Szabolcs Beke

Towards excimer-laser-based stereolithography: a rapid process to fabricate rigid biodegradable photopolymer scaffolds

We demonstrate high-resolution photocross-linking of biodegradable poly(propylene fumarate) (PPF) and diethyl fumarate (DEF) using UV excimer laser photocuring at 308 nm. The curing depth can be tuned in a micrometre range by adjusting the total energy dose (total fluence). Youngs moduli of the scaffolds are found to be a few gigapascal, high enough to support bone formation. The results presented here demonstrate that the proposed technique is an excellent tool for the fabrication of stiff and biocompatible structures on a micrometre scale with defined patterns of high resolution in all three spatial dimensions. Using UV laser photocuring at 308 nm will significantly improve the speed of rapid prototyping of biocompatible and biodegradable polymer scaffolds and enables its production in a few seconds, providing high lateral and horizontal resolution. This short timescale is indeed a tremendous asset that will enable a more efficient translation of technology to clinical applications. Preliminary cell tests proved that PPF : DEF scaffolds produced by excimer laser photocuring are biocompatible and, therefore, are promising candidates to be applied in tissue engineering and regenerative medicine.

Rapid fabrication of rigid biodegradable scaffolds by excimer laser mask projection technique: a comparison between 248 and 308 nm

High-resolution photocrosslinking of the biodegradable poly(propylene fumarate) (PPF) and diethyl fumarate (DEF), using pulsed laser light at 248 and 308 nm is presented. The curing depth can be modulated between a few hundreds of nm and a few μm when using 248 nm and ten to a hundred μm when using 308 nm. By adjusting the total fluence (pulse numbers×laser fluence) dose and the weight ratios of PPF, DEF, and the photoinitiator in the photocrosslinkable mixtures, the height of polymerized structures can be precisely tuned. The lateral resolution is evaluated by projecting a pattern of a grid with a specified line width and line spacing. Youngs modulus of the cured parts is measured and found to be several GPa for both wavelengths, high enough to support bone formation. Several 2D and 2.5D microstructures, as well as porous 3D scaffolds fabricated by a layer-by-layer method, are presented. The results demonstrate that excimer laser-based photocuring is suitable for the fabrication of stiff and biocompatible structures with defined patterns of micrometer resolution in all three spatial dimensions.

3D scaffold fabrication by mask projection excimer laser stereolithography

The production of 3D scaffolds with well-controlled architecture at the micrometer-scale is a fundamental issue for the advancement of tissue engineering towards applications in health care. Stereolithography is a highly versatile and accurate technique to fabricate 3D scaffolds with controlled architectures. Here, a scalable stereolithography method combining mask projection with excimer laser is reported. Its capability is showcased by a variety of mm-sized 3D biodegradable scaffolds patterned with a spatial resolution well-suited for tissue engineering applications. The presented method offers a concrete possibility to scale-up stereolithography-based production of 3D scaffolds to be used in regenerative medicine with potentially high-impact on health care.

Improved cell activity on biodegradable photopolymer scaffolds using titanate nanotube coatings.

The development of bioactive materials is in the premise of tissue engineering. For several years, surface functionalization of scaffolds has been one of the most promising approaches to stimulate cellular activity and finally improve implant success. Herein, we describe the development of a bioactive composite scaffold composed of a biodegradable photopolymer scaffold and titanate nanotubes (TNTs). The biodegradable photopolymer scaffolds were fabricated by applying mask-projection excimer laser photocuring at 308 nm. TNTs were synthesized and then spin-coated on the porous scaffolds. Upon culturing fibroblast cells on scaffolds, we found that nanotubes coating affects cell viability and proliferation demonstrating that TNT coatings enhance cell growth on the scaffolds by further improving their surface topography.

Titanate nanotube coatings on biodegradable photopolymer scaffolds

Rigid, biodegradable photopolymer scaffolds were coated with titanate nanotubes (TNTs) by using a spin-coating method. TNTs were synthesized by a hydrothermal process at 150 °C under 4.7 bar ambient pressure. The biodegradable photopolymer scaffolds were produced by mask-assisted excimer laser photocuring at 308 nm. For scaffold coating, a stable ethanolic TNT sol was prepared by a simple colloid chemical route without the use of any binding compounds or additives. Scanning electron microscopy along with elemental analysis revealed that the scaffolds were homogenously coated by TNTs. The developed TNT coating can further improve the surface geometry of fabricated scaffolds, and therefore it can further increase the cell adhesion.

Ultrasmall, Ligand-Free Ag Nanoparticles with High Antibacterial Activity Prepared by Pulsed Laser Ablation in Liquid

Since ancient times, silver and its compounds have been known to have a broad spectrum of antimicrobial activities for bacteria, fungi, and viruses. Due to the increasing bacterial resistance to classic antibiotics, the investigations of Ag NPs have increased. Herein, we present the preparation of ligand-free Ag NPs with 3 and 20 nm sizes by applying picosecond laser ablation in liquid at 355 and 1065 nm. Our laser processing system allows a high control on particle sizes. The produced nanoparticles were characterized by means of transmission electron microscopy, UV-Vis spectroscopy, and X-ray diffraction. The size effect on the antibacterial activity of Ag NPs was tested against E. coli and S. aureus. The growth curves of bacteria were monitored at 0–5 mg/L of Ag NPs by a multimode microplate reader. The size effects as well as the concentration of Ag NPs on their antibacterial activity are discussed.

Elastin-Coated Biodegradable Photopolymer Scaffolds for Tissue Engineering Applications

One of the main open issues in modern vascular surgery is the nonbiodegradability of implants used for stent interventions, which can lead to small caliber-related thrombosis and neointimal hyperplasia. Some new, resorbable polymeric materials have been proposed to substitute traditional stainless-steel stents, but so far they were affected by poor mechanical properties and low biocompatibility. In this respect, a new material, polypropylene fumarate (PPF), may be considered as a promising candidate to implement the development of next generation stents, due to its complete biodegradability, and excellent mechanical properties and the ease to be precisely patterned. Besides all these benefits, PPF has not been tested yet for vascular prosthesis, mainly because it proved to be almost inert, while the ability to elicit a specific biological function would be of paramount importance in such critical surgery applications. Here, we propose a biomimetic functionalization process, aimed at obtaining specific bioactivation and thus improved cell-polymer interaction. Porous PPF-based scaffolds produced by deep-UV photocuring were coated by elastin and the functionalized scaffolds were extensively characterized, revealing a stable bound between the protein and the polymer surface. Both 3T3 and HUVEC cell lines were used for in vitro tests displaying an enhancement of cells adhesion and proliferation on the functionalized scaffolds.

Fabrication of hybrid nanocomposite scaffolds by incorporating ligand-free hydroxyapatite nanoparticles into biodegradable polymer scaffolds and release studies

Summary We report on the optical fabrication approach of preparing free-standing composite thin films of hydroxyapatite (HA) and biodegradable polymers by combining pulsed laser ablation in liquid and mask-projection excimer laser stereolithography (MPExSL). Ligand-free HA nanoparticles were prepared by ultrafast laser ablation of a HA target in a solvent, and then the nanoparticles were dispersed into the liquid polymer resin prior to the photocuring process using MPExSL. The resin is poly(propylene fumarate) (PPF), a photo-polymerizable, biodegradable material. The polymer is blended with diethyl fumarate in 7:3 w/w to adjust the resin viscosity. The evaluation of the structural and mechanical properties of the fabricated hybrid thin film was performed by means of SEM and nanoindentation, respectively, while the chemical and degradation studies were conducted through thermogravimetric analysis, and FTIR. The photocuring efficiency was found to be dependent on the nanoparticle concentration. The MPExSL process yielded PPF thin films with a stable and homogenous dispersion of the embedded HA nanoparticles. Here, it was not possible to tune the stiffness and hardness of the scaffolds by varying the laser parameters, although this was observed for regular PPF scaffolds. Finally, the gradual release of the hydroxyapatite nanoparticles over thin film biodegradation is reported.

Characterization of a bioinspired elastin-polypropylene fumarate material for vascular prostheses applications

Vascular prostheses are widely used devices fundamental to avoid the effect of life-threatening diseases and defects. Besides a long experience in the fabrication of biomaterials for vascular applications, many issues still remain unattended. In particular, obtaining a bio-resorbable and bio-active scaffold is a challenge of paramount importance. We present a novel application in which a promising biodegradable polymer, poly-propylene fumarate (PPF), is printed using three dimensional laser-induced cross-linking micromachining device. To enhance the biological role of the scaffold, a bio-inspired approach was taken, by coating the surface of the PPF with elastin, the main constituent of the innermost layer of natural veins and arteries.

Characterization of the ablation of TeO2 crystals in air with femtosecond laser pulses

Femtosecond (fs) laser pulse ablation (pulse duration of 150 fs, wavelength of 775 nm, repetition rate of 1 kHz) of single-crystalline TeO2 surfaces was performed in air using the direct focusing technique. The lateral and vertical dimensions of laser ablated craters as well as the laser damage thresholds were evaluated for different pulse numbers applied to the same spot. The joint observation using optical microscopy, atomic force microscopy and scanning electron microscopy revealed the surface morphology of the ablated craters and also showed that the ablation threshold depends significantly on the number of laser pulses applied to the same spot due to incubation effects. The incubation effects change the absorption processes involved in fs-laser ablation of the transparent material from multiphoton absorption to a single-photon absorption. These results are discussed on the basis of recent models of the interaction of fs-laser pulses with dielectrics. (Some figures in this article are in colour only in the electronic version)