Investigation into the mechanisms driving cancer cell behavior and the subsequent development of novel targeted therapeutics requires comprehensive experimental models that mimic the complexity of the tumor microenvironment. Recently, our laboratories have combined a novel tissue culture model and laser direct-write, a form of bioprinting, to spatially position single or clustered cancer cells onto ex vivo microvascular networks containing blood vessels, lymphatic vessels, and interstitial cell populations. Herein, we highlight this new model as a tool for quantifying cancer cell motility and effects on angiogenesis and lymphangiogenesis in an intact network that matches the complexity of a real tissue. Application of our proposed methodology offers an innovative ex vivo tissue perspective for evaluating the effects of gene expression and targeted molecular therapies on cancer cell migration and invasion.
Current limitations to the engineering of ex vivo and in vitro neural environments are hampering the ability to understand underlying neurophysiology. High levels of spatial specificity, reproducibility and viability have been previously reported using laser direct write (LDW) to print cells. However, despite the significant need no one has yet reported laser assisted printing of primary mammalian neuronal cells, an inherently sensitive but critically important population. Herein, we describe the use of LDW to reproducibly and accurately pattern viable dorsal root ganglion (DRG) neurons and supportive cells capable of neural outgrowth and network formation. Our demonstrated ability to engineer and control distinct micro-environmental components unlocks the potential for high throughput experiments to both understand underlying physiology and investigate therapeutic interventions.
Laser-based three-dimensional (3D) printing methods, including laser direct-write cell printing and two-photon polymerization, have seen significant advances because of their unique photonic characteristics. Several mechanisms have been developed to increase the overall throughput of two-photon polymerization. Recent efforts to develop complex medically relevant structures using laser direct-write cell printing have also been demonstrated; for example, an ex vivo experimental platform for time-lapse imaging of cancer cell dynamics during angiogenesis within a microvascular network, which combines laser direct-write cell printing into the rat mesentery culture model; a model that simulates a 3D in vivo culture. Laser 3D printing methods hold significant promise for 3D printing of tissue engineering scaffolds, microstructured medical devices, and other medically relevant structures.
<p>Conventional wound healing assays are inadequate for evaluating the influence of symmetry on wound healing as they either offer poor geometrical control of the wounds created or fail to trigger a wound healing response in cell layers. To address these issues, herein we developed a novel CAD/CAM laser direct write photoablation assay that enables the generation of wounds with precisely defined and highly customizable geometries. As proof of principle, we biologically validated this assay to study the influence of symmetry and glycemic condition on dermal wound healing. We demonstrated that dermal wounds with symmetrical geometries tend to heal faster than those with asymmetrical geometries regardless of the glycemic conditions. Further, we elucidated the relative strength of the effects of symmetry and glycemic condition on healing rates in the context of the dermal wound size.</p>
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