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Long-term maintenance of functional primary human hepatocytes in 3D gelatin matrices produced by solution blow spinning.

ABSTRACT: Solution blow spinning (SBS) has recently emerged as a novel method that can produce nano- and microfiber structures suitable for tissue engineering. Gelatin is an excellent precursor for SBS as it is derived mainly from collagens that are abundant in natural extracellular matrices. Here we report, for the first time the successful generation of 3D thermally crosslinked preforms by using SBS from porcine gelatin. These SBS mats were shown to have three-dimensional fibrous porous structure similar to that of mammalian tissue extracellular matrix. In pharma industry, there is an urgent need for adequate 3D liver tissue models that could be used in high throughput setting for drug screening and to assess drug induced liver injury. We used SBS mats as culturing substrates for human hepatocytes to create an array of 3D human liver tissue equivalents in 96-well format. The SBS mats were highly cytocompatible, facilitated the induction of hepatocyte specific CYP gene expression in response to common medications, and supported the maintenance of hepatocyte differentiation and polarization status in long term cultures for more than 3 weeks. Together, our results show that SBS-generated gelatin scaffolds are a simple and efficient platform for use in vitro for drug testing applications.

SUBMITTER: Klaas M

PROVIDER: S-EPMC8505433 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Polymer nano/micro fibers have found many applications including 3D cell culture and the creation of wound dressings. The fibers can be produced by a variety of techniques that include electrospinning, the primary disadvantage of which include the requirement for a high voltage supply (which may cause issues such as polymer denaturation) and lack of portability. More recently, solution blow spinning, where a high velocity sheath gas is used instead of high voltage, has been used to generate polymer fibers. In this work, we used blow spinning to create nano/microfibers for microchip-based 3D cell culture. First, we thoroughly investigated fiber generation from a 3D printed gas sheath device using two polymers that are amenable to cell culture (polycaprolactone, PCL and polystyrene, PS) as well as the parameters that can affect PCL and PS fiber quality. Using the 3D printed sheath device, it was found that the pressure of the sheath N2 and the concentration of polymer solutions determine if fibers can be produced as well as the resulting fiber morphology. In addition, we showed how these fibers can be used for 3D cell culture by directly depositing PCL fibers in petri dishes and well plates. It is shown the fibers have good compatibility with RAW 264.7 macrophages and the PCL fiber scaffold can be as thick as 178 ± 14 μm. PCL fibers created from solution blow spinning (with the 3D printed sheath device) were then integrated with a microfluidic device for the first time to fabricate a 3D cell culture scaffold with a flow component. After culturing and stimulating macrophages on the fluidic device, it was found that the integrated 3D fibrous scaffold is a better mimic of the extracellular matrix (as opposed to a flat, 2D substrate), with enhanced nitrite accumulation (product of nitric oxide release) from macrophages stimulated with lipopolysaccharide. PS fibers were also made and integrated in a microfluidic device for 3D culture of endothelial cells, which stayed viable for at least 72 hours (48 hours under the flowing conditions). This approach will be useful for future studies involving more realistic microchip-based culture models for studying cell-to-cell communication.

Project description:Hard-to-dissolve polymers provide next-generation alternatives for high-performance filter materials owing to their intrinsically high chemical stability, superior mechanical performance, and excellent high-temperature resistance. However, the mass production of hard-to-dissolve nanofibers still remains a critical challenge. A simple, scalable, and low-cost ionic solution blow-spinning method has herein been provided for the large-scale preparation of hard-to-dissolve Nomex polymeric nanofibers with an average diameter of nearly 100 nm. After rapidly dissolving Nomex microfibers in the lithium chloride/dimethylacetamide (LiCl/DMAc) solution system, the conductive solution can be stably and conductivity-independently processed into nanofibers. The method optimizes electrospinning and avoids spinnability degradation and potential safety hazards caused by high electrical conductivity. Owing to nanofibrous structure and high dipole moment, Nomex nanofibrous filters show a stable high filtration efficiency of 99.92% for PM0.3 with a low areal density of 4.6 g m-2, as well as a low-pressure drop of 189.47 Pa. Moreover, the flame-retardant filter can work at 250 °C and 280 °C for a long and short time without shrinking or burning, respectively, exhibiting a high filtration efficiency of 99.50% for PM0.3-10.0. The outstanding properties and low cost enable the efficient capture of PM from various high-temperature exhausts, making Nomex nanofibrous membrane an even more ideal industrial-grade air filter than polypropylene, polytetrafluoroethylene, polyimide, and ceramic nanofibrous filters.Graphical abstractHard-to-dissolve nanofibers provide alternatives for high-efficiency and low-resistant air filtration but are limited by the universality and economics of fabrication methods. A scalable and efficient ionic solution blow-spinning strategy has herein been proposed in preparing hard-to-dissolve nanofibrous filters.Supplementary informationThe online version contains supplementary material available at 10.1007/s42765-022-00231-x.

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Long-term maintenance of functional primary human hepatocytes in 3D gelatin matrices produced by solution blow spinning.

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