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At least 523 records · Page 29

The fluid dynamic and shear environment in the NASA/JSC rotating-wall perfused-vessel bioreactor

The rotating-wall perfused-vessel (RWPV) bioreactor, used for both microgravity and Earth-based cell science experiments, is characterized in terms of the fluid dynamic and fluid shear stress environment. A numerical model of the flow field is developed and verified with laser Doppler velocimeter measurements. The effects of changes in operating conditions, including rotation rates and fluid perfusion rates, are investigated with the numerical model. The operating conditions typically used for ground-based experiments (equal rotation of the inner and outer cylinders) leads to flow patterns with relatively poor mass distribution characteristics. Approximately 50% of the inlet-perfused fluid bypasses the bulk of the fluid volume and flows to the perfusion exit. For operating conditions typical in microgravity, small differential rotation rates between the inner and outer cylinders lead to greatly improved flow distribution patterns and very low fluid shear stress levels over a large percentage of the fluid volume. Differences in flow patterns for the different operating conditions are explored. Large differences in the hydrodynamic environments for operating conditions typical of true microgravity and ground-based "microgravity simulations" are demonstrated.

Bioreactors↗

Lymphocyte trafficking and HIV infection of human lymphoid tissue in a rotating wall vessel bioreactor

The pathogenesis of HIV infection involves a complex interplay between both the infected and noninfected cells of human lymphoid tissue, the release of free viral particles, the de novo infection of cells, and the recirculatory trafficking of peripheral blood lymphocytes. To develop an in vitro model for studying these various aspects of HIV pathogenesis we have utilized blocks of surgically excised human tonsils and a rotating wall vessel (RWV) cell culture system. Here we show that (1) fragments of the surgically excised human lymphoid tissue remain viable and retain their gross cytoarchitecture for at least 3 weeks when cultured in the RWV system; (2) such lymphoid tissue gradually shows a loss of both T and B cells to the surrounding growth medium; however, this cellular migration is reversible as demonstrated by repopulation of the tissue by labeled cells from the growth medium; (3) this cellular migration may be partially or completely inhibited by embedding the blocks of lymphoid tissue in either a collagen or agarose gel matrix; these embedded tissue blocks retain most of the basic elements of a normal lymphoid cytoarchitecture; and (4) both embedded and nonembedded RWV-cultured blocks of human lymphoid tissue are capable of productive infection by HIV-1 of at least three various strains of different tropism and phenotype, as shown by an increase in both p24 antigen levels and free virus in the culture medium, and by the demonstration of HIV-1 RNA-positive cells inside the tissue identified by in situ hybridization. It is therefore reasonable to suggest that gel-embedded and nonembedded blocks of human lymphoid tissue, cocultured with a suspension of tonsillar lymphocytes in an RWV culture system, constitute a useful model for simulating normal lymphocyte recirculatory traffic and provide a new tool for testing the various aspects of HIV pathogenesis.

Tissue Culture/instrumentation/methods↗

Morphologic differentiation of colon carcinoma cell lines HT-29 and HT-29KM in rotating-wall vessels

A new low shear stress microcarrier culture system has been developed at NASA's Johnson Space Center that permits three-dimensional tissue culture. Two established human colon adenocarcinoma cell lines, HT-29, an undifferentiated, and HT-29KM, a stable, moderately differentiated subline of HT-29, were grown in new tissue culture bioreactors called Rotating-Wall Vessels (RWVs). RWVs are used in conjunction with multicellular cocultivation to develop a unique in vitro tissue modeling system. Cells were cultivated on Cytodex-3 microcarrier beads, with and without mixed normal human colonic fibroblasts, which served as the mesenchymal layer. Culture of the tumor lines in the absence of fibroblasts produced spheroidlike growth and minimal differentiation. In contrast, when tumor lines were co-cultivated with normal colonic fibroblasts, initial growth was confined to the fibroblast population until the microcarriers were covered. The tumor cells then commenced proliferation at an accelerated rate, organizing themselves into three-dimensional tissue masses that achieved 1.0- to 1.5-cm diameters. The masses displayed glandular structures, apical and internal glandular microvilli, tight intercellular junctions, desmosomes, cellular polarity, sinusoid development, internalized mucin, and structural organization akin to normal colon crypt development. Differentiated samples were subjected to transmission and scanning electron microscopy and histologic analysis, revealing embryoniclike mesenchymal cells lining the areas around the growth matrices. Necrosis was minimal throughout the tissue masses. These data suggest that the RWV affords a new model for investigation and isolation of growth, regulatory, and structural processes within neoplastic and normal tissue.

Colonic Neoplasms/pathology↗

Validation and Verification of Composite Pressure Vessel Design

Ten composite pressure vessels were instrumented with fiber Bragg grating sensors and pressure tested Through burst. This paper and presentation will discuss the testing methodology, the test results, compare the testing results to the analytical model, and also compare the fiber Bragg grating sensor data with data obtained against that obtained from foil strain gages.

Kreger, Stephen T.↗

Techniques for Embedding Instrumentation in Pressure Vessel Test Articles

Many interesting structural and thermal events occur in materials that are housed within a surrounding pressure vessel. In order to measure the environment during these events and explore their causes instrumentation must be installed on or in the material. Transducers can be selected that are small enough to be embedded within the test material but these instruments must interface with an external system in order to apply excitation voltages and output the desired data. The methods for installing the instrumentation and creating an interface are complicated when the material is located in a case or housing containing high pressures and hot gases. Installation techniques for overcoming some of these difficulties were developed while testing a series of small-scale solid propellant and hybrid rocket motors at Marshall Space Flight Center. These techniques have potential applications in other test articles where data are acquired from materials that require containment due to the severe environment encountered during the test process. This severe environment could include high pressure, hot gases, or ionized atmospheres. The development of these techniques, problems encountered, and the lessons learned from the ongoing testing process are summarized.

Cornelius, Michael↗

Nondestructive Methods and Special Test Instrumentation Supporting NASA Composite Overwrapped Pressure Vessel Assessments

Many aging composite overwrapped pressure vessels (COPVs), being used by the National Aeronautics and Space Administration (NASA) are currently under evaluation to better quantify their reliability and clarify their likelihood of failure due to stress rupture and age-dependent issues. As a result, some test and analysis programs have been successfully accomplished and other related programs are still in progress at the NASA Johnson Space Center (JSC) White Sands Test Facility (WSTF) and other NASA centers, with assistance from the commercial sector. To support this effort, a group of Nondestructive Evaluation (NDE) experts was assembled to provide NDE competence for pretest evaluation of test articles and for application of NDE technology to real-time testing. Techniques were required to provide assurance that the test article had adequate structural integrity and manufacturing consistency to be considered acceptable for testing and these techniques were successfully applied. Destructive testing is also being accomplished to better understand the physical and chemical property changes associated with progression toward "stress rupture" (SR) failure, and it is being associated with NDE response, so it can potentially be used to help with life prediction. Destructive work also includes the evaluation of residual stresses during dissection of the overwrap, laboratory evaluation of specimens extracted from the overwrap to evaluate physical property changes, and quantitative microscopy to inform the theoretical micromechanics.

Saulsberry, Regor↗

A Comparison of Various Stress Rupture Life Models for Orbiter Composite Pressure Vessels and Confidence Intervals

In conjunction with a recent NASA Engineering and Safety Center (NESC) investigation of flight worthiness of Kevlar Overwrapped Composite Pressure Vessels (COPVs) on board the Orbiter, two stress rupture life prediction models were proposed independently by Phoenix and by Glaser. In this paper, the use of these models to determine the system reliability of 24 COPVs currently in service on board the Orbiter is discussed. The models are briefly described, compared to each other, and model parameters and parameter uncertainties are also reviewed to understand confidence in reliability estimation as well as the sensitivities of these parameters in influencing overall predicted reliability levels. Differences and similarities in the various models will be compared via stress rupture reliability curves (stress ratio vs. lifetime plots). Also outlined will be the differences in the underlying model premises, and predictive outcomes. Sources of error and sensitivities in the models will be examined and discussed based on sensitivity analysis and confidence interval determination. Confidence interval results and their implications will be discussed for the models by Phoenix and Glaser.

Grimes-Ledesma, Lorie↗

Simulated Microgravity Regulates Gene Transcript Profiles of 2T3 Preosteoblasts: Comparison of the Random Positioning Machine and the Rotating Wall Vessel Bioreactor

Microgravity of spaceflight induces bone loss due in part to decreased bone formation by osteoblasts. We have previously examined the microgravity-induced changes in gene expression profiles in 2T3 preosteoblasts using the Random Positioning Machine (RPM) to simulate microgravity conditions. Here, we hypothesized that exposure of preosteoblasts to an independent microgravity simulator, the Rotating Wall Vessel (RWV), induces similar changes in differentiation and gene transcript profiles, resulting in a more confined list of gravi-sensitive genes that may play a role in bone formation. In comparison to static 1g controls, exposure of 2T3 cells to RWV for 3 days inhibited alkaline phosphatase activity, a marker of differentiation, and downregulated 61 genes and upregulated 45 genes by more than two-fold as shown by microarray analysis. The microarray results were confirmed with real time PCR for downregulated genes osteomodulin, bone morphogenic protein 4 (BMP4), runx2, and parathyroid hormone receptor 1. Western blot analysis validated the expression of three downregulated genes, BMP4, peroxiredoxin IV, and osteoglycin, and one upregulated gene peroxiredoxin I. Comparison of the microarrays from the RPM and the RWV studies identified 14 gravi-sensitive genes that changed in the same direction in both systems. Further comparison of our results to a published database showing gene transcript profiles of mechanically loaded mouse tibiae revealed 16 genes upregulated by the loading that were shown to be downregulated by RWV and RPM. These mechanosensitive genes identified by the comparative studies may provide novel insights into understanding the mechanisms regulating bone formation and potential targets of countermeasure against decreased bone formation both in astronauts and in general patients with musculoskeletal disorders.

Patel, Mamta J.↗

Three-Dimensional Digital Image Correlation of a Composite Overwrapped Pressure Vessel During Hydrostatic Pressure Tests

Ambient temperature hydrostatic pressurization tests were conducted on a composite overwrapped pressure vessel (COPV) to understand the fiber stresses in COPV components. Two three-dimensional digital image correlation systems with high speed cameras were used in the evaluation to provide full field displacement and strain data for each pressurization test. A few of the key findings will be discussed including how the principal strains provided better insight into system behavior than traditional gauges, a high localized strain that was measured where gages were not present and the challenges of measuring curved surfaces with the use of a 1.25 in. thick layered polycarbonate panel that protected the cameras.

Revilock, Duane M., Jr.↗

Detecting thermal discrepancies in vessel walls

An infrared, heat-sensing catheter particularly useful for identifying potentially fatal arterial plaques in patients with disease of the coronary or other arteries and its use are detailed. In one embodiment, an infrared fiberoptic system (with or without ultrasound) is employed at the tip of the catheter to locate inflamed, heat-producing, atherosclerotic plaque, which is at greater risk for rupture, fissure, or ulceration, and consequent thrombosis and occlusion of the artery. In another embodiment, a catheter with an infrared detector (with or without ultrasound) employed at its tip will likewise locate inflamed heat-producing atherosclerotic plaque. The devices and methods of the invention may be used to detect abscesses, infection, and cancerous regions by the heat such regions differentially display over the ambient temperature of immediately adjacent tissues. The methods and devices of the invention may also be used to detect regions of cooler than ambient tissue in a vessel or organ which indicate cell death, thrombosis, cell death, hemorrhage, calcium or cholesterol accumulations, or foreign materials.

Casscells, S. Ward↗

System and method of designing a load bearing layer of an inflatable vessel

A computer-implemented method is provided for designing a restraint layer of an inflatable vessel. The restraint layer is inflatable from an initial uninflated configuration to an inflated configuration and is constructed from a plurality of interfacing longitudinal straps and hoop straps. The method involves providing computer processing means (e.g., to receive user inputs, perform calculations, and output results) and utilizing this computer processing means to implement a plurality of subsequent design steps. The computer processing means is utilized to input the load requirements of the inflated restraint layer and to specify an inflated configuration of the restraint layer. This includes specifying a desired design gap between pairs of adjacent longitudinal or hoop straps, whereby the adjacent straps interface with a plurality of transversely extending hoop or longitudinal straps at a plurality of intersections. Furthermore, an initial uninflated configuration of the restraint layer that is inflatable to achieve the specified inflated configuration is determined. This includes calculating a manufacturing gap between pairs of adjacent longitudinal or hoop straps that correspond to the specified desired gap in the inflated configuration of the restraint layer.

Spexarth, Gary R.↗

High Pressure Composite Overwrapped Pressure Vessel (COPV) Development Tests at Cryogenic Temperatures

Development tests were conducted to evaluate the performance of 2 COPV designs at cryogenic temperatures. This allows for risk reductions for critical components for a Gaseous Helium (GHe) Pressurization Subsystem for an Advanced Propulsion System (APS) which is being proposed for NASA s Constellation project and future exploration missions. It is considered an advanced system since it uses Liquid Methane (LCH4) as the fuel and Liquid Oxygen (LO2) as the oxidizer for the propellant combination mixture. To avoid heating of the propellants to prevent boil-off, the GHe will be stored at subcooled temperatures equivalent to the LO2 temperature. Another advantage of storing GHe at cryogenic temperatures is that more mass of the pressurized GHe can be charged in to a vessel with a smaller volume, hence a smaller COPV, and this creates a significant weight savings versus gases at ambient temperatures. The major challenge of this test plan is to verify that a COPV can safely be used for spacecraft applications to store GHe at a Maximum Operating Pressure (MOP) of 4,500 psig at 140R to 160R (-320 F to -300 F). The COPVs for these tests were provided by ARDE , Inc. who developed a resin system to use at cryogenic conditions and has the capabilities to perform high pressure testing with LN2.

Ray, David M.↗

Neural Network Burst Pressure Prediction in Composite Overwrapped Pressure Vessels

Acoustic emission data were collected during the hydroburst testing of eleven 15 inch diameter filament wound composite overwrapped pressure vessels. A neural network burst pressure prediction was generated from the resulting AE amplitude data. The bottles shared commonality of graphite fiber, epoxy resin, and cure time. Individual bottles varied by cure mode (rotisserie versus static oven curing), types of inflicted damage, temperature of the pressurant, and pressurization scheme. Three categorical variables were selected to represent undamaged bottles, impact damaged bottles, and bottles with lacerated hoop fibers. This categorization along with the removal of the AE data from the disbonding noise between the aluminum liner and the composite overwrap allowed the prediction of burst pressures in all three sets of bottles using a single backpropagation neural network. Here the worst case error was 3.38 percent.

Hill, Eric v. K.↗

Strength, Fatigue, and Fracture Toughness of Ti-6Al-4V Liner from a Composite Over-Wrapped Pressure Vessel

It was demonstrated by way of experiment that Composite Over-wrapped Pressure Vessel (COPV) Ti-6Al-4V liner material can sustain the expected service loads and cycles. The experiments were performed as part of investigations on the residual life of COPV tanks being used in Space Shuttle Orbiters. Measured properties included tensile strength, compressive strength, reversed loading cycles to simulate liner proof strains, and cyclic fatigue loading to demonstrate the ability to sustain 1000 cycles after liner buckling. The liner material came from a salvaged 40 in. Columbia (orbiter 102) tank (SN029), and tensile strength measurements were made on both boss-transition (thick) and membrane regions (thin). The average measured yield strength was 131 ksi in the boss-transition and membrane regions, in good agreement with measurements made on 1970 s vintage forged plate stock. However, Young s modulus was 17.4+/-0.3 Msi, somewhat higher than typical handbook values (approx.16 Msi). The fracture toughness, as estimated from a failed fatigue specimen, was 74 ksi/sq in, in reasonable agreement with standardized measurements made on 1970 s vintage forged plate stock. Low cycle fatigue of a buckled test specimen implied that as-imprinted liners can sustain over 4000 load cycles.

Salem, Jonathan A.↗

Strain Measurement during Stress Rupture of Composite Over-Wrapped Pressure Vessel with Fiber Bragg Gratings Sensors

Fiber optic Bragg gratings were used to measure strain fields during Stress Rupture (SSM) test of Kevlar Composite Over-Wrapped Pressure Vessels (COPV). The sensors were embedded under the over-wrapped attached to the liner released from the Kevlar and attached to the Kevlar released from the liner. Additional sensors (foil gages and fiber bragg gratings) were surface mounted on the COPY liner.

Banks, Curtis E.↗

Overview of NASA White Sands Test Facility Composite Overwrapped Pressure Vessel Testing

This viewgraph presentation examines the White Sands Test Facility testing of Composite overwrapped pressure vessel (COPV). A COPV is typically a metallic liner overwrapped with a fiber epoxy matrix. There is a weight advantage over the traditional all metal design. The presentation shows pictures of the facilities at White Sands, and then examines some of the testing performed. The tests include fluids compatibility, and Kevlar COPV. Data for the Kevlar tests are given, and an analysis is reviewed. There is also a comparison between Carbon COPVs and the Kevlar COPVs.

Greene, Nathanael↗