Biomolecular Corona Associated with Nanostructures: The Potentially Disruptive Role of Raman Microscopy

When nanostructures and other materials are exposed to biological fluids, they are immediately covered by a layer of biological molecules, which is typically referred to as a “biomolecular corona” (BC). This represents the first component of a material that interacts with biological systems, so characterizing the composition and the dynamic evolution of BC is essential for predicting the interactions of materials and living organisms. This review provides an analysis of current BC characterization techniques, with particular attention to nanostructures involved in biomedical applications. The influence on cell–nanostructure interactions is assessed and the advantages and limitations of each technique are discussed and compared. An in-depth analysis of Raman microscopy, a relatively unexploited tool with great potential in the characterization of BC, is then conducted. Raman microscopy can be used to analyze a vast amount of specimens without the need for staining, and can provide analysis on a spatial scale of hundreds of nanometers: it may thus represent a potentially disruptive tool for the characterization of BC, as it overcomes many of the limitations posed by current techniques

Probing the Ultrastructure of Spheroids and Their Uptake of Magnetic Nanoparticles by FIB–SEM

Spheroids are 3D cellular systems largely adopted as model for high-throughput screening of molecules and diagnostics tools. Furthermore, those cellular platforms also represent a model for testing new delivery carries for selective targeting. The coupling between the 3D cell environment and the nanovectors can be explored at the macroscale by optical microscopy. However, the nanomaterial-cell interplay finds major action at the single cell and extracellular matrix level with nanoscale interactions. Electron microscopy offers the resolution to investigate those interactions; however, the specimen preparation finds major drawbacks in its operation time and preciseness. In this context, focused ion beam and scanning electron microscopy (FIB–SEM) allows for fast processing and high resolution of the cell-nanomaterial interface. Here, in fact, a novel approach is shown to prepare large-area 3D spheroid cell culture specimens for FIB–SEM. Sectioning procedures are explored to preserve the peculiar structure of spheroids and their interaction with magnetic nanovectors. The results pave the way for advanced investigations of 3D cellular systems with nano and micromaterials relevant to tissue engineering, bioelectronics, and diagnostics

Barium titanate nanoparticles and hypergravity stimulation improve differentiation of mesenchymal stem cells into osteoblasts.

BACKGROUND: Enhancement of the osteogenic potential of mesenchymal stem cells (MSCs) is highly desirable in the field of bone regeneration. This paper proposes a new approach for the improvement of osteogenesis combining hypergravity with osteoinductive nanoparticles (NPs). MATERIALS AND METHODS: In this study, we aimed to investigate the combined effects of hypergravity and barium titanate NPs (BTNPs) on the osteogenic differentiation of rat MSCs, and the hypergravity effects on NP internalization. To obtain the hypergravity condition, we used a large-diameter centrifuge in the presence of a BTNP-doped culture medium. We analyzed cell morphology and NP internalization with immunofluorescent staining and coherent anti-Stokes Raman scattering, respectively. Moreover, cell differentiation was evaluated both at the gene level with quantitative real-time reverse-transcription polymerase chain reaction and at the protein level with Western blotting. RESULTS: Following a 20 g treatment, we found alterations in cytoskeleton conformation, cellular shape and morphology, as well as a significant increment of expression of osteoblastic markers both at the gene and protein levels, jointly pointing to a substantial increment of NP uptake. Taken together, our findings suggest a synergistic effect of hypergravity and BTNPs in the enhancement of the osteogenic differentiation of MSCs. CONCLUSION: The obtained results could become useful in the design of new approaches in bone-tissue engineering, as well as for in vitro drug-delivery strategies where an increment of nanocarrier internalization could result in a higher drug uptake by cell and/or tissue constructs

Investigation of interactions between poly-l-lysine-coated boron nitride nanotubes and C2C12 cells: up-take, cytocompatibility, and differentiation

Boron nitride nanotubes (BNNTs) have generated considerable interest within the scientific community by virtue of their unique physical properties, which can be exploited in the biomedical field. In the present in vitro study, we investigated the interactions of poly-l-lysine-coated BNNTs with C2C12 cells, as a model of muscle cells, in terms of cytocompatibility and BNNT internalization. The latter was performed using both confocal and transmission electron microscopy. Finally, we investigated myoblast differentiation in the presence of BNNTs, evaluating the protein synthesis of differentiating cells, myotube formation, and expression of some constitutive myoblastic markers, such as MyoD and Cx43, by reverse transcription – polymerase chain reaction and Western blot analysis. We demonstrated that BNNTs are highly internalized by C2C12 cells, with neither adversely affecting C2C12 myoblast viability nor significantly interfering with myotube formation

Artificially altered gravity elicits cell homeostasis imbalance in planarian worms, and cerium oxide nanoparticles counteract this effect

Gravity alterations elicit complex and mostly detrimental effects on biological systems. Among these, a prominent role is occupied by oxidative stress, with consequences for tissue homeostasis and development. Studies in altered gravity are relevant for both Earth and space biomedicine, but their implementation using whole organisms is often troublesome. Here we utilize planarians, simple worm model for stem cell and regeneration biology, to characterize the pathogenic mechanisms brought by artificial gravity alterations. In particular, we provide a comprehensive evaluation of molecular responses in intact and regenerating specimens, and demonstrate a protective action from the space-apt for nanotechnological antioxidant cerium oxide nanoparticles

Delivery of thyronamines (TAMs) to the brain: A preliminary study

Recent reports highlighted the significant neuroprotective effects of thyronamines (TAMs), a class of endogenous thyroid hormone derivatives. In particular, 3-iodothyronamine (T1AM) has been shown to play a pleiotropic role in neurodegeneration by modulating energy metabolism and neurological functions in mice. However, the pharmacological response to T1AM might be influenced by tissue metabolism, which is known to convert T1AM into its catabolite 3-iodothyroacetic acid (TA1). Currently, several research groups are investigating the pharmacological effects of T1AM systemic administration in the search of novel therapeutic approaches for the treatment of interlinked pathologies, such as metabolic and neurodegenerative diseases (NDDs). A critical aspect in the development of new drugs for NDDs is to know their distribution in the brain, which is fundamentally related to their ability to cross the blood–brain barrier (BBB). To this end, in the present study we used the immortalized mouse brain endothelial cell line bEnd.3 to develop an in vitro model of BBB and evaluate T1AM and TA1 permeability. Both drugs, administered at 1 µM dose, were assayed by high-performance liquid chromatography coupled to mass spectrometry. Our results indicate that T1AM is able to efficiently cross the BBB, whereas TA1 is almost completely devoid of this property