Selenium nanoparticle fluorescent probe capable of imaging in vivo as well as preparation method and application of selenium nanoparticle fluorescent probe
By recombining selenium quantum dots of specific wavelengths with ICG, the emission wavelength of redshift ICG and enhancing tissue penetration ability, the shortcomings of existing fluorescent probes in in vivo imaging are solved, and efficient in vivo imaging and antioxidant performance are achieved, suitable for imaging and monitoring of spinal cord injury repair.
Patent Information
- Application Number
- CN202510063843.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-15
- Publication Date
- 2025-04-25
AI Technical Summary
In terms of in vivo imaging, existing fluorescent probes have problems such as low tissue penetration ability, poor solubility in water and low quantum yield. In clinical practice, near-infrared organic dyes such as ICG have disadvantages such as poor light stability, short emission peaks and liver targeting, making it difficult to achieve efficient in vivo imaging and reactive oxygen scavenging.
By preparing selenium quantum dots of specific wavelengths and recombining with ICG, the emission wavelength of redshifting ICGs can enhance tissue penetration ability, and using the reactive oxygen scavenging ability of selenium quantum dots, a fluorescent probe of selenium nanoparticles that can be imaged in vivo were prepared.
It achieves deeper imaging in vivo, enhances tissue penetration ability, has excellent antioxidant properties and excellent biocompatibility, and is suitable for imaging and real-time monitoring in spinal cord injury repair.
Smart Images
Figure CN119875641A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of biomedical materials and biomedical engineering, and relates to a selenium nanoparticle fluorescent probe capable of in vivo imaging and a preparation method and application thereof. Background Art
[0002] Traditional fluorescent probes (small molecule dyes, fluorescent proteins, etc.) have the disadvantages of single function, low resolution and low tissue penetration. Compared with the single fluorescence development function of traditional fluorescent probes, nanoparticles can be modified by functional molecules, showing specific targeting, realizing integrated diagnosis and treatment. Nanoparticles have quantum effects, so that the optical properties of materials can be regulated at the microscopic level. Selenium quantum dots play an important role in antioxidant defense and redox regulation, and have the advantages of good biocompatibility, small size, low cost and adjustable fluorescence. However, because their emission wavelength is in the visible light range, their tissue penetration ability is low, and they can only achieve intracellular imaging, not in vivo imaging. At the same time, they also have the disadvantages of poor solubility in water and low quantum yield. Studies have shown that near-infrared 700-1700nm can achieve in vivo imaging, but the existing near-infrared organic dyes such as ICG in clinical practice have the disadvantages of poor photostability, short emission peak, and liver targeting. Therefore, fluorescent probes with integrated diagnosis and treatment of near-infrared in vivo imaging and scavenging reactive oxygen species are urgently needed for clinical diagnosis and medical treatment. Summary of the invention
[0003] The purpose of the present invention is to at least partially solve the above technical problems, and to provide a selenium nanoparticle fluorescent probe capable of in vivo imaging and its preparation method and application. The selenium nanoparticle fluorescent probe prepared by the present invention has excellent fluorescence performance and good biocompatibility, and can be used for real-time monitoring in vivo.
[0004] The first aspect of the present invention provides a method for preparing a selenium nanoparticle fluorescent probe capable of in vivo imaging, comprising:
[0005] S1. Under light-proof conditions, sodium selenite pentahydrate, citric acid, and arginine are added to water, the pH is adjusted to 9-11, and the reaction is heated to 150°C-200°C. After the reaction is completed, the mixture is cooled to room temperature, dialyzed with distilled water, and then freeze-dried to obtain purified selenium quantum dots;
[0006] S2. Under light-proof conditions, the selenium quantum dots are added into water, indocyanine green is added, and a reaction is carried out. After the reaction is completed, the selenium quantum dots are washed with water, centrifuged, and freeze-dried to obtain the selenium nanoparticle fluorescent probe.
[0007] In some embodiments of the present invention, in step S1, the molar ratio of sodium selenite pentahydrate, citric acid and arginine is (0.9-1.2):(10-11):(40-41).
[0008] In some embodiments of the present invention, in step S1, the reaction time is 6-10 hours.
[0009] In some embodiments of the present invention, in step S1, the dialysis bag used for dialysis has a specification of 500D.
[0010] In some embodiments of the present invention, in step S2, the mass ratio of selenium quantum dots to indocyanine green is (10-15) mg:(1-2) mg.
[0011] In some embodiments of the present invention, in step S2, the reaction time is 10 to 12 hours.
[0012] The second aspect of the present invention provides a selenium nanoparticle fluorescent probe prepared by the above preparation method.
[0013] The third aspect of the present invention provides a use of the above-mentioned selenium nanoparticle fluorescent probe in the preparation of spinal cord injury repair biomaterials.
[0014] A fourth aspect of the present invention provides a use of the above-mentioned selenium nanoparticle fluorescent probe in the preparation of a real-time monitoring material for spinal cord injury repair.
[0015] The selenium nanoparticle fluorescent probe capable of in vivo imaging and its preparation method and application according to the embodiment of the present invention have at least one of the following advantages:
[0016] (1) The present invention prepares quantum dots of a specific wavelength and compounds them with ICG, thereby red-shifting the emission wavelength of ICG and enhancing tissue penetration, thereby achieving deeper in vivo imaging. It can be used to achieve in vivo imaging under specific conditions (high subcutaneous depth requirements), for example, imaging in spinal cord injury repair, thereby achieving real-time monitoring of the repair process.
[0017] (2) The selenium nanoparticle fluorescent probe prepared by the present invention not only realizes deeper imaging, but also utilizes the active oxygen scavenging ability of selenium quantum dots to make the material have excellent antioxidant properties and excellent biocompatibility. BRIEF DESCRIPTION OF THE DRAWINGS
[0018] These and / or other aspects and advantages of the present invention will become apparent and readily understood from the following description of the preferred embodiments in conjunction with the accompanying drawings, in which:
[0019] Figure 1 Schematic diagram of transmission electron microscopy (TEM) of fluorescent nanoparticles prepared in Example 1; A is a schematic diagram of SeQDs TEM; B is a schematic diagram of fluorescent nanoparticles (Se@ICG) TEM;
[0020] Figure 2is a schematic diagram of elemental analysis of fluorescent nanoparticles prepared in Example 1;
[0021] Figure 3 It is a schematic diagram of the fluorescence spectrum of the fluorescent nanoparticles prepared in Example 1. DETAILED DESCRIPTION
[0022] The technical solution of the present invention is further specifically described below by examples and in conjunction with the accompanying drawings. In the specification, the same or similar reference numerals indicate the same or similar components. The following description of the embodiments of the present invention with reference to the accompanying drawings is intended to explain the overall inventive concept of the present invention and should not be construed as a limitation of the present invention.
[0023] The embodiment of the present invention provides a method for preparing a selenium nanoparticle fluorescent probe capable of in vivo imaging, comprising:
[0024] S1. Under light-proof conditions, add sodium selenite pentahydrate, citric acid and arginine into water, adjust the pH to 9-11, heat to 150℃-200℃ for reaction, and after the reaction is completed, cool to room temperature, dialyze with distilled water, and then freeze-dry to obtain purified selenium quantum dots.
[0025] In a preferred embodiment, the molar ratio of sodium selenite pentahydrate, citric acid and arginine is (0.9-1.2):(10-11):(40-41).
[0026] In a preferred embodiment, the reaction time is 6-10 hours.
[0027] In a preferred embodiment, the dialysis bag used for dialysis has a specification of 500D.
[0028] The selenium quantum dots prepared in the embodiment of the present invention have maximum excitation at 590 nm and an emission wavelength of 683 nanometers, which can reach the near-infrared region and are suitable for in vivo imaging. They have a large number of carboxyl and amino groups on the surface, which are different from the selenium quantum dots sold on the market.
[0029] S2. Under light-proof conditions, the purified selenium quantum dots are added into water, indocyanine green is added, and a reaction is carried out. After the reaction is completed, the mixture is washed with water, centrifuged, and freeze-dried to obtain the selenium nanoparticle fluorescent probe.
[0030] Selenium is an essential component of glutathione peroxidase, a component of the body's antioxidant system. Appropriate selenium supplementation can increase the activity of glutathione peroxidase, thereby improving the body's antioxidant capacity.
[0031] In a preferred embodiment, the mass ratio of selenium quantum dots to indocyanine green is (10-15) mg:(1-2) mg.
[0032] After the selenium quantum dots prepared in the embodiment of the present invention are combined with indocyanine green, on the one hand, the emission wavelength of ICG is red-shifted, the tissue penetration ability is enhanced, and deeper imaging in the body is achieved (for example, the original ICG imaging at a depth of 1 cm under the skin is changed to imaging at a depth of more than 1 cm under the skin). Such a composite material realizes imaging in spinal cord injury repair, and thus realizes real-time monitoring of the repair process. On the other hand, due to the biocompatibility and active oxygen scavenging ability of selenium quantum dots, selenium quantum dots can make the composite material have excellent antioxidant properties.
[0033] Therefore, the in vivo imaging selenium nanoparticle fluorescent probe prepared by the present invention can not only be used for real-time monitoring of the spinal cord injury repair process, but also can be used for the preparation of spinal cord injury repair biomaterials.
[0034] Example 1
[0035] 1) 5.26 mg sodium selenite pentahydrate, 38.42 mg citric acid, and 139.36 mg arginine were dissolved in 40 mL distilled water, the pH was adjusted to 11, the mixed solution was added to a reactor, heated at 180° C. for 6 h, cooled to room temperature, dialyzed against distilled water for 3-5 days, and freeze-dried to obtain purified selenium quantum dots (SeQDs).
[0036] 2) Add 20 mg of purified selenium quantum dots to 50 mL of distilled water, add 2.05 mg of indocyanine green (ICG), and react for 10 to 12 hours. Wash with water, centrifuge three times, and freeze-dry to obtain the selenium nanoparticle fluorescent probe Se@ICG.
[0037] Figure 1 Figure 1 is a schematic diagram of transmission electron microscopy (TEM) of SeQDs and Se@ICG in Example 1, wherein Figure A is a TEM image of selenium quantum dots and Figure B is a TEM image of Se@ICG. Figure 1 It can be seen that SeQDs are spherical particles with a size of about 8 nm, and Se@ICG is a spherical particle with a size of about 160 nm, which proves that the fluorescent nanoparticles are successfully prepared.
[0038] Figure 2 Schematic diagram of elemental analysis of Se@ICG in Example 1, wherein the left figure is a schematic diagram of carbon element analysis, the middle figure is a schematic diagram of selenium element analysis, and the right figure is a schematic diagram of composite analysis of carbon and selenium elements. Figure 2 It can be seen that the selenium element is evenly distributed in the fluorescent nanoparticles, proving that SeQDs are successfully combined with ICG to produce a new product Se@ICG.
[0039] Figure 3 Schematic diagram of fluorescence spectra of ICG, SeQDs in Example 1, and Se@ICG in Example 1. Figure 3It can be seen that the emission wavelength of Se@ICG has a certain red shift compared with that of ICG, which proves that the incorporation of SeQDs is beneficial to deepen the tissue penetration of Se@ICG.
[0040] Example 2
[0041] 1) Dissolve 26.3 mg sodium selenite pentahydrate, 192.1 mg citric acid, and 696.8 mg arginine in 200 mL distilled water, adjust the pH to 11, add the mixed solution into a reactor, heat at 180° C. for 6 h, cool to room temperature, dialyze with distilled water for 3-5 days, and freeze-dry to obtain purified selenium quantum dots (SeQDs).
[0042] 2) Add 39.98 mg of purified selenium quantum dots to 50 mL of distilled water, add 4.01 mg of indocyanine green (ICG), react for 10 to 12 hours, wash with water, centrifuge three times, and freeze-dry to obtain Se@ICG.
[0043] Example 3
[0044] 1) Dissolve 26.5 mg sodium selenite pentahydrate, 191.8 mg citric acid, and 700 mg arginine in 200 mL distilled water, adjust the pH to 11, add the mixed solution into a reactor, heat at 180° C. for 6 h, cool to room temperature, dialyze against distilled water for 3-5 days, and freeze-dry to obtain purified selenium quantum dots (SeQDs).
[0045] 2) Add 19.68 mg of purified selenium quantum dots to 50 mL of distilled water, add 1.97 mg of indocyanine green (ICG), react for 10 to 12 hours, wash with water, centrifuge three times, and freeze-dry to obtain Se@ICG.
[0046] The selenium nanoparticle fluorescent probe capable of in vivo imaging and its preparation method and application according to the embodiment of the present invention have at least one of the following advantages:
[0047] (1) The present invention prepares quantum dots of a specific wavelength and compounds them with ICG, thereby red-shifting the emission wavelength of ICG and enhancing tissue penetration, thereby achieving deeper in vivo imaging. It can be used to achieve in vivo imaging under specific conditions (high subcutaneous depth requirements), for example, imaging in spinal cord injury repair, thereby achieving real-time monitoring of the repair process.
[0048] (2) The selenium nanoparticle fluorescent probe prepared by the present invention not only realizes deeper imaging, but also utilizes the active oxygen scavenging ability of selenium quantum dots to make the material have excellent antioxidant properties and excellent biocompatibility.
[0049] Although some embodiments of the present general inventive concept have been shown and described, it will be appreciated by those skilled in the art that changes may be made to these embodiments without departing from the principles and spirit of the present general inventive concept, the scope of which is defined by the claims and their equivalents.
Claims
1. A method for preparing a selenium nanoparticle fluorescent probe capable of in vivo imaging, characterized in that: include: S1. Under light-proof conditions, sodium selenite pentahydrate, citric acid, and arginine are added to water, the pH is adjusted to 9-11, and the reaction is heated to 150°C-200°C. After the reaction is completed, the mixture is cooled to room temperature, dialyzed with distilled water, and then freeze-dried to obtain purified selenium quantum dots; S2. Under light-proof conditions, the selenium quantum dots are added into water, indocyanine green is added, and a reaction is carried out. After the reaction is completed, the selenium quantum dots are washed with water, centrifuged, and freeze-dried to obtain the selenium nanoparticle fluorescent probe.
2. The preparation method according to claim 1, characterized in that In step S1, the molar ratio of sodium selenite pentahydrate, citric acid and arginine is (0.9-1.2):(10-11):(40-41).
3. The preparation method according to claim 1, characterized in that: In step S1, the reaction time is 6-10 hours.
4. The preparation method according to claim 1, characterized in that: In step S1, the dialysis bag used for dialysis is of 500D specification.
5. The preparation method according to claim 1, characterized in that: In step S2, the mass ratio of selenium quantum dots to indocyanine green is (10-15) mg:(1-2) mg.
6. The preparation method according to claim 1, characterized in that: In step S2, the reaction time is 10 to 12 hours.
7. The selenium nanoparticle fluorescent probe prepared by the preparation method according to any one of claims 1 to 6.
8. Use of the selenium nanoparticle fluorescent probe as claimed in claim 7 in the preparation of biomaterials for repairing spinal cord injury.
9. Use of the selenium nanoparticle fluorescent probe as claimed in claim 7 in preparing real-time monitoring materials for spinal cord injury repair.