@article {2835,
	title = {Clarifying CLARITY: Quantitative Optimization of the Diffusion Based Delipidation Protocol for Genetically Labeled Tissue},
	journal = {Frontiers in Neuroscience},
	year = {2016},
	abstract = {<p>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 {\textquotedblleft}goodness{\textquotedblright} 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.</p>
},
	keywords = {Bioengineering},
	doi = {http://dx.doi.org/10.3389/fnins.2016.00179},
	url = {http://journal.frontiersin.org/article/10.3389/fnins.2016.00179/full},
	author = {C. Magliaro and A. L. Callara and G. Mattei and M. Morcinelli and C. Viaggi and Vaglini, F and A. Ahluwalia}
}
@conference {1801,
	title = {Continuous functionally graded materials (cFGMs) for TE},
	booktitle = {TERMIS 3rd World Congress},
	year = {2012},
	address = {Vienna, Austria},
	abstract = {<p>Biological structures are not uniform but possess spatially distributed functions and properties, or functional gradients. To ensure functional, mechanical and structural integration, a tissue engineered (TE) scaffold has to reproduce these functional gradients. However the fabrication of functionally graded materials is challenging and usually an experimental trial-and-error approach is used. In this work we present a controlled method for the fabrication of cFGMs using the gravitational sedimentation of discrete solid particles within a primary fluid phase. To have an overall control over particle distribution, a time-varying dynamic viscosity solution (i.e. thermo-sensitive) was used as fluid phase. Computational fluid dynamic models were developed to have a fine control over particle distribution. Biomimetic osteochondral cFGMs scaffolds were fabricated using hydroxyapatite (HA) and gelatin. Glutaraldehyde was used to covalently bind gelatin-HA graded scaffolds. Mechanical properties were measured and correlated as a function of HA volume fraction. SEM-EDX analysis was used to further characterise HA content and its distribution within gelatin-HA cFGMs. Finally gelatin-HA cFGMs scaffold were seeded using periosteum derived progenitor cells, to investigate how the HA gradient modulates cell response. This approach represents an innovative yet simple tool for the fabrication of tailored cFGMs with biologically and physiologically relevant gradients for TE applications.</p>
},
	keywords = {Bioengineering},
	author = {G. Mattei and A. Ahluwalia and Ferretti, C and Mattioli-Belmonte, M. and Tirella, A}
}
@article {1514,
	title = {Cooperation of Biological and Mechanical Signals in Cardiac Progenitor Cell Differentiation},
	journal = {ADVANCED MATERIALS},
	year = {2011},
	keywords = {Bioengineering},
	issn = {0935-9648},
	author = {S. Pagliari and A.C. Vilela-Silva and G. Forte and F. Pagliari and C. Mandoli and G. Vozzi and S. Pietronave and M. Prat and S. Licoccia and A. Ahluwalia and E. Traversa and M. Minieri and P. Di Nardo}
}
@article {1550,
	title = {Chitosan/gelatin blends for biomedical applications},
	journal = {JOURNAL OF BIOMEDICAL MATERIALS RESEARCH. PART A},
	volume = {86},
	year = {2008},
	pages = {311{\textendash}322},
	keywords = {Bioengineering},
	issn = {1549-3296},
	author = {PULIERI, E and V. Chiono and G. Ciardelli and G. Vozzi and A. Ahluwalia and C. Domenici and Vozzi, F. and P. Giusti}
}
@article {1546,
	title = {Comparative study of porous and engineered biomaterials},
	journal = {INTERNAL MEDICINE},
	volume = {15},
	year = {2008},
	pages = {4{\textendash}8},
	keywords = {Bioengineering},
	issn = {0918-2918},
	author = {Mattioli-Belmonte, M. and K KYRIAKIDOU and LUCARINI, G and ZIZZI, A and A. Ahluwalia and G. Vozzi}
}
@article {1559,
	title = {Controlled in vitro growth of cell microtubes: towards the realisation of artificial microvessels},
	journal = {BIOMEDICAL MICRODEVICES},
	year = {2008},
	keywords = {Bioengineering},
	issn = {1387-2176},
	author = {Migliore, A. and Vozzi, F. and G. Vozzi and A. Ahluwalia}
}
@article {1549,
	title = {Criticality of the Biological and Physical Stimuli Array Inducing Resident Cardiac Stem Cell Determination},
	journal = {STEM CELLS},
	volume = {26},
	year = {2008},
	pages = {2093{\textendash}2103},
	keywords = {Bioengineering},
	issn = {1066-5099},
	author = {G. Forte and CAROTENUTO, F and F. Pagliari and S. Pagliari and COSSA, P and FIACCAVENTO, R and A. Ahluwalia and G. Vozzi and Vinci, B. and SERAFINO, A and RINALDI, A and E. Traversa and CAROSELLA, L and M. Minieri and P. Di Nardo}
}
@conference {1657,
	title = {Cardiac and Mesenchymal Stem Cell Growth and Selective Differentiation on Three Dimensional Bioerodable Scaffolds},
	booktitle = {Termis Eu-Meeting London (UK) 4-7 September 2007 Tissue Engineering 2007, 13 (7): 1647},
	year = {2007},
	pages = {{\textendash}},
	keywords = {Bioengineering},
	author = {G. Forte and CAROTENUTO, F and G. Vozzi and COSSA, P and FIACCAVENTO, R and M. Minieri and F. Pagliari and S. Pagliari and Romano, R. and A. Ahluwalia and E. Traversa and P. Di Nardo}
}
