Preparation method and application of plant cell loaded selenium disulfide compound

Abstract

The invention discloses a preparation method of a plant cell loaded selenium disulfide compound, and belongs to the technical field of nano materials and daily chemicals. The preparation method comprises the following steps: S1, preparing a plant cell suspension; s2, wall breaking treatment of plant cells; s3, preparing a selenium disulfide nano dispersion liquid; and S4, loading of selenium disulfide. The method is simple in overall preparation and easy to popularize and apply, the defects of selenium disulfide can be well overcome, meanwhile, selenium disulfide is organically combined with plants, plant cells are used as carriers of the active components, unstable active components (such as selenium disulfide) are loaded into the plant cells, and the selenium disulfide is prepared. The invention has the advantages of improving stability, reducing irritation, realizing function synergy and improving the sense of the product. The plant cell loaded selenium disulfide compound prepared by the method disclosed by the invention can be effectively applied to various personal care products, and has extremely strong market competitiveness.

Description

Technical Field

[0001] This invention belongs to the field of nanomaterials and daily chemical technology, specifically relating to a method for preparing a plant cell-loaded selenium disulfide complex and its application. Background Technology

[0002] Dandruff is the result of excessive proliferation and shedding of scalp keratinocytes. Its production is closely related to the scalp microbiota (especially the excessive proliferation of Malassezia), excessive scalp sebum secretion, and individual inflammatory responses. Severe dandruff is often accompanied by scalp itching, redness, and stinging, which not only affects personal appearance but also serves as an important signal of scalp health imbalance. Currently, commonly used chemical antidandruff agents on the market include zinc pyrithione, piroctone olamine salt, ketoconazole, clomiphene citrate, and selenium sulfide.

[0003] Selenium disulfide (SeS2) is a recognized and highly effective anti-dandruff and anti-seborrheic dermatitis ingredient. It works through multiple pathways, including inhibiting Malassezia growth, slowing epidermal cell renewal, and reducing sebum secretion, resulting in significant dandruff-reducing effects. However, selenium disulfide has the following serious drawbacks in application: 1. Its physicochemical properties are unstable, easily agglomerating and settling in formulations, leading to uneven distribution and reduced activity. 2. It has strong cytotoxicity; high concentrations of selenium disulfide are significantly toxic to scalp keratinocytes and hair follicle cells, potentially causing scalp dryness, stinging, and even hair loss, limiting its concentration and frequency of use. 3. It has a poor sensory experience; selenium disulfide has an unpleasant characteristic odor (similar to rotten eggs), and may leave an orange-red residue when it leaches into products, severely impacting the user experience. 4. It has poor biocompatibility; its strong hydrophobic properties make it difficult to coexist harmoniously with other natural active ingredients in the formulation, restricting its application in high-end, gentle shampoos and conditioners.

[0004] In response to the problems associated with selenium disulfide, several patents have proposed solutions involving the encapsulation of selenium disulfide: Patent CN118436555A discloses a method for obtaining selenium disulfide encapsulated in chitosan quaternary ammonium salt. By leveraging the adsorption capacity of quaternary ammonium salt chitosan on hair, the deposition of selenium disulfide on hair is enhanced, thereby improving the dandruff-removing effect of selenium disulfide. However, commercially available quaternary ammonium salt chitosan is generally a non-long carbon chain quaternary ammonium salt chitosan product that has undergone quaternization with trimethylammonium chloride. This technology is difficult to mass-produce in actual production. Patent CN 120267541A discloses a hair composition containing selenium disulfide inclusions, which uses cyclodextrin as a carrier and a grinding method to achieve the encapsulation of selenium disulfide. Although the encapsulation with cyclodextrin is effective, it has limited improvement in reducing irritation. Patent CN118873421A discloses a composite of selenium disulfide, wax, and adsorbent, in which selenium disulfide is dispersed in the pores of the adsorbent and uniformly coated on the surface with wax. Although the protection of wax helps to improve the stability of selenium disulfide and extend its effective period, the strong system closure caused by wax may affect the release of the efficacy of selenium disulfide. In addition, wax may bring an oily and heavy skin feel, affecting the shampooing experience. Patent CN114788788B discloses a method of treating the surface of selenium disulfide powder with a silane coupling agent to form a surface structure with external organic chain ends, which then further cross-links and encapsulates it with silicone oil. Although this effectively prevents oxidation and discoloration and exhibits good dispersion and suspension effects in subsequent formulation development, the strong closed-loop nature of the silicone oil system may affect the release of selenium disulfide's efficacy. The controversy surrounding silicone oil residue makes it unacceptable to consumers seeking "silicone-free" products. Despite the existence of various methods for treating selenium disulfide, its application still suffers from drawbacks such as high preparation costs, no significant improvement in irritation, decreased efficacy, and a disconnect from the consumer market.

[0005] Utilizing plant extracts for scalp care is a significant industry trend, with many plants proven to possess anti-dandruff and soothing activities. Among these plants, licorice and turmeric exhibit bioactivity highly aligned with the needs for dandruff removal and antibacterial action. Licorice's main active ingredient, glycyrrhizin, possesses excellent anti-inflammatory and immunomodulatory effects, effectively relieving scalp redness and itching, while also exhibiting significant antibacterial capabilities. Turmeric's core component, curcumin, possesses powerful antioxidant, anti-inflammatory, and broad-spectrum antibacterial activities, showing good inhibitory effects against Malassezia. Combining licorice, turmeric, and selenium sulfide not only leverages their synergistic effects in anti-inflammatory, antibacterial, and antioxidant properties but also conditions the scalp and improves its microenvironment.

[0006] There is a need in this field for an innovative technology that can overcome the inherent defects of selenium disulfide while integrating the natural active advantages of plants. This technology would utilize plant cells as carriers for active ingredients, loading unstable active ingredients (such as selenium disulfide) into plant cells, thus improving stability, reducing irritation, achieving functional synergy, and enhancing the sensory appeal of the product. Currently, there are no reports of using licorice or turmeric plant cells as carriers to prepare selenium disulfide complexes through specific cell wall disruption and loading processes, and applying them to personal care products. Summary of the Invention

[0007] To address the above problems, this invention provides a method for preparing a plant cell-loaded selenium disulfide complex and its application.

[0008] This invention provides the following technical solution: A method for preparing a selenium disulfide complex loaded into plant cells includes the following steps: S1. Preparation of plant cell suspension: Select licorice root or turmeric root as raw material, crush them, and disperse them in an aqueous solvent using ultrasonication to form a plant cell suspension. S2. Plant cell cell disruption treatment: The cell suspension obtained in step S1 is subjected to high-voltage electroporation and then high-voltage micro-jet instantaneous blasting to form a plant cell suspension with nanoparticles that can be introduced into the porous structure. S3. Preparation of selenium disulfide nano-dispersion: Micron-sized selenium disulfide powder is mixed and dispersed with an aqueous solvent, and selenium disulfide nano-dispersion is prepared by mechanical method. S4. Loading of selenium disulfide: The selenium disulfide nano-dispersion obtained in step S3 is added to the plant cell suspension treated in step S2, and the selenium disulfide nanoparticles are introduced into the interior of the plant cells through the pores by ultrasound.

[0009] Furthermore, the pulverization process described in step S1 is performed using ultrafine pulverization technology.

[0010] Furthermore, the percentage of cell mass in the plant cell suspension described in step S1 is 30-60%.

[0011] Furthermore, in step S2, the voltage control during the high-voltage electroporation process is 100-500V, the time is 20-100 ms, and the number of pulses is 1-8.

[0012] Furthermore, in step S2, the pressure control during the instantaneous blasting of the high-pressure microjet is 3000-30000 psi, and the number of cycles is 3-10.

[0013] Furthermore, the mechanical method mentioned in step S3 is ball milling, specifically using zirconium oxide microspheres as the grinding medium, controlling the ball milling speed at 300-800 rpm, and the ball milling time at 2-12 h; the particle size range of the selenium disulfide nano-dispersion is 300-600 nm.

[0014] Furthermore, in step S4, the ultrasonic power is controlled to be 100-500W, the treatment time is controlled to be 0.5-4h, and the temperature is 20-40℃.

[0015] Furthermore, in step S4, the mass ratio of selenium disulfide, turmeric root, and licorice root in the final composite obtained is 10-20:5-15:5-15.

[0016] An application of a plant cell-loaded selenium disulfide complex, which is then used in personal care products.

[0017] Furthermore, the personal care products include liquids, emulsions, creams, granules, tablets, and gels.

[0018] The beneficial effects of this invention are: the overall preparation method is simple and easy to promote and apply, effectively overcoming the inherent defects of selenium disulfide, and organically combining it with plants. By utilizing plant cells as carriers of active ingredients, unstable active ingredients (such as selenium disulfide) are loaded into plant cells, resulting in improved stability, reduced irritation, synergistic effects, and enhanced product sensory appeal. The plant cell-loaded selenium disulfide complex prepared by this invention can be effectively applied to various personal care products, possessing strong market competitiveness. Attached Figure Description

[0019] Figure 1 This is a particle size distribution diagram of the selenium disulfide nanodispersion prepared in Example 2 of the present invention.

[0020] Figure 2 This is a particle size distribution diagram of the plant cell-loaded selenium disulfide complex particles prepared according to Example 2 of the present invention.

[0021] Figure 3 This is a SEM image of the plant cell-loaded selenium disulfide complex prepared according to Example 2 of the present invention.

[0022] Figure 4 This is a comparison chart of the results of the corresponding Malassezia inhibition experiment in Example 6 of the present invention.

[0023] Figure 5 This is a comparison chart of the results of the corresponding Raw cell toxicity experiment in Example 7 of the present invention. Detailed Implementation

[0024] The present invention will now be described in detail with reference to specific embodiments. However, the scope of protection of the present invention is not limited to the following specific embodiments. The described embodiments are merely some embodiments of the present invention, not all embodiments, and are not intended to limit the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Unless otherwise specified, the experimental methods used in the embodiments are conventional methods; the materials, reagents, equipment, etc. used are commercially available unless otherwise specified.

[0025] Example 1 A method for preparing a selenium disulfide complex loaded into plant cells includes the following steps: (a) Preparation of plant cell suspension: Licorice root and turmeric root were pulverized and ultrasonically dispersed in an aqueous solvent to form a cell suspension, so that the percentage of cell mass in the obtained plant cell mixture was 30%; (b) Cell disruption treatment: The cell suspension obtained in step (a) was subjected to high-voltage electroporation at a voltage of 100V for 20 ms and 3 pulses; then subjected to high-voltage microjet instantaneous blasting at a pressure of 10000psi and 3 cycles to form a cell suspension with a porous structure in which nanoparticles can be introduced. (c) Preparation of selenium disulfide nano-dispersion: micron-sized selenium disulfide powder was mixed and dispersed with an aqueous solvent, and the nano-dispersion was prepared by ball milling. The grinding medium was zirconia microspheres, the ball mill speed was 800 rpm, and the ball milling time was 6 hours. The particle size range of the obtained selenium disulfide nano-dispersion was 200-600 nm.

[0026] (d) Loading of selenium disulfide: The selenium disulfide nano-dispersion obtained in step (c) is added to the plant cell suspension treated in step (b), wherein the mass ratio of selenium disulfide, turmeric root and licorice root is 20:10:10. The selenium disulfide nanoparticles are introduced into the interior of the plant cells through the pores by ultrasound to achieve loading and fixation. The ultrasonic treatment power is 120W, the temperature is 25℃ and the time is 1 hour.

[0027] Example 2 A method for preparing a selenium disulfide complex loaded into plant cells includes the following steps: (a) Preparation of plant cell suspension: Licorice root and turmeric root were pulverized and ultrasonically dispersed in an aqueous solvent to form a cell suspension, so that the percentage of cell mass in the obtained plant cell mixture was 50%; (b) Cell disruption treatment: The cell suspension obtained in step (a) was subjected to high-voltage electroporation at a voltage of 300V for 60 ms and 5 pulses; then subjected to high-voltage microjet instantaneous blasting at a pressure of 25000psi and 5 cycles to form a cell suspension with a porous structure in which nanoparticles can be introduced. (c) Preparation of selenium disulfide nano-dispersion: micron-sized selenium disulfide powder was mixed and dispersed with an aqueous solvent, and the nano-dispersion was prepared by ball milling. The grinding medium was zirconia microspheres, the ball mill speed was 800 rpm, and the ball milling time was 9 hours. The particle size range of the obtained selenium disulfide nano-dispersion was 200-600 nm.

[0028] (d) Loading of selenium disulfide: The selenium disulfide nano-dispersion obtained in step (c) is added to the plant cell suspension treated in step (b), wherein the mass ratio of selenium disulfide, turmeric root and licorice root is 20:15:5. The selenium disulfide nanoparticles are introduced into the interior of the plant cells through the pores by ultrasound to achieve loading and fixation. The ultrasonic treatment power is 240W, the temperature is 25℃ and the time is 2 hours.

[0029] Example 3 A method for preparing a selenium disulfide complex loaded into plant cells includes the following steps: (a) Preparation of plant cell suspension: Licorice root and turmeric root were pulverized and ultrasonically dispersed in an aqueous solvent to form a cell suspension, so that the percentage of cell mass in the obtained plant cell mixture was 60%; (b) Cell disruption treatment: The cell suspension obtained in step (a) was subjected to high-voltage electroporation at a voltage of 500V for 100 ms and 8 pulses; then subjected to high-voltage microjet instantaneous blasting at a pressure of 30,000 psi and 10 cycles to form a cell suspension with a porous structure in which nanoparticles can be introduced. (c) Preparation of selenium disulfide nano-dispersion: micron-sized selenium disulfide powder was mixed and dispersed with an aqueous solvent, and the nano-dispersion was prepared by ball milling. The grinding medium was zirconia microspheres, the ball mill speed was 800 rpm, and the ball milling time was 12 hours. The particle size range of the obtained selenium disulfide nano-dispersion was 200-500 nm.

[0030] (d) Loading of selenium disulfide: The selenium disulfide nano-dispersion obtained in step (c) is added to the plant cell suspension treated in step (b), wherein the mass ratio of selenium disulfide, turmeric root and licorice root is 20:5:5. The selenium disulfide nanoparticles are introduced into the interior of the plant cells through the pores by ultrasound to achieve loading and fixation. The ultrasonic treatment power is 360W, the temperature is 30℃ and the time is 4 hours.

[0031] Example 4 A method for preparing a selenium disulfide complex loaded into plant cells includes the following steps: (a) Preparation of plant cell suspension: Licorice root and turmeric root were pulverized and ultrasonically dispersed in an aqueous solvent to form a cell suspension, so that the percentage of cell mass in the obtained plant cell mixture was 50%; (b) Cell disruption treatment: The cell suspension obtained in step (a) was subjected to high-voltage electroporation at a voltage of 300V for 60 ms and 5 pulses; then subjected to high-voltage microjet instantaneous blasting at a pressure of 20,000 psi for 5 cycles, and then at a pressure of 30,000 psi for 5 cycles, to form a cell suspension with a porous structure in which nanoparticles can be introduced. (c) Preparation of selenium disulfide nano-dispersion: micron-sized selenium disulfide powder was mixed and dispersed with an aqueous solvent, and the nano-dispersion was prepared by ball milling. The grinding medium was zirconia microspheres, the ball mill speed was 800 rpm, and the ball milling time was 12 hours. The particle size range of the obtained selenium disulfide nano-dispersion was 200-500 nm.

[0032] (d) Loading of selenium disulfide: The selenium disulfide nano-dispersion obtained in step (c) is added to the plant cell suspension treated in step (b), wherein the mass ratio of selenium disulfide, turmeric root and licorice root is 20:15:5. The selenium disulfide nanoparticles are introduced into the interior of the plant cells through the pores by ultrasound to achieve loading and fixation. The ultrasonic treatment power is 120W, the temperature is 25℃ and the time is 1.5 hours.

[0033] Example 5 To further investigate and test this invention, the morphology and size of the selenium disulfide nanoparticles prepared in Example 2 and the final plant cell-loaded selenium disulfide complex were studied using a laser particle size analyzer and scanning electron microscopy (SEM). (See attached image.) Figure 1 The particle size range of the selenium disulfide nanodispersion shown is 200-600 nm, with the main particle size being 300 nm. (See attached image.) Figure 2 The particle size range shown is 400 nm–100 μm, with the predominant particle size being 10 μm–20 μm, representing a complex of plant cells loaded with selenium disulfide. (See attached image.) Figure 3 These are images of plant cells loaded with selenium disulfide, observed using scanning electron microscopy (SEM). The oval-shaped particles are selenium disulfide loaded into the plant cells.

[0034] Example 6 To further investigate and test the present invention, this embodiment conducted an experiment to inhibit Malassezia, specifically selecting a concentration of 10... 8 100 μL of a CFU / mL Malassezia bacterial suspension was added to a liquid culture medium containing an equal volume of selenium disulfide. The medium was incubated on a shaker at 37°C and 180 rpm for 5 min (simulating daily hair washing time). After incubation, 100 μL was spread onto the solidified culture medium and incubated at 37°C for 48 h. The results are attached. Figure 4 As shown, it can be observed that at a selenium disulfide concentration of 0.125 mg / mL, the selenium disulfide complex prepared in Example 2 of this technology did not grow bacteria. The control sample, 20% selenium disulfide (commercially available), grew bacteria, but slightly less than the blank control group. This indicates that commercially available selenium disulfide also has a certain inhibitory effect on Malassezia, but the effect is quite different from that of the present invention.

[0035] Example 7 To further investigate and test the present invention, a Raw cell toxicity experiment was conducted in this embodiment: RAW264.7 cells were first cultured to confluence, then blown off the culture dish and transferred to a 15 mL sterile centrifuge tube for cell counting. A cell suspension of 50,000 cells / mL was prepared using DMEM medium containing 10% fetal bovine serum (FBS). 100 μL of the cell suspension (5000 cells / well) was seeded into each well of a 96-well plate and cultured for 24 h in a 5% CO2, 37°C cell culture incubator. The culture medium was then removed. A blank control group was prepared by adding 10% FBS complete medium. Drug control groups were prepared by adding 20% ​​selenium disulfide (commercially available) and the selenium disulfide complex loaded onto the corresponding plant in Example 2. All were diluted with 10% FBS complete medium to the appropriate concentration, with 100 μL of the solution added to each well, and cultured for 24 h. After 24 hours, the contents were aspirated, and 100 μL of 0.5 mg / mL thiazolyl blue (MTT) solution was added to each well. The cells were incubated for 4 hours. After 4 hours, the contents were aspirated, and 150 μL of dimethyl sulfoxide (DMSO) was added. The cells were then incubated in a solid incubator in the dark for 15 minutes to fully dissolve the cells. The OD value was measured at 570 nm using a microplate reader after shaking for 20 seconds. The experimental data were saved. The relative cell viability (%) was determined by comparing the absorbance values ​​of treated cells and control cells. Cell viability was calculated using the following formula: Cell viability(%)=(ODsample÷ODcontrol)×100%; From the results Figure 5 It can be seen that within the concentration range of 12.5-200 μg / mL, the survival rate of Raw cells decreased with increasing selenium disulfide concentration, showing a concentration-dependent effect. The relative survival rate of Raw cells treated with commercially available 20% selenium disulfide was lower than that of the plant cells coated with selenium disulfide in Example 2, which is attributed to the inherent irritant and toxic properties of selenium disulfide. After being loaded with plant cells, the barrier effect provided by the coating reduced the direct toxicity of selenium disulfide, and the relative cell survival rate was greatly improved, especially at a concentration of 100 μg / mL, where the cell survival rate was 20.4% higher than that of the commercially available group.

[0036] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for preparing a plant cell-loaded selenium disulfide complex, characterized in that, Includes the following steps: S1. Preparation of plant cell suspension: Licorice root and turmeric root are selected as raw materials, pulverized, and dispersed in an aqueous solvent by ultrasonication to form a plant cell suspension. S2. Plant cell cell disruption treatment: The cell suspension obtained in step S1 is subjected to high-voltage electroporation and then high-voltage micro-jet instantaneous blasting to form a plant cell suspension with nanoparticles that can be introduced into the porous structure. S3. Preparation of selenium disulfide nano-dispersion: Micron-sized selenium disulfide powder is mixed and dispersed with an aqueous solvent, and selenium disulfide nano-dispersion is prepared by mechanical method. S4. Loading of selenium disulfide: The selenium disulfide nano-dispersion obtained in step S3 is added to the plant cell suspension treated in step S2, and the selenium disulfide nanoparticles are introduced into the interior of the plant cells through the pores by ultrasound.

2. The method for preparing a plant cell-loaded selenium disulfide complex according to claim 1, characterized in that, The pulverization process described in step S1 is performed using ultrafine pulverization technology.

3. The method for preparing a plant cell-loaded selenium disulfide complex according to claim 1, characterized in that, The percentage of cell mass in the plant cell suspension described in step S1 is 30-60%.

4. The method for preparing a plant cell-loaded selenium disulfide complex according to claim 1, characterized in that, In step S2, the voltage control during high-voltage electroporation is 100-500V, the time is 20-100 ms, and the number of pulses is 1-8.

5. The method for preparing a plant cell-loaded selenium disulfide complex according to claim 1, characterized in that, The pressure control during the instantaneous blasting of the high-pressure microjet in step S2 is 3000-30000 psi, and the number of cycles is 3-10.

6. The method for preparing a plant cell-loaded selenium disulfide complex according to claim 1, characterized in that, The mechanical method mentioned in step S3 is ball milling, specifically using zirconium oxide microspheres as the grinding medium, controlling the ball milling speed at 300-800 rpm, and the ball milling time at 2-12 h; the particle size range of the selenium disulfide nano-dispersion is 300-600 nm.

7. The method for preparing a plant cell-loaded selenium disulfide complex according to claim 1, characterized in that, In step S4, the ultrasonic power is controlled to be 100-500W, the treatment time is controlled to be 0.5-4h, and the temperature is controlled to be 20-40℃.

8. The method for preparing a plant cell-loaded selenium disulfide complex according to claim 1, characterized in that, In step S4, the mass ratio of selenium disulfide, turmeric root, and licorice root in the final composite obtained is 10-20:5-15:5-15.

9. An application of a plant cell-loaded selenium disulfide complex, characterized in that, The selenium disulfide complex prepared by any one of claims 1-8 can be used in personal care products.

10. The application of a plant cell-loaded selenium disulfide complex according to claim 9, characterized in that, The personal care products include liquids, emulsions, creams, granules, tablets, and gels.

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