Modellizzazione numerica e caratterizzazione sperimentale di macchine e componenti oleodinamici

Nella prima parte di questa tesi di dottorato sono presentate le attività svolte, di carattere numerico, ai fini della modellizzazione di macchine volumetriche ad ingranaggi esterni. In particolare viene dapprima presentato un modello a parametri concentrati utilizzato per l’analisi dei fenomeni che coinvolgono l’area di ingranamento della macchina; un codice di calcolo associato al modello è stato sviluppato ed utilizzato per la determinazione dell’influenza delle condizioni di funzionamento e delle caratteristiche geometriche della macchina sulle sovra-pressioni e sull’eventuale instaurarsi della cavitazione nei volumi tra i denti che si trovano nell’area di ingranamento. In seguito vengono presentati i risultati ottenuti dall’analisi del bilanciamento assiale di diverse unità commerciali, evidenziando l’influenza delle caratteristiche geometriche delle fiancate di bilanciamento; a questo proposito, viene presentato anche un semplice modello a parametri concentrati per valutare il rendimento volumetrico della macchina ad ingranaggi esterni, con l’intenzione di usare tale parametro quale indice qualitativo della bontà del bilanciamento assiale. Infine, viene presentato un modello completo della macchina ad ingranaggi esterni, realizzato in un software commerciale a parametri concentrati, che permette di analizzare nel dettaglio il funzionamento della macchina e di studiare anche l’interazione della stessa con il circuito idraulico in cui è inserita. Nella seconda parte della tesi si presentano le attività legate alla messa in funzione di due banchi prova idraulici per la caratterizzazione sperimentale di macchine volumetriche e componenti di regolazione, con particolare attenzione dedicata alla messa a punto del sistema di acquisizione e gestione dei dati sperimentali; si presentano infine i risultati di alcune prove eseguite su componenti di regolazione e macchine volumetriche.

Studio Sperimentale e Modellazione della Separazione di Proteine con Membrane di Affinità

Chromatography represents one of the most important and widely used unit operation in the biotechnology industry. However this technique suffers from several limitations such as high pressure drop, slow mass transfer through the diffusive pores and strong dependence of the binding capacity on flow rate. In this work, affinity membranes with improved capacity have been considered as an alternative technology for the capturing step in antibody manufacturing. Several affinity membranes have been prepared starting from various membrane supports. Different affinity ligands have been utilized like Protein A, the natural ligand of choice for antibodies, as well as synthetic ligands that exhibit affinity for the Fc portion of antibodies. The membranes have been characterized in detail: binding and elution performance was evaluated in adsorption experiments using pure IgG solutions, while membrane selectivity was evaluated using complex solutions like a cell culture supernatant. The most promising affinity membranes were extensively tested in dynamic experiments. The effects of operating parameters like feed concentration and flow rate on separation performances like binding capacity, selectivity and process yield have been studied in detail in order to find the optimal conditions for binding and elution steps. The membranes have been used over several complete chromatographic cycles to evaluate the effects of ageing and of membrane regeneration on dynamic binding capacity. A novel mathematical model is proposed that can describe all the chromatographic steps involved in the membrane affinity chromatography process for protein purification. The mathematical description is based on the species continuity equation coupled with a proper binding kinetic equation, and suitable to describe adequately the dispersion phenomena occurring both in the micro-porous membranes as well as in the extra-column devices used in the system. The model considers specifically all the different chromatographic steps, namely adsorption, washing and elution. The few relevant fitting parameters of the model were derived from a calibration with the experimental affinity cycles performed with pure IgG solutions, then the model is used to describe experimental data obtained in chromatographic cycles carried out with complex feeds as the cell culture supernatant. Simulations reveal a good agreement with experimental data in all the chromatography steps, both in the case of pure IgG solutions and for the cell culture supernatant considered.

3D nanostructured microcarriers for cell therapy in regenerative medicine

Supercritical Emulsion Extraction technology (SEE-C) was proposed for the production of poly-lactic-co-glycolic acid microcarriers. SEE-C operating parameters as pressure, temperature and flow rate ratios were analyzed and the process performance was optimized in terms of size distribution and encapsulation efficiency. Microdevices loaded with bovine serum insulin were produced with different sizes (2 and 3 µm) or insulin charges (3 and 6 mg/g) and with an encapsulation efficiency of 60%. The microcarriers were characterized in terms of insulin release profile in two different media (PBS and DMEM) and the diffusion and degradation constants were also estimated by using a mathematical model. PLGA microdevices were also used in a cultivation of embryonic ventricular myoblasts (cell line H9c2 obtained from rat) in a FBS serum free medium to monitor cell viability and growth in dependence of insulin released. Good cell viability and growth were observed on 3 µm microdevices loaded with 3 mg/g of insulin. PLGA microspheres loaded with growth factors (GFs) were charged into alginate scaffold with human Mesenchimal Steam Cells (hMSC) for bone tissue engineering with the aim of monitoring the effect of the local release of these signals on cells differentiation. These “living” 3D scaffolds were incubated in a direct perfusion tubular bioreactor to enhance nutrient transport and exposing the cells to a given shear stress. Different GFs such as, h-VEGF, h-BMP2 and a mix of two (ratio 1:1) were loaded and alginate beads were recovered from dynamic (tubular perfusion system bioreactor) and static culture at different time points (1st, 7th, 21st days) for the analytical assays such as, live/dead; alkaline phosphatase; osteocalcin; osteopontin and Van Kossa Immunoassay. The immunoassay confirmed always a better cells differentiation in the bioreactor with respect to the static culture and revealed a great influence of the BMP-2 released in the scaffold on cell differentiation.

Refined Estimation of Earthquake Source Parameters: Methods, Applications and Scaling Relationships

The objective of this work of thesis is the refined estimations of source parameters. To such a purpose we used two different approaches, one in the frequency domain and the other in the time domain. In frequency domain, we analyzed the P- and S-wave displacement spectra to estimate spectral parameters, that is corner frequencies and low frequency spectral amplitudes. We used a parametric modeling approach which is combined with a multi-step, non-linear inversion strategy and includes the correction for attenuation and site effects. The iterative multi-step procedure was applied to about 700 microearthquakes in the moment range 1011-1014 N•m and recorded at the dense, wide-dynamic range, seismic networks operating in Southern Apennines (Italy). The analysis of the source parameters is often complicated when we are not able to model the propagation accurately. In this case the empirical Green function approach is a very useful tool to study the seismic source properties. In fact the Empirical Green Functions (EGFs) consent to represent the contribution of propagation and site effects to signal without using approximate velocity models. An EGF is a recorded three-component set of time-histories of a small earthquake whose source mechanism and propagation path are similar to those of the master event. Thus, in time domain, the deconvolution method of Vallée (2004) was applied to calculate the source time functions (RSTFs) and to accurately estimate source size and rupture velocity. This technique was applied to 1) large event, that is Mw=6.3 2009 L’Aquila mainshock (Central Italy), 2) moderate events, that is cluster of earthquakes of 2009 L’Aquila sequence with moment magnitude ranging between 3 and 5.6, 3) small event, i.e. Mw=2.9 Laviano mainshock (Southern Italy).