Application of a DNA tetrahedral nanostructure as a drug carrier for inner ear diseases

A nanostructure and tetrahedron technology, applied in the field of biomedicine, can solve the problems of biological toxicity and high cost of carriers, and achieve the effects of reducing costs, low synthesis costs, and maintaining structural integrity and stability

Active Publication Date: 2022-05-13
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To effectively solve the problems of high cost and biological toxicity of existing vectors

Method used

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  • Application of a DNA tetrahedral nanostructure as a drug carrier for inner ear diseases
  • Application of a DNA tetrahedral nanostructure as a drug carrier for inner ear diseases
  • Application of a DNA tetrahedral nanostructure as a drug carrier for inner ear diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Round window injection of neonatal mice

[0034] Neonatal mice from P2 to P3 were anesthetized with hypothermia for 2-3 minutes until they lost consciousness, and then moved to an ice pad for subsequent operations. The operation time was limited to 5-10min, and the operation was performed on the left ear of each animal. After anesthesia, a retroauricular incision was made to expose the bulla and the cochlea was observed. According to the relative position of temporal bone and facial nerve, the round window is exposed. Special care should be taken during the operation to avoid damage to the facial nerve. Using a glass microtube (25 μm) controlled by an ultramicropump, the DNA tetrahedral nanostructure solution was injected through the round window. The injection volume was controlled at 1.5-2 μL / cochlea. The negative control was injected with the same amount of Cy5 fluorophore solution. After injection, the skin incision is closed using veterinary tissue ...

Embodiment 2

[0035] Example 2: Cochlear tissue preparation and confocal imaging

[0036]Mice were killed by cervical dislocation after anesthesia, and the inner ear was carefully removed after decapitation, and transferred to a fixative solution containing 4% paraformaldehyde. The excess tissue of the inner ear was removed under a stereomicroscope, and the round window, oval window, and cochlear top bone wall were opened with a needle. Suction and blow gently into the middle cochlea. Fix at room temperature for 2 hours, rinse twice with PBS solution to remove residual fixative solution, transfer to 10% EDTA-2Na decalcification solution for 72 hours at 4°C, and change the solution every day. After the inner ear of the mouse was fully decalcified, it was transferred to a glass dish filled with PBS buffer, and the decalcified volute was carefully removed under a stereomicroscope, and the basement membrane was gradually separated from the apical gyrus to the apical gyrus, and the surrounding i...

Embodiment 3

[0045] Example 3: Preparation of DNA tetrahedron nanostructure The framework nucleic acid tetrahedron provided by the present invention is synthesized from 4 DNA single strands, which were synthesized by Shanghai Shenggong Company and named T1, T2, T3, T4-Cy5 respectively. Among them, T1, T2, and T3 are ordinary nucleotide sequences without any chemical group modification. The 5' port of T4-Cy5 is modified with a Cy5 fluorophore to allow visualization of the tetrahedral position using confocal fluorescence microscopy. The numbers of DNA single strands are T1, T2, T3, T4-Cy5 respectively, and the specific sequences are shown in Table 1:

[0046] Table 1: DNA tetrahedron sequence design

[0047]

[0048] Concrete preparation method is:

[0049] (1) Dissolve and measure the concentration of DNA single strands ordered in dry powder state with numbers T1, T2, T3, T4-Cy5.

[0050] First, centrifuge the dry-fractionated DNA molecules numbered T1, the specific operation is to ce...

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Abstract

The invention provides the application of a DNA tetrahedron nanostructure as a carrier of inner ear disease medicine; wherein, the DNA tetrahedron nanostructure reaches functional cells of the inner ear through injection through a round window. In the present invention, it is successfully confirmed that the DNA tetrahedral nanostructure can enter the functional cells of the inner ear through the mouse round window drug delivery technique. Therefore, for the first time, the present invention realizes the enrichment of the hearing hair cells of the inner ear with the DNA tetrahedron nanostructure, and provides a new way for delivering drugs for inner ear diseases. At the same time, nucleic acid molecules naturally have the advantages of low toxicity, excellent biocompatibility, etc., and the synthesis of framework nucleic acids is cheap (the cost of each injection is about a few yuan), so the DNA tetrahedron nanostructure provided by the present invention is ideal for treating inner ear diseases. ideal drug carrier.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of a DNA tetrahedron nanostructure as a carrier of inner ear disease medicine. Background technique [0002] According to WHO statistics, more than 5% of the world's population suffers from hearing loss, most of which involve damage to inner ear tissues such as auditory neurons or cochlear sensory cells. Therefore, the research on inner ear diseases is the key research field of otological diseases. [0003] In the current research, the treatment of inner ear diseases mostly adopts the way of surgical administration. The use of drug delivery vehicles to deliver targeted drugs to the inner ear through the vasculature has become a common method for the treatment of inner ear diseases. Among them, the development of synthetic drug delivery carriers provides a new way for the treatment of inner ear tissue damage diseases. For example, nanoparticles modified with...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/26A61K9/00A61K45/00A61P27/16B82Y5/00B82Y40/00
CPCA61K47/26A61K9/0019A61K45/00B82Y5/00B82Y40/00A61P27/16
Inventor 潘林强孙宇徐飞陈森陈奎廷谢乐金媛
Owner HUAZHONG UNIV OF SCI & TECH
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