Acute myeloid leukemia (AML) is a highly aggressive type of cancer caused by the uncontrolled proliferation of undifferentiated myeloblasts, affecting the bone marrow and blood. Systemic chemotherapy is considered the primary treatment strategy; unfortunately, healthy cells are also affected to a large extent, leading to severe side effects of this treatment. Targeted drug therapies are becoming increasingly popular in modern medicine, as they bypass normal tissues and cells. Two-dimensional MoS2-based nanomaterials have attracted attention in the biomedical field as promising agents for cancer diagnosis and therapy. Cancer cells typically (over)express distinctive cytoplasmic membrane-anchored or -spanning protein-based structures (e.g., receptors, enzymes) that distinguish them from healthy, non-cancerous cells. Targeting cancer cells via tumor-specific markers using MoS2-based nanocarriers loaded with labels or drugs can significantly improve specificity and reduce side effects of such treatment. SKM-1 is an established AML cell line that has been employed in various bio-research applications. However, to date, it has not been used as the subject of studies on selective cancer targeting by inorganic nanomaterials. Here, we demonstrate an efficient targeting of AML cells using MoS2 nanoflakes prepared by a facile exfoliation route and functionalized with anti-CD33 antibody that binds to CD33 receptors expressed by SKM-1 cells. Microscopic analyses by confocal laser scanning microscopy supplemented by label-free confocal Raman microscopy proved that (anti-CD33)-MoS2 conjugates were present on the cell surface and within SKM-1 cells, presumably having been internalized via CD33-mediated endocytosis. Furthermore, the cellular uptake of SKM-1 specific (anti-CD33)-MoS2 conjugates assessed by flow cytometry analysis was significantly higher compared with the cellular uptake of SKM-1 nonspecific (anti-GPC3)-MoS2 conjugates. Our results indicate the importance of appropriate functionalization of MoS2 nanomaterials by tumor-recognizing elements that significantly increase their specificity and hence suggest the utilization of MoS2-based nanomaterials in the diagnosis and therapy of AML.
Gel filtration is a versatile technique employed for biological molecules and nanoparticles, offering their reproducible classification based on size and shape. Colloidal nanoparticles are of significant interest in biomedical applications due to a large number of solution-based bioconjugation procedures. Nevertheless, the inherent polydispersity of the nanoparticles produced by various techniques necessitates the employment of high yield separation and purification techniques. Here we demonstrate the employment of gel filtration on non-stoichiometric plasmonic MoO x nanoparticles, prepared by an oxidation process during liquid-phase exfoliation of few-layer MoS2 nanosheets. This resulted in the separation of two types of MoO x particles, in the form of two different chromatographic fractions. They showed different sizes, morphological and optical properties. The fraction containing smaller particles with diameters of 1–4 nm, exhibited an increased absorbance peak in the near IR region and responded with a significant temperature increase to laser irradiation at the wavelength close to the maximal absorption. The fraction with the larger particles from 3 up to 10 nm, showed weak photoluminescence and a preferred orientation upon the deposition on a planar substrate. However, it had no absorbance in the near IR compared to the former fraction. According to our knowledge, this is the first time that the gel filtration was applied to the separation of molybdenum oxide nanomaterials. This step ensured the isolation of plasmonic MoO x nanoparticles suitable for further bioconjugation and target photothermal treatment.
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