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In a recent discovery, coaxial electrospinning was explored to encapsulate living organisms within a continuous bio-polymeric microthread from which active biological scaffolds were fabricated (Townsend-Nicholson and Jayasinghe, Biomacromolecules 2006, 7, 3364). The cells were demonstrated to have gone through all expected cellular activity without their viability being compromised. These biologically active threads and scaffolds have direct and tremendous applicability from regenerative to therapeutic medicine. Currently these post-processed cells as composite threads and scaffolds are being investigated indepth at a cellular level to establish if the processing methodology has any affect on the cellular make-up. We now demonstrate a competing non-electric field driven approach for fabricating composite threads and scaffolds influenced only by a differential pressure. We refer to this novel composite thread to scaffold fabrication methodology as coaxial aerodynamically assisted bio-threading (CAABT).Our investigations firstly, demonstrate that this technique can process handle living organisms without biologically perturbing them in anyway. Secondly the process is elucidated as possessing the ability to form composite active threads from which biologically viable scaffolds are formed. Finally our study employs florescent activated cell sorting (FACScan), a method by which the cellular dynamics and viability are quantified on control and threaded cellular samples at two prescribed time points. In parallel with FACScan, optical comparison of cellular morphology at three time points within a period of three weeks is carried out to photographically observe any changes in the post-processed cellular phenotype. Our developmental investigations into this novel aerodynamically assisted threading methodology has unearthed a unique biomicrofabrication approach, which joins cell electrospinning in the cell threading to scaffold fabrication endeavor.

We recently demonstrated the ability to use electrified jets under stable conditions for the generation of cell-bearing droplets to the formation of composite threads which are biologically active. Our studies established that processed cells were viable over several generations post-jetting and -threading. These harmless and successful techniques for jet-based cell handling to deployment for precision deposition have great potential and widespread applications in bioengineering and biotechnology. Nonetheless, our investigations into 'bio-electrosprays' and 'cell electrospinning' have elucidated these jets having direct applicability in regenerative and therapeutic medicine to studies in developmental biology. For these very reasons, jet methodologies having the capability to safely handle living organisms for drop and placing are increasingly gaining the interests of life scientists. We now demonstrate yet another technique (a non-electric field-driven approach, previously never explored with jetting living cells), possessing the ability to directly handle the processing of primary living organisms by means of the flow of a cell suspension within a needle placed in a pressure chamber in the presence of an applied pressure difference. The technique we introduce here is referred to as 'aerodynamically assisted bio-jets/-jetting' which is driven completely by aerodynamic forces applied over an exit orifice by way of a differential pressure. Our investigations present an operational window in which stable jetting conditions are achieved for the formation of a near-monodispersed distribution of cell-bearing droplets and droplet residues. Finally, the aerodynamically bio-jetted living primary organisms are assessed (over both short and long time points) for cellular viability by means of FACScan, a flow cytometry technology which quantifies the percentage of living and dead cells. These advanced biophysical and bioengineering studies elucidate the emergence of a non-electric field-driven bio-jetting technology which now joins the cell jetting race.

The physical sciences have increasingly demonstrated a significant influence on the life sciences. Engineering in particular has shown its input through the development of novel medical devices and processes having significance to the biomedical field. This review introduces and discusses several fiber generation protocols, which have recently undergone development and exploration for directly handling living cells from which continuous cell-bearing or living threads to scaffolds and membranes have been fabricated. In doing so these protocols have not only demonstrated their versatility but also opened several unique possibilities for the use of these scaffolds in a plethora of biological and medical applications. In particular, these living fibrous structural units could be explored for regeneration purposes, e.g., from accelerated wound healing to combating a wide range of pathologies when coupled with gene therapy. Thus, "living entities" such as these scaffolds could be most useful in surgery/medicine, including its exploration with stem cells for the preparation of unspecialized living scaffolds and membranes.

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