Burr-like, laser-made 3D microscaffolds for tissue spheroid encagement.

The modeling, fabrication, cell loading, and mechanical and in vitro biological testing of biomimetic, interlockable, laser-made, concentric 3D scaffolds are presented. The scaffolds are made by multiphoton polymerization of an organic-inorganic zirconium silicate. Their mechanical properties are theoretically modeled using finite elements analysis and experimentally measured using a Microsquisher(®). They are subsequently loaded with preosteoblastic cells, which remain live after 24 and 72 h. The interlockable scaffolds have maintained their ability to fuse with tissue spheroids. This work represents a novel technological platform, enabling the rapid, laser-based, in situ 3D tissue biofabrication.

Adhesion and growth of human bone marrow mesenchymal stem cells on precise-geometry 3D organic-inorganic composite scaffolds for bone repair.

Engineering biomaterial scaffolds that promote attachment and growth of mesenchymal stem cells in three dimensions is a crucial parameter for successful bone tissue engineering. Towards this direction, a lot of research effort has focused recently into the development of three-dimensional porous scaffolds, aiming to elicit positive cellular behavior. However, the fabrication of three-dimensional tissue scaffolds with a precise geometry and complex micro- and nano-features, supporting cell in-growth remains a challenge. In this study we report on a positive cellular response of human bone marrow-derived (BM) mesenchymal stem cells (MSCs) onto hybrid material scaffolds consisting of methacryloxypropyl trimethoxysilane, zirconium propoxide, and 2-(dimethylamino)ethyl methacrylate (DMAEMA). First, we use Direct fs Laser Writing, a 3D scaffolding technology to fabricate the complex structures. Subsequently, we investigate the morphology, viability and proliferation of BM-MSCs onto the hybrid scaffolds and examine the cellular response from different donors. Finally, we explore the effect of the materials' chemical composition on cell proliferation, employing three different material surfaces: (i) a hybrid consisting of methacryloxypropyl trimethoxysilane, zirconium propoxide and 50mol% DMAEMA, (ii) a hybrid material comprising methacryloxypropyl trimethoxysilane and zirconium propoxide, and (iii) a purely organic polyDMAEMA. Our results show a strong adhesion of BM-MSCs onto the hybrid material containing 50% DMAEMA from the first 2h after seeding, and up to several days, and a proliferation increase after 14 and 21days, similar to the polystyrene control, independent of cell donor. These findings support the potential use of our proposed cell-material combination in bone tissue engineering.

Redox multiphoton polymerization for 3D nanofabrication.

We report for the first time on the redox multiphoton polymerization of an organic-inorganic composite material, in which one of the components, a vanadium metallo-organic complex, initiates the polymerization. The composite employs multiphoton absorption to self-generate radicals by photoinduced reduction of the metal species from vanadium (V) to vanadium (IV). We exploit this material for the fabrication of fully 3D structures by multiphoton polymerization with 200 nm resolution, employing a femtosecond laser operating at 800 nm, in the absence of a photoinitiator. Nonlinear absorption measurements indicate that the use of an 800 nm laser initiates the photopolymerization due to three-photon absorption of the vanadium alkoxide. The laser power required to induce this three-photon polymerization is comparable to what is required for inducing two-photon polymerization in materials using standard two-photon absorbers, most likely due to the high content of vanadium in the final composite (up to 50% mole).

Micro-structured polymer scaffolds fabricated by direct laser writing for tissue engineering.

This work presents the latest results on direct laser writing of polymeric materials for tissue engineering applications. A femtosecond Yb:KGW laser (300 fs, 200 kHz, 515 nm) was used as a light source for non-linear lithography. Fabrication was implemented in various photosensitive polymeric materials, such as: hybrid organic-inorganic sol-gel based on silicon-zirconium oxides, commercial ORMOCER® class photoresins. These materials were structured via multi-photon polymerization technique with submicron resolution. Porous three-dimensional scaffolds for artificial tissue engineering were fabricated with constructed system and were up to several millimeters in overall size with 10 to 100 µm internal pores. Biocompatibility of the used materials was tested in primary rabbit muscle-derived stem cell culture in vitro and using laboratory rats in vivo. This interdisciplinary study suggests that proposed technique and materials are suitable for tissue engineering applications.

Three-dimensional micro-/nano-structuring via direct write polymerization with picosecond laser pulses.

We demonstrate capability to structure photo-polymers with sub-wavelength resolution, ∼200-500 nm, and retrieve three-dimensional (3D) structures using a picosecond laser exposure. This alternative to commonly used ultra-short femtosecond lasers extends accessibility of 3D direct write. A popular hybrid sol-gel resist SZ2080 was used for quantitative determination of structuring resolution at 1064 nm and 532 nm wavelengths and for pulses of 8-25 ps duration at the repetition rates of 0.2-1 MHz. Systematic study of feature size dependence of 3D suspended nano-rods shows that linear power dependence of photopolymerization on the dose-per-pulse becomes dominant at higher repetition rates (≥0.5 MHz) while the two-photon nonlinear absorption is still distinguishable at rates lower than 0.2 MHz and shorter pulses (≤8 ps). Thermal accumulation defines polymerization when cooling time of the focal volume is larger than separation between pulses. Photopolymerization and its scaling mechanisms, quality, and fidelity at tight focusing of fs-, ps-, and cw-laser radiation are revealed and explained. 3D scaffolds for biomedicine and microlenses for optical applications are fabricated by the ps-laser direct write.