Publications

Introduction: In recent years there has been ample interest in nanoscale modifications of synthetic biomaterials to understand fundamental aspects of cell-surface interactions towards improved biological outcomes. In this study, we aimed at closing in on the effects of nanotubular TiO2 surfaces with variable nanotopography on the response on human mesenchymal stem cells (hMSCs). Although the influence of TiO2 nanotubes on the cellular response, and in particular on hMSC activity, has already been addressed in the past, previous studies overlooked critical morphological, structural and physical aspects that go beyond the simple nanotube diameter, such as spatial statistics. Methods: To bridge this gap, we implemented an extensive characterization of nanotubular surfaces generated by anodization of titanium with a focus on spatial structural variables including eccentricity, nearest neighbour distance (NND) and Voronoi entropy, and associated them to the hMSC response. In addition, we assessed the biological potential of a two-tiered honeycomb nanoarchitecture, which allowed the detection of combinatory effects that this hierarchical structure has on stem cells with respect to conventional nanotubular designs. We have combined experimental techniques, ranging from Scanning Electron (SEM) and Atomic Force (AFM) microscopy to Raman spectroscopy, with computational simulations to characterize and model nanotubular surfaces. We evaluated the cell response at 6 hrs, 1 and 2 days by fluorescence microscopy, as well as bone mineral deposition by Raman spectroscopy, demonstrating substrate-induced differential biological cueing at both the short- and long-term. Results: Our work demonstrates that the nanotube diameter is not sufficient to comprehensively characterize nanotubular surfaces and equally important parameters, such as eccentricity and wall thickness, ought to be included since they all contribute to the overall spatial disorder which, in turn, dictates the overall bioactive potential. We have also demonstrated that nanotubular surfaces affect the quality of bone mineral deposited by differentiated stem cells. Lastly, we closed in on the integrated effects exerted by the superimposition of two dissimilar nanotubular arrays in the honeycomb architecture. Discussion: This work delineates a novel approach for the characterization of TiO2 nanotubes which supports the incorporation of critical spatial structural aspects that have been overlooked in previous research. This is a crucial aspect to interpret cellular behaviour on nanotubular substrates. Consequently, we anticipate that this strategy will contribute to the unification of studies focused on the use of such powerful nanostructured surfaces not only for biomedical applications but also in other technology fields, such as catalysis.

JTD Keywords: Nanotubes, Nanotopography, Spatial statistics, Stem cells, Bone quality

Cardiorespiratory Phase Synchronization (CRPS) is a manifestation of coupling between cardiac and respiratory systems complementary to Respiratory Sinus Arrhythmia. In this work, we investigated CRPS during wake and sleep stages in Polysomnographic (PSG) recordings of 30 subjects suspected from Obstructive Sleep Apnea (OSA). The population was classified into three severity groups according to the Apnea Hypopnea Index (AHI): G1 (AHI<;15), G2 (15<;=AHI<;30) and G3 (AHI>30). The synchrogram between single lead ECG and respiratory abdominal band signals from PSG was computed with the Hilbert transform technique. The different phase locking ratios (PLR) m:n were monitored throughout the night. Ratio 4:1 was the most frequent and it became more dominant as OSA severity increased. CRPS was characterized by the percentage of synchronized time (%Sync) and the average duration of synchronized epochs (AvDurSync) using three different thresholds. Globally, we observed that %Sync significantly decreased and AvDurSync slightly increased with OSA severity. A high synchronization threshold enhanced these population differences. %Sync was significantly higher in NREM than in REM sleep in G2 and G3 groups. Population differences observed during sleep did not translate to the initial wake state. Reduced CRPS could be an early marker of OSA severity during sleep, but further studies are needed to determine whether CRPS is also present during wakefulness.

JTD Keywords: Band-pass filters, Electrocardiography, Heart beat, Sleep apnea, Sociology, Statistics, Synchronization

The relationship between physicochemical properties of odor molecules and perceived odor quality is arguably one of the most important issues in olfaction and the rules governing this relationship remain unknown. Any given odor molecule will stimulate more than one type of receptor in the nose, perhaps hundreds, and this stimulation reflects itself in the neural code of the olfactory nervous system. We present a method to investigate neural coding at the glomerular level of the olfactory bulb, the first relay for olfactory processing in the brain. Our results give insights into localization of coding sites, relevance of odorant properties for information processing, and the size of coding zones.

JTD Keywords: Classification, Glomeruli, Non-parametric statistics, Odorants, Olfactory bulb, Olfactory coding, Property-activity relationship

Shape-dependent local differentials in cell proliferation are considered to be a major driving mechanism of structuring processes in vivo, such as embryogenesis, wound healing, and angiogenesis. However, the specific biophysical signaling by which changes in cell shape contribute to cell cycle regulation remains poorly understood. Here, we describe our study of the roles of nuclear volume and cytoskeletal mechanics in mediating shape control of proliferation in single endothelial cells. Micropatterned adhesive islands were used to independently control cell spreading and elongation. We show that, irrespective of elongation, nuclear volume and apparent chromatin decondensation of cells in G1 systematically increased with cell spreading and highly correlated with DNA synthesis (percent of cells in the S phase). In contrast, cell elongation dramatically affected the organization of the actin cytoskeleton, markedly reduced both cytoskeletal stiffness (measured dorsally with atomic force microscopy) and contractility (measured ventrally with traction microscopy), and increased mechanical anisotropy, without affecting either DNA synthesis or nuclear volume. Our results reveal that the nuclear volume in G1 is predictive of the proliferative status of single endothelial cells within a population, whereas cell stiffness and contractility are not. These findings show that the effects of cell mechanics in shape control of proliferation are far more complex than a linear or straightforward relationship. Our data are consistent with a mechanism by which spreading of cells in G1 partially enhances proliferation by inducing nuclear swelling and decreasing chromatin condensation, thereby rendering DNA more accessible to the replication machinery.

JTD Keywords: Cell Line, Cell Nucleus/ physiology, Cell Proliferation, Cell Size, Computer Simulation, Endothelial Cells/ cytology/ physiology, G1 Phase/ physiology, Humans, Mechanotransduction, Cellular/ physiology, Models, Biological, Statistics as Topic