A 3D scanning system for biomedical purposes

The use of three-dimensional (3D) scanning systems for acquiring the external shape features of biological objects has recently been gaining popularity in the biomedical field. A simple, low cost, 3D scanning syste

MicroRNAs delivery into human cells grown on 3D-printed PLA scaffolds coated with a novel fluorescent PAMAM dendrimer for biomedical applications

Many advanced synthetic, natural, degradable or non-degradable materials have been employed to create scaffolds for cell culture for biomedical or tissue engineering applications. One of the most versatile material is poly-lactide (PLA), commonly used as 3D printing filament. Manufacturing of multifunctional scaffolds with improved cell growth proliferation and able to deliver oligonucleotides represents an innovative strategy for controlled and localized gene modulation that hold great promise and could increase the number of applications in biomedicine. Here we report for the first time the synthesis of a novel Rhodamine derivative of a poly-amidoamine dendrimer (G = 5) able to transfect cells and to be monitored by confocal microscopy that we also employed to coat a 3D-printed PLA scaffold. The coating do not modify the oligonucleotide binding ability, toxicity or transfection properties of the scaffold that is able to increase cell proliferation and deliver miRNA mimics (i.e., pre-mir-503) into human cells. Although further experiments are required to optimize the dendrimer/miRNA ratio and improve transfection efficiency, we demonstrated the effectiveness of this promising and innovative 3D-printed transfection system to transfer miRNAs into human cells for future biomedical applications. © 2018, The Author(s)

Phase-Retrieved Tomography enables imaging of a Tumor Spheroid in Mesoscopy Regime

Optical tomographic imaging of biological specimen bases its reliability on the combination of both accurate experimental measures and advanced computational techniques. In general, due to high scattering and absorption in most of the tissues, multi view geometries are required to reduce diffuse halo and blurring in the reconstructions. Scanning processes are used to acquire the data but they inevitably introduces perturbation, negating the assumption of aligned measures. Here we propose an innovative, registration free, imaging protocol implemented to image a human tumor spheroid at mesoscopic regime. The technique relies on the calculation of autocorrelation sinogram and object autocorrelation, finalizing the tomographic reconstruction via a three dimensional Gerchberg Saxton algorithm that retrieves the missing phase information. Our method is conceptually simple and focuses on single image acquisition, regardless of the specimen position in the camera plane. We demonstrate increased deep resolution abilities, not achievable with the current approaches, rendering the data alignment process obsolete.Comment: 21 pages, 5 figure

Restriction landmark genomic scanning (RLGS) spot identification by second generation virtual RLGS in multiple genomes with multiple enzyme combinations.

BackgroundRestriction landmark genomic scanning (RLGS) is one of the most successfully applied methods for the identification of aberrant CpG island hypermethylation in cancer, as well as the identification of tissue specific methylation of CpG islands. However, a limitation to the utility of this method has been the ability to assign specific genomic sequences to RLGS spots, a process commonly referred to as "RLGS spot cloning."ResultsWe report the development of a virtual RLGS method (vRLGS) that allows for RLGS spot identification in any sequenced genome and with any enzyme combination. We report significant improvements in predicting DNA fragment migration patterns by incorporating sequence information into the migration models, and demonstrate a median Euclidian distance between actual and predicted spot migration of 0.18 centimeters for the most complex human RLGS pattern. We report the confirmed identification of 795 human and 530 mouse RLGS spots for the most commonly used enzyme combinations. We also developed a method to filter the virtual spots to reduce the number of extra spots seen on a virtual profile for both the mouse and human genomes. We demonstrate use of this filter to simplify spot cloning and to assist in the identification of spots exhibiting tissue-specific methylation.ConclusionThe new vRLGS system reported here is highly robust for the identification of novel RLGS spots. The migration models developed are not specific to the genome being studied or the enzyme combination being used, making this tool broadly applicable. The identification of hundreds of mouse and human RLGS spot loci confirms the strong bias of RLGS studies to focus on CpG islands and provides a valuable resource to rapidly study their methylation

Characterisation of periodically poled materials using nonlinear microscopy

Periodically poled crystalline materials are extremely attractive for processes such as second harmonic generation and optical parametric generation due to their very high conversion efficiency. For optimal performance, fabrication of poled regions with sub-micron tolerance is required. In this paper we introduce multi-photon laser scanning luminescence microscopy as a powerful minimally-invasive measurement technique which provides information about internal device structure with high spatial resolution that cannot be easily obtained with existing methods. A comparative study of confocal and multi-photon imaging of periodically poled crystalline materials is also performed

Measuring the shape. Performance evaluation of a photogrammetry improvement applied to the Neanderthal skull Saccopastore 1

Several digital technologies are nowadays developed and applied to the study of the human fossil record. Here, we present a low-cost hardware implementation of the digital acquisition via photogrammetry, applied to a specimen of paleoanthropological interest: the Neanderthal skull Saccopastore 1. Such implementation has the purpose to semi-automatize the procedures of digital acquisition, by the introduction of an automatically rotating platform users can easily build on their own with minimum costs. We provide all the technical specifications, mostly based on the Arduino UNO™ microcontroller technology, and evaluate the performance and the resolution of the acquisition by comparing it with the CT-scan of the same specimen through the calculation of their shape differences. In our opinion, the replication of the automatic rotating platform, described in this work, may contribute to the improvement of the digital acquisition processes and may represent, in addition, a useful and affordable tool for both research and dissemination

Yeast-derived biosynthesis of silver/silver chloride nanoparticles and their antiproliferative activity against bacteria

Here, we provide the first evidence of yeast strains assisted Ag/AgCl-NPs production in vitro. The formed nanoparticles were characterized by spectroscopic and electron microscopy approaches. UV-vis supported the biosynthesis. TEM analysis evidenced that the nanoparticles mainly presented a circular shape and their diameters varied mostly being in the range 2 to 10 nm. XRD analysis showed a crystalline structure, with diffraction peaks corresponding to metallic silver and silver chloride nanoparticles, and when analyzed by high-resolution transmission electron microscopy (HRTEM), instead of being round, (111) (octahedral) and (200) (cubic) symmetry facets appeared systematically in one side of the nanoparticles. Analysis of ultra-thin sections by TEM indicated that the domain of the synthesis of Ag/AgCl-NPs was mainly between the cell wall and the plasma membrane. By using 3D reconstruction obtained from focused ion beam scanning electron microscopy (FIB/SEM) the spatial distribution of the domains of nanoparticle synthesis was mapped and nanoaggregates of Ag/AgCl-NPs up 35 nm in diameter were observed. Extracellular synthesis also occurred; in accordance with the fact that conditioned media from yeast isolates were as efficient at producing Ag/AgCl-NPs as live-cell cultures. Exposure of Gram-positive Staphylococcus aureus and Gram-negative Klebsiella pneumoniae cultures to Ag/AgCl-NPs led to a strong growth inhibition as shown by optical density measurements. The Ag/AgCl-NPs described here have characteristics compatible with a strong potential for use in the biotechnology industry, particularly for biomedical applications

Nanogels for pharmaceutical and biomedical applications and their fabrication using 3D printing technologies

Nanogels are hydrogels formed by connecting nanoscopic micelles dispersed in an aqueous medium, which give an opportunity for incorporating hydrophilic payloads to the exterior of the micellar networks and hydrophobic payloads in the core of the micelles. Biomedical and pharmaceutical applications of nanogels have been explored for tissue regeneration, wound healing, surgical device, implantation, and peroral, rectal, vaginal, ocular, and transdermal drug delivery. Although it is still in the early stages of development, due to the increasing demands of precise nanogel production to be utilized for personalized medicine, biomedical applications, and specialized drug delivery, 3D printing has been explored in the past few years and is believed to be one of the most precise, efficient, inexpensive, customizable, and convenient manufacturing techniques for nanogel production