Acute kidney injury (AKI) is an emergent public health problem that affects millions of patients worldwide. Several pharmacologic therapies that can accelerate recovery and improve survival were successful in experimental models but failed to manifest any significant beneficial effect in clinical practice [1]. Recent studies have indicated that adult renal stem/progenitor cells (ARPCs) are able to repair damaged renal proximal tubular epithelial cells (RPTECs) in AKI induced by toxic agents, via the secretion of both inhibin-A (INHB-A), FGF2 chemokines and specific microvesicle-vehicled mRNA [2]. Our goal was to synthesize a natural polymer-based nanosystem for efficacious delivery of INHB-A. Alginate (AL) and chitosan (CS), due to their promising properties are being exploited for the development of drug delivery systems [3]. INHB-Aloaded Polysaccharides Synthetic Vesicles (INHB-A-PSSV) were synthesized by two steps methods: ionotropic pre-gelation of AL core, followed by CS polyelectrolyte complexation. A microfluidic device was appositely fabricated in order to optimize the INHB-PSSV -at interface-assembly process, in terms of polymers and INHB-A working amount as well as vesicles size distribution. TEM and DLS characterization showed highly mono-disperse spherical PSSV ((157 ± 30) nm in diameter) and ?-potential measurement ((+56 ± 4) mV) confirmed the CS coating stability. Cellular uptake and INHB-A-PSSV effectiveness were tested in an in vitro model of cisplatin (CisPt) induced cell toxicity. RPTECs were exposed to 2.5 µmol/l CisPt for 6 h and, after drug withdrawal, cell viability was performed at 3 days after treatment. CisPt treated cell viability significantly decreased, compared with healthy control. We showed that addition of INHB-A-PSSV to CisPt -treated RPTECs led to a substantial increase in cell number and viability after 3 days of culture. Remarkably, a very low dosage of functional loaded protein (8 ng/25 µl) was sufficient to induce cell regeneration and the percentage of viable cells was similar to that of RPTECs without CisPt treatment.

Carnosine is an endogenous dipeptide abundant in the central nervous system, where by acting as intracellular pH buffering molecule, Zn/Cu ion chelator, antioxidant and anti-crosslinking agent, it exerts a well-recognized multi-protective homeostatic function for neuronal and non-neuronal cells. Carnosine seems to counteract proteotoxicity and protein accumulation in neurodegenerative conditions, such as Alzheimer's Disease (AD). However, its direct impact on the dynamics of AD-related fibril formation remains uninvestigated. We considered the effects of carnosine on the formation of fibrils/aggregates of the amyloidogenic peptide fragment A beta 1-42, a major hallmark of AD injury. Atomic force microscopy and thioflavin T assays showed inhibition of A beta 1-42 fibrillogenesis in vitro and differences in the aggregation state of A beta 1-42 small pre-fibrillar structures (monomers and small oligomers) in the presence of carnosine. in silico molecular docking supported the experimental data, calculating possible conformational carnosine/A beta 1-42 interactions. Overall, our results suggest an effective role of carnosine against A beta 1-42 aggregation

Alginate (ALG) and chitosan (CS) have been extensively used for biomedical applications; however, data relative to immune responses exerted by them are scarce. We synthesized a submicron vesicle system (SV) displaying a CS shell over an ALG core. Intravenous injection of these promising carriers could be a possible route of delivery; therefore, we evaluated their impact on human peripheral blood mononuclear cells (PBMCs). By this ex vivo approach, we established how SV chemical-physical characteristics affected the immune cells in terms of cellular uptake, viability, and state of activation. By flow cytometry, we demonstrated that SVs were internalized by PBMCs with differential trends. No substantial necrotic and apoptotic signals were recorded, and SVs weakly affected activation status of PBMCs (concerning the markers CD69, CD25, CD80, and the cytokines TNF-? and IL-6), showing high immune biocompatibility and low immunomodulating properties. Our findings gain particular value toward the biomedical applications of SVs and make these polymer-based structures more attractive for translation into clinical uses.

Acute kidney injury (AKI) is a public health problem worldwide. Several therapeutic strategies have been made to accelerate recovery and improve renal survival. Recent studies have shown that human adult renal progenitor cells (ARPCs) participate in kidney repair processes, and may be used as a possible treatment to promote regeneration in acute kidney injury. Here, we show that human tubular ARPCs (tARPCs) protect physically injured or chemically damaged renal proximal tubular epithelial cells (RPTECs) by preventing cisplatin-induced apoptosis and enhancing proliferation of survived cells. tARPCs without toll-like receptor 2 (TLR2) expression or TLR2 blocking completely abrogated this regenerative effect. Only tARPCs, and not glomerular ARPCs, were able to induce tubular cell regeneration process and it occurred only after damage detection. Moreover, we have found that ARPCs secreted inhibin-A and decorin following the RPTEC damage and that these secreted factors were directly involved in cell regeneration process. Polysaccharide synthetic vesicles containing these molecules were constructed and co-cultured with cisplatin damaged RPTECs. These synthetic vesicles were not only incorporated into the cells, but they were also able to induce a substantial increase in cell number and viability. The findings of this study increase the knowledge of renal repair processes and may be the first step in the development of new specific therapeutic strategies for renal repair.

In the field on nanomedicine, superparamagnetic nanoparticles are one of the moststudied nanomaterials for theranostics. In this paper, a one-pot synthesis of magneticnanoparticles is presented, with elevated control on particles size from 10 to 40 nm. Themonitoring of vacuum level is here introduced as a crucial parameter for achieving a fine particlemorphology. Magnetic properties of these nanoparticles are highly affected by disorders ormismatches in crystal structure. A prolonged oxidation step is applied to the obtainednanoparticles to transform the magnetic phases into a pure maghemite one, confirmed by a highresolution XPS analysis, by Mössbauer spectrometry and, indirectly, by increased performancesin magnetization curves and in relaxation times. Afterward, the attained nanoparticles aretransferred in water by a non-derivatized dextran coating. The thermogravimetric analysisconfirms that the polysaccharide molecules replace the oleic acid on the surface by stabilizingthe particles in aqueous phase and culture media. Preliminary in vitro test reveals as the dextrancoated nanoparticles are not passively internalized from the cells. As proof of concept, asecondary layer of chitosan assures a positive charge to the nanoparticle surface, thus enhancingthe cellular internalization.

Cytokine proteins are known as biomarker molecules, characteristic of a disease or specific bodycondition. Monitoring of the cytokine pattern in body fluids contributes to the diagnosis of rheumatic diseases associated with rheumatoid factor. Here, the proinflammatory cytokine Tumor Necrosis Factor alpha (TNF-?) was chosen as the first target of interest. We report on the development of a fast sandwich immunosorbent assay: anti-TNF-? antibody (Ab) immobilization on a pre-activated 3D micro-structured glass slide, coupled with direct fluorescence readout method. Two differently functionalized glass - Ab-immobilization substrates - were compared: O2 PLASMA/APTESactivated glass - having a two-dimensional structure-, and the polystyrene (PS) beads coated glass, -having a 3D structure-. The 3D structured substrate was aimed to increase antibody anchorage sites. Then, the optimization of buffer media was carefully investigated with respect to non-specific protein binding. As a first step towards real sample analysis, a proof of principle of on/ off cytokine detection has been analyzed in the presence of human serum. High-density one-step immobilizationof anti-TNF-? antibody onto nanobead-coated glass slide chip has been demonstrated to be a promising device for application in in vitro diagnostic and profiling of cytokines with potential application toward personalized therapeutic interventions.

Oxygen consumption rate (OCR) is a significant parameter helpful to determine in vitro respiratory efficiency of living cells. Oxygen is an excellent oxidant and its electrocatalytic reduction on a noble metal allows accurately detecting it. By means of microfabrication technologies, handy, low-cost, and disposable chip can be attained, minimizing working volumes and improving sensitivity and response time. In this respect, here is presented a microoxygraph device (MOD), based on Clark's electrode principle, displaying many advantageous features in comparison to other systems. This lab-on-chip platform is composed of a three-microelectrode detector equipped with a microgrooved electrochemical cell, sealed with a polymeric reaction chamber. Au working/counter electrodes and Ag/AgCl reference electrode were fabricated on a glass slide. A microchannel was realized by photoresist lift-off technique and a polydimethylsiloxane (PDMS) nanoporous film was integrated as oxygen permeable membrane (OPM) between the probe and the microreaction chamber. Electrochemical measurements showed good reproducibility and average response time, assessed by periodic injection and suction of a reducing agent. OCR measurements on 3T3 cells, subjected, in real time, to chemical stress on the respiratory chain, were able to show that this chip allows performing consistent metabolic analysis.

The aim of this study is to investigate the effect of a non-invasive physical treatment on activated microglia of a well-established mouse model of Parkinson disease (PD), based on intoxication with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). To this aim, we proposed a Radio electric asymmetric conveyer (REAC) technology, with its specific therapeutic protocols of regenerative medicine: tissue optimization - regenerative (TO-RGN), as an innovative therapeutic strategy for the treatment of PD. We observed that REAC TO-RGN exposition can attenuate the damage of nigrostriatal pathway induced by MPTP treatment in mice, decreasing levels of pro-inflammatory mediators in the substantia nigra pars compacta (SNpc) of MPTP-mice. Besides, TH immunostaining in MPTP-treated mice exposed to REAC TO-RGN resulted more pronounced in both substantia nigra and striatum in comparison to animals that received only MPTP treatment. Overall, these data suggest that REAC TO-RGN treatment can have neuroprotective effects in MPTP-induced PD mice model, which may be related to reduced inflammatory reaction.

Novel synthetic peptides represent smart molecules for antigen antibody interactions in several bioanalytics applications, from purification to serum screening. Their immobilization onto a solid phase is considered a key point for sensitivity increasing. In this view, we exploited Quartz Crystal Microbalance with simultaneous frequency and dissipation monitoring (QCM-D) with a double aim, specifically, as investigative tool for spacers monolayer assembling and its functional evaluation, as well as high sensitive method for specific immunosorbent assays. The method was applied to pancreatic ductal adenocarcinoma (PDAC) detection by studying the interactions between synthetic phosphorylated and un-phosphorylated alpha-enolase peptides with sera of healthy and PDAC patients. The synthetic peptides were immobilized on the gold surface of the QCM-D sensor via a self-assembled alkanethiol monolayer. The presented experimental results can be applied to the development of surfaces less sensitive to non-specific interactions with the final target to suggest specific protocols for detecting PDAC markers with un-labeled biosensors.

In this study, the effects of Radio Electric Asymmetric Conveyer (REAC), a non-invasive physical treatment, on neuroinflammatory responses in a mouse model of parkinsonism induced by intoxication with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), were investigated in vivo. We found that the REAC tissue optimization treatment specific for neuro-regenerative purposes (REAC TO-RGN-N) attenuated the inflammatory picture evoked by MPTP-induced nigro-striatal damage in mice, decreasing the levels of pro-inflammatory molecules and increasing anti-inflammatory mediators. Besides, there was a significant reduction of both astrocyte and microglial activation in MPTP-treated mice exposed to REAC TO-RGN-N. These results indicated that REAC TO-RGN-N treatment modulates the pro-inflammatory responses and reduces neuronal damage in MPTP-induced parkinsonism.

The present invention refers to a micro culture device (5) to be inserted in a chamber (4) of an oxygraph (7). The micro culture device is divided into an upper microchamber (2) and a lower microchamber (3) by a semi-permeable nanoporous membrane (1), suitable for acting as adhering support for cells. The upper microchamber (2) presents at its upper side either an inlet (2.1) coaxially aligned with an oxygraph plunger precision bore (6), or a semi-permeable membrane. The lower microchamber (3) presents at its bottom side either an inlet (3.1) coaxially aligned with an oxygraph piston bore (8) or a semi-permeable membrane (3.2).