01836nas a2200205 4500008004100000245007900041210006900120260001200189520120800201653001901409100001801428700001401446700001401460700001701474700001501491700001901506700001501525700001301540856007701553 2018 eng d00aSystemic and vascular inflammation in an in-vitro model of central obesity0 aSystemic and vascular inflammation in an invitro model of centra c02/20183 a

Metabolic disorders due to over-nutrition are a major global health problem, often associated with obesity and related morbidities. Obesity is peculiar to humans, as it is associated with lifestyle and diet, and so difficult to reproduce in animal models. Here we describe a model of human central adiposity based on a 3-tissue system consisting of a series of interconnected fluidic modules. Given the causal link between obesity and systemic inflammation, we focused primarily on pro-inflammatory markers, examining the similarities and differences between the 3-tissue model and evidence from human studies in the literature. When challenged with high levels of adiposity, the in-vitro system manifests cardiovascular stress through expression of E-selectin and von Willebrand factor as well as systemic inflammation (expressing IL-6 and MCP-1) as observed in humans. Interestingly, most of the responses are dependent on the synergic interaction between adiposity and the presence of multiple tissue types. The set-up has the potential to reduce animal experiments in obesity research and may help unravel specific cellular mechanisms which underlie tissue response to nutritional overload.

10aBioengineering1 aAhluwalia, A.1 aMisto, A.1 aVozzi, G.1 aMagliaro, C.1 aMattei, G.1 aMC, Marescotti1 aAVOGARO, A1 aIori, E. uhttp://journals.plos.org/plosone/article?id=10.1371/journal.pone.019282402116nas a2200193 4500008004100000245009500041210006900136260001200205520148600217653001901703100001501722700001701737700001401754700002501768700001401793700002001807700001801827856007701845 2017 eng d00aOn the adhesion-cohesion balance and oxygen consumption characteristics of liver organoids0 aadhesioncohesion balance and oxygen consumption characteristics  c03/20173 a

Liver organoids (LOs) are of interest in tissue replacement, hepatotoxicity and pathophysiological studies. However, it is still unclear what triggers LO self-assembly and what the optimal environment is for their culture. Hypothesizing that LO formation occurs as a result of a fine balance between cell-substrate adhesion and cell-cell cohesion, we used 3 cell types (hepatocytes, liver sinusoidal endothelial cells and mesenchymal stem cells) to investigate LO self-assembly on different substrates keeping the culture parameters (e.g. culture media, cell types/number) and substrate stiffness constant. As cellular spheroids may suffer from oxygen depletion in the core, we also sought to identify the optimal culture conditions for LOs in order to guarantee an adequate supply of oxygen during proliferation and differentiation. The oxygen consumption characteristics of LOs were measured using an O2 sensor and used to model the O2 concentration gradient in the organoids. We show that no LO formation occurs on highly adhesive hepatic extra-cellular matrix-based substrates, suggesting that cellular aggregation requires an optimal trade-off between the adhesiveness of a substrate and the cohesive forces between cells and that this balance is modulated by substrate mechanics. Thus, in addition to substrate stiffness, physicochemical properties, which are also critical for cell adhesion, play a role in LO self-assembly.

10aBioengineering1 aMattei, G.1 aMagliaro, C.1 aGiusti, S1 aRamachandran, S., D.1 aHeinz, S.1 aBraspenning, J.1 aAhluwalia, A. uhttp://journals.plos.org/plosone/article?id=10.1371/journal.pone.017320602255nas a2200157 4500008004100000245012100041210006900162260001200231520172100243653001901964100001501983700001701998700001502015700001802030856004902048 2017 eng d00aDecellularized Human Liver Is Too Heterogeneous for Designing a Generic Extracellular Matrix Mimic Hepatic Scaffold.0 aDecellularized Human Liver Is Too Heterogeneous for Designing a  c05/20173 a

Decellularized human livers are considered the perfect extracellular matrix (ECM) surrogate because both three-dimensional architecture and biological features of the hepatic microenvironment are thought to be preserved. However, donor human livers are in chronically short supply, both for transplantation or as decellularized scaffolds, and will become even scarcer as life expectancy increases. It is hence of interest to determine the structural and biochemical properties of human hepatic ECM to derive design criteria for engineering biomimetic scaffolds. The intention of this work was to obtain quantitative design specifications for fabricating scaffolds for hepatic tissue engineering using human livers as a template. To this end, hepatic samples from five patients scheduled for hepatic resection were decellularized using a protocol shown to reproducibly conserve matrix composition and microstructure in porcine livers. The decellularization outcome was evaluated through histological and quantitative image analyses to evaluate cell removal, protein, and glycosaminoglycan content per unit area. Applying the same decellularization protocol to human liver samples obtained from five different patients yielded five different outcomes. Only one liver out of five was completely decellularized, while the other four showed different levels of remaining cells and matrix. Moreover, protein and glycosaminoglycan content per unit area after decellularization were also found to be patient- (or donor-) dependent. This donor-to-donor variability of human livers thus precludes their use as templates for engineering a generic "one-size fits all" ECM-mimic hepatic scaffold.

10aBioengineering1 aMattei, G.1 aMagliaro, C.1 aPirone, A.1 aAhluwalia, A. uhttps://www.ncbi.nlm.nih.gov/pubmed/2854340302803nas a2200181 4500008004100000245012600041210006900167520217400236653001902410100001702429700002002446700001502466700001902481700001502500700001502515700001802530856007302548 2016 eng d00aClarifying CLARITY: Quantitative Optimization of the Diffusion Based Delipidation Protocol for Genetically Labeled Tissue0 aClarifying CLARITY Quantitative Optimization of the Diffusion Ba3 a

Tissue clarification has been recently proposed to allow deep tissue imaging without light scattering. The clarification parameters are somewhat arbitrary and dependent on tissue type, source and dimension: every laboratory has its own protocol, but a quantitative approach to determine the optimum clearing time is still lacking. Since the use of transgenic mouse lines that express fluorescent proteins to visualize specific cell populations is widespread, a quantitative approach to determine the optimum clearing time for genetically labeled neurons from thick murine brain slices using CLARITY2 is described. In particular, as the main objective of the delipidation treatment is to clarify tissues, while limiting loss of fluorescent signal, the “goodness” of clarification was evaluated by considering the bulk tissue clarification index (BTCi) and the fraction of the fluorescent marker retained in the slice as easily quantifiable macroscale parameters. Here we describe the approach, illustrating an example of how it can be used to determine the optimum clearing time for 1 mm-thick cerebellar slice from transgenic L7GFP mice, in which Purkinje neurons express the GFP (green fluorescent protein) tag. To validate the method, we evaluated confocal stacks of our samples using standard image processing indices (i.e., the mean pixel intensity of neurons and the contrast-to-noise ratio) as figures of merit for image quality. The results show that detergent-based delipidation for more than 5 days does not increase tissue clarity but the fraction of GFP in the tissue continues to diminish. The optimum clearing time for 1 mm-thick slices was thus identified as 5 days, which is the best compromise between the increase in light penetration depth due to removal of lipids and a decrease in fluorescent signal as a consequence of protein loss: further clearing does not improve tissue transparency, but only leads to more protein removal or degradation. The rigorous quantitative approach described can be generalized to any clarification method to identify the moment when the clearing process should be terminated to avoid useless protein loss.

10aBioengineering1 aMagliaro, C.1 aCallara, A., L.1 aMattei, G.1 aMorcinelli, M.1 aViaggi, C.1 aVaglini, F1 aAhluwalia, A. uhttp://journal.frontiersin.org/article/10.3389/fnins.2016.00179/full00587nas a2200157 4500008004100000245009000041210006900131260001200200653001900212100001700231700001500248700001500263700001500278700001800293856011800311 2015 eng d00a HisTOOLogy: an open-source tool for quantitative analysis of histological sections 0 aHisTOOLogy an opensource tool for quantitative analysis of histo c08/201510aBioengineering1 aMagliaro, C.1 aTirella, A1 aMattei, G.1 aPirone, A.1 aAhluwalia, A. uhttp://118699.obes.asia/publications/histoology-open-source-tool-quantitative-analysis-histological-sections.html00631nas a2200169 4500008004100000245016100041210006900202260001200271300001000283490001000293653001900303100001500322700001700337700001800354700001800372856007100390 2015 eng d00aParvalbumin expression in the claustrum of human dog. An immunohistochemical and topographical study with comparative notes of the structure of the nucleus0 aParvalbumin expression in the claustrum of human dog An immunohi c03/2015 a33-420 v64-6510aBioengineering1 aPirone, A.1 aMagliaro, C.1 aGiannessi, E.1 aAhluwalia, A. uhttp://www.sciencedirect.com/science/article/pii/S089106181500014900511nas a2200157 4500008004100000245009700041210006900138260001200207490001100219653001900230100001500249700001800264700001700282700001800299856003600317 2015 eng d00aProfile analysis of hepatic porcine and murine brain tissue slices obtained with a vibratome0 aProfile analysis of hepatic porcine and murine brain tissue slic c04/20150 v3:e93210aBioengineering1 aMattei, G.1 aCristiani, I.1 aMagliaro, C.1 aAhluwalia, A. uhttps://peerj.com/articles/932/00621nas a2200169 4500008004100000245010000041210006900141490000600210653001900216100001500235700001700250700001400267700001700281700001600298700001800314856011900332 2014 eng d00aSphyga: a multiparameter open source tool for fabricating smart and tunable hydrogel microbeads0 aSphyga a multiparameter open source tool for fabricating smart a0 v610aBioengineering1 aTirella, A1 aMagliaro, C.1 aPenta, M.1 aTroncone, M.1 aPimentel, R1 aAhluwalia, A. uhttp://118699.obes.asia/publications/sphyga-multiparameter-open-source-tool-fabricating-smart-and-tunable-hydrogel00565nas a2200145 4500008004100000245009100041210006900132490000600201653001900207100001500226700001700241700001500258700001800273856012800291 2013 eng d00aNEuronMOrphological analysis tool: open-source software for quantitative morphometrics0 aNEuronMOrphological analysis tool opensource software for quanti0 v710aBioengineering1 aBilleci, L1 aMagliaro, C.1 aPioggia, G1 aAhluwalia, A. uhttp://118699.obes.asia/publications/neuronmorphological-analysis-tool-open-source-software-quantitative-morphometrics.html