Development and validation of low-intensity pulsed ultrasound systems for highly controlled in vitro cell stimulation

This work aims to describe the development and validation of two low-intensity pulsed ultrasound stimulation systems able to control the dose delivered to the biological target. Transducer characterization was performed in terms of pressure field shape and intensity, for a high-frequency range (500 kHz to 5 MHz) and for a low-frequency value (38 kHz). This allowed defining the distance, on the beam axis, at which biological samples should be placed during stimulation and to exactly know the intensity at the target. Carefully designed retaining systems were developed, for hosting biological samples. Sealing tests proved their impermeability to external contaminants. The assembly/de-assembly time of the systems resulted ~3 min. Time-domain acoustic simulations allowed to precisely estimate the ultrasound beam within the biological sample chamber, thus enabling the possibility to precisely control the pressure to be transmitted to the biological target, by modulating the transducer's input voltage. Biological in vitro tests were also carried out, demonstrating the sterility of the system and the absence of toxic and inflammatory effects on growing cells after multiple immersions in water, over seven days

Development and validation of low-intensity pulsed ultrasound systems for highly controlled in vitro cell stimulation

This work aims to describe the development and validation of two low-intensity pulsed ultrasound stimulation systems able to control the dose delivered to the biological target. Transducer characterization was performed in terms of pressure field shape and intensity, for a high-frequency range (500 kHz to 5 MHz) and for a low- frequency value (38 kHz). This allowed defining the distance, on the beam axis, at which biological samples should be placed during stimulation and to exactly know the intensity at the target. Carefully designed retaining systems were developed, for hosting biological samples. Sealing tests proved their impermeability to external contaminants. The assembly/de-assembly time of the systems resulted ~3 min. Time-domain acoustic simula- tions allowed to precisely estimate the ultrasound beam within the biological sample chamber, thus enabling the possibility to precisely control the pressure to be transmitted to the biological target, by modulating the trans- ducer’s input voltage. Biological in vitro tests were also carried out, demonstrating the sterility of the system and the absence of toxic and inflammatory effects on growing cells after multiple immersions in water, over seven day

Ultrasound Stimulation of Piezoelectric Nanocomposite Hydrogels Boosts Chondrogenic Differentiation in Vitro, in Both a Normal and Inflammatory Milieu

The use of piezoelectric nanomaterials combined with ultrasound stimulation is emerging as a promising approach for wirelessly triggering the regeneration of different tissue types. However, it has never been explored for boosting chondrogenesis. Furthermore, the ultrasound stimulation parameters used are often not adequately controlled. In this study, we show that adipose-tissue-derived mesenchymal stromal cells embedded in a nanocomposite hydrogel containing piezoelectric barium titanate nanoparticles and graphene oxide nanoflakes and stimulated with ultrasound waves with precisely controlled parameters (1 MHz and 250 mW/cm2, for 5 min once every 2 days for 10 days) dramatically boost chondrogenic cell commitment in vitro. Moreover, fibrotic and catabolic factors are strongly down-modulated: proteomic analyses reveal that such stimulation influences biological processes involved in cytoskeleton and extracellular matrix organization, collagen fibril organization, and metabolic processes. The optimal stimulation regimen also has a considerable anti-inflammatory effect and keeps its ability to boost chondrogenesis in vitro, even in an inflammatory milieu. An analytical model to predict the voltage generated by piezoelectric nanoparticles invested by ultrasound waves is proposed, together with a computational tool that takes into consideration nanoparticle clustering within the cell vacuoles and predicts the electric field streamline distribution in the cell cytoplasm. The proposed nanocomposite hydrogel shows good injectability and adhesion to the cartilage tissue ex vivo, as well as excellent biocompatibility in vivo, according to ISO 10993. Future perspectives will involve preclinical testing of this paradigm for cartilage regeneration

3D MODEL VISUALIZATION ENHANCEMENTS IN REAL-TIME GAME ENGINES

This paper describes two procedures used to disseminate tangible cultural heritage through real-time 3D simulations providing accurate-scientific representations. The main idea is to create simple geometries (with low-poly count) and apply two different texture maps to them: a normal map and a displacement map. There are two ways to achieve models that fit with normal or displacement maps: with the former (normal maps), the number of polygons in the reality-based model may be dramatically reduced by decimation algorithms and then normals may be calculated by rendering them to texture solutions (baking). With the latter, a LOD model is needed; its topology has to be quad-dominant for it to be converted to a good quality subdivision surface (with consistent tangency and curvature all over). The subdivision surface is constructed using methodologies for the construction of assets borrowed from character animation: these techniques have been recently implemented in many entertainment applications known as “retopology”. The normal map is used as usual, in order to shade the surface of the model in a realistic way. The displacement map is used to finish, in real-time, the flat faces of the object, by adding the geometric detail missing in the low-poly models. The accuracy of the resulting geometry is progressively refined based on the distance from the viewing point, so the result is like a continuous level of detail, the only difference being that there is no need to create different 3D models for one and the same object. All geometric detail is calculated in real-time according to the displacement map. This approach can be used in Unity, a real-time 3

Pulmonary alveolar microlithiasis with concurrent pleural mesothelioma in a dog

Pulmonary alveolar microlithiasis (PAM) is a rare pulmonary disorder characterized by the accumulation of calcium phosphate microliths within the alveoli, with only a few cases described in animals. A 10-year-old female Bulldog was euthanized due to history of dyspnea and recurrent pleural and pericardial effusions. At necropsy, numerous multifocal to coalescent protruding nodules of 1–5 mm in diameter were scattered throughout the thoracic serosal surfaces. Moreover, lungs showed a diffuse pale gray color and had a generalized fine grainy consistency. Histological investigations revealed abundant intra-alveolar laminated microliths that stained positive with periodic acid–Schiff and von Kossa stains. The pulmonary interstitium showed multifocal, mild to moderate thickening, due to collagen deposition and mild hyperplasia of type 2 pneumocytes. The pulmonary lesion was not associated with any inflammatory response, and mineral deposition was not observed in any other organ or tissue. In addition, pulmonary, pericardial, and pleural surfaces were extensively infiltrated by an epithelioid mesothelioma. Immunohistochemical staining revealed neoplastic cells that strongly coexpressed vimentin and cytokeratin, supporting the diagnosis of mesothelioma. An overview of PAM, including pathogenesis and histological characteristics, are discussed in relation to the concurrent pleural mesothelioma. The potential cause and effect relationship between the 2 conditions could neither be established nor ruled out