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New biological application of black phosphorus nanosheet

A nanosheet and black phosphorus technology, applied in the field of biomedicine, can solve the problems of limiting the survival rate of transplanted cells and nerve tissue damage, and achieve the effects of improving survival rate, resisting oxidative damage, and improving antioxidant capacity

Pending Publication Date: 2022-06-03
SUN YAT SEN UNIV SHENZHEN +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Oxidative damage caused by ischemia-reperfusion not only severely damages the neural tissue, but also greatly limits the cell survival rate of transplantation therapy

Method used

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  • New biological application of black phosphorus nanosheet
  • New biological application of black phosphorus nanosheet
  • New biological application of black phosphorus nanosheet

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] In this embodiment, a black phosphorus nanosheet is prepared, and the specific process is as follows:

[0045] Disperse ≤30mg of black phosphorus powder in 50mL of N-methylpyrrolidone solution, use a probe-type ultrasonic crusher (parameter setting: ultrasonic for 5s, stop for 5s, 50% power) in an ice-water bath for 24h, and centrifuge for 1000 ~ 4000r segment precipitated; washed twice with ultrapure water.

[0046] Concentration test: Disperse the product again in an appropriate amount of pure water. The concentration of black phosphorus nanosheets was determined by measuring the absorbance at a wavelength of 460 nm with a UV spectrophotometer. Sonicate in a water bath before use.

[0047] Characterization: The size of the black phosphorus nanosheets was measured using transmission electron microscopy and a particle sizer. The result is as figure 1 shown, where figure 1 (a) is the TEM image of the black scale nanosheets, scale bar: 2 μm; figure 1(b) is the parti...

Embodiment 2

[0050] This embodiment cultivates a kind of neural precursor cells, and the specific process is:

[0051] figure 2 (a) A flow chart is shown for differentiation from iPSCs to NPCs with black phosphorus nanosheets added on day 8 of differentiation. mTeSR is iPSC medium; NIM1 is NPC induction medium 1, the specific components are 50% DMEM / F12, 50% Neurobasal, 1x N2, 1x B27, 1X GlutaMAX, 10ng / mL hLIF, 4μM CHIR99021, 3μM SB431542, 2μM Dorsomorphin, and 0.1μM Compound E; NIM2 is NPC induction medium 2, the specific components are 50% DMEM / F12, 50% Neurobasal, 1x N2, 1x B27, 1X GlutaMAX, 10ng / mL hLIF, 4μM CHIR99021, 3μM SB431542, and 0.1μM Compound E; NSMM is the maintenance medium for NPC, the specific components are 50% DMEM / F12, 50% Neurobasal, 1x N2, 1x B27, 1X GlutaMAX, 10ng / mL hLIF, 3μM CHIR99021, and 2μMSB431542. like figure 2 As shown in (a), on the 8th day of differentiation from induced human-derived induced pluripotent stem cells (iPSCs) to neural precursor cells (NP...

Embodiment 3

[0057] This example compares the effects of the normal control group and the black phosphorus nanosheet group on the expression of antioxidant genes in NPC cells. The specific process is as follows:

[0058] According to the differentiation method of Example 2, NPC cells of P3 generation (normal control group and black phosphorus-treated group) were collected, and total RNA was extracted, and then qPCR experiment was performed. The result is as image 3 shown. in, image 3 (a) qPCR detects the expression of glutamate transporter and related antioxidant genes in NPC; image 3 (b) WB (Western blot) to detect the expression of antioxidant-related proteins and image 3 (c) Quantification. Among them, GAPDH is the internal reference protein of total protein, and Lamin-B1 is the internal reference protein of nuclear protein. C-Nrf2 is the Nrf2 protein in the cytoplasm, and N-Nrf2 is the Nrf2 protein in the nucleus.

[0059] from image 3 (a) The gene expression results showed...

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Abstract

The invention provides novel biological application of black phosphorus nanosheets, and provides a neural precursor cell culture medium which comprises the black phosphorus nanosheets and can improve the oxidation resistance of neural precursor cells, so that oxidative damage caused by stripping of hydrogen peroxide and sugar oxygen is effectively resisted; and the survival rate of the neural precursor cells in an in-vivo or in-vitro stress environment is improved.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and particularly relates to a new biological application of black phosphorus nanosheets. Background technique [0002] Stroke is a malignant disease caused by cerebral vascular lesions, among which ischemic stroke accounts for more than 80% of stroke types, and has the characteristics of high mortality and high disability rate. Timely thrombus removal after ischemia occurs can greatly improve patient outcomes. However, after restoring blood perfusion, excess oxidative substances can trigger oxidative stress. Excessive reactive oxygen species directly attack lipids, proteins, and nucleic acids, causing them to oxidize, leading to cell damage and secondary damage to nerve tissue. Oxidative stress and a series of excessive inflammatory responses are the main obstacles to neurological recovery after ischemic stroke. [0003] Animal models and clinical trials have demonstrated that transplantati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0797C12N5/10A61K33/42A61P25/00A61P9/10A61P39/06
CPCC12N5/0618A61K33/42A61P25/00A61P9/10A61P39/06C12N2500/05C12N2506/45C12N2510/00
Inventor 邓文斌和福美刘赣刘泽琦熊月齐军阳林珣
Owner SUN YAT SEN UNIV SHENZHEN
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