Comparative cytotoxicity evaluation of different size gold nanoparticles in human dermal fibroblasts

The aim of this work was to compare the effects of three commercially available gold nanoparticles (AuNPs) of different sizes (30, 50 and 90 nm) on the viability of normal human dermal fibroblasts (NHDF). In addition, we evaluated protective effect of N-Acetyl-L-cysteine (NAC), total glutathione content (GSH/GSSG), superoxide dismutase (SOD) activity and reactive oxygen species (ROS) production to investigate if oxidative stress was involved in the cytotoxic response of these AuNPs. Although AuNP-induced cytotoxicity was dose and time dependent, nanoparticle size slightly influenced the cytotoxic response of AuNPs assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase. Regarding oxidative parameters, NAC produced no significant protection of NHDF cells against treatment with any of the three AuNPs. Independently on nanoparticle size, GSH/GSSG content was drastically depleted after 24 h of incubation with the three AuNPs (less than 15% in all cases), while no statistically significant changes on SOD activity were reported (～90% of activity). The three AuNPs also caused a notable increase in the ROS production of NHDF cells. In conclusion, our data suggest that AuNP-induced cytotoxicity in NHDF is mediated by oxidative stress and it is independent of nanoparticle size.     

Differential cytotoxicity of metal oxide nanoparticles

Concerns about the potential health hazards of nanomaterials are growing. To determine the potential toxicity of metal oxide nanoparticles, human SH-SY5Y neuroblastoma and H4 neuroglioma cells were exposed to Fe₂O₃, CuO and ZnO nanoparticles and their metal ion counterparts (Fe³⁺, Cu²⁺ and Zn²⁺) at a concentration range of 0.01–100 µM for 48 h, under the cell culture conditions: 95% O₂, 5% CO₂, 85% humidity, 37°C. Their ensemble cell viability was determined by MTS cell proliferation assay. A live/dead cell assay was also performed, and cellular images were acquired by a high-content fluorescence microscope and quantified by a novel computerised image analysis protocol. Our data indicated that exposure of these nanoparticles induced differential toxic effects in both SH-SY5Y and H4 cells, and the cells had dose-dependent toxic responses to the CuO nanoparticle insult. In conclusion, the toxic responses of the nanoparticles are complex, and they warrant further in vivo studies. However, it remains to be determined if these nanopartilces have synergistically enhancing or cancelling toxic effects upon both SH-SY5Y and H4 cells.     

Zn1-xCdxTe三元量子点的微波合成及其生物相容性研究

采用微波辐射加热的方法,在水相中一步合成出高质量的谷胱甘肽修饰的Zn1-xCdxTe三元量子点,利用X射线粉末衍射(XRD)、高分辨透射电镜(HRTEM)、紫外-可见吸收光谱(UV-Vis)和荧光发射光谱(PL)等技术表征产物的物相结构和光学性质。研究了不同反应条件(如反应时间、Cd2+/Zn2+投料比、反应温度、前驱体溶液pH值)对量子点荧光性能的影响。采用MTT法研究Zn1-xCdxTe三元量子点的细胞毒性,进一步将制备好的三元量子点与肌动蛋白抗体结合作为荧光探针初步应用于细胞标记。     

Preparation and Characterization of Highly Fluorescent, Glutathione-coated Near Infrared Quantum Dots for in Vivo Fluorescence Imaging

Fluorescent probes that emit in the near-infrared (NIR, 700–1,300 nm) region are suitable as optical contrast agents for in vivo fluorescence imaging because of low scattering and absorption of the NIR light in tissues. Recently, NIR quantum dots (QDs) have become a new class of fluorescent materials that can be used for in vivo imaging. Compared with traditional organic fluorescent dyes, QDs have several unique advantages such as size- and composition-tunable emission, high brightness, narrow emission bands, large Stokes shifts, and high resistance to photobleaching. In this paper, we report a facile method for the preparation of highly fluorescent, water-soluble glutathione (GSH)-coated NIR QDs for in vivo imaging. GSH-coated NIR QDs (GSH-QDs) were prepared by surface modification of hydrophobic CdSeTe/CdS (core/shell) QDs. The hydrophobic surface of the CdSeTe/CdS QDs was exchanged with GSH in tetrahydrofuran-water. The resulting GSH-QDs were monodisperse particles and stable in PBS (phosphate buffered saline, pH = 7.4). The GSH-QDs (800 nm emission) were highly fluorescent in aqueous solutions (quantum yield = 22% in PBS buffer), and their hydrodynamic diameter was less than 10 nm, which is comparable to the size of proteins. The cellular uptake and viability for the GSH-QDs were examined using HeLa and HEK 293 cells. When the cells were incubated with aqueous solutions of the GSH-QDs (10 nM), the QDs were taken into the cells and distributed in the perinuclear region of both cells. After 12 hrs incubation of 4 nM of GSH-QDs, the viabilities of HeLa and HEK 293 cells were ca. 80 and 50%, respectively. As a biomedical utility of the GSH-QDs, in vivo NIR-fluorescence imaging of a lymph node in a mouse is presented.     

Peptide-grafted nanodiamonds: preparation, cytotoxicity and uptake in cells

Nanodiamonds that were prepared by high pressure/high temperature were functionalized with biomolecules for biological applications. Nanodiamonds (NDs, < or =35 nm) that were coated by silanization or with polyelectrolyte layers were grafted with a fluorescent thiolated peptide via a maleimido function; this led to an aqueous colloidal suspension that was stable for months. These substituted NDs were not cytotoxic for CHO cells. Their capacity to enter mammalian cells, and their localisation inside were ascertained after labelling the nucleus and actin, by examining the cells by confocal, reflected light and fluorescence microscopy.     

Cytotoxic salan-titanium(IV) complexes: high activity toward a range of sensitive and drug-resistant cell lines, and mechanistic insights

The cytotoxicities of highly efficient salan-Ti(IV) complexes toward a range of cell lines, including drug-resistant cells, are reported along with preliminary mechanistic insights. Five salan-Ti(IV) complexes were investigated toward eight different human and murine cancer-derived cell lines, including colon, ovarian, lung, cervical, pancreatic, leukemic, skin, and breast. The salan complexes are more active toward the cells analyzed than cisplatin and the known titanium compound (bzac)(2) Ti(OiPr)(2) , and no cell line resistant to the salan complexes was identified. Moreover, the salan-Ti(IV) complexes are highly active toward both cisplatin-sensitive (A2780) and cisplatin-resistant (A2780CisR) human ovarian cancer cell lines. Similarly, the salan complexes are cytotoxic toward multi-drug-resistant (ABCB1-expressing) mouse lymphoma cell lines HU-1 and HU-2. Importantly, minimal or no activity was observed toward primary murine cells (bone marrow, heart, liver, kidney, spleen, and lung), supporting selectivity for cancer cells. Additionally, the salan complexes maintain high cytotoxicity for up to 24 h following exposure to cell culture medium, whereas reference complexes (bzac)(2) Ti(OiPr)(2) and Cp(2) TiCl(2) rapidly lose much of their activity upon exposure to medium, within ~1 h. The upregulation of p53 followed by cell-cycle arrest in G(1) phase is likely one mechanism of action of the salan complexes. Taken together, the results indicate that these compounds are selectively toxic to cancer cells and are able to circumvent two independent mechanisms of drug resistance, thus expanding the scope of their potential medicinal utility.     

Preliminary study of the anticancer applications of mesoporous materials functionalized with the natural product betulinic acid

Dehydrated MCM-41 (S1) was functionalized under nitrogen with 3-chloropropyltriethoxysilane (CPTS) and 3-aminopropyltriethoxysilane (APTS) by grafting in toluene at 80 °C over 48 h to give the corresponding materials S2 and S3, respectively. Subsequently, S2 and S3 were suspended in methanol and reacted in a nitrogen atmosphere with betulinic acid (BA) for 48 h at 65 °C (in the presence of the triethylamine of S2) to give the BA-functionalized materials S4 and S5. All materials studied were characterized by powder X-ray diffraction, X-ray fluorescence, nitrogen gas sorption, multinuclear MAS NMR spectroscopy, thermogravimetry, UV spectroscopy, IR, SEM, and TEM. To study the release of BA, S4 and S5 were suspended in solutions simulating various body pH conditions (pH 7.4, 5.5, and 3.0). Results of the quantification of BA release by HPLC for S4 show a pH-dependent and very slow BA release following a logarithmic tendency, while S5 behaves differently, also pH-dependent but, in this case, fast release of BA which requires only days for total release of the therapeutic compound. In addition, the cytotoxic activity of all synthesized materials against various cancer cell lines was studied. The results show the absence of an antiproliferative effect on the surfaces without BA S1-S3, while an antiproliferative effect was observed with S4 and S5 and was attributed to the release of BA in the medium.     

Cell interaction study of amyloid by using luminescent conjugated polythiophene: implication that amyloid cytotoxicity is correlated with prolonged cellular binding

Needles and noodles: Studying amyloid toxicity is important for understanding protein misfolding diseases. Using a luminescent conjugated polythiophene, we found that cell binding of nontoxic filamentous amyloids of insulin and β2-microglobulin was less efficient than that of toxic fibrillar amyloids; this suggests a correlation between amyloid toxicity and cell binding.     

Synthesis and Biological Studies of Pyrazolyl‐Diamine PtII Complexes Containing Polyaromatic DNA‐Binding Groups

New [PtCl(pz*NN)]ⁿ⁺ complexes anchored by pyrazolyl‐diamine (pz*NN) ligands incorporating anthracenyl or acridine orange DNA‐binding groups have been synthesized so as to obtain compounds that would display synergistic effects between platination and intercalation of DNA. Study of their interaction with supercoiled DNA indicated that the anthracenyl‐containing complex L²Pt displays a covalent type of binding, whereas the acridine orange counterpart L³Pt shows a combination of intercalative and covalent binding modes with a strong contribution from the former. L²Pt showed a very strong cytotoxic effect on ovarian carcinoma cell lines A2780 and A2780cisR, which are, respectively, sensitive to and resistant to cisplatin. In these cell lines, L²Pt is nine to 27 times more cytotoxic than cisplatin. In the sensitive cell line, L³Pt showed a cytotoxic activity similar to that of cisplatin, but like L²Pt was able significantly to overcome cisplatin cross‐resistance. Cell‐uptake studies showed that L²Pt accumulates preferentially in the cytoplasm, whereas L³Pt reaches the cell nucleus more easily, as clearly visualized by time‐lapse confocal imaging of live A2870 cells. Altogether, these findings seem to indicate that interaction with biological targets other than DNA might be involved in the mechanism of action of L²Pt because this compound, despite having a weaker ability to target the cell nucleus than L³Pt , as well as an inferior DNA affinity, is nevertheless more cytotoxic. Furthermore, ultrastructural studies of A2870 cells exposed to L²Pt and L³Pt revealed that these complexes induce different alterations in cell morphology, thus indicating the involvement of different modes of action in cell death.     

Anti-melanoma, tyrosinase inhibitory and anti-microbial activities of gold nanoparticles synthesized from aqueous leaf extracts of Teraxacum officinale

There has been a tenacious search for pharmaceuticals of natural origin, as they are cost-effective and are noted for having little or no side effects. The rate at which diseases are developing resistance to synthetic drugs is quite alarming, and the side effects of these drugs remain an excruciating agony to the pharmaceutical industry. Gold nanoparticles (AuNPs) have wide applications in current technology. However, their use in medicine has not been adequately explored. Chemical methods for the synthesis are associated with environmental benignity and tissue toxicity on in vivo administration. For the first time, we have synthesized AuNPs from leaf extracts of Teraxacum officinale that were found to have significant anti-melanoma, tyrosinase inhibitory and anti-microbial effects, and hence stand as promising candidates for use in cosmetics medical and food industries.