We have developed a laser-based printing technique, called biological laser printing (BioLP). BioLP is a non-contact, orifice-free technique that rapidly deposits fL to nL scale volumes of biological material with spatial accuracy better than 5 microm. The printer's orifice-free nature allows for transfer of a wide range of biological material onto a variety of substrates. Control of transfer is performed via a computer-aided design/computer-aided manufacturing (CAD/CAM) system which allows for deposition rates up to 100 pixels of biological material per second using the current laser systems. In this article, we present a description of the apparatus, a model of the transfer process, and a comparison to other biological printing techniques. Further, examples of current system capabilities, such as adjacent deposition of multiple cell types, large-scale cell arrays, and preliminary experiments on creating multi-layer cell constructs are presented. These cell printing experiments not only demonstrate near 100% viability, they also are the first steps toward using BioLP to create heterogeneous 3-dimensional constructs for use in tissue engineering applications.
We have investigated the electrochemical, spectroscopic, and electroluminescent properties of a family of diimine complexes of Ru featuring various aliphatic side chains as well as a more extended pi-conjugated system. The performance of solid-state electroluminescent devices fabricated from these complexes using indium tin oxide (ITO) and gold contacts appears to be dominated by ionic space charge effects. Their electroluminescence efficiency was limited by the photoluminescence efficiency of the Ru films and not by charge injection from the contacts. The incorporation of di-tert-butyl side chains on the dipyridyl ligand was found to be the most beneficial substitution in terms of reducing self-quenching of luminescence.
A technique by which to print patterns and multilayers of scaffolding and living cells could be used in tissue engineering to fabricate tissue constructs with cells, materials, and chemical diversity at the micron scale. We describe here studies using a laser forward transfer technology to print single-layer patterns of pluripotent murine embryonal carcinoma cells. This report focuses on verifying cell viability and functionality as well as the ability to differentiate cells after laser transfer. We find that when cells are printed onto model tissue scaffolding such as a layer of hydrogel, greater than 95% of the cells survive the transfer process and remain viable. In addition, alkaline comet assays were performed on transferred cells, showing minimal single-strand DNA damage from potential ultraviolet-cell interaction. We also find that laser-transferred cells express microtubular associated protein 2 after retinoic acid stimulus and myosin heavy chain protein after dimethyl sulfoxide stimulus, indicating successful neural and muscular pathway differentiation. These studies provide a foundation so that laser printing may next be used to build heterogeneous multilayer cellular structures, enabling cell growth and differentiation in heterogeneous three-dimensional environments to be uniquely studied.
Cell printing has been popularized over the past few years as a revolutionary advance in tissue engineering has potentially enabled heterogeneous 3-D scaffolds to be built cell-by-cell. This review article summarizes the state-of-the-art cell printing techniques that utilize fluid jetting phenomena to deposit 2- and 3-D patterns of living eukaryotic cells. There are four distinct categories of jetbased approaches to printing cells. Laser guidance direct write (LG DW) was the first reported technique to print viable cells by forming patterns of embryonic-chick spinal-cord cells on a glass slide (1999). Shortly after this, modified laser-induced forward transfer techniques (LIFT) and modified ink jet printers were also used to print viable cells, followed by the most recent demonstration using an electrohydrodynamic jetting (EHDJ) method. The low cost of some of these printing technologies has spurred debate as to whether they could be used on a large scale to manufacture tissue and possibly even whole organs. This review summarizes the published results of these cell printers (cell viability, retained genotype and phenotype), and also includes a physical description of the various jetting processes with a discussion of the stresses and forces that may be encountered by cells during printing. We conclude the review by comparing and contrasting the different jet-based techniques, while providing a map for future experiments that could lead to significant advances in the field of tissue engineering.
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