Innovative chitin-glucan based material obtained from mycelium of wood decay fungal strains.

Fungi are an alternative source to animal-based chitin. In fungi, chitin fibrils are strongly interconnected and bound with glucans that justify the unique matrix. The present study aimed to extract chitin and glucans from the mycelium of several wood decay fungal strains in order to obtain flexible materials and to check correlations between chitin content and the mechanical properties of these materials. Five strains were chosen in consideration of their different cell wall chemical composition (high content of α-glucans, ß-glucans or chitin) to evaluate how these differences could influence the mechanical and chemical characteristics of the material. The fungal strains were cultivated in liquid-submerged dynamic fermentation (both flasks and bioreactor). Chitin and glucans were crosslinked with acetic acid and plasticized with glycerol to obtain flexible sheets. Abortiporus biennis, Fomitopsis iberica and Stereum hirsutum strains were found to adapt to produce material with adequate flexibility. The obtained materials were characterized by Thermogravimetric analysis (TGA) for the understanding of the material composition. The material obtained from each species was mechanically tested in terms of tear strength, elongation at break, and Young's modulus.

Effect of Micromixer Design on Lipid Nanocarriers Manufacturing for the Delivery of Proteins and Nucleic Acids.

Lipid-based nanocarriers have emerged as helpful tools to deliver sensible biomolecules such as proteins and oligonucleotides. To have a fast and robust microfluidic-based nanoparticle synthesis method, the setup of versatile equipment should allow for the rapid transfer to scale cost-effectively while ensuring tunable, precise and reproducible nanoparticle attributes. The present work aims to assess the effect of different micromixer geometries on the manufacturing of lipid nanocarriers taking into account the influence on the mixing efficiency by changing the fluid-fluid interface and indeed the mass transfer. Since the geometry of the adopted micromixer varies from those already published, a Design of Experiment (DoE) was necessary to identify the operating (total flow, flow rate ratio) and formulation (lipid concentration, lipid molar ratios) parameters affecting the nanocarrier quality. The suitable application of the platform was investigated by producing neutral, stealth and cationic liposomes, using DaunoXome®, Myocet®, Onivyde® and Onpattro® as the benchmark. The effect of condensing lipid (DOTAP, 3-10-20 mol%), coating lipids (DSPE-PEG550 and DSPE-PEG2000), as well as structural lipids (DSPC, eggPC) was pointed out. A very satisfactory encapsulation efficiency, always higher than 70%, was successfully obtained for model biomolecules (myoglobin, short and long nucleic acids).

Type III aortic arch angulation increases aortic stiffness: Analysis from an ex vivo porcine model.

Objective: The relationship among increased aortic arch angulation, aortic flow dynamics, and vessel wall stiffness remains unclear. This experimental ex vivo study investigated how increased aortic arch angulation affects aortic stiffness and stent-graft induced aortic stiffening, assessed by pulse wave velocity (PWV). Methods: Porcine thoracic aortas were connected to a circulatory mock loop in a Type I and Type III aortic arch configuration. Baseline characteristics and blood pressures were measured. Proximal and distal flow curves were acquired to calculate PWV in both arch configurations. After that, a thoracic stent-graft (VAMF2626C100TU) was deployed in aortas with adequate proximal landing zone diameters to reach 10% t0 20% oversizing. Acquisitions were repeated for both arch configurations after stent-graft deployment. Results: Twenty-four aortas were harvested, surgically prepared, and mounted. Cardiac output was kept constant for both arch configurations (Type I: 4.74 ± 0.40 and Type III: 4.72 ± 0.38 L/minute; P = .703). Compared with a Type I arch, aortic PWV increased significantly in the Type III arch (3.53 ± 0.40 vs 3.83 ± 0.40 m/second; P < .001), as well as blood pressures. A stent-graft was deployed in 15 aortas. After deployment, Type I arch PWV increased (3.55 ± 0.39 vs 3.81 ± 0.44 m/second; P < .001) and Type III arch PWV increased although not significantly (3.86 ± 0.42 vs 4.03 ± 0.46 m/second; P = .094). Type III arch PWV resulted the highest and significantly higher compared with the Type I arch after stent-graft deployment (3.81 ± 0.44 vs 4.03 ± 0.46 m/second; P = .023). Conclusions: Increased aortic arch angulation-as in a Type III arch-is associated with higher aortic PWV and blood pressures and this may negatively influence cardiovascular health.

Design and development of a hepatic lyo-dECM powder as a biomimetic component for 3D-printable hybrid hydrogels.

Bioprinting offers new opportunities to obtain reliable 3Din vitromodels of the liver for testing new drugs and studying pathophysiological mechanisms, thanks to its main feature in controlling the spatial deposition of cell-laden hydrogels. In this context, decellularized extracellular matrix (dECM)-based hydrogels have caught more and more attention over the last years because of their characteristic to closely mimic the tissue-specific microenvironment from a biological point of view. In this work, we describe a new concept of designing dECM-based hydrogels; in particular, we set up an alternative and more practical protocol to develop a hepatic lyophilized dECM (lyo-dECM) powder as an 'off-the-shelf' and free soluble product to be incorporated as a biomimetic component in the design of 3D-printable hybrid hydrogels. To this aim, the powder was first characterized in terms of cytocompatibility on human and porcine mesenchymal stem cells (MSCs), and the optimal powder concentration (i.e. 3.75 mg ml-1) to use in the hydrogel formulation was identified. Moreover, its non-immunogenicity and capacity to reactivate the elastase enzyme potency was proved. Afterward, as a proof-of-concept, the powder was added to a sodium alginate/gelatin blend, and the so-defined multi-component hydrogel was studied from a rheological point of view, demonstrating that adding the lyo-dECM powder at the selected concentration did not alter the viscoelastic properties of the original material. Then, a printing assessment was performed with the support of computational simulations, which were useful to definea priorithe hydrogel printing parameters as window of printability and its post-printing mechanical collapse. Finally, the proposed multi-component hydrogel was bioprinted with cells inside, and its post-printing cell viability for up to 7 d was successfully demonstrated.

Lumped-parameter model as a non-invasive tool to assess coronary blood flow in AAOCA patients.

Anomalous aortic origin of the coronary artery (AAOCA) is a rare disease associated with sudden cardiac death, usually related to physical effort in young people. Clinical routine tests fail to assess the ischemic risk, calling for novel diagnostic approaches. To this aim, some recent studies propose to assess the coronary blood flow (CBF) in AAOCA by computational simulations but they are limited by the use of data from literature retrieved from normal subjects. To overcome this limitation and obtain a reliable assessment of CBF, we developed a fully patient-specific lumped parameter model based on clinical imaging and in-vivo data retrieved during invasive coronary functional assessment of subjects with AAOCA. In such a way, we can estimate the CBF replicating the two hemodynamic conditions in-vivo analyzed. The model can mimic the effective coronary behavior with high accuracy and could be a valuable tool to quantify CBF in AAOCA. It represents the first step required to move toward a future clinical application with the aim of improving patient care. The study was registered at Clinicaltrial.gov with (ID: NCT05159791, date 2021-12-16).

Hybrid 3D-Printed and Electrospun Scaffolds Loaded with Dexamethasone for Soft Tissue Applications.

BACKGROUND: To make the regenerative process more effective and efficient, tissue engineering (TE) strategies have been implemented. Three-dimensional scaffolds (electrospun or 3D-printed), due to their suitable designed architecture, offer the proper location of the position of cells, as well as cell adhesion and the deposition of the extracellular matrix. Moreover, the possibility to guarantee a concomitant release of drugs can promote tissue regeneration. METHODS: A PLA/PCL copolymer was used for the manufacturing of electrospun and hybrid scaffolds (composed of a 3D-printed support coated with electrospun fibers). Dexamethasone was loaded as an anti-inflammatory drug into the electrospun fibers, and the drug release kinetics and scaffold biological behavior were evaluated. RESULTS: The encapsulation efficiency (EE%) was higher than 80%. DXM embedding into the electrospun fibers resulted in a slowed drug release rate, and a slower release was seen in the hybrid scaffolds. The fibers maintained their nanometric dimensions (less than 800 nm) even after deposition on the 3D-printed supports. Cell adhesion and proliferation was favored in the DXM-loading hybrid scaffolds. CONCLUSIONS: The hybrid scaffolds that were developed in this study can be optimized as a versatile platform for soft tissue regeneration.

Aortic wall thickness in dilated ascending aorta: Comparison between tricuspid and bicuspid aortic valve.

BACKGROUND: Bicuspid aortic valve (BAV) is frequently associated with dilatation of the thoracic aorta. Peculiar anatomical, histological and mechanical changes of the aortic wall in BAV aortopathy have been hypothesized to suggest an increased risk of acute aortic complications in patients with BAV. AIM: In this study we tried to clarify any differences in the adaptability of the aortic wall to the mechanism of dilatation between patients with BAV and those with TAV. METHODS: In total, 354 samples were taken from 71 patients undergoing elective aortic surgery and divided into two groups: BAV group (n=16; 101 samples); and TAV group (n=55; 253 samples). Aortic wall thickness was measured with a dedicated caliper. The relationship between aortic wall thickness and aortic dilatation and demographic variables was evaluated cumulatively and comparatively (BAV versus TAV). In patients with more than three samples available, intrapatient variability was also studied. Finally, potential risk factors for severely reduced aortic wall thickness were also assessed. RESULTS: Analysis of preoperative characteristics revealed significant differences in patient age (54±16years for BAV and 66±11years for TAV; P=0.0011), with no differences in variables related to aortic dilatation (including phenotype). Cumulative aortic wall thickness was significantly thinner in the anterior than in the posterior wall. In the comparative analysis, aortic wall thickness was significantly thinner in patients with BAV in both the anterior and posterior regions. Furthermore, in patients with BAV, dilatation>51mm was a significant predictor of severely reduced aortic wall thickness. CONCLUSIONS: In our experience, patients with BAV aortopathy reached the cut-off for the surgical indication at an early age. Careful monitoring in patients with BAV is mandatory when aortic dilatation has reached 51mm, as it is related to significant anatomical changes.

Elaboration and development of a realistic 3D printed model for training in ultrasound-guided placement of peripheral central venous catheter in children.

BACKGROUND: Simulation for training is becoming a trend topic worldwide, even if its applications are commonly limited to adulthood. Ultrasound-guided procedures require practice and experience-especially in the pediatric field, where the small size of the involved anatomical structures poses major problems. In this context, a realistic 3D printed pediatric phantom for training of the ultrasound-guided placement of peripheral central venous catheters in children was developed. MATERIALS AND METHODS: Starting from Computed Tomography scans of an 8 years-old girl, her left arm was virtually reconstructed-including bones, arteries, and veins-through a semi-automatic segmentation process. According to preliminary results, the most suitable 3D printing technologies to reproduce the different anatomical structures of interest were selected, considering both direct and indirect 3D printing techniques. Experienced operators were asked to evaluate the efficacy of the final model through a dedicated questionnaire. RESULTS: Vessels produced through indirect 3D printing latex dipping technique exhibited the best echogenicity, thickness, and mechanical properties to mimic real children's venous vessels, while arteries-not treated and/or punctured during the procedure-were directly 3D printed through Material Jetting technology. An external mold-mimicking the arm skin-was 3D printed and a silicone-based mixture was poured to reproduce real patient's soft tissues. Twenty expert specialists were asked to perform the final model's validation. The phantom was rated as highly realistic in terms of morphology and functionality for the overall simulation, especially for what concerns vessels and soft tissues' response to puncturing. On the other hand, the involved structures' US appearance showed the lower score. CONCLUSIONS: The present work shows the feasibility of a patient-specific 3D printed phantom for simulation and training in pediatric ultrasound-guided procedures.

Morphological Changes of Anomalous Coronary Arteries From the Aorta During the Cardiac Cycle Assessed by IVUS in Resting Conditions.

BACKGROUND: Anomalous aortic origin of coronary artery (AAOCA) with intramural segment is associated with risk of sudden cardiac death, probably related to a compressive mechanism exerted by the aorta. However, the intramural compression occurrence and magnitude during the cardiac cycle remain unknown. We hypothesized that (1) in end diastole, the intramural segment is narrower, more elliptic, and has greater resistance than extramural segment; (2) the intramural segment experiences a further compression in systole; and (3) morphometry and its systolic changes vary within different lumen cross-sections of the intramural segment. METHODS: Phasic changes of lumen cross-sectional coronary area, roundness (minimum/maximum lumen diameter), and hemodynamic resistance (Poiseuille law for noncircular sections) were derived from intravascular ultrasound pullbacks at rest for the ostial, distal intramural, and extramural segments. Data were obtained for 35 AAOCA (n=23 with intramural tract) after retrospective image-based gating and manual lumen segmentation. Differences between systolic and end-diastolic phases in each section, between sections of the same coronary, and between AAOCA with and without intramural tract were assessed by nonparametric statistical tests. RESULTS: In end diastole, both the ostial and distal intramural sections were more elliptical (P<0.001) than the reference extramural section and the correspondent sections in AAOCA without intramural segment. In systole, AAOCA with intramural segment showed a flattening at the ostium (-6.76% [10.82%]; P=0.024) and a flattening (-5.36% [16.56%]; P=0.011), a narrowing (-4.62% [11.38%]; P=0.020), and a resistance increase (15.61% [30.07%]; P=0.012) at the distal intramural section. No-intramural sections did not show morphological changes during the entire cardiac cycle. CONCLUSIONS: AAOCA with intramural segment has pathological segment-specific dynamic compression mainly in the systole under resting conditions. Studying AAOCA behavior with intravascular ultrasound during the cardiac cycle may help to evaluate and quantify the severity of the narrowing.

Assessment of different manufacturing techniques for the production of bioartificial scaffolds as soft organ transplant substitutes.

Introduction: The problem of organs' shortage for transplantation is widely known: different manufacturing techniques such as Solvent casting, Electrospinning and 3D Printing were considered to produce bioartificial scaffolds for tissue engineering purposes and possible transplantation substitutes. The advantages of manufacturing techniques' combination to develop hybrid scaffolds with increased performing properties was also evaluated. Methods: Scaffolds were produced using poly-L-lactide-co-caprolactone (PLA-PCL) copolymer and characterized for their morphological, biological, and mechanical features. Results: Hybrid scaffolds showed the best properties in terms of viability (>100%) and cell adhesion. Furthermore, their mechanical properties were found to be comparable with the reference values for soft tissues (range 1-10 MPa). Discussion: The created hybrid scaffolds pave the way for the future development of more complex systems capable of supporting, from a morphological, mechanical, and biological standpoint, the physiological needs of the tissues/organs to be transplanted.

3D Printing of Copper Using Water-Based Colloids and Reductive Sintering.

Copper was manufactured by using a low-cost 3D printing device and copper oxide water-based colloids. The proposed method avoids the use of toxic volatile solvents (used in metal-based robocasting), adopting copper oxide as a precursor of copper metal due to its lower cost and higher chemical stability. The appropriate rheological properties of the colloids have been obtained through the addition of poly-ethylene oxide-co-polypropylene-co-polyethylene oxide copolymer (Pluronic P123) and poly-acrylic acid to the suspension of the oxide in water. Mixing of the components of the colloidal suspension was performed with the same syringes used for the extrusion, avoiding any material waste. The low-temperature transition of water solutions of P123 is used to facilitate the homogenization of the colloid. The copper oxide is then converted to copper metal through a reductive sintering process, performed at 1000°C for a few hours in an atmosphere of Ar-10%H2. This approach allows the obtainment of porous copper objects (up to 20%) while retaining good mechanical properties. It could be beneficial for many applications, for example current collectors in lithium batteries.

Comparison of Two Generations of Thoracic Aortic Stent Grafts and Their Impact on Aortic Stiffness in an Ex Vivo Porcine Model.

Objective: Little is known about the cardiovascular changes after TEVAR and regarding the impact on aortic stiffness for different stent graft generations specifically, following changes in device design. The present study evaluated the stent graft induced aortic stiffening of two generations of the Valiant thoracic aortic stent graft. Methods: This was an ex vivo porcine investigation using an experimental mock circulatory loop. Thoracic aortas of young healthy pigs were harvested and connected to the mock circulatory loop. At a 60 bpm heart rate and stable mean arterial pressure, baseline aortic characteristics were obtained. Pulse wave velocity (PWV) was calculated before and after stent graft deployment. Paired and independent sample t tests or their non-parametric alternatives were performed to test for differences where appropriate. Results: Twenty porcine thoracic aortas were divided into two equal subgroups, in which a Valiant Captivia or a Valiant Navion stent graft was deployed. Both stent grafts were similar in diameter and length. Baseline aortic characteristics did not differ between the subgroups. Mean arterial pressure values did not change after either stent graft, while pulse pressures increased statistically significantly after Captivia (mean 44 ± 10 mmHg to 51 ± 13 mmHg, p = .002) but not after Navion. Mean baseline PWV increased after both Captivia (4.4 ± 0.6 m/s to 4.8 ± 0.7 m/s, p = .007) and Navion (4.6 ± 0.7 m/s to 4.9 ± 0.7 m/s, p = .002). There was no statistically significant difference in the mean percentage increase in PWV for either subgroup (8 ± 4% vs. 6 ± 4%, p = .25). Conclusion: These experimental findings showed no statistically significant difference in the percentage increase of aortic PWV after either stent graft generation and confirm that TEVAR increases aortic PWV. As a surrogate for aortic stiffness, this calls for further improvements in future thoracic aortic stent graft designs regarding device compliance.

Prediction of the mechanical response of a 3D (bio)printed hybrid scaffold for improving bone tissue regeneration by structural finite element analysis.

Scaffolds for bone tissue engineering should be osteoinductive, osteoconductive, biocompatible, biodegradable, and, at the same time, exhibit proper mechanical properties. The present study investigated the mechanical properties of a coprinted hybrid scaffold made of polycaprolactone (PCL) and an alginate-based hydrogel, which was conceived to possess a double function of in vivo bio-integration (due to the ability of the hydrogel to release lyosecretome, a freeze-dried formulation of mesenchymal stem cell secretome with osteoinductive and osteoconductive properties) and withstanding loads (due to the presence of polycaprolactone, which provides mechanical resistance). To this end, an in-silico study was conducted to predict mechanical properties. Structural finite element analysis (FEA) of the hybrid scaffold under compression was performed to compare the numerical results with the corresponding experimental data. The impact of alginate inclusion and infill patterns on scaffold stiffness was investigated. Results show an increase in mechanical properties by changing the scaffold infill pattern (linear: 145.38±28.90 vs. honeycomb: 278.96±50.19, mean and standard deviation, n = 8), while alginate inclusion does not always impact the mechanical performance of the hybrid scaffold (stiffness: 145.38±28.90 vs. 195.42±38.68 N/mm, with vs without hydrogel inclusion, respectively). This is confirmed by FEA analysis, in which a good correspondence between experimental and numerical stiffness is shown (142±28.94 vs. 117.18, respectively, linear scaffold with hydrogel inclusion). In conclusion, the computational framework is a valid tool for predicting the mechanical performance of scaffolds and is promising for future clinical applications in the maxillofacial field.

3D bioprinted osteosarcoma model for experimental boron neutron capture therapy (BNCT) applications: Preliminary assessment.

Osteosarcoma is the most frequently primary malignant bone tumor characterized by infiltrative growth responsible for relapses and metastases. Treatment options are limited, and a new therapeutic option is required. Boron neutron capture therapy (BNCT) is an experimental alternative radiotherapy able to kill infiltrative tumor cells spearing surrounding healthy tissues. BNCT studies are performed on 2D in vitro models that are not able to reproduce pathological tumor tissue organization or on in vivo animal models that are expensive, time-consuming and must follow the 3R's principles. A 3D in vitro model is a solution to better recapitulate the complexity of solid tumors meanwhile limiting the animal's use. Objective of this study is to optimize the technical assessment for developing a 3D in vitro osteosarcoma model as a platform for BNCT studies: printing protocol, biomaterial selection, cell density, and crosslinking process. The best parameters that allow a fully colonized 3D bioprinted construct by rat osteosarcoma cell line UMR-106 are 6 × 106 cells/ml of hydrogel and 1% CaCl2 as a crosslinking agent. The proposed model could be an alternative or a parallel approach to 2D in vitro culture and in vivo animal models for BNCT experimental study.

Androgens and NGF Mediate the Neurite-Outgrowth through Inactivation of RhoA.

Steroid hormones and growth factors control neuritogenesis through their cognate receptors under physiological and pathological conditions. We have already shown that nerve growth factor and androgens induce neurite outgrowth of PC12 cells through a reciprocal crosstalk between the NGF receptor, TrkA and the androgen receptor. Here, we report that androgens or NGF induce neuritogenesis in PC12 cells through inactivation of RhoA. Ectopic expression of the dominant negative RhoA N19 promotes, indeed, the neurite-elongation of unchallenged and androgen- or NGF-challenged PC12 cells and the increase in the expression levels of ßIII tubulin, a specific neuronal marker. Pharmacological inhibition of the Ser/Thr kinase ROCK, an RhoA effector, induces neuritogenesis in unchallenged PC12 cells, and potentiates the effect of androgens and NGF, confirming the role of RhoA/ROCK axis in the neuritogenesis induced by androgen and NGF, through the phosphorylation of Akt. These findings suggest that therapies based on new selective androgen receptor modulators and/or RhoA/ROCK inhibitors might exert beneficial effects in the treatment of neuro-disorders, neurological diseases and ageing-related processes.

Impact of thoracic endovascular aortic repair on aortic biomechanics: Integration of in silico and ex vivo analysis using porcine model.

Thoracic endovascular aortic repair (TEVAR) is widespread in clinical practice for treating aortic diseases but it has relevant systemic complications, such as increase of the cardiac workload due to post-TEVAR aortic stiffening, and local issues such as re-entry tears due to the tissue damage caused by endograft interaction. The present study aims to elucidate these aortic biomechanical mechanisms by coupling ex vivo and in silico analysis. By ex vivo tests, the pulse wave velocity before and after TEVAR is measured. Uni-axial tensile tests are performed to measure regional mechanical response of tissue samples, supplied as input data for the in silico analysis. Numerical analysis is finally performed to compute the wall stress induced by the stent-graft deployment and the arterial pressurization. The ex vivo results highlight an increase of baseline PWV by a mean .78 m/s or 12% after TEVAR with a 100 mm stent-graft (p <.013). In the in silico analysis, the average von Mises stress in the landing zone increases of about 15% and 20% using, respectively stent-graft with radial oversizing of 10% and 20%. This work shows the effectiveness of integrated framework to analyze the biomechanical post TEVAR mechanisms. Moreover, the obtained results quantify the effect of prosthesis selection on the stiffening of the aorta after TEVAR and on the local increase of the aortic wall stress that is proportional to the stent-graft oversizing.

Computer-aided engineering and additive manufacturing for bioreactors in tissue engineering: State of the art and perspectives.

Two main challenges are currently present in the healthcare world, i.e., the limitations given by transplantation and the need to have available 3D in vitro models. In this context, bioreactors are devices that have been introduced in tissue engineering as a support for facing the mentioned challenges by mimicking the cellular native microenvironment through the application of physical stimuli. Bioreactors can be divided into two groups based on their final application: macro- and micro-bioreactors, which address the first and second challenge, respectively. The bioreactor design is a crucial step as it determines the way in which physical stimuli are provided to cells. It strongly depends on the manufacturing techniques chosen for the realization. In particular, in bioreactor prototyping, additive manufacturing techniques are widely used nowadays as they allow the fabrication of customized shapes, guaranteeing more degrees of freedom. To support the bioreactor design, a powerful tool is represented by computational simulations that allow to avoid useless approaches of trial-and-error. In the present review, we aim to discuss the general workflow that must be carried out to develop an optimal macro- and micro-bioreactor. Accordingly, we organize the discussion by addressing the following topics: general and stimulus-specific (i.e., perfusion, mechanical, and electrical) requirements that must be considered during the design phase based on the tissue target; computational models as support in designing bioreactors based on the provided stimulus; manufacturing techniques, with a special focus on additive manufacturing techniques; and finally, current applications and new trends in which bioreactors are involved.

Development and optimization of microfluidic assisted manufacturing process to produce PLGA nanoparticles.

Nanomedicine consists in the application of nanotechnology in medicine to revolutionize the healthcare sector through transformative new diagnostic and therapeutic tools. In this field, nanostructures or nanocarriers (i.e., nanoparticles) are extensively used as a drug delivery system. Despite the well-defined profits offered by nanomedicines based on poly (lactic-co-glycolic acid) (PLGA), the major barriers hampering the launch of a nanoparticles-based product on the market are batch-to-batch variations and its lack of reproducibility from the benchtop to an industrial scale production. Currently, microfluidics technology has emerged as potential tool to achieve a continuous manufacturing with a precise control over fluids mixing and particles quality attributes. This work aims at defining a tailored strategy to produce PLGA NPs, exploiting a new microfluidic device. Moreover, Design of Experiments (DoE) and computational fluid dynamics approaches were exploited to understand the main process parameters and material attributes affecting the quality of the final product as well as the NPs manufacturing process. Finally, the ability to incorporate a drug into the PLGA nanoparticles was investigated by using Curcumin as model payload reaching encapsulation efficiency in the rank 28-44%. This paper is proposed as useful guide for the preparation of PLGA NPs by microfluidic technique.

Mechanosensor YAP cooperates with TGF-ß1 signaling to promote myofibroblast activation and matrix stiffening in a 3D model of human cardiac fibrosis.

Cardiac fibrosis is characterized by a maladaptive remodeling of the myocardium, which is controlled by various inflammatory pathways and cytokines. This remodeling is accompanied by a significant stiffening of the matrix, which may contribute to further activate collagen synthesis and scar formation. Evidence suggests that TGF-ß1 signaling, the main pro-fibrotic pathway in cardiac fibrosis, might cooperates with the Hippo transcriptional pathway by activating YAP. To directly test the cooperation of mechanical cues and paracrine signaling in cardiac fibrosis, we developed a 3D model of cardiac extracellular matrix remodeling by generating tissue blocks with Gelatin Methacrylate, a bioink with tunable stiffness, and human cardiosphere-derived stromal cells. Using this strategy, we assessed the cooperation of TGF-ß1 and YAP transcriptional factor to matrix compaction. Using mechanical compression tests, Masson's trichrome staining, immunofluorescence, and RT-qPCR, we demonstrate that pharmacological inhibition of YAP complex reverts almost completely the pro-compaction phenotype and the matrix-remodeling activity of cells treated with TGF-ß1. Our data show a direct connection between the classical pro-fibrotic signaling driven by TGF-ß1 and the mechanically activated pathways under the control of YAP in cardiac remodeling. Treatment with the elective drug targeting YAP is sufficient to override this cooperation with potential benefits for anti-fibrotic therapeutic applications. STATEMENT OF SIGNIFICANCE: Heart failure is a pathology in continuous growth worldwide, characterized by a progressive fibrosis, which decreases the pumping efficiency of the heart. Experimental evidences suggest that fibroblasts, normally responsible for the turnover of the cardiac matrix, are involved in myocardial fibrosis by differentiating into 'myofibroblasts'. These cells remodel extensively the cardiac extracellular matrix and deposit abundant collagen with a consequent increase in stiffness. In the present contribution, we propose a new 3D model of cell-mediated cardiac extracellular matrix stiffening to investigate the mechano-chemical mechanisms underlying the onset of the pathology. We also consolidate a pharmacological treatment able to prevent the pathological activation of fibroblasts with potential benefits for anti-fibrotic treatment of the failing heart.

Photocatalyzed Functionalization of Alkenoic Acids in 3D-Printed Reactors.

The valorization of alkenoic acids possibly deriving from biomass (fumaric and citraconic acids) was carried out through conversion in important building blocks, such as γ-keto acids and succinic acid derivatives. The functionalization was carried out by addition onto the C=C double bond of radicals generated under photocatalyzed conditions from suitable hydrogen donors (mainly aldehydes) and by adopting a decatungstate salt as the photocatalyst. Syntheses were performed under batch (in a glass vessel) and flow (by using 3D-printed reactors) conditions. The design of the latter reactors allowed for an improved yield and productivity.