Search

The aim of this study is to demonstrate the accuracy required for the investigation of the role of solid scaffolds’ porosity in cell proliferation. We therefore present a qualitative investigation into the effect of porosity on MC3T3-E1 pre-osteoblastic cell ingrowth of three-dimensional (3D) scaffolds fabricated by direct femtosecond laser writing. The material we used is a purpose made photosensitive pre-polymer based on polylactide. We designed and fabricated complex, geometry-controlled 3D scaffolds with pore sizes ranging from 25 to 110 μm, representing porosities 70%, 82%, 86%, and 90%. The 70% porosity scaffolds did not support cell growth initially and in the long term. For the other porosities, we found a strong adhesion of the pre-osteoblastic cells from the first hours after seeding and a remarkable proliferation increase after 3 weeks and up to 8 weeks. The 86% porosity scaffolds exhibited a higher efficiency compared to 82% and 90%. In addition, bulk material degradation studies showed that the employed, highly-acrylated polylactide is degradable. These findings support the potential use of the proposed material and the scaffold fabrication technique in bone tissue engineering.

Peptide-based arrays and patterns have provided a powerful tool in the study of protein recognition and function. A variety of applications have been identified, including the interactions between peptides–enzymes, peptides–proteins, peptides–DNA, peptides–small molecules and peptides–cells. One of the main and most critical unresolved issues is the generation of high-density arrays which maintain the biological function of the peptides. In this study, we employ nanosecond laser-induced forward transfer for the generation of high-density peptide arrays and patterns on modified glass surfaces. We show that peptide-based microarrays can be fabricated on solid surfaces and specifically recognized by appropriate fluorescent tags, with the transfer not affecting the ability of the peptides to form fibrils. These initial results are poised to the construction of larger peptide patterns as scaffolds for the incorporation and display of ligands critical for cell attachment and growth, or for the templating of inorganic materials.

Laser-induced forward transfer has been used for the deposition of photoactive biotin in micron-scale patterns. The process uses a 500 fs pulsed KrF laser beam to transfer small amounts of a liquid solution target as micron-size droplets to a substrate placed parallel and in close proximity to it. The biomolecules remain active after the transfer; this is demonstrated through fluorescence assays. In addition to the laser parameters, those regarding the target composition and the receiving surface for the miniaturization of the transferred patterns have been studied and optimized. Droplets as small as 5 μm have been obtained by reducing the target thickness and transfer energy; by increasing the percentage of glycerol added in the biomolecules solution and by using hydrophobic surfaces as receiving substrates. The influence of the glycerol addition and the hydrophobicity of the receiving surfaces on the activity of the transferred biomolecules have also been studied.

A new method is demonstrated, where three-dimensional protein structures are made by employing multi-photon polymerization and photobiotin photolysis. The technique enables the construction of arbitrary two- and three-dimensional shapes with submicron resolution. The integrity of the immobilized biotin is confirmed by derivatization with fluorescently labeled streptavidin. Fluorescence microscopy is used in order to visualize the distribution of fluorescent streptavidin on the 3D structure.

We have performed a comparative study of UV laser ablation of SrTiO3 with nanosecond- and sub-picosecond sources, respectively. The experiments were performed with lasers at a wavelength of 248 nm and pulse durations of 34 ns and 500 fs. Femtosecond ablation turns out to be more efficient by one order of magnitude and eliminated the known problem of cracking of SrTiO3 during laser machining with longer pulses. In addition, the cavities ablated with femtosecond pulses display a smoother surface with no indication of melting and well-defined, sharp edges. These effects can be explained by the reduced thermal shock effect on the material by using ultrashort pulses.