itching.

[0027] 3. The preparation method provided by the present invention is simple and easy to implement, and the overall parameters and process are mature, which is suitable for large-scale industrial production. DETAILED DESCRIPTION

[0028] The present invention is described in detail below with reference to specific examples. The examples described below are only preferred embodiments of the present invention. It should be noted that the following description is only for the purpose of explaining the present invention and is not intended to limit the present invention in any form. Any simple modifications, equivalent changes and modifications made to the embodiments based on the technical essence of the present invention are within the scope of the technical solution of the present invention.

[0029] In the following examples, the experimental methods used are conventional methods unless otherwise specified. The reagents used in the examples are all commercially available products.

[0030] The microfluidic device structure used in the present invention has been disclosed in Chinese patent document CN117695227A-A method for preparing PDRN liposomes based on a microfluidic process. The inner diameter of the lipid phase channel is 0.5-1 mm, and the inner diameter of the aqueous phase channel is 1.5-3 mm.

[0031] The recombinant collagen described in the examples was produced by fermentation using the genetically engineered bacteria SMD1163 / pPIC9K-AF2-COIII as disclosed in Chinese patent document CN117025655A.

[0032] Example 1

[0033] A method for preparing a composite soluble microneedle gel patch mask comprises the following steps:

[0034] (1) Accurately weigh 60 mg of lecithin and 20 mg of cholesterol and dissolve them in 5 mL of ethanol. Ultrasonic dissolution was performed in a water bath at 50°C to obtain a lipid oil phase solution. 10 mg of Prinus utilis exosomes and 2 mg of sodium deoxycholate were dissolved in 10 mL of PBS buffer to obtain an aqueous phase solution. The lipid oil phase solution and the aqueous phase solution were then added to a microfluidic device, and the flow rate ratio of the two phase solutions was adjusted to 15:1 (lipid oil phase solution 15 mL / min, aqueous phase solution 1 mL / min). After mixing, the mixture was centrifuged at 10,000 rpm for 15 min to remove unencapsulated exosomes and obtain a liposome suspension.

[0035] (2) 200 mg of recombinant collagen and 200 mg of hydroxypropyl methylcellulose (HPMC) were dissolved in 10 mL of NaOH solution (0.1 M), stirred and mixed evenly, and then 1 mg of tetrakis(hydroxymethyl)phosphonium chloride (THPC) was added. The mixture was magnetically stirred at 4°C for 2 h to allow cross-linking reaction to obtain recombinant collagen hydrogel.

[0036] (3) adding 10 mg of tea polyphenols and 5 mL of the liposome suspension obtained in step (1) to 100 mg of the recombinant collagen hydrogel obtained in step (2), and homogenizing at 10,000 rpm for 5 min to obtain a composite gel;

[0037] (4) The composite gel was injected into a PDMS microneedle mold (needle length 300 μm, needle tip diameter 45 μm), vacuum dried at -0.1 MPa and 25°C for 12 h, and then demolded to obtain a composite soluble microneedle gel patch mask.

[0038] Example 2

[0039] A method for preparing a composite soluble microneedle gel patch mask comprises the following steps:

[0040] (1) Accurately weigh 80 mg of lecithin and 20 mg of cholesterol and dissolve them in 5 mL of ethanol. Ultrasonic dissolution was performed in a water bath at 50°C to obtain a lipid oil phase solution. 12 mg of Centella asiatica exosomes and 3 mg of sodium deoxycholate were dissolved in 10 mL of PBS buffer to obtain an aqueous phase solution. The lipid oil phase solution and the aqueous phase solution were then added to a microfluidic device, and the flow rate ratio of the two phase solutions was adjusted to 20:1 (lipid oil phase solution 20 mL / min, aqueous phase solution 1 mL / min). After mixing, the mixture was centrifuged at 10,000 rpm for 15 min to remove unencapsulated exosomes and obtain a liposome suspension.

[0041] (2) 100 mg of recombinant collagen and 100 mg of hydroxypropyl methylcellulose (HPMC) were dissolved in 5 mL of NaOH solution (0.1 M), stirred and mixed evenly, and then 0.1 mg of tetrakis(hydroxymethyl)phosphonium chloride (THPC) was added. The mixture was magnetically stirred at 25°C for 3 h to cross-link and obtain a recombinant collagen hydrogel.

[0042] (3) adding 10 mg of tea polyphenols and 5 mL of the liposome suspension obtained in step (1) to 60 mg of the recombinant collagen hydrogel obtained in step (2), and homogenizing at 10,000 rpm for 5 min to obtain a composite gel;

[0043] (4) The composite gel was injected into a PDMS microneedle mold (needle length 300 μm, needle tip diameter 45 μm), vacuum dried at -0.1 MPa and 25°C for 12 h, and then demolded to obtain a composite soluble microneedle gel patch mask.

[0044] Example 3

[0045] A method for preparing a composite soluble microneedle gel patch mask comprises the following steps:

[0046] (1) Accurately weigh 40 mg of lecithin and 20 mg of cholesterol and dissolve them in 5 mL of ethanol. Ultrasonic dissolution was performed in a water bath at 50°C to obtain a lipid oil phase solution. 8 mg of Centella asiatica exosomes and 3 mg of sodium deoxycholate were dissolved in 10 mL of PBS buffer to obtain an aqueous phase solution. The lipid oil phase solution and the aqueous phase solution were then added to a microfluidic device, and the flow rate ratio of the two phase solutions was adjusted to 10:1 (lipid oil phase solution 10 mL / min, aqueous phase solution 1 mL / min). After mixing, the mixture was centrifuged at 10,000 rpm for 15 min to remove unencapsulated exosomes and obtain a liposome suspension.

[0047] (2) 150 mg of recombinant collagen and 50 mg of polyethylene glycol (PEG) were dissolved in 30 mL of NaOH solution (0.1 M), stirred and mixed evenly, and then 1.2 mg of tetrakis(hydroxymethyl)phosphonium chloride (THPC) was added. The cross-linking reaction was carried out under magnetic stirring at 37°C for 1 h to obtain a recombinant collagen hydrogel;

[0048] (3) adding 10 mg of tea polyphenols and 5 mL of the liposome suspension obtained in step (1) to 100 mg of the recombinant collagen hydrogel obtained in step (2), and homogenizing at 10,000 rpm for 5 min to obtain a composite gel;

[0049] (4) The composite gel was injected into a PDMS microneedle mold (needle length 300 μm, needle tip diameter 45 μm), vacuum dried at -0.1 MPa and 25°C for 12 h, and then demolded to obtain a composite soluble microneedle gel patch mask.

[0050] Comparative Example 1

[0051] A method for preparing a composite soluble microneedle gel patch mask, the specific steps of which are as described in Example 1, except that, in step (2), the mass of the recombinant collagen is 50 mg, and the mass of hydroxypropyl methylcellulose (HPMC) is 200 mg.

[0052] Comparative Example 2

[0053] A method for preparing a composite soluble microneedle gel patch mask, the specific steps of which are as described in Example 1, except that in step (2), glutaraldehyde is used instead of tetrakis(hydroxymethyl)phosphonium chloride (THPC) as a cross-linking agent, and the added amount is 0.1% of the weight of the recombinant collagen.

[0054] Comparative Example 3

[0055] A method for preparing a composite soluble microneedle gel patch mask, the specific steps of which are as described in Example 1, except that, in step (1), an ultrasonic method is used instead of a microfluidic device method to prepare a liposome suspension, and the ultrasonic parameters are: 200W, 10min.

[0056] Test example

[0057] 1. Determine the encapsulation efficiency (%) of the liposome suspensions prepared in Examples 1 to 3 and Comparative Examples 1 to 3.

[0058] The determination method is as follows: the liposome suspension obtained in step (1) is filtered through a 0.22 μm filter membrane to collect the unencapsulated exosomes, and the unencapsulated exosomes are quantitatively analyzed by HPLC to calculate the encapsulation efficiency (%) = (total exosome amount - unencapsulated amount) / total exosome amount × 100%.

[0059] 2. Determine the elastic modulus G' (Pa) of the recombinant collagen hydrogels prepared in Examples 1 to 3 and Comparative Examples 1 to 3.

[0060] The determination method is: using a rotational rheometer at a frequency of 1 Hz, a strain of 1%, and a temperature of 25° C. to measure the elastic modulus G′ (Pa) of the recombinant collagen hydrogel obtained in step (2).

[0061] 3. Determine the microneedle breaking strength (N / mm) of the composite soluble microneedle gel patch mask prepared in Examples 1 to 3 and Comparative Examples 1 to 3. 2 ), dissolution time (min), transdermal efficiency (μg / cm 2 / h) and exosome activity retention rate (%).

[0062] The determination method is:

[0063] ① Microneedle fracture strength: A universal material testing machine (Instron 5943) was used to apply vertical pressure to the composite soluble microneedle gel patch mask at a rate of 0.1 mm / s, and the maximum force at the moment of fracture (N) and the cross-sectional area of ​​the needle tip (mm) were recorded. 2 ) ratio.

[0064] ② Dissolution time: Apply the composite soluble microneedle gel patch mask to the isolated pig skin (thickness 1mm) and observe the complete dissolution time of the microneedles at 37°C.

[0065] ③. Transdermal efficiency: Franz diffusion cell method was used, with recombinant collagen as the marker and PBS (pH 7.4) as the receiving solution. The cumulative permeation of the composite soluble microneedle gel patch mask was measured after 24 hours (μg / cm 2 / h)

[0066] ④. Exosome activity retention rate: The CCK-8 method was used to detect the proliferation promotion rate of exosomes in the composite soluble microneedle gel patch mask on human skin fibroblasts (HSF), and the retention rate (%) was calculated by comparing it with the activity of fresh exosomes.

[0067] The above encapsulation rate (%), elastic modulus G' (Pa), microneedle breaking strength (N / mm 2 ), dissolution time (min), transdermal efficiency (μg / cm 2 The results of the activity retention rate (%) and exosome activity retention rate (%) are shown in Table 1.

[0068] Table 1

[0069] Test indicators Example 1 Example 2 Example 3 Comparative Example 1 Comparative Example 2 Comparative Example 3 Encapsulation efficiency (%) 89 92 86 88 90 65 Elastic modulus G'(Pa) 480 580 510 220 150 420 <![CDATA[Microneedle breaking strength (N / mm 2 )]]> 12.8 14.2 13.1 5.8 4.3 12.5 Dissolution time (min) 25 20 30 45 60 28 <![CDATA[Transdermal efficiency (μg / cm 2 / h)]]> 38.7 42.5 35.2 12.3 9.8 18.6 Exosome activity retention rate (%) 95 97 93 91 90 72

[0070] As shown in Table 1, the elastic modulus G', microneedle breaking strength, transdermal efficiency and exosome activity retention rate of Examples 1 to 3 of the present invention are significantly higher than those of Comparative Examples 1 to 3, and the dissolution time is shorter than that of Comparative Examples 1 to 3. In particular, the performance of the composite soluble microneedle gel patch mask prepared in Example 2 is the best, with an elastic modulus G' of 580 Pa and a microneedle breaking strength of 14.2 N / mm 2 , the transdermal efficiency reached 42.5μg / cm 2 / h and the exosome activity retention rate reached 97%, and the dissolution time was only 20min.

[0071] This is because: in Comparative Example 1, the ratio of recombinant collagen to HPMC is unbalanced (50:200), resulting in insufficient cross-linking degree of the hydrogel, elastic modulus (220Pa) and transdermal efficiency (12.3μg / cm 2 / h) dropped significantly. In Comparative Example 2, the use of glutaraldehyde crosslinking agent residue resulted in the hydrogel elastic modulus (150Pa) and transdermal efficiency (9.8μg / cm2 / h) deteriorated, and the exosome activity (90%) decreased due to toxicity. In Comparative Example 3, due to the destruction of the liposome structure by ultrasound, the encapsulation efficiency (65%) and exosome activity (72%) were significantly lower than those in the embodiment (≥85%, ≥93%), confirming the key role of microfluidic technology in stability. Finally, it should be noted that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention can be modified and varied in various ways. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing a composite soluble microneedle gel patch mask, characterized in that: The steps are as follows: (1) dissolving lecithin and cholesterol in ethanol to obtain a lipid oil phase solution; dissolving plant exosomes and sodium deoxycholate in PBS buffer to obtain an aqueous phase solution; and then mixing the lipid oil phase solution with the aqueous phase solution through a microfluidic device to obtain a liposome suspension; (2) dissolving the recombinant collagen and the high molecular weight polymer in an alkaline solution, stirring and mixing uniformly, then adding tetrakis(hydroxymethyl)phosphonium chloride, and magnetically stirring the solution at 4-37°C for a cross-linking reaction for 0.5-4 hours to obtain a recombinant collagen hydrogel; (3) adding the tea polyphenols and the liposome suspension obtained in step (1) to the recombinant collagen hydrogel obtained in step (2), and homogenizing to obtain a composite gel; (4) The composite gel is injected into a mold, vacuum dried, and then demoulded to obtain a composite soluble microneedle gel patch mask.

2. The preparation method according to claim 1, wherein In step (1), the mass volume ratio of lecithin, cholesterol and ethanol is (40-80): (15-25): (3-8), unit: mg / mg / mL.

3. The preparation method according to claim 1, wherein In step (1), the mass volume ratio of the plant exosomes, sodium deoxycholate and PBS buffer is (0.5-1.5): (0.1-0.3): (8-12), unit: mg / mg / mL; the plant exosomes are prickle fruit exosomes or Centella asiatica exosomes.

4. The preparation method according to claim 1, wherein In step (1), the flow rate ratio of the lipid oil phase solution to the aqueous phase solution in the microfluidic device is (10-20):

1.

5. The preparation method according to claim 1, wherein In step (2), the high molecular weight polymer is hyaluronic acid, hydroxypropyl methylcellulose (HPMC) or polyethylene glycol (PEG); and the alkaline solution is a 0.0.5-0.15M NaOH solution.

6. The preparation method according to claim 1, wherein In step (2), the mass volume ratio of the recombinant collagen, high molecular polymer and alkaline solution is (100-200): (50-100): (500-1000), unit: mg / mg / mL.

7. The preparation method according to claim 1, wherein In step (2), the amount of tetrakis(hydroxymethyl)phosphonium chloride added is 0.02-1% of the mass of the recombinant collagen.

8. The preparation method according to claim 1, wherein In step (3), the mass volume ratio of the tea polyphenols, liposome suspension and recombinant collagen hydrogel is (8-12): (3-8): (40-60), unit: mg / mL / mg.

9. The preparation method according to claim 1, wherein In step (3), the homogenization parameters are: homogenization at 8000-12000 rpm for 3-8 minutes.

10. A composite soluble microneedle gel patch mask, characterized in that: It is prepared according to the method according to any one of claims 1 to 9.