PF127修饰的石墨烯复合材料的制备及其载药系统研究

实验制备负载阿霉素的PF127修饰的还原态石墨烯复合材料,并对其性能进行研究和评价。其方法是首先制备由弗朗尼克F127非共价功能化修饰的还原氧化石墨烯材料,记为PF127/GN,然后将该复合物对药物阿霉素进行负载,记为PF127/GN/DOX,并与未经阿霉素负载的纳米药物载体（PF127/GN）进行对比分析。观察其药物释放行为,细胞内递送过程和细胞毒性。研究发现,PF127/GN粒子大小为80μm左右,且粒子分布较均匀,厚度约增加至9.646μm;利用阿霉素对纳米载体进行负载之后,负载效果随药物浓度的增加而不断提高,负载在氧化石墨烯表面的阿霉素的释放行为可通过改变体系的pH值进行调节。负载了阿霉素的纳米药物载体能够发挥良好的释放作用,具有相对较强的生物毒性。因此经过阿霉素负载的PF127修饰的还原态石墨烯复合材料性能良好,具有广阔的应用前景。     

纳米银/碳质球复合抗菌材料的制备及其抗菌性能研究

本文研究了纳米银/碳质球复合抗菌材料的制备及其形貌与结构,以大肠杆菌为代表菌株,研究了复合抗菌材料的抗菌活性,获得了抗菌性能良好的复合抗菌材料,所制备的抗菌材料有望在有害微生物防治方面获得广泛应用。     

基于石墨烯复合材料的电化学传感器在基因检测中的研究进展

近年来,石墨烯由于其优异的物理化学性能,被广泛应用于电化学传感领域。为了解石墨烯复合材料在基因检测中的应用研究现状,笔者首先介绍了石墨烯组成结构、理化特性和制备方法;然后,重点综述了基于石墨烯复合材料(石墨烯/无机复合材料、石墨烯/有机复合材料、石墨烯/有机/无机复合材料以及其他石墨烯复合材料)修饰的电化学传感器在基因检测应用中取得的最新进展;最后,总结了石墨烯复合材料在基因检测研究应用中的优缺点并对其进一步研究提出一些建议。     

基于石墨烯及其相关材料高分析性能传感器研究进展

基于石墨烯优异的导电性能、大的比表面积、良好的生物相容性,在传感器的构建方面,表现出比其他材料更加优良的性能。石墨烯在传感领域中的应用一般通过功能化来实现,石墨烯与聚合物或纳米粒子的结合可以显著增强传感器的响应,提高检测的灵敏性。综述了近年来石墨烯及其相关材料在临床分析、环境监测和食品安全控制等传感领域中的应用研究进展,通过灵敏度、检测限等分析数据对具有良好水分散性和生物相容性,比表面大,表面修饰灵活以及制备简单的氧化石墨烯及其衍生物(含氧基团)为基础的传感器的分析性能进行了综合评价。同时对石墨烯及其衍生物这一新型传感材料在未来的研究趋势进行了展望:精确控制石墨烯单分散片的尺寸、形状;研究催化作用下的石墨烯与分析物分子电极反应间的传感机制;减少石墨烯片的聚集,精确控制石墨烯基传感系统微结构。未来可望发展具有更加强大特性的便携式、芯片化传感器,实现更短时间内复杂环境样品的多重分析,进一步提高检测灵敏性和选择性,增强传感器的稳定性和重复使用性,克服毒性和生物不容性。     

医用纳米金属及其氧化物的制备、性能与抗菌应用研究进展

抗生素在临床抗菌中发挥越来越重要作用,然而,其滥用也带来了毒副反应、出现耐药病原、免疫力降低等问题,临床亟需新的抗菌方案。近年来,纳米金属及其氧化物由于广谱抗菌活性而受到广泛关注,纳米银、纳米铜、纳米锌及其氧化物等逐渐应用于生物医用领域。本文介绍了纳米金属材料分类和导电、超塑延展、催化、抗菌等基本性能;概述了物理法、化学法和生物法等常见制备技术;总结了细胞膜、氧化应激、破坏DNA和降低细胞呼吸等4种主要抗菌机理;并综述了纳米金属及其氧化物的尺寸、形状、浓度和表面化学特性对抗菌有效性的影响以及细胞毒性、遗传毒性、生殖毒性等生物安全性的研究现状。尽管目前纳米金属及其氧化物已在医用抗菌、癌症治疗等临床领域得到应用,但诸如绿色制备工艺开发、抗菌机理完善、生物安全性改进以及应用领域拓展仍有待深入探索。     

纳米银/植物源复合抗菌剂的制备及其抑菌性能的研究

以硝酸银作为银源,水溶性淀粉作保护剂,丙酮酸钠作还原剂,氨水提供碱性环境来制备纳米银胶,并以聚乙烯吡咯烷酮(PVP)作分散稳定剂,复配红景天提取液和无患子提取液制备出纳米银/植物源复合抗菌剂。实验结果表明,纳米银胶或植物提取液仅对部分细菌或霉菌有较强抑制效果,而复合抗菌剂对细菌、霉菌均有很强抑制效果。在湿巾液中添加0.5%复合抗菌剂时,其对大肠杆菌,金黄色葡萄球菌和白色念珠菌的抗菌效率可达99%,且经过常温六个月、高温55℃一个月保存后,其抗菌活性分别可达到95%、90%左右,表明复合抗菌剂具有较强的抗菌效率及抗菌稳定性。     

载ZnO/活性碳复合材料抗菌性能及安全性研究

废催化剂微波烧结法制备的ZnO/活性碳复合材料,在多孔活性炭表面附载有ZnO,是一种新型的环保材料。为评价其抗菌活性和使用安全性,本研究进行了材料对金黄色葡萄球菌、大肠杆菌、枯草芽孢杆菌及酵母菌的抑菌试验,结果表明ZnO/活性碳复合材料有很好的抑制细菌增殖效果,低浓度下(0.1μg/mL)作用2h的抑菌率能达到90%以上,对真菌-酵母菌的抑制活性相对较低;经细胞毒性试验、动物口服急性毒性试验,皮肤过敏性试验发现,材料对小鼠经口灌喂半致死剂量(LD50)2000mg/kg,对兔皮肤刺激性积分为0,对细胞生长无影响,属于实际生物无毒级。试验结果表明ZnO/活性碳复合材料具有很好的抗菌效果及使用安全性。     

超高含量的α-酮戊二酸和谷胱甘肽促成HeLa细胞耐受纳米银

纳米银（silver nanoparticles，AgNPs）兼有优良的抗菌和抗癌作用，但病原体或癌细胞对其的耐受作用将影响其临床应用，前者耐受纳米银已见报道。本研究以HeLa细胞为模型，探讨癌细胞耐受纳米银的可能性及耐受机制。将HeLa细胞代谢物与纳米银混合后测定其抗菌活性和细胞毒性变化，并用紫外-可见（ultraviolet visible，UV-Vis）分光光度计、粒度仪和透射电镜等检测混合体系中纳米银的理化特征。用非靶向代谢组学分析纳米银结合的代谢物种类，经腹腔注射HeLa细胞建立荷瘤小鼠，并分析血清对纳米银稳定性的影响。结果表明HeLa细胞代谢物可抑制纳米银的抗癌和抗菌作用，这种抑制作用表现出剂量依赖性，对纳米银生物学活性产生抑制作用的效应代谢物耐热、不溶于氯仿、含硫元素，分子量小于1 kDa。抑制作用的实质是使纳米银发生了聚集，筛选得到有115种代谢物能结合纳米银。进一步探究发现仅当α-酮戊二酸（α-ketoglutarate，AKG）和谷胱甘肽（glutathione，GSH）的浓度共同达到一定阈值才导致纳米银聚集，而HeLa细胞代谢物中两者的浓度分别是正常宫颈上皮细胞的10倍和6倍，达到了该阈值。动物实验结果显示荷瘤小鼠血清导致纳米银聚集率显著高于健康鼠血清（P<0.05）。本研究揭示HeLa细胞中具有超常含量的α-酮戊二酸和谷胱甘肽，两者协同破坏纳米银的胶体稳定性，从而实现逃逸纳米银的抗癌作用。     

离子液体修饰碳复合材料固定化脱氢酶

为了探究离子液体修饰的氧化石墨烯-聚乙烯亚胺(GO-PEI)材料对苯丙氨酸脱氢酶固定化的影响,首先采用不同咪唑基离子液体对游离苯丙氨酸脱氢酶进行修饰,其次利用咪唑基离子液体[BMIM]Cl修饰GO-PEI,分别考察GO、GO-PEI和GO-PEI-[BMIM]Cl固定化苯丙氨酸脱氢酶的性能,然后利用扫描电镜(SEM)以及红外光探究酶固定化的机制。结果表明：咪唑基离子液体中[BMIM]Cl对酶有激活作用,其相对酶活为110.0%;GO-PEI-PheDH-[BMIM]Cl固定化酶的酶活可达93.5%,高于GO-PheDH和GO-PEI-PheDH固定化酶,且温度耐受性和pH耐受性均比游离酶有所提高;GO-PEI-PheDH-[BMIM]Cl固定化酶经过6次重复利用,仍可保持82.7%的酶活,而对照的GO-PEI固定化酶的酶活为69.7%。离子液体修饰的碳复合材料具有良好的生物相容性和导电性,在非水相酶催化和生物电催化领域具有广阔的应用前景。     

猪脱细胞真皮基质与RV-23抗菌肽复合材料的制备及生物学特性初步研究

目的:制备低免疫原性猪脱细胞真皮基质(PADM)与抗菌肽RV-23的复合材料,并对其生物学特性进行初步评价。方法:将抗菌肽RV-23分别以1、5、20μmol/L的浓度加到直径6 mm、厚约1 mm的低免疫原性PADM上,制备复合材料;菌落计数分析实验检测复合材料的抗菌能力;溶血实验检测复合材料对红细胞膜的裂解能力;CCK-8实验检测复合材料对真核细胞的细胞毒性;Tricine-SDS-PAGE检验RV-23的稳定性。结果:制备了PADM与抗菌肽RV-23复合材料;活菌计数实验表明复合材料对大肠杆菌有很强的抗菌活性,并随着抗菌肽浓度的升高不断增强;溶血实验表明PADM能有效降低RV-23裂解红细胞膜的能力,增加血液相容性;CCK-8实验显示PADM能够有效降低RV-23的细胞毒性,复合材料对人表皮角化细胞HaCaT和小鼠成纤维细胞NIH-3T3几乎没有毒性;Tricine-SDSPAGE实验结果显示复合材料抗菌肽RV-23稳定性较好。结论:PADM/RV-23复合材料比较稳定,不仅具有较强的抗菌性,而且有良好的血液相容性和极低的细胞毒性,有望成为新型创伤修复材料。     

纳米银胶/壳聚糖复合抗菌剂的制备及其在洗涤产品中抗菌性能

本文研究了纳米银胶/壳聚糖抗菌剂的制备及其形貌的表征分析,以大肠杆菌为代表菌株,研究了复合抗菌剂在洗涤产品中的抗菌效率及抗菌的稳定性,结果说明复合抗菌剂在洗涤产品中添加1.0%时,其抗菌效率达99%,经180 d长期分析,其抗菌活性仍保持95%左右。此外,复合抗菌剂对不同菌株的抗菌性能也均较强。     

Nanoparticles functionalized with ampicillin destroy multiple-antibiotic-resistant isolates of Pseudomonas aeruginosa and Enterobacter aerogenes and methicillin-resistant Staphylococcus aureus

We show here that silver nanoparticles (AgNP) were intrinsically antibacterial, whereas gold nanoparticles (AuNP) were antimicrobial only when ampicillin was bound to their surfaces. Both AuNP and AgNP functionalized with ampicillin were effective broad-spectrum bactericides against Gram-negative and Gram-positive bacteria. Most importantly, when AuNP and AgNP were functionalized with ampicillin they became potent bactericidal agents with unique properties that subverted antibiotic resistance mechanisms of multiple-drug-resistant bacteria.     

Synthesis and Optical Properties of Highly Stabilized Peptide-Coated Silver Nanoparticles

The interaction between peptide and silver nanoparticle surfaces has been increasingly of interest for bionanotechnology applications. To fully understand how to control such interactions, we have studied the optical properties of peptide-modified silver nanoparticles. However, the impacts of peptide binding motif upon the surface characteristics and physicochemical properties of nanoparticles remain not yet fully understood. Here, we have prepared sodium citrate-stabilized silver nanoparticles and coated with peptide IVD (ID₃). These nanomaterials were characterized by UV-visible, transmission electron microscopy (TEM), and z-potential measurement. The results indicate that silver nanoparticles (AgNP)-peptide interface is generated using ID₃ peptide and suggested that the reactivity of peptide is governed by the conformation of the bound peptide on the nanoparticle surface. The peptide-nanoparticle interactions could potentially be used to make specific functionality into the peptide capped nanomaterials and antibacterial applications.     

Upregulation of Metallothioneins After Exposure of Cultured Primary Astrocytes to Silver Nanoparticles

To test for the prolonged consequences of a short transient exposure of astrocytes to silver nanoparticles (AgNP), cultured primary astrocytes were incubated for 4 h in the presence of AgNP and the cell viability as well as various metabolic parameters were investigated during a subsequent incubation in AgNP-free medium. Acute exposure of astrocytes to AgNP led to a concentration-dependent increase in the specific cellular silver content to up to 46 nmol/mg protein, but did not compromise cell viability. During a subsequent incubation of the cells in AgNP-free medium, the cellular silver content of AgNP-treated astrocytes remained almost constant for up to 7 days. The cellular presence of AgNP did neither induce any delayed cell toxicity nor were alterations in cellular glucose consumption, lactate production or in the cellular ratio of glutathione to glutathione disulfide observed. However, Western blot analysis and immunocytochemical staining revealed that AgNP-treated astrocytes strongly upregulated the expression of metallothioneins. These results demonstrate that a prolonged presence of accumulated AgNP does not compromise the viability and the basal metabolism of cultured astrocytes and suggest that the upregulation of metallothioneins may help to prevent silver-mediated toxicity that could be induced by AgNP-derived silver ions.     

The synthesis of chitosan-based silver nanoparticles and their antibacterial activity

Chitosan-based silver nanoparticles were synthesized by reducing silver nitrate salts with nontoxic and biodegradable chitosan. The silver nanoparticles thus obtained showed highly potent antibacterial activity toward both Gram-positive and Gram-negative bacteria, comparable with the highly active precursor silver salts. Silver-impregnated chitosan films were formed from the starting materials composed of silver nitrate and chitosan via thermal treatment. Compared with pure chitosan films, chitosan films with silver showed both fast and long-lasting antibacterial effectiveness against Escherichia coli. The silver antibacterial materials prepared in our present system are promising candidates for a wide range of biomedical and general applications.     

Synthesis and comparative study on the antimicrobial activity of hybrid materials based on silver nanoparticles (AgNps) stabilized by polyvinylpyrrolidone (PVP)

Hybrid materials based on polyvinylpyrrolidone (PVP) with silver nanoparticles (AgNps) were synthesized applying two different strategies based on thermal or chemical reduction of silver ions to silver nanoparticles using PVP as a stabilizer. The formation of spherical silver nanoparticles with diameter ranging from 9 to 16 nm was confirmed by TEM analysis. UV-vis and FTIR spectroscopy were also applied to confirm the successful formation of AgNps. The antibacterial activity of the synthesized AgNPs/PVP against etalon strains of three different groups of bacteria—Staphylococcus aureus (S. aureus; gram-positive bacteria), Escherichia coli (E. coli; gram-negative bacteria), Pseudomonas aeruginosa (P. aeruginosa; non-ferment gram-negative bacteria), as well as against spores of Bacillus subtilis (B. subtilis) was studied. AgNps/PVP were tested for the presence of fungicidal activity against different yeasts and mold such as Candida albicans, Candida krusei, Candida tropicalis, Candida glabrata, and Aspergillus brasiliensis. The hybrid materials showed a strong antimicrobial effect against the tested bacterial and fungal strains and therefore have potential applications in biotechnology and biomedical science.     

Mechanism of Silver Nanoparticles Action on Insect Pigmentation Reveals Intervention of Copper Homeostasis

Silver nanoparticles (AgNPs), like almost all nanoparticles, are potentially toxic beyond a certain concentration because the survival of the organism is compromised due to scores of pathophysiological abnormalities past that concentration. However, the mechanism of AgNP toxicity remains undetermined. Instead of applying a toxic dose, we attempted to monitor the effects of AgNPs at a nonlethal concentration on wild type Drosophila melanogaster by exposing them throughout their development. All adult flies raised in AgNP doped food showed that up to 50 mg/L concentration AgNP has no negative influence on median survival; however, these flies appeared uniformly lighter in body color due to the loss of melanin pigments in their cuticle. Additionally, fertility and vertical movement ability were compromised due to AgNP feeding. Determination of the amount of free ionic silver (Ag⁺) led us to claim that the observed biological effects have resulted from the AgNPs and not from Ag⁺. Biochemical analysis suggests that the activity of copper dependent enzymes, namely tyrosinase and Cu-Zn superoxide dismutase, are decreased significantly following the consumption of AgNPs, despite the constant level of copper present in the tissue. Consequently, we propose a mechanism whereby consumption of excess AgNPs in association with membrane bound copper transporter proteins cause sequestration of copper, thus creating a condition that resembles copper starvation. This model also explains the cuticular demelanization effect resulting from AgNP since tyrosinase activity is essential for melanin biosynthesis. Finally, we claim that Drosophila, an established genetic model system, can be well utilized for further understanding of the biological effects of nanoparticles.     

Extracellular biosynthesis of silver nanoparticles using a novel and non-pathogenic fungus,Neurospora intermedia: controlled synthesis and antibacterial activity

In the present study, the biosynthesis of silver nanoparticles (AgNPs) using Neurospora intermedia, as a new non-pathogenic fungus was investigated. For determination of biomass harvesting time, the effect of fungal incubation period on nanoparticle formation was investigated using UV–visible spectroscopy. Then, AgNPs were synthesized using both culture supernatant and cell-free filtrate of the fungus. Two different volume ratios (1:100 and 1:1) of the culture supernatant to the silver nitrate were employed for AgNP synthesis. In addition, cell-free filtrate and silver nitrate were mixed in presence and absence of light. Smallest average size and highest productivity were obtained when using equal volumes of the culture supernatant and silver nitrate solution as confirmed by UV–visible spectra of colloidal AgNPs. Comparing the UV–visible spectra revealed that using cell-free filtrate for AgNP synthesis resulted in the formation of particles with higher stability and monodispersity than using culture supernatant. The absence of light in cell-free filtrate mediated synthesis led to the formation of nanoparticles with the lowest rate and the highest monodispersity. The presence of elemental silver in all prepared samples was confirmed using EDX, while the crystalline nature of synthesized particles was verified by XRD. FTIR results showed the presence of functional groups which reduce Ag⁺ and stabilize AgNPs. The presence of nitrate reductase was confirmed in the cell-free filtrate of the fungus suggesting the potential role of this enzyme in AgNP synthesis. Synthesized particles showed significant antibacterial activity against E. coli as confirmed by examining the growth curve of bacterial cells exposed to AgNPs.     

Multifunctional bionanocomposite films of poly(lactic acid), cellulose nanocrystals and silver nanoparticles

Nanocomposite films were prepared by the addition of cellulose nanocrystals (CNCs) eventually surfactant modified (s-CNC) and silver (Ag) nanoparticles in the polylactic acid (PLA) matrix using melt extrusion followed by a film formation process. Multifunctional composite materials were investigated in terms of morphological, mechanical, thermal and antibacterial response. The nanocomposite films maintained the transparency properties of the PLA matrix. Thermal analysis showed increased values of crystallinity in the nanocomposites, more evident in the s-CNC based formulations that had the highest tensile Young modulus. The presence of surfactant favoured the dispersion of cellulose nanocrystals in the polymer matrix and the nucleation effect was remarkably enhanced. Moreover, an antibacterial activity against Staphylococcus aureus and Escherichia coli cells was detected for ternary systems, suggesting that these novel nanocomposites may offer good perspectives for food packaging applications which require an antibacterial effect constant over time.