Nanoworld Institute

Genoa, Italy

Nanoworld Institute

Genoa, Italy
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Barone A.,Versilia Hospital | Barone A.,University of Genoa | Ricci M.,Nanoworld Institute | Calvo-Guirado J.L.,University of Murcia | And 2 more authors.
Clinical Oral Implants Research | Year: 2011

Introduction: After a tooth extraction, the height of the buccal wall tends to decrease. The literature indicates that regenerative techniques (guided bone regenerative [GBR] techniques) have succeeded in improving the bone levels. Therefore, this experiment set out to compare the physiological bone remodelling in Beagle dog models after implant placement in a fresh extraction socket, with and without the application of regenerative procedure. Materials and methods: Five dogs were used in this study. Test and control sites were randomly selected. The experimental teeth (fourth pre-molar and first molar) were hemi-sected removing the distal roots and placing implants. Porcine bone was placed to fill the gap around the implant on the test sites and a reabsorbable membrane was used to cover the area. The dogs were put down at different times (2 weeks, 1 month and 3 months). The measurements were taken immediately and at 2, 4, 12 weeks after implant placement. Student's test for paired data was used to compare the means of the clinical measurements. Results: At 2 weeks: On the control sites, few signs of resorption were detected at the first molar only, while at the test sites bone levels were placed at the implant shoulder or above. At 4 weeks: On the control site, slight bone remodelling was observed, while on the test site minor signs of resorption or an increase of bone levels were detected. At 12 weeks: The alveolar crest on the control sites showed various degrees of remodelling. On the test sites stable bone levels or an increase of bone crest was observed. Conclusion: With the limits of this study, the findings showed that GBR techniques were able to limit resorption of the alveolar crest after tooth extraction. A pattern of bone remodelling after tooth extraction and implant placement was observed in the control sites (no GBR) as well as in test sites (GBR), and although the exact cause of this is unclear, surgical trauma could play a role. Further studies are necessary to confirm these results and to clarify the precise causes of bone remodelling in fresh extraction sockets. © 2011 John Wiley & Sons A/S.

Nicolini C.,University of Genoa | Nicolini C.,Nanoworld Institute | Nicolini C.,Arizona State University | Bragazzi N.,University of Genoa | And 2 more authors.
Advanced Drug Delivery Reviews | Year: 2012

Nucleic Acid Programmable Protein Arrays utilize a complex mammalian cell free expression system to produce proteins in situ. In alternative to fluorescent-labeled approaches a new label free method, emerging from the combined utilization of three independent and complementary nanotechnological approaches, appears capable to analyze protein function and protein-protein interaction in studies promising for personalized medicine. Quartz Micro Circuit nanogravimetry, based on frequency and dissipation factor, mass spectrometry and anodic porous alumina overcomes indeed the limits of correlated fluorescence detection plagued by the background still present after extensive washes. This could be further optimized by a homogeneous and well defined bacterial cell free expression system capable to realize the ambitious objective to quantify the regulatory protein networks in humans. Implications for personalized medicine of the above label free protein array using different test genes proteins are reported. © 2012 Elsevier B.V.

Nicolini C.,Nanoworld Institute | Bragazzi N.,Nanoworld Institute | Pechkova E.,Nanoworld Institute
Journal of Bioanalysis and Biomedicine | Year: 2013

This overview describes the optimal implementation and utilization of different, newly conceived nanosensors for human biosafety purposes, exploiting a variety of methods (amperometric, conductometric, spectrometric and nanogravimetric), and a wide range of nanocomposites, genes and recombinant enzymes. Namely, while biological nanosensors were designed based on Nucleic Acid Programmable Protein Arrays (NAPPA), with or without SNAPtag, and on Langmuir-Blodgett (LB) thin films of recombinant laccase (Rigidoporus lignosus, formerly known as Rigidoporus microporus), organic nanosensors were based on matrices of calcium oxide (CaO) and on carbon nanotubes-either multi-walled (MWNTs) or single-walled (SWNTs)- embedded in poly(o-methylaniline) (POTO). Special attention was paid both to detecting useful and relevant substances (such as carbon dioxide, phenols and phenolic derivatives and compounds) and designing devices and molecules for human biosafety like vaccines and others, by means of amperometry, conductimetry, mass spectrometry (MS) and other label-free technologies, such as quartz crystal microbalance with dissipation monitoring (QCM_D). © 2013 Nicolini C, et al.

Pechkova E.,Nanoworld Institute | Gebhardt R.,European Synchrotron Radiation Facility | Riekel C.,European Synchrotron Radiation Facility | Nicolini C.,Nanoworld Institute
Biophysical Journal | Year: 2010

In this study, we used microbeam grazing-incidence small-angle x-ray scattering (mGISAXS) to investigate in situ protein nucleation and crystal growth assisted by a protein nanotemplate, and introduced certain innovations to improve the method. Our aim was to understand the protein nanotemplate method in detail, as this method has been shown to be capable of accelerating and increasing crystal size and quality, as well as inducing crystallization of proteins that are not crystallizable by classical methods. The nanotemplate experimental setup was used for drops containing growing protein crystals at different stages of nucleation and growth. Two model proteins, lysozyme and thaumatin, were used under unique flow conditions to differentially probe protein crystal nucleation and growth. © 2010 by the Biophysical Society.

Gebhardt R.,European Synchrotron Radiation Facility | Gebhardt R.,TU Munich | Pechkova E.,Nanoworld Institute | Riekel C.,European Synchrotron Radiation Facility | Nicolini C.,Nanoworld Institute
Biophysical Journal | Year: 2010

The formation of thaumatin crystals by Langmuir-Blodgett (LB) film nanotemplates was studied by the hangingdrop technique in a flow-through cell by synchrotron radiation micrograzing-incidence small-angle x-ray scattering. The kinetics of crystallization was measured directly on the interface of the LB film crystallization nanotemplate. The evolution of the micro-grazing-incidence small-angle x-ray scattering patterns suggests that the increase in intensity in the Yoneda region is due to protein incorporation into the LB film. The intensity variation suggests several steps, which were modeled by system dynamics based on first-order differential equations. The kinetic data can be described by two processes that take place on the LB film, a first, fast, process, attributed to the crystal growth and its detachment from the LB film, and a second, slower process, attributed to an unordered association and conversion of protein on the LB film. © 2010 by the Biophysical Society.

Pechkova E.,Nanoworld Institute | Nicolini C.,Fondazione ELBA
Journal of Synchrotron Radiation | Year: 2011

Ultrasmall lysozyme microcrystals are grown by classical hanging-drop vapor diffusion and by its modification using a homologous protein thin-film template displaying long-range order. The nucleation and growth mechanisms of lysozyme microcrystals are studied at the thin lysozyme film surface using a new in situ μGISAXS (microbeam grazing-incidence small-angle X-ray scattering) technique recently developed at the microfocus beamline of the ESRF in Grenoble, France. New insight on the nucleation and crystallization processes appear to emerge. © 2011 International Union of Crystallography.

Nicolini C.,Nanoworld Institute
Bioengineered | Year: 2013

This paper investigates the application of anodic porous alumina as an advancement on chip laboratory for gene expressions. The surface was prepared by a suitable electrolytic process to obtain a regular distribution of deep micrometric holes and printed bypen robot tips under standard conditions. The gene expression within the Nucleic Acid Programmable Protein Array (NAPPA) is realized in a confined environment of 16 spots, containing circular DNA plasmids expressed using rabbit reticulocyte lysate. Authors demonstrated the usefulness of APA in withholding the protein expression by detecting with a CCD microscope the photoluminescence signal emitted from the complex secondary antibody anchored to Cy3 and confined in the pores. Friction experiments proved the mechanical resistance under external stresses by the robot tip pens printing. So far, no attempts have been made to directly compare APA with any other surface/substrate; the rationale for pursuing APA as a potential surface coating is that it provides advantages over the simple functionalization of a glass slide, overcoming concerns about printing and its ability to generate viable arrays.

Nicolini C.,Nanoworld Institute
Wiley Interdisciplinary Reviews: Nanomedicine and Nanobiotechnology | Year: 2010

This review presents the status of technological developments of nanogenomics and its applications to medicine. Even if particular emphasis is placed on what has been accomplished in our laboratory in the last few years in the area of genes microarrays, significant reference to the recent activity of numerous other groups can be found in Refs 1,2. © 2009 John Wiley & Sons, Inc.

Bragazzi N.L.,Nanoworld Institute | Sivozhelezov V.,Nanoworld Institute | Nicolini C.,Nanoworld Institute
Journal of Proteomics and Bioinformatics | Year: 2011

DNA microarrays are one of the most promising methods for molecular genomics, but this technique is often associated with experimental complications and difficulties in the analysis. Moreover, the greatest part of genes displayed on an array is often not directly involved in the cellular process being studied. Recently, we proposed a data mining algorithm, based on the identification of genes involved in a given process, the calculation of interactions among them and their ranking according to number of interactions. Genes in the highest cluster are defined as "leader genes". These findings may lead to an ad hoc and therefore more significant experimentation. However, at present this complex process is performed manually. In this work, we present the general architecture of LeaderGene, an automated tool for ab-initio molecular genomics. Three different and independent parts: (1) Identification of gene list; (2) Calculation of weighted number of links; (3) Genes clustering. Initial inputs are provided by user; then, output of part 1 and part 2, respectively, become inputs of parts 2 and 3. The development of an user-friendly software capable to automatically compute leader genes in a given cellular system will allow further progresses in this field of molecular genomics. © 2011 Bragazzi NL, et al.

Pechkova E.,Nanoworld Institute | Nicolini C.,Nanoworld Institute
Anticancer Research | Year: 2010

New topographic details appeared evident in protein crystal buffered with glycerol solution native on mica by atomic force microscopy and after laser irradiation on glass by light microscopy. This observation indicates the existence of distinct domains in the 3D crystal organisation that are quite different in size and number between the lysozyme crystals grown by Langmuir-Blodgett (LB) nanotemplate with respect to traditional hanging-drop vapour diffusion. Nanodiffraction by highly focused synchrotron radiation of laser cut submicron crystals confirmed the atomic structure of all residues of LB lysozyme crystals as being the most resistant to radiation damage. Crystals grown by LB nanotemplate still diffracted at good resolution after several steps of X-ray 'burning', while the classical crystals decayed very quickly at the same exposure.

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