Unraveling the role of mechanical stimulation on smooth muscle cells: A comparative study between 2D and 3D models.

A thorough understanding of cell response to combined culture configuration and mechanical cues is of paramount importance in vascular tissue engineering applications. Herein, we investigated and compared the response of vascular smooth muscle cells (vSMCs) cultured in different culture environments (2D cell monolayers and 3D cellularized collagen-based gels) in combination with mechanical stimulation (7% uniaxial cyclic strain, 1 Hz) for 2 and 5 days. When cyclic strain was applied, two different responses, in terms of cell orientation and expression of contractile-phenotype proteins, were observed in 2D and 3D models. Specifically, in 2D configuration, cyclic strain caused ∼50% of cell population to align nearly perpendicular (80-90 degrees) to the strain direction, while not influencing the contractile-phenotype protein expression, as compared to the 2D static controls. Conversely, the application of uniaxial strain to 3D constructs induced a ∼60% cell alignment almost parallel (0-10 degrees) to the strain direction. Moreover, 3D mechanical stimulation applied for 5 days induced a twofold increase of SM α-actin level and a 14-fold increase of calponin expression as compared to 3D static controls. Altogether these findings provide a new insight into the potential to drive cell behavior by modulating the extracellular matrix and the biomechanical environment. Biotechnol. Bioeng. 2016;113: 2254-2263. © 2016 Wiley Periodicals, Inc.

A dynamic distention protocol for whole-organ bladder decellularization: histological and biomechanical characterization of the acellular matrix.

A combined physical-chemical protocol for whole full-thickness bladder decellularization is proposed, based on organ cyclic distention through repeated infusion/withdrawal of the decellularization agents through the urethra. The dynamic decellularization was intended to enhance cell removal efficiency, facilitating the delivery of detergents within the inner layers of the tissue and the removal of cell debris. The use of mild chemical detergents (hypotonic solution and non-ionic detergent) was employed to limit adverse effects upon matrix 3D ultrastructure. Inspection of the presence of residual DNA and RNA was carried out on decellularized matrices to verify effective cell removal. Histological investigation was focused on assessing the retention of adequate structural and functional components that regulate the biomechanical behaviour of the acellular tissue. Biomechanical properties were evaluated through uniaxial tensile loading tests of tissue strips and through ex vivo filling cystometry to evaluate the whole-organ mechanical response to a physiological-like loading state. According to our results, a dynamic decellularization protocol of 17 h duration with a 5 ml/min detergent infusion flow rate revealed higher DNA removal efficiency than standard static decellularization, resulting in residual DNA content < 50 ng/mg dry tissue weight. Furthermore, the collagen network and elastic fibres distribution were preserved in the acellular ECM, which exhibited suitable biomechanical properties in the perspective of its future use as an implant for bladder augmentation.

A novel passive left heart platform for device testing and research.

Integration of biological samples into in vitro mock loops is fundamental to simulate real device's operating conditions. We developed an in vitro platform capable of simulating the pumping function of the heart through the external pressurization of the ventricle. The system consists of a fluid-filled chamber, in which the ventricles are housed and sealed to exclude the atria from external loads. The chamber is connected to a pump that drives the motion of the ventricular walls. The aorta is connected to a systemic impedance simulator, and the left atrium to an adjustable preload. The platform reproduced physiologic hemodynamics, i.e. aortic pressures of 120/80 mmHg with 5 L/min of cardiac output, and allowed for intracardiac endoscopy. A pilot study with a left ventricular assist device (LVAD) was also performed. The LVAD was connected to the heart to investigate aortic valve functioning at different levels of support. Results were consistent with the literature, and high speed video recordings of the aortic valve allowed for the visualization of the transition between a fully opening valve and a permanently closed configuration. In conclusion, the system showed to be an effective tool for the hemodynamic assessment of devices, the simulation of surgical or transcatheter procedures and for visualization studies.

Electrical conditioning of adipose-derived stem cells in a multi-chamber culture platform.

In tissue engineering, several factors play key roles in providing adequate stimuli for cells differentiation, in particular biochemical and physical stimuli, which try to mimic the physiological microenvironments. Since electrical stimuli are important in the developing heart, we have developed an easy-to-use, cost-effective cell culture platform, able to provide controlled electrical stimulation aimed at investigating the influence of the electric field in the stem cell differentiation process. This bioreactor consists of an electrical stimulator and 12 independent, petri-like culture chambers and a 3-D computational model was used to characterize the distribution and the intensity of the electric field generated in the cell culture volume. We explored the effects of monophasic and biphasic square wave pulse stimulation on a mouse adipose-derived stem cell line (m17.ASC) comparing cell viability, proliferation, protein, and gene expression. Both monophasic (8 V, 2 ms, 1 Hz) and biphasic (+4 V, 1 ms and -4 V, 1 ms; 1 Hz) stimulation were compatible with cell survival and proliferation. Biphasic stimulation induced the expression of Connexin 43, which was found to localize also at the cell membrane, which is its recognized functional mediating intercellular electrical coupling. Electrically stimulated cells showed an induced transcriptional profile more closely related to that of neonatal cadiomyocytes, particularly for biphasic stimulation. The developed platform thus allowed to set-up precise conditions to drive adult stem cells toward a myocardial phenotype solely by physical stimuli, in the absence of exogenously added expensive bioactive molecules, and can thus represent a valuable tool for translational applications for heart tissue engineering and regeneration.

In vitro study of the aortic interleaflet triangle reshaping.

Aortic interleaflet triangle reshaping (AITR) is a surgical approach to aortic valve incontinence that involves placing three stitches at half of the interleaflet triangles height. In this work, the relationship between the actual stitch height and valve functioning, and the safety margin that the surgeon can rely on in applying the stitches were systematically investigated in vitro. AITR surgery was applied to six swine aortic roots placing the stitches empirically at 50%, 60% and 75% of the triangle heights. Then the actual stitch heights were measured and the hydrodynamic performances were evaluated with a pulsatile hydrodynamic mock loop. Actual stitch heights were 45±2%, 61±4% and 79±6%. As compared to untreated conditions, the 50% configuration induced a significant variation in the effective orifice area. With stitches placed at 60%, the mean systolic pressure drop increased significantly with respect to the untreated case, but no significant changes were recorded with respect to the 50% configuration. At 75%, all the hydrodynamic parameters of systolic valve functioning worsened significantly. Summarizing, the AITR technique, when performed in a conservative manner did not induce significant alterations in the hydrodynamics of the aortic root in vitro, while more aggressive configurations did. The absence of a statistically significant difference between the 50% and 60% configurations suggests that there is a reasonably limited risk of inducing valve stenosis in the post-op scenario due to stitch misplacement.

A novel environmental chamber for neuronal network multisite recordings.

Environmental stability is a critical issue for neuronal networks in vitro. Hence, the ability to control the physical and chemical environment of cell cultures during electrophysiological measurements is an important requirement in the experimental design. In this work, we describe the development and the experimental verification of a closed chamber for multisite electrophysiology and optical monitoring. The chamber provides stable temperature, pH and humidity and guarantees cell viability comparable to standard incubators. Besides, it integrates the electronics for long-term neuronal activity recording. The system is portable and adaptable for multiple network housings, which allows performing parallel experiments in the same environment. Our results show that this device can be a solution for long-term electrophysiology, for dual network experiments and for coupled optical and electrical measurements.

In vitro hemodynamics and valve imaging in passive beating hearts.

Due to their high complexity, surgical approaches to valve repair may benefit from the use of in vitro simulators both for training and for the investigation of those measures which can lead to better clinical results. In vitro tests are intrinsically more effective when all the anatomical substructures of the valvular complexes are preserved. In this work, a mock apparatus able to house an entire explanted porcine heart and subject it to pulsatile fluid-dynamic conditions was developed, in order to enable the hemodynamic analysis of simulated surgical procedures and the imaging of the valvular structures. The mock loop's hydrodynamic design was based on an ad-hoc defined lumped-parameter model. The left ventricle of an entire swine heart was dynamically pressurized by an external computer-controlled pulse duplicator. The ascending aorta was connected to a hydraulic circuit which simulated the input impedance of the systemic circulation; a reservoir passively filled the left atrium. Accesses for endoscopic imaging were located in the apex of the left ventricle and in the aortic root. The experimental pressure and flow tracings were comparable with the typical in vivo curves; a mean flow of 3.5±0.1l pm and a mean arterial pressure of 101±2 mmHg was obtained. High-quality echographic and endoscopic video recordings demonstrated the system's excellent potential in the observation of the cardiac structures dynamics. The proposed mock loop represents a suitable in vitro system for the testing of minimally-invasive cardiovascular devices and surgical procedures for heart valve repair.

In vitro study of aortic valves treated with neo-chordae grafts: hydrodynamics and tensile force measurements.

Reparative surgery of the aortic root functional unit (ARFU) allows for a better preservation of the functionality of the native structure compared to prosthesis implantation. Post-operative results are satisfactory, whereas mid- and long-term results are challenging, for example in terms of cusps prolapse recurrence. At the Cardiothoracic Surgery Unit of the Sacco Hospital, a new surgical technique aimed at the stabilization in time of the results of standard ARFU repair operations has been applied. This technique, inspired by the mitral neo-chordae (NC) implantation, consists of implanting an e-PTFE suture thread between the prolapsed cusp and the sinotubular junction. Aim of this study was to assess the influence of NC implantation on the ARFU functioning by evaluating with an owned pulsatile in vitro apparatus the force magnitude acting on the NC and the dynamic behavior of porcine ARFUs treated according to the operating-room procedures. The maximum recorded values of the mean and peak diastolic forces were 0.064 and 0.186 N, respectively, and were linearly dependent upon the mean diastolic pressure across the valve. In addition, the measurements of the opening-closing times and valve leakage volumes, performed at pre- and post-surgeries, yielded that the valve functionality was not significantly influenced by NC implantation.

Mechanisms of polymyxin B endotoxin removal from extracorporeal blood flow: molecular interactions.

The outer leaflet of Gram-negative bacteria membrane contains a great amount of lipopolysaccharides, also known as endotoxins, which play a central role in the pathogenesis of septic shock. It has been demonstrated that the polymyxin B (PMB) molecule has both antibacterial and antiendotoxin capabilities; in fact, it is able to compromise the bacterial outer membrane and bind lipopolysaccharides, thereby neutralizing its toxic effects. Extracorporeal hemoperfusion treatments based on cartridges containing PMB-immobilized fibers (Toraymyxin PMX-F; Toray Industries, Tokyo, Japan) are used to remove endotoxins circulating in the blood flow. In this study, we focused on the characterization of the interactions occurring in the formation of the PMB-endotoxin complex at the molecular level. In particular, the molecular mechanics approach was used to evaluate the interaction energy and eventually the interaction force between the two molecules. PMB was faced with five molecular portions of lipopolysaccharides differing in their structure. The interaction energy occurring for each molecular complex was calculated at different intermolecular distances and the binding forces were estimated by fitting interaction energy data. Results show that the short-range interactions between PMB and endotoxins are mediated mainly by hydrophobic forces, while in the long term, the complex formation is driven by ionic forces only. Maximum binding forces calculated via molecular mechanics for the PMB-endotoxin complex are in the range of 1.39-3.79 nN. Understanding the interaction mechanism of the single molecular complex is useful both in order to figure out the molecular features of such interaction and to perform higher scale level analysis, where such nanoscale detail is impractical but could be used to account for molecular behavior at a coarse level of discretization.

Mechanisms of polymyxin B endotoxin removal from extracorporeal blood flow: hydrodynamics of sorption.

The removal of blood endotoxins with the Toraymyxin extracorporeal sorption device exploits the capability of immobilized polymyxin B (PMB) to bind endotoxins stably with a high specificity. Although adsorption is a molecular-scale mechanism, it involves hydrodynamic phenomena in the whole range from the macroscopic down to the supramolecular scales. In this paper we summarize our experience with a computational, multiscale investigation of this device's hydrodynamic functionality. 3D computational fluid dynamics models were developed for the upper-scale studies. The flow behavior in the sorbent material was either modeled as a homogeneous Darcy's flow (macroscale study), or described as the flow through realistic geometrical models of its knitted fibers (mesoscale study). In the microscale study, simplified 2D models were used to track the motion of modeled endotoxin particles subjected to the competition of flow drag and molecular attraction by the fiber-grafted PMB. The results at each scale level supplied worst-case input data for the subsequent study. The macroscale results supplied the peak velocity of the flow field that develops in the sorbent. This was used in the mesoscale analysis, yielding a realistic range for the shear stresses in the fluid next to the fiber surface. With wall shear stresses in this range, endotoxin particle tracking was studied both in the vicinity of a single immobilized PMB molecule, and in the presence of a layer of PMB molecules evenly distributed at the fiber surface. Results showed that the capability to seize endotoxin molecules extends at least at a distance of 10-20 nm from the surface, which is one order of magnitude greater than the stable intermolecular bond characteristic distance. We conclude that a multiscale approach has the power to provide a comprehensive understanding, shedding light both upon the physics involved at each scale level and the mutual interactions of phenomena occurring at different scales.

Multi-scale analysis of the toraymyxin adsorption cartridge. Part I: molecular interaction of polymyxin B with endotoxins.

Endotoxins or lipopolysaccharides are the main constituents of the outer leaflet of Gram-negative bacteria membrane and play a central role in the pathogenesis of the septic shock. Polymyxin B has both antibacterial and antiendotoxin capability; indeed it is able to destroy the bacterial outer membrane and bind endotoxin neutralizing its toxic effects. Cartridges containing polymyxin B-immobilized fibers (Toraymyxin PMX-F, Toray Industries, Japan) are used in extracorporeal hemoperfusion to remove circulating endotoxin. The aim of this study is the characterization of the polymyxin B-endotoxin system at the molecular level, thus providing quantitative evaluation of the binding forces exerted in the molecular complex. Polymyxin B was interfaced with five molecular models of lipopolysaccharides differing in their structure and molecular mechanics simulations were performed at different intermolecular distances aimed at calculating the interaction energies of the complex. Binding forces were calculated by fitting interaction energies data. Results show that in the short range the polymyxin B-endotoxin complex is mediated by hydrophobic forces and in the long range the complex is driven by ionic forces only. From a mechanical standpoint, polymyxin B-endotoxin complex is characterized by maximum binding forces ranging between 1.39 nN to 3.79 nN. The knowledge of the binding force behavior at different intermolecular distances allows further investigations at higher scale level (Part II).

A mechanical model lung for hydraulic testing of total liquid ventilation circuits.

A new model lung (ML), designed to reproduce the tracheal pressure vs. fluid flow relationship in animals undergoing total liquid ventilation (TLV) trials, was developed to be used as a mock bench test for neonatal TLV circuits. The ML is based on a linear inertance-resistance-compliance (LRC) lumped-parameter model of the respiratory system with different resistance values for inspiration (R insp ) or expiration (R exp ). The resistant element was set up using polypropylene hollow fibres packed inside a tube. A passive one-way valve was used to control the resistance cross-section area provided for the liquid to generate different values for R insp or R exp , each adjustable by regulating the active length of the respective fibre pack. The compliant element consists of a cylindrical column reservoir, in which bars of different diameter were inserted to adjust compliance (C). The inertial phenomena occurring in the central airways during TLV were reproduced by specifically dimensioned conduits into which the endotracheal tube connecting the TLV circuit to the ML was inserted. A number of elements with different inertances (L) were used to simulate different sized airways. A linear pressure drop-to-flow rate relationship was obtained for flow rates up to 5 l/min. The measured C (0.8 to 1.3 mL cmH2O (-1) kg(-1)), R insp (90 to 850 cmH2O s l(-1)), and R exp (50 to 400 cmH2O s l(-1)) were in agreement with the literature concerning animals weighing from 1 to 12 kg. Moreover, features observed in data acquired during in vivo TLV sessions, such as pressure oscillations due to fluid inertia in the upper airways, were similarly obtained in vitro thanks to the inertial element in the ML.

A numerical and experimental study of compliance and collapsibility of preterm lamb tracheae.

Knowledge of the mechanical behaviour of immature tracheae is crucial in order to understand the effects exerted on central airways by ventilatory treatments, particularly of Total Liquid Ventilation. In this study, a combined experimental and computational approach was adopted to investigate the compliance and particularly collapsibility of preterm lamb tracheae in the range of pressure likely applied during Total Liquid Ventilation (-30 to 30 cmH2O). Tracheal samples of preterm lambs (n = 5; gestational age 120-130 days) were tested by altering transmural pressure from -30 to 30 cmH2O. Inflation (Si) and collapsing (Sc) compliance values were calculated in the ranges 0 to 10 cmH2O and -10 to 0 cmH2O, respectively. During the tests, an asymmetric behaviour of the DeltaV/V0 vs. P curves at positive and negative pressure was observed, with mean Si = 0.013 cmH2O(-1) and Sc = 0.053 cmH2O(-1). A different deformed configuration of the sample regions was observed, depending on the posterior shape of cartilaginous ring. A three-dimensional finite-element structural model of a single tracheal ring, based on histology measurements of the tested samples was developed. The model was parameterised in order to represent rings belonging to three different tracheal regions (craniad, median, caudal) and numerical analyses replicating the collapse test conditions were performed to evaluate the ring collapsibility at pressures between 0 and -30 cmH2O. Simulation results were compared to experimental data to verify the model's reliability. The best model predictions occurred at pressures -30 to -10 cmH2O. In this range, a model composed of median rings best interpreted the experimental data, with a maximum error of 2.7%; a model composed of an equal combination of all rings yielded an error of 12.6%.

Clinical evaluation of internal hemodiafiltration (iHDF): a diffusive-convective technique performed with internal filtration enhanced high-flux dialyzers.

AIM: Efficiency in removing middle molecules such as beta2-microglobulin (beta2-MG) is one of the main purposes of modern dialytic therapy. In order to achieve this, techniques requiring complex machines and substitution fluid have been developed over recent years. Alternatively, the internal filtration/back filtration phenomenon can be used. The recent development of a so-called "internal filtration enhanced dialyser" prompted us to compare the removal of beta2-MG together with other small molecules when the dialyser was used either in standard hemodiafiltration (HDF) or internal hemodiafiltration (iHDF). METHODS: Ten stable, anuric, hemodialysis (HD) patients treated by thrice weekly standard bicarbonate HD using low-flux synthetic membrane entered the study. A new high-flux polysulfone dialyser designed with the specific aim of enhancing internal filtration (BS-1.6 UL, 1.6 m2, Toray Industries) was used. Post dilution HDF (2.5 l/hour of substitution fluid, dialysate flow 500 ml/min) was compared with iHDF (dialysate flow 750 ml/min), with blood flow at 300 ml/min. Samples were obtained at the start and at the end of the session in order to measure the % removal of urea, creatinine, uric acid, phosphate and beta2-MG (corrected for total protein concentration). In addition, after 20 min of dialysis the clearances of the same molecules were measured. A mathematical model has been developed for the description of the hydrodynamic phenomena taking place within the dialyser and of fluid filtration across the membrane. RESULTS: No significant differences have been observed in removal rate switching from HDF to iHDF except for beta2-MG removal, which was slightly higher in HDF than in iHDF Phosphate clearance is significantly higher than those obtained with creatinine in both HDF (p<0.005) and iHDF (p<0.01) modalities. The total convection calculated with the model is reduced with respect to HDF only by 24% (4100 ml/h vs. 5400 ml/h on the average). CONCLUSIONS: iHDF is a high flux dialysis method, which, if performed with a dialyser designed to enhance internal filtration, obtains a much higher removal rate in comparison with dialysers in traditional high flux dialysis, as previously reported in the literature. Provided that the dialyser is used on a dialysis machine working with ultra pure dialysate and UF control, this dialyser line can perform reliable internal HDF without the need for replacement solution. Considering the narrow difference in performance observed between iHDF and HDF, and the increasing number (and age) of patients leading to higher dialysis costs, iHDF represents a cost-effective alternative to other diffusive-convective techniques.

Pressure drop vs flow relationship in isolated preterm lamb tracheae.

Knowledge of immature tracheae mechanical behavior is fundamental in understanding the effects exerted on the upper airways by tidal liquid ventilation (TLV). Particularly, negative pressure can take place along the airways during expiration, which can cause airway collapse and flow limitation; therefore, representing a critical issue in preterm infant patients, whose airways are less stiff than adult ones. In this study, we investigated the expiratory pressure drop vs flow relationship of isolated preterm lamb tracheal samples to determine their hydraulic resistance, collapse pressure and collapse flow rate; a liquid flow through the samples was obtained by applying negative pressure at the outlet (cephalad) extremity of the tra-cheal sample, while keeping the inlet (caudal) extremity at atmospheric pressure. Histological analyzes were performed on the tracheal samples after each test session, in order to examine the morphological structure of the tracheal wall. Flow resistance tests demonstrated progressive lumen narrowing at increasing pressure drop (∆P=P in -P out ). The flow rate increased with ∆P un-til a plateau was reached, and then decreased, describing the onset of a collapse phenomenon; however, complete occlusion was not reached. The tracheal samples demonstrated a similar behavior to that of a Starling resistor during the collapse phase: when a critical ∆P was reached, collapse was observed starting at the outlet region, which was subjected to the greatest negative pressure, then propagating towards the caudal direction. (Journal of Applied Biomaterials & Biomechanics 2004; 2: 177-82).

Perfluorocarbons attenuate oxidative lung damage.

The aim of this study was to investigate the effect of tidal liquid ventilation (TLV) compared to conventional mechanical ventilation (CMV) on oxidative lung damage in the setting of acute respiratory distress syndrome (ARDS). After repeated lung lavages, 10 minipigs were treated with CMV or TLV for 4 hr before the animals were sacrificed. Samples for blood gas analysis and bronchial aspirate samples were withdrawn before the induction of lung injury, and at 10 min, 2 hr, and 4 hr after the beginning of ventilatory support. To assess lung oxidative damage, total hydroperoxide (TH) and advanced oxidation protein product (AOPP) concentrations were measured in bronchial aspirate samples. After 2 and 4 hr of ventilatory support, partial oxygen tension (PaO(2)) and base excess (BE) were significantly higher in the TLV group than in the CMV group, while PaCO(2) was slightly higher, but with no statistical significance. In the CMV group, the AOPP level was significantly higher at 4 hr than at baseline. TH and AOPP bronchial aspirate concentrations were higher in the CMV group than in the TLV group at 2 and 4 hr of ventilation. We conclude that animals treated with TLV showed lower oxidative lung damage compared to animals treated with CMV.

Hydraulic functional characterisation of aortic mechanical heart valve prostheses through lumped-parameter modelling.

Lumped-parameter modelling techniques are proposed as a method for studying the hydraulic characteristics of mechanical prosthetic heart valves (PHVs). The global hydraulic behaviour of PHVs in the open position was modelled by taking into account the (nonlinear) resistive and (linear) inertial factors governing the time-dependent relationship between transvalvular pressure drop and fluid flow rate, and neglecting the leaflets' opening and closure transient phenomena. Statistically defined indices associated to the parameters' values attest how properly the model describes PHV hydraulic behaviour. Local fluid dynamics is not modelled with this approach. The proposed method was implemented in a software program and applied to the characterisation of the aortic StJude Medical, StJude Medical Hemodynamic Plus and CarboMedics PHVs, basing on steady- and pulsatile-flow hydraulic-bench experimental data. The results showed that reliable parameters expressing hydraulic resistance can be derived from steady-flow data (R(2)>0.995). Inertance parameters derived from pulsatile-flow experiments are liable to a degree of uncertainty (confidence intervals up to 17%), however, comparing the reconstructed vs. measured pressure drop during systolic time demonstrates that this deficiency is mostly due to the missing description of initial, transient oscillations presumably related to the leaflets' opening (not modelled).

Mechanobiology of engineered cartilage cultured under a quantified fluid-dynamic environment.

Natural cartilage remodels both in vivo and in vitro in response to mechanical forces and hence mechanical stimulation is believed to have a potential as a tool to modulate extra-cellular matrix synthesis in tissue-engineered cartilage. Fluid-induced shear is known to enhance chondrogenesis on animal cells. A well-defined hydrodynamic environment is required to study the biochemical response to shear of three-dimensional engineered cell systems. We have developed a perfused-column bioreactor in which the culture medium flows through chondrocyte-seeded porous scaffolds, together with a computational fluid-dynamic model of the flow through the constructs' microstructure. A preliminary experiment of human chondrocyte growth under static versus dynamic conditions is described. The median shear stress imposed on the cells in the bioreactor culture, as predicted by the CFD model, is 3 x 10(-3) Pa (0.03 dyn/cm(2)) at a flow rate of 0.5 ml/min corresponding to an inlet fluid velocity of 44.2 mum/s. Providing a fluid-dynamic environment to the cells yielded significant differences in cell morphology and in construct structure.

A model of neonatal tidal liquid ventilation mechanics.

Tidal liquid ventilation (TLV) with perfluorocarbons (PFC) has been proposed to treat surfactant-deficient lungs of preterm neonates, since it may prevent pulmonary instability by abating saccular surface tension. With a previous model describing gas exchange, we showed that ventilator settings are crucial for CO(2) scavenging during neonatal TLV. The present work is focused on some mechanical aspects of neonatal TLV that were hardly studied, i.e. the distribution of mechanical loads in the lungs, which is expected to differ substantially from gas ventilation. A new computational model is presented, describing pulmonary PFC hydrodynamics, where viscous losses, kinetic energy changes and lung compliance are accounted for. The model was implemented in a software package (LVMech) aimed at calculating pressures (and approximately estimate shear stresses) within the bronchial tree at different ventilator regimes. Simulations were run taking the previous model's outcomes into account. Results show that the pressure decrease due to high saccular compliance may compensate for the increased pressure drops due to PFC viscosity, and keep airway pressure low. Saccules are exposed to pressures remarkably different from those at the airway opening; during expiration negative pressures, which may cause airway collapse, are moderate and appear in the upper airways only. Delivering the fluid with a slightly smoothed square flow wave is convenient with respect to a sine wave. The use of LVMech allows to familiarize with LV treatment management taking the lungs' mechanical load into account, consistently with a proper respiratory support.