Laser Microprinting of Transparent and Weakly Absorbing Solutions

A laser-based technique for printing transparent andweakly absorbing liquids is developed. Its principleof operation relies in the tight focusing of shortlaser pulses inside the liquid and close to its freesurface, in such a way that the laser radiation isabsorbed in a tiny volume around the beam waist,with practically no absorption in any other locationalong the beam path. If the absorbed energyovercomes the optical breakdown threshold, acavitation bubble is generated, and its expansionresults in the propulsion of a small fraction of liquidwhich can be collected on a substrate, leading to theprinting of a microdroplet for each laser pulse. Thetechnique does not require the preparation of theliquid in thin film form, and its forward mode ofoperation imposes no restriction concerning theoptical properties of the substrate. We demonstratethat the technique is capable of printingmicrodroplets with good resolution, reproducibilityand control, and analyze the influence of the mainprocess parameters. The mechanisms of liquidprinting are also investigated: time-resolvedimaging provides a clear picture of the dynamics ofliquid transfer which allows understanding the mainfeatures observed in the printed droplets

Survival and differentiation of embryonic neural explants on different biomaterials

Biomaterials prepared from polyacrylamide, ethyl acrylate (EA), and hydroxyethyl acrylate (HEA) in various blend ratios, methyl acrylate and chitosan, were tested in vitro as culture substrates and compared for their ability to be colonized by the cells migrating from embryonic brain explants. Neural explants were isolated from proliferative areas of the medial ganglionic eminence and the cortical ventricular zone of embryonic rat brains and cultured in vitro on the different biomaterials. Chitosan, poly(methyl acrylate), and the 50% wt copolymer of EA and HEA were the most suitable substrates to promote cell attachment and differentiation of the neural cells among those tested. Immunofluorescence microscopy analysis showed that progenitor cells had undergone differentiation and that the resulting glial and neuronal cells expressed their intrinsic morphological characteristics in culture

Water-induced (nano) organization in poly(ethyl acrylate-co-hydroxyethyl acrylate) networks

The conformational changes in poly(ethyl acrylate-co-hydroxyethyl acrylate), P(EA-co-HEA) chains, which constitute a copolymer network hydrogel, induced by the presence of water are investigated by different experimental techniques and compared with the behaviour of the corresponding xerogel. The mechanical relaxation spectrum shows the presence of a new water-induced relaxation, the water content dependence of the glass transition is measured by DSC, and the dielectric relaxation assesses the effect of water for the lower concentrations. Hydrophilic and hydrophobic monomeric units in the P(EA-co-HEA) network are able to aggregate to form two separated (nano)phases in the presence of water due to hydrophobic interaction. Phase separation takes place when the water content of the sample is higher than a critical value estimated as two water molecules per –OH group in the copolymer chain. The existence of the hydrophobic domains is detected by their glass transition being nearly independent on the water content of the sample. Phase separation is also clearly revealed by phase angle measurements in AFM experiments

Analysis of the biological response of endothelial and fibroblast cells cultured on synthetic scaffolds with various hydrophilic/hydrophobic ratios: influence of fibronectin adsorption and conformation

In this study we developed polymer scaffolds intended as anchorage rings for cornea prostheses among other applications, and examined their cell compatibility. In particular, a series of interconnected porous polymer scaffolds with pore sizes from 80 to 110 microns were manufactured varying the ratio of hydrophobic to hydrophilic monomeric units along the polymer chains. Further, the effects of fibronectin precoating, a physiological adhesion molecule, were tested. The interactions between the normal human fibroblast cell line MRC-5 and primary human umbilical vein endothelial cells (HUVECs) with the scaffold surfaces were evaluated. Adhesion and growth of the cells was examined by confocal laser scanning microscopy. Whereas MRC-5 fibroblasts showed adhesion and spreading to the scaffolds without any precoating, HUVECs required a fibronectin precoating for adhesion and spreading. Although both cell types attached and spread on scaffold surfaces with a content of up to a 20% hydrophilic monomers, cell adhesion, spreading, and proliferation increased with increasing hydrophobicity of the substrate. This effect is likely due to better adsorption of serum proteins to hydrophobic substrates, which then facilitate cell adhesion. In fact, atomic force microscopy measurements of fibronectin on surfaces representative of our scaffolds revealed that the amount of fibronectin adsorption correlated directly with the hydrophobicity of the surface. Besides cell adhesion we also examined the inflammatory state of HUVECs in contact with the scaffolds. Typical patterns of platelet/endothelial cell adhesion molecule-1 expression were observed at intercellular boarders. HUVECs adhering on the scaffolds retained their proinflammatory response potential as shown by E-selectin mRNA expression after stimulation with lipopolyssacharide (LPS). The proinflammatory activation occurred in most of the cells, thus confirming the presence of a functionally intact endothelium. Little or no expression of the proinflammatory activation markers in the absence of LPS stimulation was observed for HUVECs growing on scaffolds with up to a 20% of hydrophilic component, whereas activation of these markers was observed after stimulation. In conclusion, scaffolds containing up to 20% hydrophilic monomers exhibited excellent cell compatibility toward human fibroblast cell line MRC-5 and human endothelial cells. Atomic force microscopy confirmed that adsorbed serum proteins such as fibronectin probably accounted for the positive correlation of HUVEC adhesion and surface hydrophobicity

Deposition and characterization of lines printed through laser-induced forward transfer

The possibility of printing two-dimensional micropatterns of biomolecule solutions is of great interest in many fields of research in biomedicine, from cell-growth and development studies to the investigation of the mechanisms of communication between cells. Although laser-induced forward transfer (LIFT) has been extensively used to print micrometric droplets of biological solutions, the fabrication of complex patterns depends on the feasibility of the technique to print micron-sized lines of aqueous solutions. In this study we investigate such a possibility through the analysis of the influence of droplet spacing of a water and glycerol solution on the morphology of the features printed by LIFT. We prove that it is indeed possible to print long and uniform continuous lines by controlling the overlap between adjacent droplets. We show how, depending on droplet spacing, several printed morphologies are generated, and we offer, in addition, a simple explanation of the observed behavior based on the jetting dynamics characteristic of the LIFT of liquids