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Method for promoting nanoparticles to enter cells efficiently based on nanosecond pulsed electric field

A nanoparticle and pulsed electric field technology, applied in the field of biology, can solve the problems of limiting clinical application and lack of uniform use standards

Inactive Publication Date: 2016-08-31
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, there is no unified standard for the use of different cells, which will seriously limit the future clinical application

Method used

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  • Method for promoting nanoparticles to enter cells efficiently based on nanosecond pulsed electric field
  • Method for promoting nanoparticles to enter cells efficiently based on nanosecond pulsed electric field
  • Method for promoting nanoparticles to enter cells efficiently based on nanosecond pulsed electric field

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Human cervical cancer HeLa cells in the logarithmic growth phase with an adhesion rate of 80% were digested, centrifuged and resuspended into a single cell suspension, and the cell concentration was adjusted to 6×10 by cell counting. 5 cells / mL, proceed as follows:

[0032] 1) Blank control group: without any treatment;

[0033] 2) Individual pulse group: apply nsPEF: parameters are selected as 100ns, 20kV / cm, 10Pulses;

[0034] 3) Magnetic particle treatment group alone: ​​co-incubate with magnetic nanoparticles (final concentration: 4 mg / mL) with an average particle size of about 10 nm in a 6-well plate for 8 hours;

[0035] 4) Combined treatment group: mixed with magnetic nanoparticles (final concentration: 4 mg / mL) with an average particle size of about 10 nm, and after the above nanosecond short pulse treatment, the cells were seeded into a 6-well plate and incubated for 8 hours.

[0036] The cell culture medium inoculated on the 6-well plate was discarded, gentl...

Embodiment 2

[0038] Human normal HaCaT cells in logarithmic growth phase with 80% adherence rate were digested, centrifuged and resuspended into a single cell suspension, and the cell concentration was adjusted to 6×10 by cell counting. 5 cells / mL, proceed as follows:

[0039] 1) Blank control group: without any treatment;

[0040] 2) Individual pulse group: apply nsPEF: parameters are selected as 100ns, 20kV / cm, 10Pulses;

[0041] 3) Magnetic particle treatment group alone: ​​co-incubate with magnetic nanoparticles (final concentration: 4 mg / mL) with an average particle size of about 10 nm in a 6-well plate for 8 hours;

[0042] 4) Combined treatment group: mixed with magnetic nanoparticles (final concentration: 4 mg / mL) with an average particle size of about 10 nm, and after the above nanosecond short pulse treatment, the cells were seeded into a 6-well plate and incubated for 8 hours.

[0043]The cell culture medium inoculated on the 6-well plate was discarded, gently washed with PBS ...

Embodiment 3

[0045] Collect human cervical cancer cell HeLa cells in the logarithmic growth phase with an adhesion rate of 80%, digest, centrifuge and resuspend into a single cell suspension, and adjust the cell concentration to 6×10 for cell counting. 5 Cells / mL were grouped as follows. After the treatment, 3000 cells per well were seeded in 96-well plates, with 3 replicate wells in each group. Place the 96-well plate at 37°C, 5% CO 2 , in an incubator with saturated humidity, after culturing for 24 hours, 10 μL of CCK-8 solution per well was added to a 96-well plate for cell viability detection.

[0046] Experimental group:

[0047] 1) Blank control group: without any treatment;

[0048] 2) Pure alternating magnetic field treatment group: the magnetic field frequency is 287kHz, the current is 35A, and the treatment time is 50min;

[0049] 3) Individual pulse group: apply nsPEF processing with different parameters: 100ns, 10 / 20 / 30kV / cm, 10 / 30Pulses;

[0050] 4) Individual heating grou...

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Abstract

The invention discloses a method for promoting nanoparticles to enter cells. The method for promoting the nanoparticles to enter the cells has the advantages that the cells are subjected to pretreatment by the nanosecond pulsed electric field to promote the number of the nanoparticles entering the cells to be increased remarkably and are heated uniformly and effectively under the action of an alternating magnetic field, the pure nanosecond pulsed electric field has little effect on activity of the cells, and accordingly the method verifies that the nanosecond pulsed electric field is capable of promoting the nanoparticles to enter the cells effectively as a safe physical pretreatment means and is of potential clinical application value on cell imaging and treatment.

Description

technical field [0001] The invention belongs to the field of biology and relates to a method for efficiently promoting the entry of nanoparticles into cells by using a nanosecond pulsed electric field. Background technique [0002] Nanoscale materials are materials ranging from 1-100nm in at least one dimension. Compared with bulk materials, nanomaterials have unique optical, electrical, mechanical and chemical properties, and can be effectively used in imaging and other fields. [0003] Nanomaterials have a high specific surface area due to their small particle size, and are widely used in the research of dyes, drugs or gene carriers, for the diagnosis and treatment of diseases, and for drug delivery, such as magnetic nanoparticles loaded with anti-tumor drugs, entering cells Afterwards, the treatment of tumors can be realized, and at the same time, magnetic nanoparticles can be used as magnetic resonance contrast agents for magnetic resonance imaging to monitor the therape...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K41/00C12N5/09C12N5/071
CPCA61K41/00C12N5/0602C12N5/0629C12N5/0693
Inventor 左莎莎张锐王瑞雪张珏方竞
Owner PEKING UNIV
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