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Nano composite material based on enzyme degradation enhanced photo-thermal removal of bacterial biofilms, and preparation method and application of nano composite material

A nanocomposite material and bacterial biofilm technology, applied in the field of nanocomposite materials and its preparation, can solve the problems of lack of biofilm targeting ability, large side effects, poor curative effect, etc., and achieve the problem of not easy bacterial resistance, low potential toxicity, The effect of low toxicity

Active Publication Date: 2020-04-10
NANJING UNIV OF POSTS & TELECOMM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current nanotechnology has the following problems: lack of biomembrane targeting ability, large side effects, and poor curative effect due to EPS hindrance

Method used

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  • Nano composite material based on enzyme degradation enhanced photo-thermal removal of bacterial biofilms, and preparation method and application of nano composite material
  • Nano composite material based on enzyme degradation enhanced photo-thermal removal of bacterial biofilms, and preparation method and application of nano composite material
  • Nano composite material based on enzyme degradation enhanced photo-thermal removal of bacterial biofilms, and preparation method and application of nano composite material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] MnO 2 - Preparation of amylase-PEG-ICG (MAPI) nanosheets

[0031] 1. MnO 2 Preparation of nanosheets

[0032] Take 0.6 g MnCl 2 4H 2 O (3 mM) crystals in a 50 mL reaction flask, add 10 mL H 2 O. Take 12 mL of TMA OH (tetramethylammonium hydroxide, 12 mM) solution, add 2 mL of H 2 o 2 (30%wt), and diluted with water to 20 mL. The above two solutions were mixed, the solution turned from colorless and transparent to brown, and the reaction was stirred on a magnetic stirrer for 12 h. After the reaction, the solution was taken out, centrifuged at 3000 rpm for 10 min, and the lower layer of precipitate was taken. Wash with ethanol and water three times respectively under the above centrifugation conditions, and finally add aqueous solution to dilute to 30 mL. Take 10 mL of cleaned MnO 2 , add 20 mL H 2 O, use a probe sonicator to sonicate for 10 h (power: 40%, time interval: 5 s), centrifuge the obtained solution at 12000 rpm for 45 min, discard the lower sediment...

Embodiment 2

[0041] Biocompatibility of MAPI Nanosheets

[0042] HeLa cells were cultured in DMEM medium containing FBS (10%), penicillin (80 U / mL) and streptomycin (0.08 mg / mL). Add the collected logarithmic phase cells into a 96-well plate (100 μL per well, adjust the cell density to 10 3 -10 4 / hole), placed in a 5% CO 2 Incubate in a 37°C incubator. After the cells adhered to the wall, the culture medium was removed, and 100 μL of MAP (MnO 2 : 0, 5, 10, 20, 40, 80, 160 μg / mL; amylase: 0, 18, 36, 72,144, 288, 576 μg / mL) and MAPI (MnO 2 : 0, 5, 10, 20, 40, 80, 160 μg / mL; amylase: 0,18, 36, 72, 144, 288, 576 μg / mL; ICG: 0, 1.6, 3.2, 6.4, 12.8, 25.6 , 51.2 μg / mL) of DMEM medium (5 replicate wells for each concentration), and then placed in a 5% CO 2 Incubate for 24 h in a 37°C incubator in the dark. Afterwards, 20 μL of MTT solution (5 mg / mL, 0.5% MTT) was added to each well, and the incubation was continued for 4 h in an incubator protected from light. The incubation was terminate...

Embodiment 3

[0045] The effect of MAPI nanosheets on removing bacterial biofilm

[0046] Stretch out a single colony of methicillin-resistant Staphylococcus aureus (MRSA, ATCC43300) in 5 mL of liquid LB medium, incubate and grow in a constant temperature shaker for 10-12 h (rotational speed: 220 rpm, temperature: 37°C), and obtain the Count the MRSA suspension in growth phase. Wash the bacterial solution three times with normal saline (centrifugation condition: 10000 rpm, 3 min), and quantify it with a microplate reader (OD 600 = 0.1, the bacterial solution concentration is 1×10 7 CFU / mL). Dilute the washed bacterial solution to 1×10 with LB medium containing 1% glucose 8 CFU / mL was added to a 96-well plate (200 μL per well) and a 6-well plate containing silicon chips (3 mL per well), and incubated in a 37°C incubator for 2 days to obtain MRSA biofilms.

[0047] (A) Crystal violet staining method

[0048] refer to figure 2 a and 2c, remove the supernatant from the biofilm grown in...

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Abstract

The invention discloses a nano composite material based on enzyme degradation enhanced photo-thermal removal of bacterial biofilms, and a preparation method and application of the nano composite material. The nano composite material is a MnO2 nano sheet (namely MnO2-amylase-PEG-ICG NSs, MAPI for short) modified by amylase, polyethylene glycol (PEG) and indocyanine green (ICG). The material can degrade extracellular polymers (EPS) of a biofilm through the amylases, damage the structure of the biofilm, and further enhance the photo-thermal sterilization effect of the ICG. The MAPI provided by the invention has the advantages of good stability, excellent bacteria killing effect in a biofilm and small cytotoxicity, and can meet basic conditions of biological medicine application.

Description

technical field [0001] The invention belongs to the technical field of nano anti-biofilm, and in particular relates to a nanocomposite material based on enzymatic degradation enhanced photothermal removal of bacterial biofilm, a preparation method and application thereof. Background technique [0002] Bacterial infections pose a serious threat to human health, causing more than 2 million illnesses and more than 23,000 deaths in the United States each year. Among them, about 80% of human bacterial infections are related to biofilms formed on living tissues. In biofilms, bacteria are held together by self-synthesized EPS, which typically consists of polysaccharides, proteins, and extracellular DNA (eDNA). Protected by this sticky and firm framework, bacteria within biofilms are significantly more resistant to antibiotics, host immune defenses, and environmental stress than free bacteria. Bacterial biofilms, if not completely eradicated, often lead to persistent infection and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K41/00A61K38/47A61K47/60A61K47/69A61P31/04B82Y5/00B82Y20/00B82Y30/00B82Y40/00
CPCA61K47/60A61K47/6923A61K38/47A61K41/0052A61P31/04B82Y5/00B82Y20/00B82Y30/00B82Y40/00A61K2300/00
Inventor 宇文力辉汪联辉甘思钰修尉峻仇球
Owner NANJING UNIV OF POSTS & TELECOMM
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