Screening the Cytotoxicity of Single-Walled Carbon Nanotubes Using Novel 3D Tissue-Mimetic Models

Single-walled carbon nanotubes (SWNTs) are promising candidates for a wide range of biomedical applications due to their fascinating properties. However, safety concerns are raised on their potential human toxicity and on the techniques that need to be used to assess such toxicity. Here, we integrate for the first time 3D tissue-mimetic models in the cytotoxicity assessment of purified (p-) and oxidized (o-) SWNTs. An established ultrasound standing wave trap was used to generate the 3D cell aggregates, and results were compared with traditional 2D cell culture models. Protein-based (bovine serum albumin) and surfactant-based (Pluronic F68) nanotube dispersions were tested and compared to a reference suspension in dimethyl sulfoxide. Our results indicated that p- and o-SWNTs were not toxic in the 3D cellular model following a 24 h exposure. In contrast, 2D cell cultures were significantly affected by exposure to p- and o-SWNTs after 24 h, as assessed by high-content screening and analysis (HCSA). Finally, cytokine (IL-6 and TNF-α) secretion levels were elevated in the 2D but remained essentially unchanged in the 3D cell models. Our results strongly indicate that 3D cell aggregates can be used as alternative <i>in vitro</i> models providing guidance on nanomaterial toxicity in a tissue-mimetic manner, thus offering future cost-effective solutions for toxicity screening assays under the experimental conditions more closely related to the physiological scenario in 3D tissue microenvironments

Nanotechnology in pulmonary medicine

Nanotechnology in medicine—nanomedicine—is extensively employed to diagnose, treat, and prevent pulmonary diseases. Over the last few years, this brave new world has made remarkable progress, offering opportunities to address historical clinical challenges in pulmonary diseases including multidrug resistance, adverse side effects of conventional therapeutic agents, novel imaging, and earlier disease detection. Nanomedicine is also being applied to tackle the new emerging infectious diseases, including severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East Respiratory Syndrome Coronavirus (MERS-CoV), influenza A virus subtype H1N1 (A/H1N1), and more recently, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this review we provide both a historical overview of the application of nanomedicine to respiratory diseases and more recent cutting-edge approaches such as nanoparticle-mediated combination therapies, novel double-targeted nondrug delivery system for targeting, stimuli-responsive nanoparticles, and theranostic imaging in the diagnosis and treatment of pulmonary diseases.</div

RAW data manuscript

Figure raw data and statitistical analysi

Supplementary Information from Silver nanoparticles as a medical device in healthcare settings: a five-step approach for candidate screening of coating agents

Additional information on: Ag+ collection and quantification; HF5 separation and characterisation; Cytokine secretion

Highly Sensitive Single Domain Antibody–Quantum Dot Conjugates for Detection of HER2 Biomarker in Lung and Breast Cancer Cells

Despite the widespread availability of immunohistochemical and other methodologies for screening and early detection of lung and breast cancer biomarkers, diagnosis of the early stage of cancers can be difficult and prone to error. The identification and validation of early biomarkers specific to lung and breast cancers, which would permit the development of more sensitive methods for detection of early disease onset, is urgently needed. In this paper, ultra-small and bright nanoprobes based on quantum dots (QDs) conjugated to single domain anti-HER2 (human epidermal growth factor receptor 2) antibodies (sdAbs) were applied for immunolabeling of breast and lung cancer cell lines, and their performance was compared to that of anti-HER2 monoclonal antibodies conjugated to conventional organic dyes Alexa Fluor 488 and Alexa Fluor 568. The sdAbs–QD conjugates achieved superior staining in a panel of lung cancer cell lines with differential HER2 expression. This shows their outstanding potential for the development of more sensitive assays for early detection of cancer biomarkers

Assessment of the volume of SWCNT aggregates in the lung tissue sections from w/t and MPO k/o mice.

<p>a. Representative images of the lung tissue sections. Insert - higher magnifigation (2.5× zoom) of a field illustrating the presence of SWCNT (green punctuate spots pointed by white arrows). b. Quantitation of SWCNT aggregates (SWCNT volume/total lung volume) using their specific absorbance (750–850 nm), * p<0.05, <i>vs</i> w/t 1 day post exposure, # p<0.05, <i>vs</i> w/t 28 days post exposure. c. Assessment of SWCNT aggregates - number per microscopic field - using an automated IN Cell Analyser 1000 microscope, * p<0.05, <i>vs</i> w/t 1 day post exposure, # p<0.05, <i>vs</i> w/t 28 days post exposure.</p

Characterization of pulmonary inflammatory responses to SWCNT in w/t and MPO k/o mice at day 1 after pharyngeal aspiration exposure.

<p>a–c. Levels of pro-inflammatory cytokines (a - TNF-α; b - IL-6; c – MCP-1) in BAL fluid of w/t and MPO k/o mice. Mean ± SEM (n = 6 mice/group). *p<0.05, <i>vs</i> control PBS-exposed mice. d. Content of PMNs in BAL fluid of w/t and MPO k/o mice. Mean ± SEM (n = 6 mice/group). *p<0.05, <i>vs</i> control PBS-exposed mice. e. Typical microscopic images of inflammatory cells in BAL fluid with SWCNT inclusions (red arrows). f. Content of PMNs with engulfed SWCNT in BAL fluid of w/t and MPO k/o mice. Mean ± SEM (n = 6 mice/group). *p<0.05, <i>vs</i> w/t mice.</p

Changes in the content of collagen and average thickness of alveolar connective tissue in the lungs of w/t and MPO k/o mice at days 1 and 28 after pharyngeal aspiration of SWCNT.

<p>a. Accumulation of collagen in the lung of w/t or MPO k/o mice. Mean ± SEM (n = 6 mice/group). *p<0.05, <i>vs</i> control PBS-exposed mice, # p<0.05, <i>vs</i> w/t mice 28 days post exposure. b. Morphometric assessments of average thickness of alveolar connective tissue in the lung of w/t or MPO k/o mice. Mean ± SEM (n = 6 mice/group). *p<0.05, <i>vs</i> control PBS-exposed mice, # p<0.05, <i>vs</i> w/t 28 days post exposure.</p

Evaluation of SWCNT size distribution in solubilized lungs of w/t and MPO k/o mice at days 1 and 28 post exposure by t

<p>ransmission electron <b>microscopy.</b> a. Typical TEM images of SWCNT after solubilization of the lung tissue. b. Size distribution of SWCNT present in the lung at days 1 and 28 post exposure. * p<0.05, <i>vs</i> w/t 1 day post exposure, # p<0.05, <i>vs</i> w/t 28 days post exposure. c. Changes in size distribution of SWCNT at day 28 post exposure compared to day 1 post exposure.</p