Sponge bone needle with nano-particles adsorbed on surface as well as preparation method and application of sponge bone needle

Abstract

The invention discloses a sponge spicule with nanoparticles adsorbed on the surface and a preparation method and application thereof, the sponge spicule comprises a modified bee sponge spicule and mesoporous silica nanoparticles, the modified bee sponge spicule is used as a carrier, the surface of the modified bee sponge spicule is provided with active silicon hydroxyl, the mesoporous silica nanoparticles are loaded on the modified bee sponge spicule through the active silicon hydroxyl, and the average particle size of the mesoporous silica nanoparticles is 90-1000nm. The nano-particles can pierce into the stratum corneum to open the skin barrier and stay in the stratum corneum for a long time to form a large number of continuously existing micro-channels (not less than 72 hours), and the nano-particles carrying more drugs are released and enter the skin to release the drugs to play a role.

Description

Technical Field

[0001] The present invention relates to the technical field of sponge spicules, and specifically to a sponge spicule with nanoparticles adsorbed on its surface, and a preparation method and use thereof. Background Art

[0002] With the growing demand for patient-friendly therapies, transdermal drug delivery, a forward-looking drug delivery strategy, has been continuously developed to make up for the limitations of oral and injection drug delivery. However, due to the presence of the stratum corneum of the skin, transdermal drug delivery is not easy. It is generally believed that only a very small number of drug molecules with a moderate oil-water partition coefficient (molecular weight not higher than 500 Daltons) can be effectively absorbed through the skin. Therefore, skin permeation enhancement technology has also become a hot topic in the field of skin drug delivery.

[0003] The current skin permeation enhancement technologies are divided into physical permeation enhancement and chemical permeation enhancement. Microneedle technology is a physical permeation enhancement technology that has been widely popular in recent years. Microneedle technology can open a large number of microchannels on the skin surface with minimal damage, allowing drugs and drug carriers to enter the skin through the microchannels.

[0004] Sponge Haliclona sp. Spicules (SHS) is a new type of silicon microneedle that can be used for transdermal drug delivery. The morphological structure of the sponge spicule is a free double needle tip with a sharp end. It has a smooth surface, and its size does not change easily. It has high mechanical strength, which can keep the skin channel open for large-area use. It can still maintain good efficacy after acting on the skin surface for 72 hours. At the same time, the trauma of the sponge spicule to the skin is almost negligible, so it can effectively avoid problems such as bleeding and infection. While significantly enhancing the transdermal absorption capacity of hydrophilic macromolecules, it can also ensure the safety and controllability of the drug delivery process. It is a penetration-enhancing technology with great application prospects. However, SHS also has certain limitations: for hydrophilic drugs, as the molecular weight of the drug increases, the SHS enhancement effect on its transdermal penetration will gradually decrease, while lipophilic drugs cannot penetrate into the deep skin through the hydrophilic channel opened by SHS. Summary of the invention

[0005] The present invention aims to overcome the defects of the prior art and provide a sponge spicule with nanoparticles adsorbed on its surface.

[0006] Another object of the present invention is to provide a method for preparing the sponge spicules with nanoparticles adsorbed on the surface.

[0007] Another object of the present invention is to provide the use of the sponge spicule with nanoparticles adsorbed on its surface.

[0008] The technical solution of the present invention is as follows:

[0009] A sponge spicule with nanoparticles adsorbed on the surface, comprising a modified bee sponge spicule with active silanol groups on the surface as a carrier and mesoporous silica nanoparticles loaded thereon by the active silanol groups, wherein the average particle size of the mesoporous silica nanoparticles is 90-1000nm;

[0010] The modified bee sponge spicules are prepared by washing the bee sponge spicules and reacting them with NaOH solution and hydrochloric acid solution in sequence;

[0011] The above mesoporous silica nanoparticles are prepared by reacting CTAB, NaOH and TEOS and modified with APTES;

[0012] The modified bee sponge spicules and mesoporous silica nanoparticles are connected by reacting with a citric acid-trisodium citrate aqueous solution.

[0013] In a preferred embodiment of the present invention, the pH of the citric acid-trisodium citrate aqueous solution is 4.

[0014] The method for preparing the sponge spicule with nanoparticles adsorbed on the surface comprises the following steps:

[0015] (1) Preparation of mesoporous silica nanoparticles using CTAB, NaOH and TEOS;

[0016] (2) modifying the above mesoporous silica nanoparticles with APTES to obtain APTES-modified mesoporous silica nanoparticles;

[0017] (3) treating the cleaned bee sponge spicules with a NaOH solution and a hydrochloric acid solution in sequence to obtain modified bee sponge spicules with active silanol groups on the surface;

[0018] (4) reacting the APTES-modified mesoporous silica nanoparticles obtained in step (2) and the modified bee sponge spicules obtained in step (3) with a citric acid-trisodium citrate aqueous solution, and then washing and drying to obtain the product.

[0019] In a preferred embodiment of the present invention, the step (1) is as follows: CTAB and NaOH are added to distilled water, mixed evenly, heated to 80°C and stirred continuously, wherein the ratio of CTAB, NaOH and distilled water is 1 g:0.25-0.5 g:480 mL; then 5-10 times the mass of TEOS of CTAB is added dropwise thereto, and stirring is continued at 80°C and 1300-1500 rpm for 2-3 hours, and the obtained white precipitate is washed with distilled water and methanol in turn, and then dried, and finally CTAB is removed by calcination to obtain mesoporous silica nanoparticles.

[0020] In a preferred embodiment of the present invention, the step (2) is: adding the mesoporous silica nanoparticles prepared in step (1) to distilled water to obtain a MSNs suspension; then adding 175 μL / mL of an APTES aqueous solution to the MSNs suspension under a dry nitrogen purge, and then stirring and refluxing the mixture under a nitrogen atmosphere at 77°C. The reaction product is washed with ethanol and then dried to obtain APTES-modified mesoporous silica nanoparticles.

[0021] In a preferred embodiment of the present invention, the step (3) is: ultrasonically clean the bee sponge spicules with water, ethanol and acetone in sequence, and obtain SHS with a clean surface after drying, then add it to a 5 mol / L NaOH solution and stir for 30 minutes, then wash it with distilled water, then add it to a 10% hydrochloric acid solution and stir for 30 minutes, then wash it with distilled water, and after drying, obtain modified bee sponge spicules with active silanol groups on the surface.

[0022] In a preferred embodiment of the present invention, the step (4) is: preparing a citric acid-trisodium citrate aqueous solution with a pH of 4, adding the APTES-modified mesoporous silica nanoparticles obtained in step (2) and the modified bee sponge spicules obtained in step (3) in a mass ratio of 7:1, stirring the reaction, washing the reaction product with distilled water and drying to obtain the sponge spicules with nanoparticles adsorbed on the surface.

[0023] Further preferably, the citric acid-trisodium citrate aqueous solution is prepared from a 0.1 mol / L citric acid aqueous solution and a 0.1 mol / L trisodium citrate aqueous solution.

[0024] The sponge bone needle with nanoparticles adsorbed on the surface is used as a carrier for transdermal drug delivery.

[0025] A transdermal drug delivery carrier comprises the sponge bone needles with the nanoparticles adsorbed on the surface.

[0026] The beneficial effects of the present invention are:

[0027] 1. The present invention can penetrate the stratum corneum to open the skin barrier, and stay in the stratum corneum of the skin for a long time to form a large number of persistent microchannels (not less than 72 hours), releasing nanoparticles carrying more drugs, entering the skin to release drugs and exert their effects.

[0028] 2. The present invention can utilize the ability of mesoporous silica nanoparticles to load multiple drugs simultaneously, so as to carry different types of drugs for simultaneous administration, and is expected to achieve synergistic treatment of diseases with multiple drugs.

[0029] 3. The present invention can load different types of drug molecules into mesoporous silica nanoparticles to improve the solubility or stability of the drugs. The drugs will penetrate the skin along with the present invention and will not be affected by the physical and chemical properties of the drugs themselves, and then be released from the mesopores of the mesoporous silica nanoparticles.

[0030] 4. After the present invention acts on the skin, it will automatically detach from the skin as the keratinocytes physiologically peel off, and the skin barrier can recover by itself without causing serious trauma to the skin.

[0031] 5. The present invention is different from the previously prepared mesoporous bee sponge spicules (mSHS) in that after the mSHS penetrates the skin, the drug is released from its mesoporous layer, while after the present invention penetrates the skin, the nanoparticles can first diffuse into the deeper layers of the skin and then release the drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 These are scanning electron microscope photos of the modified bee sponge spicules in Example 1 of the present invention, wherein: a is the morphology of the modified bee sponge spicules under a scanning electron microscope with a magnification of 500 times, b is the morphology of the modified bee sponge spicules under a scanning electron microscope with a magnification of 5000 times, and c is the morphology of the modified bee sponge spicules under a scanning electron microscope with a magnification of 10000 times.

[0033] Figure 2 This is a scanning electron microscope photograph of the mesoporous silica nanoparticles in Example 1 of the present invention, wherein: a is MSNs prepared under condition A, with an average diameter of 90 nm; b is MSNs prepared under condition B, with an average diameter of 120 nm; c is MSNs prepared under condition C, with an average diameter of 500 nm; d is MSNs prepared under condition D, with an average diameter of 1000 nm.

[0034] Figure 3 The scanning electron microscope photographs of the mesoporous silica nanoparticles before and after modification in Example 1 of the present invention, specifically, the MSNs (i.e., MSNs500) successfully prepared under experimental condition C were amino-modified using the post-grafting method, a is MSNs500 before modification, and b is MSNs500 after modification.

[0035] Figure 4 This is a scanning electron microscope photograph of the bee sponge spicule (nSHS) with mesoporous nanoparticles adsorbed on the surface prepared in Example 1 of the present invention.

[0036] Figure 5 This is a graph showing the changes in the content of coumarin 6 in different cortical layers after 16 hours of in vitro transdermal penetration of nSHS loaded with coumarin 6 in Example 2 of the present invention.

[0037] Figure 6It shows that the nSHS in Example 3 of the present invention can carry two different drugs at the same time. DETAILED DESCRIPTION

[0038] The technical solution of the present invention is further illustrated and described below through specific implementation modes in combination with the accompanying drawings.

[0039] Example 1

[0040] (1) Preparation of mesoporous silica nanoparticles (MSNs): Sodium hydroxide and N-hexadecyltrimethylammonium bromide (CTAB) were added to 480 mL of distilled water according to Table 1 below, and the resulting solution was heated to 80°C and stirred continuously. Then tetraethyl orthosilicate (TEOS) was added dropwise to the above solution, and the mixture was stirred vigorously at 80°C for 2-3 h. The resulting white precipitate was washed three times with distilled water and methanol, respectively, and then dried in an oven for 8 h. Finally, the product was placed in a muffle furnace and heated to 550°C at 4°C / min for 3 h to remove CTAB, and MSNs (such as Figure 2 and Figure 3 shown).

[0041] The experimental conditions and material amounts of this step are shown in Table 1 Experimental conditions and material amounts for MSNs preparation.

[0042] Table 1 Experimental conditions and material dosage for MSNs preparation

[0043]

[0044]

[0045] (2) MSNs modification: Add 70 mg of MSNs obtained in step (1) to 75 mL of distilled water to prepare a MSNs suspension, which was then added to a three-necked flask. Prepare 2 mL of 175 μL / mL APTES aqueous solution and add it to the MSNs suspension under a dry nitrogen purge. Then, reflux for 14 h at 77 °C under nitrogen conditions with magnetic stirring (500 rpm), wash with ethanol three times, and dry at 60 °C for 8 h. Figure 3 As shown, the modification process in this step does not change the shape and average diameter of MSNs.

[0046] (3) SHS modification: Use 500 mL of water, 500 mL of ethanol and 400 mL of acetone to ultrasonically clean SHS in turn, and dry it to obtain SHS with a clean surface. Add 8 g of SHS with a clean surface into 400 mL of 5 mol / L NaOH solution and stir for 30 min, then wash it with distilled water. Then add the product into 400 mL of 10% hydrochloric acid solution and stir for 30 min. Wash the obtained product with distilled water and dry it to obtain modified SHS with active silanol groups on the surface. Figure 1 As shown in Figure 3, the surface of the modified SHS is smooth, but a pore structure with a diameter of about 100 nm appears.

[0047] (4) Combination: Using citric acid monohydrate, trisodium citrate and distilled water as raw materials, 0.1 mol / L citric acid aqueous solution and 0.1 mol / L trisodium citrate aqueous solution were prepared. 32.75 mL of 0.1 mol / L citric acid aqueous solution was added to 17.25 mL of 0.1 mol / L trisodium citrate aqueous solution to prepare 50 mL of citric acid-trisodium citrate aqueous solution with a pH of 4. 70 mg of modified MSN-NH2 and 10 mg of modified SHS were added to 50 mL of citric acid-trisodium citrate aqueous solution with a pH of 4, stirred for 8 h (200 rpm), and the obtained product was washed 3 times with distilled water and dried at 60 ° C for 8 h. The obtained product was constructed as shown in FIG. Figure 4 Shown are the surface of bee sponge spicules (nSHS) with adsorbed mesoporous silica nanoparticles.

[0048] Table 2 Results of MSNs prepared under different experimental conditions and material dosages

[0049]

[0050]

[0051] From the above comparative experimental results, it can be seen that in the process of preparing MSNs, increasing the amount of NaOH and TEOS can increase the size of MSNs. Because in the preparation of MSNs, TEOS is used as a silicon source, increasing the amount of TEOS will provide more silicon and produce larger MSNs. At the same time, it can be found that as the size of MSNs increases, its shape will change from spherical to round.

[0052] Some studies have mentioned that the increase in CTAB concentration is accompanied by an increase in the size of the synthesized silica particles. As an acid-base regulator, when the alkalinity is weak, the number of negative charges adsorbed by the hydrolyzed silicon precursor during the reaction is small, so it is poorly combined with the surfactant micelles, and it is easy to generate indeterminate silica products; however, when the alkalinity is strong, the hydrolysis rate of TEOS is too fast, which is also one of the reasons why it is difficult to form long-chain oligomeric silica particles. This may lead to a considerable number of silicon sources in more silicon precursors being unable to fully combine with the CTAB micelles to generate solid silica. In addition, in the reaction system, the interaction between CTAB micelles of different forms and silicon precursors is less likely to reach equilibrium in a short period of time, so particles with different morphological characteristics will be generated. As the reaction time increases, the hydrophobic properties of the micelles will play a modifying role on the nanoparticles. In addition, there is sufficient time for dense accumulation inside the nanoparticles, so mesoporous silica nanoparticles with uniform morphology can be formed.

[0053] Therefore, the amount of CTAB, NaOH and TEOS, as well as the reaction temperature and time, are very important for the preparation of MSNs. In order to prepare MSNs with uniform morphology, stable size and large specific surface area, it is necessary to pay close attention to each preparation condition.

[0054] Example 2

[0055] In this example, coumarin 6 was used as a fat-soluble model drug to study the drug loading and transdermal administration effects of the nSHS prepared in Example 1 on coumarin 6.

[0056] (1) nSHS drug loading method: Weigh 10 mg of nSHS and place it in a 1.5 mL EP tube, then add 600 μL of 40 mg / mL coumarin 6 solution (solvent is dichloromethane), vortex and oscillate to mix thoroughly, then wrap the centrifuge tube with aluminum foil and place it on a rotary mixer for 12 hours (70 rpm). Remove the centrifuge tube from the rotary mixer and let it stand for 5 minutes to allow nSHS to naturally settle at the bottom of the centrifuge tube. Place the centrifuge tube in a centrifuge for centrifugation (1500 rpm, 5 minutes), and use a pipette to remove the supernatant as completely as possible after centrifugation. Wrap the centrifuge tube after removing the supernatant drug with aluminum foil and place it in a freeze dryer for freeze drying (or place it in a fume hood for natural air drying) to obtain nSHS@coumarin 6.

[0057] (2) Quantitative determination of drug loading of nSHS@coumarin 6: nSHS@coumarin 6 was placed in a certain amount of ethanol solution and shaken in a shaker (180 rpm, 28°C) overnight. 1 mL of the extracted solution was taken and the fluorescence value was measured using an ELISA instrument and substituted into the standard curve (the standard curve of coumarin 6 in ethanol solution: y = 15468x + 282.68, R 2 =0.999. y is the fluorescence value, x is the concentration of coumarin 6, in μg / mL), and then the drug loading of the corresponding nSHS is calculated according to the content of coumarin 6 in the ethanol solution (as shown in Table 3).

[0058] Table 3 Particle size of nanoparticles carried by nSHS and corresponding drug loading

[0059]

[0060] It can be found that as the particle size of the nanoparticles carried by nSHS increases, the drug loading capacity also increases accordingly.

[0061] (3) nSHS@Coumarin 6 was applied to the study of the effect of percutaneous penetration enhancement in vitro:

[0062] In this example, ex vivo pig skin was used as a skin model for experiments.

[0063] First, select pigskin without obvious trauma on the back of the pig, then carefully remove the subcutaneous fat tissue of the pigskin with a scalpel, and further use an electric shaver to shave the hair on the pigskin to a length of less than 5 mm, then use ultrapure water to wash the pigskin treated above and store it at -20°C for use. Thaw the skin at room temperature before use. Use a circular punch with a diameter of 40 mm to drill pigskin of the same diameter and install it on the Franz diffusion transdermal device.

[0064] The skin conductivity was measured by ex vivo transcutaneous resistance test to ensure the integrity of the skin barrier. That is, a waveform generator was used to measure the current passing through the pig skin at 100mV and 100Hz. Pig skin with a current less than 5A was considered intact skin and the next step of the experiment could be carried out. The effective penetration area of ​​the Franz diffusion cell was 1.77cm2 and the receptor volume was 12mL.

[0065] The skin was placed on the top of a vertical Franz diffusion cell, and the receptor cell was filled with 0.2M PBS solution (pH=7.4) containing 1% Tween80, and a small rotor was added to the receptor cell. The in vitro transdermal experiment was divided into 6 groups, each with 3 parallels:

[0066] nSHS@Coumarin 6 group: Take a certain amount of nSHS drug-loaded powder (according to the above step (1) of this example), add 1 mL of 0.2 M PBS to dissolve, take 200 μL and add it to the drug delivery cavity, so that the dosage of Coumarin 6 is 200 μg, and evenly distribute it on the surface of pig skin; use a household electric massager (apply a force of about 0.3 N, a speed of about 300 rpm) to topically apply nSHS to the skin for 2 minutes. Then seal the drug delivery cavity with a sealing film.

[0067] SHS+MSNs@Coumarin 6 group: 100 μL of PBS (0.2M, pH=7.4) solution containing 10 mg SHS was placed in the administration cavity and evenly distributed on the pig skin surface; SHS was topically applied to the skin for 2 min using a household electric massager (applying a force of about 0.3 N and a rotation speed of about 300 rpm). After application, the skin was washed 3 times with 0.2 M PBS solution to remove the residual SHS on the skin. Then 200 μL of MSNs1000 powder suspension loaded with coumarin 6 (with a coumarin 6 content of 200 μg) was added to the receptor cavity and evenly distributed on the pig skin surface, and then the administration cavity was sealed with a sealing film.

[0068] mSHS@Coumarin 6 group: 100 μL of PBS (0.2M, pH=7.4) solution containing 10 mg of mSHS@Coumarin 6 was placed in the drug administration cavity to evenly distribute it on the pig skin surface; mSHS@Coumarin 6 was topically applied to the skin for 2 min using a household electric massager (applying a force of about 0.3 N and a rotation speed of about 300 rpm / min) to massage the mSHS@Coumarin 6 as evenly as possible on the skin surface, and finally the drug administration cavity was sealed with a sealing film; the mSHS is a mesoporous bee sponge spicule, see CN 112237567 A "A mesoporous sponge spicule, its preparation method and use".

[0069] SHS+Coumarin 6 group: 100 μL of PBS (0.2M, pH=7.4) solution containing 10 mg SHS was placed in the administration cavity and evenly distributed on the pig skin surface; SHS was topically applied to the skin for 2 min using a household electric massager (applying a force of about 0.3 N and a rotation speed of about 300 rpm). After application, the skin was washed 3 times with 0.2 M PBS solution to remove the residual SHS on the skin. Then 200 μL of saturated Coumarin 6 solution (solvent was 30% ethanol) was added to the receptor cavity, and the Coumarin 6 solution was evenly distributed on the pig skin surface, and then the administration cavity was sealed with a sealing film.

[0070] Control group: 200 μL of saturated coumarin 6 solution (solvent: 30% ethanol) was added into the administration cavity, and the coumarin 6 solution was evenly distributed on the surface of the pig skin, and then the administration cavity was sealed with a sealing film.

[0071] Blank group: 200 μL of 0.2 M PBS solution (pH=7.4) containing 1% Tween 80 was added into the administration cavity and evenly distributed on the surface of the pig skin, and then the administration cavity was sealed with a sealing film.

[0072] The Franz diffusion cells of the above groups were placed in a water bath at 37±0.5℃, and the speed was set to 600rpm / min, so that the drugs in the receptor pool could be evenly dispersed. Finally, the outlet of the receptor pool was sealed with a sealing film, and the permeation was carried out in the dark for 16 hours, and each group was repeated at least three times. After the transdermal permeation, the drugs remaining in the drug delivery cavity were washed 5 times with PBS solution (control group) or ethanol solution, and then the skin was removed, wrapped with aluminum foil in the dark, and kept for use. 1mL of the solution in the receptor pool was taken to determine the content of coumarin 6 in the receptor pool.

[0073] The liquid in the receptor pool was taken out to determine the content of coumarin 6 that had penetrated into the subcutaneous tissue, and then the content of coumarin 6 in each layer of the skin was determined by the tape stripping method.

[0074] The content of coumarin 6 in each layer of skin was determined by tape stripping method. The skin removed after transdermal penetration was fixed on the stripping device, and then the skin stratum corneum (SC) was stripped 10 times continuously with Scotch@ transparent tape. The stripped tapes were collected and placed in a brown glass bottle according to the following scheme: the first strip was recorded as stratum corneum 1, the second to fifth strips were recorded as stratum corneum 2-5, and the sixth to tenth strips were recorded as stratum corneum 6-10. After the stratum corneum was collected, the active epidermis was collected, that is, the active epidermis was separated from the dermis with a scalpel, and the separated active epidermis was placed in a brown glass bottle. Finally, two dermis layers were drilled with a punch (d = 5 mm), chopped with a scalpel, and placed in a brown glass bottle. 4 mL of methanol solution was added to each of the above 5 brown glass bottles, and then placed in a shaker for overnight shaking (25°C, 180 rpm) to fully extract coumarin 6 in the skin. After the extraction, the extract in the glass bottle was centrifuged (5000 rpm, 5 min), and the supernatant was used to determine the coumarin 6 content.

[0075] The extracts of the stratum corneum, active epidermis, dermis and the solution in the receiving pool were placed in an ELISA instrument to measure the corresponding fluorescence values ​​(excitation wavelength 466nm, emission wavelength 504nm), and then substituted into the standard curve to obtain the corresponding coumarin 6 content in different cortical layers.

[0076] The experimental results are shown in Figure 5 ,From the quantitative analysis of the experimental results, it can be found that the transdermal amount of nSHS90@Coumarin 6 group (31.26±5.29μg), nSHS500@Coumarin 6 group (35.41±5.42μg) and nSHS1000@Coumarin 6 group (59.76±21.13μg) are all higher than that of SHS+Coumarin 6 group (1.88±0.59μg) and control group (1.19±0.17μg). The total amount of nSHS@Coumarin 6 group showed an increasing trend with the increase of the particle size of the adsorbed MSNs when the dosage of Coumarin 6 was controlled to be the same. The amount of nSHS1000@Coumarin 6 group permeated was about 50.22 times that of the control group. The drug permeation rate of nSHS1000@Coumarin 6 group was 29.88%±10.57%, while the drug permeation rate of the control group was only 0.59%±0.08%. There was no significant difference in the total amount of permeation of SHS+Coumarin 6 group and the control group, which shows that it is difficult for general lipophilic drugs to directly enter the skin, and SHS alone has no significant permeation-enhancing effect when used to deliver lipophilic drugs.

[0077] From the experimental results, it can be found that compared with the two groups of direct application and SHS application, nSHS carrying coumarin 6 significantly increased the total amount of coumarin 6 permeated through the skin, but it was still concentrated in the stratum corneum and epidermis, but its effect in promoting the transdermal delivery of coumarin 6 was still relatively significant.

[0078] Secondly, this example compares SHS adsorbed MSNs with SHS mixed MSNs, that is, the total permeation amount of the nSHS@coumarin 6 group is higher than that of the SHS+MSNs@coumarin 6 group (19.69±5.66 μg). It can be seen that the administration method of first applying SHS massage and then adding MSNs@coumarin 6 cannot achieve the permeation-enhancing effect of nSHS.

[0079] At the same time, this example compares nSHS with the previously prepared mesoporous spicules (mSHS), and finds that nSHS can greatly increase the drug loading of coumarin 6 and increase the transdermal delivery amount of coumarin 6.

[0080] Therefore, the experiment proves that nSHS can improve the solubility of drugs, increase the drug loading, and promote the percutaneous absorption of poorly soluble drugs. In addition, the total amount of poorly soluble drugs permeated will increase with the increase of the particle size of the adsorbed MSNs.

[0081] Example 3

[0082] In this example, fluorescein isothiocyanate (FITC) and protoporphyrin IX (PpIX) were used as model drugs to study the feasibility of the nSHS prepared in Example 1 to carry multiple drugs simultaneously.

[0083] (1) Drug delivery

[0084] Weigh 70 mg of MSNs1000 and place it in a 2 mL EP tube, then add 1.5 mL of an aqueous solution containing 5 mg FITC; vortex to mix it thoroughly, then wrap the centrifuge tube with aluminum foil and place it on a rotary mixer for rotation mixing for 12 h (70 rpm); remove the centrifuge tube from the rotary mixer and let it stand for 5 min to allow the MSNs to naturally settle at the bottom of the centrifuge tube; place the centrifuge tube in a centrifuge for centrifugation (1500 rpm, 5 min), and after centrifugation, use a pipette to completely remove the supernatant as much as possible; wrap the centrifuge tube after removing the supernatant drug with aluminum foil and place it in a freeze dryer for freeze drying (or place it in a fume hood for natural drying) to obtain MSNs@FITC.

[0085] Weigh 70 mg of MSNs1000 and place it in a 2 mL EP tube, then add 1 mL of 5 mg / mL PpIX solution (solvent is DMSO); vortex to mix thoroughly, then wrap the centrifuge tube with aluminum foil and place it on a rotary mixer for rotation mixing for 12 h (70 rpm); remove the centrifuge tube from the rotary mixer and let it stand for 5 min to allow the MSNs to naturally settle at the bottom of the centrifuge tube; place the centrifuge tube in a centrifuge for centrifugation (1500 rpm, 5 min), and after centrifugation, use a pipette to remove the supernatant as completely as possible; wrap the centrifuge tube after removing the supernatant drug with aluminum foil and place it in a freeze dryer for freeze drying (or place it in a fume hood for natural drying) to obtain MSNs@PpIX.

[0086] (2) MSNs modification

[0087] Add 30 mg MSNs@FITC and 30 mg MSNs@PpIX to 75 mL distilled water to prepare MSNs suspension, which was added to a three-necked flask. Prepare 2 mL of 175 μL / mL APTES aqueous solution and add it to the MSNs suspension under dry nitrogen purge. Then, reflux for 14 h at 77 ° C under nitrogen conditions with magnetic stirring (500 rpm), wash with ethanol three times, and freeze-dry in a freeze dryer.

[0088] (3) SHS modification

[0089] 0.5g SHS was ultrasonically cleaned with 50mL water, 50mL ethanol and 50mL acetone in sequence and dried to obtain SHS with a clean surface. 0.5g SHS with a clean surface was added to 200mL 5mol / L NaOH solution and stirred for 30min, and then washed with distilled water. Then the product was added to 200mL 10% hydrochloric acid solution and stirred for 30 minutes, and the obtained product was washed with distilled water and dried to obtain modified SHS with active silanol groups on the surface.

[0090] (4) Combination

[0091] Using citric acid monohydrate, trisodium citrate and distilled water as raw materials, 0.1 mol / L citric acid aqueous solution and 0.1 mol / L trisodium citrate aqueous solution were prepared. 32.75 mL of 0.1 mol / L citric acid aqueous solution was added to 17.25 mL of 0.1 mol / L trisodium citrate aqueous solution to prepare 50 mL of citric acid-trisodium citrate aqueous solution with a pH of 4. 60 mg of modified MSN-NH2 and 10 mg of modified SHS were added to 50 mL of citric acid-trisodium citrate aqueous solution with a pH of 4, stirred for 8 h (200 rpm), and the obtained product was washed 3 times with distilled water and freeze-dried in a freeze dryer to obtain MSNs-NH2-SHS (abbreviated as nSHS).

[0092] The nSHS that successfully carried the two drugs was placed under a fluorescence confocal microscope for observation, and it was found that the surface of the bone needles in different channels showed different colors ( Figure 6 , FITC in green, PpIX in red), demonstrating that nSHS can carry two different drugs simultaneously.

[0093] The above description is only a preferred embodiment of the present invention, and therefore cannot be used to limit the scope of the present invention. That is, equivalent changes and modifications made according to the patent scope of the present invention and the contents of the specification should still fall within the scope of the present invention.

Claims

1. A sponge spicule with nanoparticles adsorbed on its surface, characterized in that: The invention comprises a modified bee sponge spicule with active silanol groups on the surface as a carrier and mesoporous silica nanoparticles loaded thereon by the active silanol groups, wherein the average particle size of the mesoporous silica nanoparticles is 90-1000nm; The modified bee sponge spicules are prepared by washing bee sponge spicules and reacting them with NaOH solution and hydrochloric acid solution in sequence; the mesoporous silica nanoparticles are prepared by reacting CTAB, NaOH and TEOS and modified with APTES; The modified bee sponge spicules and mesoporous silica nanoparticles are connected by reacting with a citric acid-trisodium citrate aqueous solution.

2. The sponge spicule with nanoparticles adsorbed on the surface as claimed in claim 1, characterized in that: The pH of the citric acid-trisodium citrate aqueous solution is 4.

3. The method for preparing the sponge spicule with surface adsorbed nanoparticles according to claim 1 or 2, characterized in that: The steps include: (1) Preparation of mesoporous silica nanoparticles using CTAB, NaOH and TEOS; (2) modifying the above mesoporous silica nanoparticles with APTES to obtain APTES-modified mesoporous silica nanoparticles; (3) treating the cleaned bee sponge spicules with a NaOH solution and a hydrochloric acid solution in sequence to obtain modified bee sponge spicules with active silanol groups on the surface; (4) reacting the APTES-modified mesoporous silica nanoparticles obtained in step (2) and the modified bee sponge spicules obtained in step (3) with a citric acid-trisodium citrate aqueous solution, and then washing and drying to obtain the product.

4. The preparation method according to claim 3, characterized in that: The step (1) comprises: adding CTAB and NaOH into distilled water, mixing them evenly, heating them to 80° C. and stirring them continuously, wherein the ratio of CTAB, NaOH and distilled water is 1 g: 0.25-0.5 g: 480 mL; then adding 5-10 times the mass of TEOS of CTAB dropwise thereto, stirring at 80° C. and 1300-1500 rpm for 2-3 hours, washing the obtained white precipitate with distilled water and methanol in turn, drying the precipitate, and finally removing the CTAB by calcination to obtain mesoporous silica nanoparticles.

5. The preparation method according to claim 3, characterized in that: The step (2) is as follows: adding the mesoporous silica nanoparticles prepared in step (1) into distilled water to obtain a MSNs suspension; then adding 175 μL / mL of an APTES aqueous solution to the MSNs suspension under a dry nitrogen purge, and then stirring and refluxing the mixture under a nitrogen atmosphere at 77° C., washing the reaction product with ethanol and then drying the reaction product to obtain APTES-modified mesoporous silica nanoparticles.

6. The preparation method according to claim 3, characterized in that: The step (3) is: ultrasonically cleaning the bee sponge spicules with water, ethanol and acetone in sequence, obtaining SHS with a clean surface after drying, then adding it to a 5 mol / L NaOH solution, stirring for 30 minutes, and then washing it with distilled water, then adding it to a 10% hydrochloric acid solution, stirring for 30 minutes, and then washing it with distilled water, and after drying, a modified bee sponge spicule with active silanol groups on the surface is obtained.

7. The preparation method according to claim 3, characterized in that: The step (4) comprises: preparing a citric acid-trisodium citrate aqueous solution with a pH of 4, adding the APTES-modified mesoporous silica nanoparticles obtained in step (2) and the modified bee sponge spicules obtained in step (3) in a mass ratio of 7:1, stirring for reaction, washing the obtained reaction product with distilled water and drying, thereby obtaining the sponge spicules with nanoparticles adsorbed on the surface.

8. The preparation method according to claim 7, characterized in that: The citric acid-trisodium citrate aqueous solution is prepared from a 0.1 mol / L citric acid aqueous solution and a 0.1 mol / L trisodium citrate aqueous solution.

9. Use of the sponge spicule with nanoparticles adsorbed on the surface as claimed in claim 1 or 2 as a carrier for transdermal drug delivery.

10. A transdermal drug delivery carrier, characterized in that: A sponge spicule having surface-adsorbed nanoparticles as claimed in claim 1 or 2.

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