CN114569718B - Preparation method of nano material for imaging and tumor treatment - Google Patents

Preparation method of nano material for imaging and tumor treatment Download PDF

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CN114569718B
CN114569718B CN202111392247.9A CN202111392247A CN114569718B CN 114569718 B CN114569718 B CN 114569718B CN 202111392247 A CN202111392247 A CN 202111392247A CN 114569718 B CN114569718 B CN 114569718B
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CN114569718A (en
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刘伟生
俞彬
王畇鉴
王文杰
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Xinyi Yilan Green Material Industry Research Institute Co ltd
Lanzhou University
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Abstract

The invention designs a composite nano material for biological imaging and tumor treatment by combining a Cu-MOF material and a long afterglow material, and the chemical formula of the composite nano material is SiO 2 @Zn 1+x Ga 1.9‑x‑y O 4 : cr (y) @ HKUST-1, wherein x is more than or equal to 0 and less than or equal to 0.2, and y is more than or equal to 0.001 and less than or equal to 0.015. The prepared composite nano material can show good near-infrared luminescence in a mouse body, and can achieve the effect of completely curing the mouse tumor through the synergistic treatment of chemodynamic therapy and photothermal therapy.

Description

Preparation method of nano material for imaging and tumor treatment
Technical Field
The invention belongs to the field of biomedicine, and particularly relates to a nano material for combining MOF and a long afterglow material and being used for biological imaging and tumor treatment and a preparation method thereof.
Background
Cancer is one of the major diseases threatening human life at present, because of its complicated etiology, high incidence rate and high mortality rate. The traditional cancer treatment methods mainly comprise operations, medicines, radiation therapy and the like. However, most treatments damage healthy tissues and are affected by hypoxia, low pH and high intracellular expression of substances in the tumor microenvironment, and the treatment effect is unsatisfactory. Therefore, it is of great interest to search for new therapeutic approaches that avoid these disadvantages.
In order to solve the problems, the design and research are carried out, the long afterglow material is combined with the MOFs material, and the Cu-MOFs material (HKUST) is loaded by the non-toxic long afterglow nano material with good near infrared luminescence property, so that the near infrared afterglow luminescence and the targeted synergistic treatment of the tumor are realized. Experiments carried out in cells and mice prove that the prepared material (HSZGO) can carry out high-efficiency biological imaging and shows excellent near-infrared afterglow luminescence characteristics. The nano material can effectively inhibit the growth of tumors, can effectively cure tumor mice when experiments are carried out in the bodies of the mice, avoids side effects and drug resistance of the tumors, can pertinently inhibit the proliferation of tumor cells, and has high utilization value.
Disclosure of Invention
Aiming at the problems of single effect, side effect, tumor drug resistance and the like of the existing drugs, the invention aims to provide a preparation method of a composite material for biological imaging and tumor treatment, and adopts the following technical scheme:
a composite structured nanomaterial: siO 2 2 @Zn 1+x Ga 1.9-x-y O 4 ：Cr(y)@HKUST-1
Wherein x is more than or equal to 0 and less than or equal to 0.2, y is more than or equal to 0.001 and less than or equal to 0.015
Further optimal concentration: x is 0.1, y is 0.005
A preparation method of a composite nano material combining a long afterglow material and an MOF material comprises the following steps:
1. with Zn (NO) 3 ) 2 ·6H 2 O,Ga(NO 3 ) 3 H2O and Cr (NO) 3 ) 2 9H20 is taken as a raw material, is weighed according to a certain stoichiometric ratio, is dissolved in a mixed solution of an aqueous solution and an ethanol solution, and is added with mesoporous silica for ultrasonic mixing;
2. drying the mixed solution in an oven at 60 ℃ for 12h, fully grinding the obtained solid powder in a mortar, then putting the mortar into a muffle furnace in an air atmosphere, heating to 800 ℃, sintering and calcining for 5h.
3. Adding CuCl 2 Dissolving in ultrapure water to prepare solution A, and completely dissolving trimesic acid in ethanol to prepare solution B.
4. And (3) adding the material obtained in the step (3) into the solution A, fully stirring for ultrasonic reaction for 1 hour, then adding the solution B, fully stirring the mixed solution at room temperature for reaction for 1 hour, and pouring the mixed solution into a polytetrafluoroethylene lining.
5. And (3) placing the polytetrafluoroethylene lining into an autoclave, placing the autoclave into a muffle furnace with an air atmosphere, heating to 120 ℃, and preserving heat for 12 hours.
6. Centrifugally collecting and grinding the material obtained in the step 5 to obtain the composite nano material combining the long afterglow material and the MOF material
Nitrate which is easy to dissolve in water in the step (1) is mixed according to a proportion, and mesoporous silicon dioxide is used as a template agent.
And (3) in the step (2), a long afterglow structure is formed through high temperature calcination in an air atmosphere.
In the step (5), the MOF material is promoted to be formed through high-temperature reaction in a muffle furnace.
The composite nano material combining the long afterglow material and the MOF material prepared by the invention has the following advantages:
(1) The invention provides a preparation method of a composite material combining a nano long afterglow material and an MOFs material, which is simple to operate and controllable in size;
(2) The prepared composite material has complete shape, the size of the nano particles can be observed through detection, and the composite material can be applied to the biological field;
(3) The prepared composite material can stably carry out high-quality imaging in organisms;
(4) The prepared composite material can specifically inhibit tumor cells, and mouse tumors can be completely cured;
(5) The prepared composite material can avoid the damage of toxic and side effects to normal tissues and organs.
Drawings
FIG. 1 is an X-ray diffraction pattern (XRD) of a composite material.
FIG. 2 is a Scanning Electron Microscope (SEM) of the composite material.
FIG. 3 shows cytotoxicity (a) and cytotherapeutic effect (b) of the composite material.
Figure 4 is mouse in vivo imaging (a, b) and organ in vitro imaging (c, d) of the composite.
FIG. 5 is Cu of composite material 2+ Distribution in organ (a) and tumor site retention assay (b) (ICP-MS).
Figure 6 is tumor mouse photothermal imaging of the composite material.
FIG. 7 is experimental data for HSZGO treated tumor mice.
FIG. 8 is the data of the toxicity and side effects of the samples on normal tissue cells.
The specific implementation method comprises the following steps:
the structural formula of the composite nano material combining the long afterglow material and the MOF material is SiO 2 @Zn 1+x Ga 1.9-x-y O 4 ：Cr(y)
Wherein x is more than or equal to 0 and less than or equal to 0.2, y is more than or equal to 0.001 and less than or equal to 0.015
When the value of X is 0.1, the phase change of the long-afterglow material is not caused, the stability of the crystal structure can be well maintained, and the afterglow time of the composite material is effectively prolonged.
When the value of Y is 0.005, the phase change of the long afterglow material can not be caused, the stability of the crystal structure can be well maintained, and the composite material shows good near infrared luminescence and better afterglow performance.
When the composite nano material is prepared, the nano material with the near-infrared long-afterglow luminescence property is prepared by adopting a layer-by-layer synthesis method for biological imaging, and the MOF structure coated on the outer layer of the nano material can be effectively activated in a targeted manner in tumor cells and is used for tumor treatment.
The present invention is further illustrated below by specific examples.
Embodiment 1 preparation method of MOF and long afterglow composite material
Preparing the nano afterglow material:
taking Zn (NO) 3 ) 2 ·6H 2 O,Ga(NO 3 ) 3 ·H 2 O and Cr (NO) 3 ) 3 ·9H 2 0 is mixed and dissolved in a mixed solution of 0.5mL of water and 0.5mL of ethanol, 100mg of nano mesoporous silica powder is added, and ultrasonic vibration is carried out for 2 hours, so that the mixed solution and the silica are fully mixed to obtain a colloidal mixture. The mixture was dried in an oven at 60 ℃ for 12h. The resulting white powder was ground thoroughly in a mortar, transferred into a corundum crucible, placed in a muffle furnace and calcined at 800 ℃ for 3h. And grinding and storing the powder to obtain the nano-particle SZGO after the end.
MOFs material loaded on surface of nanoparticle
200mg of CuCl 2 The solution A was prepared by dissolving in ultrapure water, and the solution B was prepared by completely dissolving 174mg of trimesic acid in ethanol. The resulting SZGO nanoparticles were placed in a round bottom flask and stirred thoroughly with solution a for 1h. Solution B was then added to the round bottom flask and stirred well for 1 hour, then transferred to a teflon liner. The lining is placed in an autoclave and put into a muffle furnace, and the temperature is kept at 120 ℃ for 12h. The resulting pale blue solid powder was collected by centrifugation and then dried in a muffle oven at 150 ℃ for 12h to give the desired sample named HSZGO.
Bioimaging of HSZGO:
the nanoparticle (12 mg/mL,100 μ L) aqueous solution was irradiated with 254nm UV light for 10 min and injected into mice for near infrared luminescence imaging by IVIS II Lumina system. Then, the nano particles are irradiated for 3min in vitro by using a 650nm LED lamp, and luminescence images are captured again to verify the charging imaging capability of the nano particles in vivo.
H22 tumor-bearing mice are euthanized after being injected with HSZGO for 6H, then main organs (heart, liver, spleen, lung and kidney) and tumors are dissected and collected, and luminescent signals are collected by using an IVIS II Lumia system for imaging after being irradiated for 5min by a 254nm ultraviolet lamp.
Mouse tumor treatment with HSZGO:
female KM mice weighing about 18 g (4-6 weeks) were obtained from the laboratory animal center of Lanzhou university, and H22 tumor cells (3.5X 10) were cultured for 1 week 5 Per 100 uL/site) is implanted into the right underarm, and a tumor-bearing mouse model is established. When the tumor grows to 50-100 mm on average 3 On the left and right, all tumor-bearing mice were randomly divided into 5 groups (n = 10/group) (a) physiological saline (100 uL); (B) normal saline +808nm illumination; (C) HSZGO (20 mg/kg,100 uL); (D) HSZGO (20 mg/kg,100 uL) +808nm illumination; (E) s-adenosylmethionine (SAM, 40mg/kg,100 uL) + HSZGO + laser. Group E, injected with SAM 12H before treatment, can activate over-expressed CBS in tumor cells and catalyze L-cysteine to generate H 2 And S. Groups of tumor-bearing mice implanted with H22 tumors were injected with samples every 7 days. The treatment group requiring laser irradiation irradiated the tumor site of the mouse with 808nm laser (2W/cm 2) 26h after the injection of the sample and compared with the non-irradiated control groups A, C to evaluate the effect of the synergistic treatment. To record the change in tumor volume, tumor size was measured daily with digital calipers, as calculated by V = Length × Width 2 /2. Relative tumor volume in V/V 0 Calculation of (V) 0 Untreated initial tumor volume). During the treatment, the body weight of the mice was measured daily with a laboratory balance to obtain the relative body weight. Finally, all mice were euthanized, tumors dissected, tumor size recorded, paraffin embedded and H&E and TUNEL staining, and observing the apoptosis condition in the tumor.
Evaluation of HSZGO toxicity:
the major organs of each group of mice were dissected and fixed with 4% formaldehyde solution. The major organs were sectioned, paraffin embedded, H & E and TUNEL stained, imaged using an optical microscope, histologically analyzed, and examined for damage to normal tissue organs.
As can be seen from the materials in the drawings, FIG. 1 is an X-ray diffraction pattern (XRD) of the composite material, and the composite material simultaneously has diffraction peaks of various angles of the long afterglow material and the MOF material, has higher intensity and shows good crystallinity;
FIG. 2 is a Scanning Electron Microscope (SEM) of the composite material, and the nano particles show good monodispersity and are about 150nm in size;
FIG. 3 shows cytotoxicity (a) and cytotherapeutic effect (b) of the composite material, the material has no toxicity to normal cell GES-1, and has obvious proliferation inhibition effect on two tumor cells H22 and HepG 2;
FIG. 4 is a graph of in vivo mouse imaging (a, b) and in vitro organ imaging (c, d) of composite materials showing good near infrared imaging ability and good luminescent signals observed in both in vivo and in vitro mice;
FIG. 5 is Cu of composite material 2+ Distribution in organs (a) and tumor site retention detection (b) (ICP-MS), the material has better accumulation capacity in tumor tissues than in other organ tissues, and can be retained in the tumor for a longer time.
FIG. 6 shows the photothermal imaging of the tumor mouse made of the composite material, wherein the material shows excellent photothermal performance in the mouse body, and can be rapidly heated within three minutes under 808nm illumination for tumor treatment;
fig. 7 is experimental data of tumor mice treated with HSZGO sample, the body weight of the mice did not change abnormally during the treatment process, the tumor volume of the mice in the control group increased to about 10 times, and the treated mice did not increase any more, even cured directly.
FIG. 8 is the data of the toxicity and side effects of the sample on normal tissue cells, which indicates that the material does not cause damage to other organs during the tumor treatment process.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention shall fall within the protection scope of the present invention.

Claims (8)

1. A composite nano material is characterized in that: the chemical formula is as follows:
SiO 2 @Zn 1+x Ga 1.9-x-y O 4 ：Cr（y）@HKUST-1
0≤x≤0.2，0.001≤y≤0.015。
2. the composite nanomaterial of claim 1, wherein: x is 0.1 and y is 0.005.
3. Composite nanomaterial according to claim 1, characterized in that: the preparation method comprises the following steps:
(1) With Zn (NO) 3 ) 2 ·6H 2 O, Ga (NO 3 ) 3 ·H 2 O and Cr (NO) 3 ) 2 ·9 H 2 Weighing O as a raw material, dissolving the O in a mixed solution of an aqueous solution and an ethanol solution, and adding mesoporous silica for ultrasonic mixing;
(2) Drying the mixed solution in an oven, fully grinding the obtained solid powder in a mortar, then putting the ground solid powder into a muffle furnace in an air atmosphere, sintering and calcining to obtain a material named as SZGO;
(3) Adding CuCl 2 Dissolving in ultrapure water to prepare a solution A, and completely dissolving trimesic acid in ethanol to prepare a solution B;
(4) Adding the material SZGO obtained in the step (2) into the solution A, fully stirring for ultrasonic reaction for 1h, then adding the solution B, fully stirring the mixed solution at room temperature for reaction, and pouring the reaction mixture into a polytetrafluoroethylene lining;
(5) Placing the polytetrafluoroethylene lining in a high-pressure kettle, putting the high-pressure kettle in a muffle furnace in an air atmosphere, heating and preserving heat;
(6) And (4) centrifugally collecting and grinding the material obtained in the step (5) to obtain the composite nano material named as HSZGO.
4. Composite nanomaterial according to claim 3, characterized in that: in the preparation method, zn (NO) 3 ) 2 ·6H 2 O, Ga (NO 3 ) 3 •H 2 O and Cr (NO) 3 ) 2 ·9 H 2 O mixed and dissolved in a mixed solution of water and ethanol of 1.
5. The composite nanomaterial according to claim 3, wherein the sintering process in the step (2) is high-temperature calcination at 800 ℃ for 3h, and a temperature programming of 5 ℃/min is required.
6. The composite nanomaterial of claim 3, wherein the HSZGO nanoparticles obtained in step (6) are maintained in a muffle furnace at 120 ℃ for 12 hours.
7. Use of a composite nanomaterial according to any of claims 1 to 6 in the preparation of a bioimaging material.
8. Composite nanomaterial according to any of claims 1 to 6, characterized by the use in the preparation of a material for the treatment of tumors.
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