3D打印技术制备的含银纳米复合材料的抗菌和抗病毒活性

Q4 Biochemistry, Genetics and Molecular Biology Mikrobiolohichnyi zhurnal Pub Date : 2022-12-17 DOI:10.15407/microbiolj84.03.069
N. Rybalchenko, К.S. Naumenko, L. Artiukh, V. Demchenko, О.P. Masiuchok, V. Ovsyankina, T. Rybalchenko, V.V. Yurzhenko
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The aim of the work was to study the antimicrobial and antiviral activity of silver-containing nanocomposites formed on the basis of PLA with the addition of chitosan or polyethyleneimine (PEI) by 3D printing technology. Methods. Peculiarities of the structural organization of silver-containing materials were investigated by the method of wide-angle radiography on an XRD-7000 diffractometer. The morphology of the samples was studied by transmission electron microscopy on a JEM-1230 instrument. Filament formation proceeded through melting PLA-AgPalm-PEI or PLA-AgPalm-chitosan films in an extruder at a temperature of T=160±1°С. Samples were formed from the obtained filaments using a 3D printer. Th e antimicrobial activity of silvercontaining nanocomposites was determined by the disk diffusion method against opportunistic pathogens S. aureus and E. coli. Cytotoxicity and antiviral activity were investigated using the MTT method and staining by gentian violet. Results. Analysis of wide-angle X-ray diffraction patterns of products formed by 3D printing technology at 160°C showed that PLA-Ag-PEI and PLA-Ag-chitosan materials contain metallic silver. Analysis of microphotographs of PLA-4%Ag-PEI and PLA-4%Ag-chitosan products formed by 3D printing technology showed that silver nanoparticles formed by adding chitosan as a reducing agent and stabilizer to the polymer matrix of PLA are much smaller than when using PEI. Silver-containing nanocomposite samples, such as filaments and products formed from PLA-4%Ag-PEI and PLA-4%Agchitosan films subjected to heat treatment and without heat treatment, show antimicrobial activity against test cultures of S. aureus and E. coli. It was found that nanocomposites based on PEI and chitosan do not show cytotoxic effects in MDCK and HEP-2 cultures. Nanocomposites of both types show a weak antiviral effect against adenovirus serotype 2; the reduction of infectious titer was 0.5 lgTCID50/mL. 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引用次数: 0

摘要

银纳米颗粒具有药理作用显著、对人体和环境毒性低、在极端条件下具有较高的稳定性等特点,已成为众多研究人员开发高效抗菌和抗病毒药物的热点。要用银纳米颗粒制造抗菌药物,需要使用合成聚合物和天然聚合物的基质。生物聚合物聚乳酸(PLA)由于其物理化学和工艺性能而成为3D打印(增材生产)最有前途的材料之一。本工作的目的是通过3D打印技术研究在聚乳酸的基础上添加壳聚糖或聚乙烯亚胺(PEI)形成的含银纳米复合材料的抗菌和抗病毒活性。方法。用XRD-7000衍射仪广角射线照相法研究了含银材料的结构组织特性。在JEM-1230型透射电镜上观察样品的形貌。在T=160±1°С的温度下,将PLA-AgPalm-PEI或pla - agpalm -壳聚糖薄膜熔化成丝。用3D打印机将获得的细丝制成样品。采用纸片扩散法测定含银纳米复合材料对条件致病菌金黄色葡萄球菌和大肠杆菌的抑菌活性。采用MTT法和龙胆紫染色法研究其细胞毒性和抗病毒活性。结果。在160℃下对3D打印技术形成的产品进行广角x射线衍射图分析,发现PLA-Ag-PEI和pla - ag -壳聚糖材料中含有金属银。对3D打印技术制备的PLA-4% ag -PEI和PLA-4% ag -壳聚糖产品的显微照片分析表明,在PLA的聚合物基体中加入壳聚糖作为还原剂和稳定剂形成的银纳米粒子比使用PEI时要小得多。含银纳米复合材料样品,如由PLA-4%Ag-PEI和pla -4% ag壳聚糖薄膜经热处理和不经热处理形成的细丝和产物,对金黄色葡萄球菌和大肠杆菌的试验培养物显示出抗菌活性。我们发现PEI和壳聚糖纳米复合材料在MDCK和HEP-2培养中没有细胞毒性作用。两种类型的纳米复合材料对血清2型腺病毒的抗病毒作用较弱;感染滴度降低0.5 lgTCID50/mL。所研究的纳米复合材料均未对流感病毒模型显示抗病毒作用。结论。采用3D打印技术,将PLA-4%Ag-PEI和pla -4% ag -壳聚糖薄膜制成银浓度为重量比4%的含银纳米复合材料,对金黄色葡萄球菌和大肠杆菌试验培养物具有抗菌活性,对甲型流感病毒和人类腺病毒具有抗病毒活性,并且对细胞没有细胞毒性作用。所得数据表明,所研究的含银纳米复合材料是一种很有前途的抗菌剂,可用于医药和食品工业的各个领域。
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Antimicrobial and Antiviral Activity of Silver-Containing Nanocomposites Formed by 3D Printing Technology
Silver nanoparticles have become the focus of numerous researchers to create efficient antimicrobial and antiviral agents due to their pronounced pharmacological effects, low toxicity to the human body and the environment, and high stability in extreme conditions. To create antimicrobial drugs with silver nanoparticles, matrices from polymers of both synthetic and natural origin are used. Biopolymer polylactide (PLA) is one of the most promising materials for 3D printing (additive production) due to its physicochemical and technological properties. The aim of the work was to study the antimicrobial and antiviral activity of silver-containing nanocomposites formed on the basis of PLA with the addition of chitosan or polyethyleneimine (PEI) by 3D printing technology. Methods. Peculiarities of the structural organization of silver-containing materials were investigated by the method of wide-angle radiography on an XRD-7000 diffractometer. The morphology of the samples was studied by transmission electron microscopy on a JEM-1230 instrument. Filament formation proceeded through melting PLA-AgPalm-PEI or PLA-AgPalm-chitosan films in an extruder at a temperature of T=160±1°С. Samples were formed from the obtained filaments using a 3D printer. Th e antimicrobial activity of silvercontaining nanocomposites was determined by the disk diffusion method against opportunistic pathogens S. aureus and E. coli. Cytotoxicity and antiviral activity were investigated using the MTT method and staining by gentian violet. Results. Analysis of wide-angle X-ray diffraction patterns of products formed by 3D printing technology at 160°C showed that PLA-Ag-PEI and PLA-Ag-chitosan materials contain metallic silver. Analysis of microphotographs of PLA-4%Ag-PEI and PLA-4%Ag-chitosan products formed by 3D printing technology showed that silver nanoparticles formed by adding chitosan as a reducing agent and stabilizer to the polymer matrix of PLA are much smaller than when using PEI. Silver-containing nanocomposite samples, such as filaments and products formed from PLA-4%Ag-PEI and PLA-4%Agchitosan films subjected to heat treatment and without heat treatment, show antimicrobial activity against test cultures of S. aureus and E. coli. It was found that nanocomposites based on PEI and chitosan do not show cytotoxic effects in MDCK and HEP-2 cultures. Nanocomposites of both types show a weak antiviral effect against adenovirus serotype 2; the reduction of infectious titer was 0.5 lgTCID50/mL. None of the studied nanocomposites showed antiviral action on the influenza virus model. Conclusions. The investigated silver-containing nanocomposites with a silver concentration of 4% by weight, formed by 3D printing technology from PLA-4%Ag-PEI and PLA-4%Ag-chitosan films, show antimicrobial activity against S. aureus and E. coli test cultures and antiviral activity on influenza A virus and human adenovirus and do not show a cytotoxic effect on cells. The obtained data allow us to state that the studied silver-containing nanocomposites are promising antimicrobial agents for use in various fields of medicine and the food industry.
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Mikrobiolohichnyi zhurnal
Mikrobiolohichnyi zhurnal Medicine-Microbiology (medical)
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