Results for “RCCS”

NASA NTRS2001

Characteristics of human dendritic cells generated in a microgravity analog culture system

Generation of an effective immune response requires that antigens be processed and presented to T lymphocytes by antigen-presenting cells, the most efficient of which are dendritic cells (DC). Because of their influence on both the innate and the acquired arms of immunity, a defect in DC would be expected to result in a broad impairment of immune function, not unlike that observed in astronauts during or after space flight. In the study reported here, we investigated whether DC generation and function are altered in a culture environment that models microgravity, i.e., the rotary-cell culture system (RCCS). We observed that RCCS supported the generation of DC identified by morphology, phenotype (HLA-DR+ and lacking lineage-associated markers), and function (high allostimulatory activity). However, the yield of DC from RCCS was significantly lower than that from static cultures. RCCS-generated DC were less able to phagocytose Aspergillus fumigatus conidia and expressed a lower density of surface HLA-DR. The proportion of DC expressing CD80 was also significantly reduced in RCCS compared to static cultures. When exposed to fungal antigens, RCCS-generated DC produced lower levels of interleukin-12 and failed to upregulate some costimulatory/adhesion molecules involved in antigen presentation. These data suggest that DC generation, and some functions needed to mount an effective immune response to pathogens, may be disturbed in the microgravity environment of space.

Non-NASA Center↗

NASA NTRS2001

Microarray analysis of genes differentially expressed in HepG2 cells cultured in simulated microgravity: preliminary report

Developed at NASA, the rotary cell culture system (RCCS) allows the creation of unique microgravity environment of low shear force, high-mass transfer, and enables three-dimensional (3D) cell culture of dissimilar cell types. Recently we demonstrated that a simulated microgravity is conducive for maintaining long-term cultures of functional hepatocytes and promote 3D cell assembly. Using deoxyribonucleic acid (DNA) microarray technology, it is now possible to measure the levels of thousands of different messenger ribonucleic acids (mRNAs) in a single hybridization step. This technique is particularly powerful for comparing gene expression in the same tissue under different environmental conditions. The aim of this research was to analyze gene expression of hepatoblastoma cell line (HepG2) during early stage of 3D-cell assembly in simulated microgravity. For this, mRNA from HepG2 cultured in the RCCS was analyzed by deoxyribonucleic acid microarray. Analyses of HepG2 mRNA by using 6K glass DNA microarray revealed changes in expression of 95 genes (overexpression of 85 genes and downregulation of 10 genes). Our preliminary results indicated that simulated microgravity modifies the expression of several genes and that microarray technology may provide new understanding of the fundamental biological questions of how gravity affects the development and function of individual cells.

Non-NASA Center↗

NASA NTRS1999

Induction of three-dimensional assembly of human liver cells by simulated microgravity

The establishment of long-term cultures of functional primary human liver cells (PHLC) is formidable. Developed at NASA, the Rotary Cell Culture System (RCCS) allows the creation of the unique microgravity environment of low shear force, high-mass transfer, and 3-dimensional cell culture of dissimilar cell types. The aim of our study was to establish long-term hepatocyte cultures in simulated microgravity. PHLC were harvested from human livers by collagenase perfusion and were cultured in RCCS. PHLC aggregates were readily formed and increased up to 1 cm long. The expansion of PHLC in bioreactors was further evaluated with microcarriers and biodegradable scaffolds. While microcarriers were not conducive to formation of spheroids, PHLC cultured with biodegradable scaffolds formed aggregates up to 3 cm long. Analyses of PHLC spheroids revealed tissue-like structures composed of hepatocytes, biliary epithelial cells, and/or progenitor liver cells that were arranged as bile duct-like structures along nascent vascular sprouts. Electron microscopy revealed groups of cohesive hepatocytes surrounded by complex stromal structures and reticulin fibers, bile canaliculi with multiple microvilli, and tight cellular junctions. Albumin mRNA was expressed throughout the 60-d culture. A simulated microgravity environment is conducive to maintaining long-term cultures of functional hepatocytes. This model system will assist in developing improved protocols for autologous hepatocyte transplantation, gene therapy, and liver assist devices, and facilitate studies of liver regeneration and cell-to-cell interactions that occur in vivo.

NASA Center JSC↗

NASA NTRS2008

Taking the Heat: Handling the Shuttle's RCC Wing Panels

Innovative inspection technology was developed to inspect the Reinforced Carbon-Carbon (RCC) wing panels on the vehicle, thus eliminating need to remove/reinstall all 44 RCC panels for inspections per processing flow. Manually holding inspection tools up to the RCC panels was a 3-person job with high risk of personnel injury and flight hardware damage. To further enhance ergonomics, reduce personnel/flight hardware risks, and improve repeatability, an inspection cart and fixture were constructed to physically secure the instruments for Inspectors during 652 inspection points per flow. The electric lift used to handle RCCs was also utilized to raise the heavy, bulky inspection equipment up to the wing leading edge.

Stegles, Katrine S.↗

NASA NTRS2013

Ceramic Adhesive and Methods for On-Orbit Repair of Re-Entry Vehicles

This adhesive is capable of repairing damaged leading edge components of reentry vehicles while in space, and is novel with regard to its ability to be applied in the vacuum of space, and in a microgravity environment. Once applied, the adhesive provides thermal and oxidation protection to the substrate (in this case, reinforced carbon/carbon composites, RCCs) during re-entry of a space vehicle. Although there may be many formulations for repair adhesives, at the time of this reporting, this is the first known adhesive capable of an on-orbit repair. The adhesive is an engineered ceramic material composed of a pre-ceramic polymer and refractory powders in the form of a paste or putty that can be applied to a scratched, cracked, or fractured composite surface, covering and protecting the damaged area. The adhesive is then "cured" with a heat cycle, thereby cross-linking the polymer into a hardened material and bonding it to the substrate. During the heat of reentry, the material is converted to a ceramic coating that provides thermal and oxidative stability to the repaired area, thus allowing the vehicle to pass safely from space into the upper atmosphere. Ceramic powders such as SiC, ZrB2 and Y2O3 are combined with allylhydridopolycarbosilane (AHPCS) resin, and are mixed to form a paste adhesive. The material is then applied to the damaged area by brush, spatula, trowel, or other means to fill cracks, gaps, and holes, or used to bond patches onto the damaged area. The material is then cured, in a vacuum, preferably at 250F (approximately equal to 121C) for two hours. The re-entry heating of the vehicle at temperatures in excess of 3,000F (approximately equal to 1,650C) then converts this material into a ceramic coating. This invention has demonstrated advantages in resistance to high temperatures, as was demonstrated in more than 100 arc-jet tests in representative environments at NASA. Extensive testing verified oxidation protection for the repaired substrate (RCC), and confirmed that the microstructure of the resulting repair leads to durability and resistance to melting or flow. Its processability and working life in a vacuum was demonstrated by NASA astronauts in glovebox processing studies, as well as on-orbit in the open space shuttle bay. All of these advantages increase the working life of NASA vehicles, as well as improve safety for any crew on a manned vehicle. The adhesive, trademarked NOAX(TM). or Non-Oxide Adhesive Experimental, flew on all space shuttle missions from Return To Flight (STS-114) until the final flight (STS- 135) as a crack repair material for the leading edges and nose cap of the vehicle. NOAX(TM) was patented under U.S. Patents 7,628,878 and 7,888,277.

Riedell, James A.↗

NASA NTRS2019

The Effects of Low-Shear Simulated Microgravity on Soil Bacterium Pantoea Agglomerans

As NASA's effort to establish a permanent residence in space continues, research on the effects of microgravity onbiological microorganisms is vital to protect or promote the health of plants and their astronaut counterparts. The purpose of this study is to determine the effects of microgravity on Pantoea agglomerans (P. agglomerans), using an analog microgravity simulator; the Rotary Cell Culture System (RCCS) developed at Johnson Space Center (JSC) in Houston, TX. P. agglomerans has been shown to be a plant growth promoter (PGPR) in ground based studies, but has also been shown to be a pathogen in both plants and immunocompromised patients. In this study, we will determine changes in the growth rate and antibiotic susceptibility of P. agglomerans when exposed to simulated microgravity.

HARV↗

NASA NTRS2023

Orbital Debris Shape Effect Investigations for Mitigating Risk

NASA’s Orbital Debris Program Office (ODPO) and Hypervelocity Impact Technology (HVIT) team have coordinated to better understand the risks to upper stages and spacecraft from non-spherical orbital debris. It is well understood that fragmentation (collision or explosion) events in orbit produce fragments of various materials, sizes, and shapes. To further characterize these parameters, the ODPO is developing the next-generation Orbital Debris Engineering Model (ORDEM) version 4.0 to include orbital debris shape distributions. Ground-based assets, such as radar and optical sensors, can provide size estimates and some insight into material based on radar return or optical filter photometry/spectroscopy, respectively. Characterizing an object’s shape requires more laboratory analyses to infer how shape affects these measurements. More importantly, in addition to size and material/density, the shape of fragments in orbit will alter the ballistic limit equations used in orbital debris risk assessments with NASA’s Bumper Code. The ODPO plans to release ORDEM 4.0 in the coming years. Performing ground-based laboratory impact tests on high-fidelity spacecraft mockups provides the means to directly measure size, mass, material/density, and shape of fragments, all key parameters needed to characterize real-world break up events. The DebriSat test, the results of which are provided, showcases the details of this type of experiment. The goal of this collaborative research between the ODPO and the HVIT team is to include a shape parameter in the environmental and breakup models used to assess risk for various space structures. This paper examines ground-based laboratory impact tests and the associated fragment shape categories. Provided these defined shapes, the approach is simplified by assuming a right circular cylinder (RCC) approximation with varying length-to-diameter ratios. Highlights of impact tests conducted by the HVIT team using non-spherical projectiles based on the RCC approximation are presented. Hydrocode simulations have also been performed to expand on the complexity of variations with non-spherical projectiles. Lastly, ray-tracing simulations of various RCCs of known material are provided to support the ongoing research on optical reflectance distributions with known shapes and to highlight how this may modify the current optical size estimation model. The status and plan forward are outlined for NASA's orbital debris shape effect investigation using a multidisciplinary approach by the ODPO and the HVIT team.

Heather Cowardin↗

NASA NTRS2023

Orbital Debris Shape Effect Investigations for Mitigating Risk

NASA’s Orbital Debris Program Office (ODPO) and Hypervelocity Impact Technology (HVIT) team have coordinated to better understand the risks to upper stages and spacecraft from non-spherical orbital debris. It is well understood that fragmentation (collision or explosion) events in orbit produce fragments of various materials, sizes, and shapes. To further characterize these parameters, the ODPO is developing the next-generation Orbital Debris Engineering Model (ORDEM) version 4.0 to include orbital debris shape distributions. Ground-based assets, such as radar and optical sensors, can provide size estimates and some insight into material based on radar return or optical filter photometry/spectroscopy, respectively. Characterizing an object’s shape requires more laboratory analyses to infer how shape affects these measurements. More importantly, in addition to size and material/density, the shape of fragments in orbit will alter the ballistic limit equations used in orbital debris risk assessments with NASA’s Bumper Code. The ODPO plans to release ORDEM 4.0 in the coming years. Performing ground-based laboratory impact tests on high-fidelity spacecraft mockups provides the means to directly measure size, mass, material/density, and shape of fragments, all key parameters needed to characterize real-world break up events. The DebriSat test, the results of which are provided, showcases the details of this type of experiment. The goal of this collaborative research between the ODPO and the HVIT team is to include a shape parameter in the environmental and breakup models used to assess risk for various space structures. This paper examines ground-based laboratory impact tests and the associated fragment shape categories. Provided these defined shapes, the approach is simplified by assuming a right circular cylinder (RCC) approximation with varying length-to-diameter ratios. Highlights of impact tests conducted by the HVIT team using non-spherical projectiles based on the RCC approximation are presented. Hydrocode simulations have also been performed to expand on the complexity of variations with non-spherical projectiles. Lastly, ray-tracing simulations of various RCCs of known material are provided to support the ongoing research on optical reflectance distributions with known shapes and to highlight how this may modify the current optical size estimation model. The status and plan forward are outlined for NASA's orbital debris shape effect investigation using a multidisciplinary approach by the ODPO and the HVIT team.

Heather Cowardin↗