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NIH PHASE I FINAL REPORT: FIBROUS SUBSTRATES FOR CELL CULTURE (R3RR03544A)
Abstract
THIS PHASE I SBIR FINAL REPORT DESCRIBES THE PRODUCTION AND TESTING OF FIBROUS POLYMERIC SUBSTRATES PRODUCED BY A NOVEL TECHNIQUE [ELECTROSTATIC SPINNING]. DESIGN AND DEVELOPMENT OF AN ELECTROSTATIC SPINNING DEVICE (ESD) IS DESCRIBED. THE ESD IS CAPABLE OF PRODUCING GEOMETRICALLY DEFINED, NONWOVEN MATS COMPOSED OF 0.01 TO 1.0 MICRON DIAMETER FIBRILS. UNLIKE CONVENTIONAL FIBRILLATION TECHNIQUES, THE ESD CAN UTILIZE A LARGE VARIETY OF POLYMERIC SOLUTIONS AS THE FIBRILLATION MEDIUM AND IS ABLE TO INDUCE POLAR MOLECULAR ENRICHMENT ON THE FIBRIL SURFACE. STUDIES ON SURFACE CHEMISTRY OF THE FIBRILS WERE PERFORMED VIA ESCA (XPS). FURTHER CHARACTERIZATION OF THE FIBROUS MATS WERE PERFORMED VIA THE CULTURE OF SEVERAL CELL TYPES AND SCANNING ELECTRON MICROSCOPY.
... Electrospinning was initially identified in the 1980s by the National Institute of Health (NIH) as a potential method for generating nanoscale fibrous substrates suitable for cell culture. In 1988, Simon reported the successful production of nanoscale non-woven mats of polystyrene (PS) and polycarbonate (PC) via electrospinning [4] . By 1995, Reneker demonstrated that the electrospinning process could be used with a wide variety of polymers to consistently produce polymer fibers with diameters of the order of 50 nm [ 5 , 6 ]. ...
... In its most basic arrangement, the electrospinning process generates nanofibers that tend to be a continuous fiber which accumulates in a non-woven fiber mat or mesh on the collector, as shown in Fig. 1 a. This potential for creating a nanofibrous mesh was the driving force behind the fundamental research into this topic by the NIH, which wished to explore the potential for such materials in cell culture [4] . Randomly oriented non-woven meshes have been aggressively investigated for their potential in many different biomedical applications, from cell culture to tissue scaffolding and wound dressing. ...
The application of fibrous materials can be traced back throughout human history. Widespread adoption of polymers in the 20th century has rapidly driven the development of processes for producing polymer fibers to satisfy the demands of industries and consumers. The rise of nanomaterials has opened new worlds of possibility in terms of material properties and functionality, and polymer nanofibers are no exception. Their unique combination of viscoelastic behavior and incredibly high specific surface area have created applications for polymer nanofibers in areas such as biomedical engineering, smart materials, and energy storage, all of which project to be crucial research sectors in the next few decades. Polymer nanofibers have an important role to play in the technology that will shape our future. This manuscript provides a comprehensive review of the conventional methods used for manufacturing polymer nanofibers, the properties and behavior of polymer nanofibers and nanofiber composites, and the diverse applications of polymer nanofiber systems.
... To achieve a more equivalent situation to the human body 3D culturing was introduced. This type of cell culturing, often seen as biology's new dimension, started with varying combinations of single or multiple cell structures in 2D [96]. The first researchers to develop a method for the initiation and growth of multicell spheroids were R.M. Sutherland and R.E. ...
In this thesis the design, fabrication, characterization and in vitro application of a multiparametric, electrochemical microsensor platform is described. The system was designed to investigate cell metabolism, by monitoring oxygen consumption, lactate and oxidative stress. The lack of regulatory proliferative signals in cancer, cells results in the modification of their metabolic pathways. Electrochemical sensors allow a fast and precise in vitro measurement of extracellular analytes. The introduced sensor system consists of a glass chip with integrated electrodes, enabling the integration of various electrochemical measurement principles. On top of the glass chip a PMMA tube is fixed, building the cell culture well. Microsensor chips were fabricated in an efficient full-wafer process. Nine platinum working electrodes with a size of 200 μm were integrated together with one common counter and two Ag/AgCl reference electrodes. All electrodes were covered with a hydrogel membrane. Biosensors were integrated by the immobilization of enzymes inside the membrane. On some chips a temperature sensor was integrated. The sensors were characterized by thorough studies of electrode characteristics and sensor performance. Satisfying sensitivity and linearity for both biosensors and oxygen sensors was shown. Lactate sensors reached linearity up to 5 mM with a sensitivity around 2 μA/cm²/mM. Oxygen sensors exhibited a linear behaviour across the entire atmospheric range with a defined zero-point. The linearity of the hydrogen peroxide sensors was investigated up to 200 μM showing a sensitivity around −0.02 μA/cm²/μM. Long-term stability over several days under continuous measurement, without the need for recalibration, was verified for all sensors. The sensor system was applied by monitoring two cell culture models during treatment. Oxygen consumption changes, as indicator for effective treatment, were detected under normoxic and hypoxic conditions. Lactate production, under hypoxic conditions, was successfully monitored to indicate a possible shift to the anaerobic pathway. The monitoring of oxidative stress was performed using the integrated hydrogen peroxide sensors. Additionally to single parameter measurements, the introduced platform enabled the monitoring of oxygen gradients during partial treatment. The microsensor system was successfully applied to gain new insights into the cancer-specific pathways and the resulting metabolic changes. The benefit of metabolic monitoring for cancer cell therapy models was elucidated with different therapy strategies and culture formats. In the future, these findings can be employed for metabolic monitoring in organ-on-chip systems.
... In 1988, Simon reported that electrospinning could possibly be used to manufacture nanoscale and submicron-scale fibrous mats of polystyrene and polycarbonate and precisely proposed in vitro cell substrates be used. He further reported that the changes in the fibers' surface chemistry depend on the electric field polarity during electrospinning [38]. In the 1990s, Reneker and Rutledge confirmed that through electrospinning, various organic polymers could form nanofibers [39,40]. ...
Electrospun nanomaterials and their applications have increasingly gained interest over the last decade. Nanofibers are known for their exceptional surface area and wide opportunities for their functionalization. These properties have been attractive for various sensing applications; however, mostly electric sensing principles have been reported. An overview of most frequently studied concepts will be presented. A novel approach based on optical detection will be described. Various functionalized nanofiber materials have been used to demonstrate feasibility of realization of miniature sensors of biomedical and chemical values (enzymes reactions, metal ions content, concentration, etc.). Compactness and sensitivity of the sensors are significantly enhanced through original hybrid fiber-optic/nanofiber design. The potential of the new detection principle for various applications (bio-medical, chemical, forensic, automotive, etc.) will be discussed.
Electrospinning (ES) is a flexible and straightforward strategy that permits the creation of ultrathin fibers of various compositions. The need for helpful advancements in filtration ability has led to little thought of cutting-edge materials, for example, electrospun nanofibers (NFs) for wastewater treatment. Electrospun NFs play a significant part in numerous fields because of their high surface area, high porosity, and good functional capabilities. These properties make them encouraging materials for a range of applications, most explicitly water treatment. Furthermore, electrospun nanofibers can be simply functionalized by joining multifunctional materials to meet extraordinary water treatment effects. This chapter focuses on the most recent progress of electrospun NFs with special attention on wastewater treatment applications. In particular, we discuss the synthesis and functionalization of electrospun NFs. The various process parameters involved during ES have also been discussed. Finally, the points of view are introduced in regards to difficulties, openings, and new prospects for the use of electrospun NFs.
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