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The Effect of Gravity Fields on Cellular Gene Expression

Early theoretical analysis predicted that microgravity effects on the isolated cell would be minuscule at the subcellular level; however, these speculations have not proven true in the real world. Astronauts experience a significant bone and muscle loss in as little as 2 weeks of spaceflight and changes are seen at the cellular level soon after exposure to microgravity. Changes in biological systems may be primarily due to the lack of gravity and the resulting loss of mechanical stress on tissues and cells. Recent ground and flight studies examining the effects of gravity or mechanical stress on cells demonstrate marked changes in gene expression when relatively small changes in mechanical forces or gravity fields were made. Several immediate early genes (IEG) like c-fos and c-myc are induced by mechanical stimulation within minutes. In contrast, several investigators report that the absence of mechanical forces during space flight result in decreased sera response element (SRE) activity and attenuation of expression of IEGs such as c-fos, c-jun and cox-2 mRNAs. Clearly, these early changes in gene expression may have long term consequences on mechanically sensitive cells. In our early studies on STS-56, we reported four major changes in the osteoblast; 1) prostaglandin synthesis in flight, 2) changes in cellular morphology, 3) altered actin cytoskeleton and 4) reduced osteoblast growth after four days exposure to microgravity. Initially, it was believed that changes in fibronectin (FN) RNA, FN protein synthesis or subsequent FN matrix formation might account for the changes in cytoskeleton and/ or reduction of growth. However our recent studies on Biorack (STS-76, STS-81 and STS-84), using ground and in-flight 1-G controls, demonstrated that fibronectin synthesis and matrix formation were normal in microgravity. In addition, in our most recent Biorack paper, our laboratory has documented that relative protein synthesis and mRNA synthesis are not changed after 24 hours exposure to microgravity. We did, however, find significant changes in osteoblast gene expression of IEGs, c-fos and cox-2 in microgravity exposure as compared to ground and in-flight 1-G controls. Subsequent ground studies suggest that the molecular mechanism underlying these changes may involve prostaglandin c-AMP receptors (EPs) and/or subsequent alteration of intracellular signaling in the absence of gravity.

Hughes-Fulford, Millie↗

Microgravity Science and Applications: Program Tasks and Bibliography for Fiscal Year 1996

NASA's Microgravity Science and Applications Division (MSAD) sponsors a program that expands the use of space as a laboratory for the study of important physical, chemical, and biochemical processes. The primary objective of the program is to broaden the value and capabilities of human presence in space by exploiting the unique characteristics of the space environment for research. However, since flight opportunities are rare and flight research development is expensive, a vigorous ground-based research program, from which only the best experiments evolve, is critical to the continuing strength of the program. The microgravity environment affords unique characteristics that allow the investigation of phenomena and processes that are difficult or impossible to study an Earth. The ability to control gravitational effects such as buoyancy driven convection, sedimentation, and hydrostatic pressures make it possible to isolate phenomena and make measurements that have significantly greater accuracy than can be achieved in normal gravity. Space flight gives scientists the opportunity to study the fundamental states of physical matter-solids, liquids and gasses-and the forces that affect those states. Because the orbital environment allows the treatment of gravity as a variable, research in microgravity leads to a greater fundamental understanding of the influence of gravity on the world around us. With appropriate emphasis, the results of space experiments lead to both knowledge and technological advances that have direct applications on Earth. Microgravity research also provides the practical knowledge essential to the development of future space systems. The Office of Life and Microgravity Sciences and Applications (OLMSA) is responsible for planning and executing research stimulated by the Agency's broad scientific goals. OLMSA's Microgravity Science and Applications Division (MSAD) is responsible for guiding and focusing a comprehensive program, and currently manages its research and development tasks through five major scientific areas: biotechnology, combustion science, fluid physics, fundamental physics, and materials science. FY 1996 was an important year for MSAD. NASA continued to build a solid research community for the coming space station era. During FY 1996, the NASA Microgravity Research Program continued investigations selected from the 1994 combustion science, fluid physics, and materials science NRAS. MSAD also released a NASA Research Announcement in microgravity biotechnology, with more than 130 proposals received in response. Selection of research for funding is expected in early 1997. The principal investigators chosen from these NRAs will form the core of the MSAD research program at the beginning of the space station era. The third United States Microgravity Payload (USMP-3) and the Life and Microgravity Spacelab (LMS) missions yielded a wealth of microgravity data in FY 1996. The USMP-3 mission included a fluids facility and three solidification furnaces, each designed to examine a different type of crystal growth.

Source record↗

Stationary Apparatus Would Apply Forces of Walking to Feet

A proposed apparatus would apply controlled cyclic forces to both feet for the purpose of preventing the loss of bone density in a human subject whose bones are not subjected daily to the mechanical loads of normal activity in normal Earth gravitation. The apparatus was conceived for use by astronauts on long missions in outer space; it could also be used by bedridden patients on Earth, including patients too weak to generate the necessary forces by their own efforts. The apparatus (see figure) would be a modified version of a bicycle-like exercise machine, called the cycle ergometer with vibration isolation system (CEVIS), now aboard the International Space Station. Attached to each CEVIS pedal would be a computer-controlled stress/ vibration exciter connected to the heel portion of a special-purpose pedal. The user would wear custom shoes that would amount to standard bicycle shoes equipped with cleats for secure attachment of the balls of the feet to the special- purpose pedals. If possible, prior to use of the apparatus, the human subject would wear a portable network of recording accelerometers, while walking, jogging, and running. The information thus gathered would be fed to the computer, wherein it would be used to make the exciters apply forces and vibrations closely approximating the forces and vibrations experienced by that individual during normal exercise. It is anticipated that like the forces applied to bones during natural exercise, these artificial forces would stimulate the production of osteoblasts (bone-forming cells), as needed to prevent or retard loss of bone mass. In addition to helping to prevent deterioration of bones, the apparatus could be used in treating a person already suffering from osteoporosis. For this purpose, the magnitude of the applied forces could be reduced, if necessary, to a level at which weak hip and leg bones would still be stimulated to produce osteoblasts without exposing them to the full stresses of walking and thereby risking fracture.

Hauss, Jessica↗

Behavioral Health Benefits of the ISS Pick-and-Eat Crop Growth System

Summary: We summarize findings related to assessing the pick-and-eat salad-crop productivity and acceptability of the ISS food system as a countermeasure to maintain behavioral health in long-duration spaceflight. BACKGROUND Plants are a potential countermeasure for the stresses of living in space. Caring for plants and eating fresh food have been shown to serve as a psychological benefit for previous astronauts and for others in analogous environments such as Antarctica [1,2]. Gardening can be therapeutic in terrestrial settings [3]. These effects may carry over or be more pronounced in austere environments such as spaceflight. It can reduce stress and increase sensory stimulation and enjoyment [2]. Growing crops may offer astronauts meaningful and engaging work as they care for living things in an austere environment away from Earth’s nature and as they provide sustenance for their crew. We examined the extent to which interacting with plants (e.g., tending to, consuming) was related to behavioral health outcomes in long-duration space missions. METHOD Participants were 27 astronauts that interacted with the plant system VEG-04, VEG-05, and HRF-VEG (VEG-03 I-L, PH-04). In total, there were 106 in-flight observations. Participants completed monthly measures of mood and well-being, enjoyment and time spent performing specific crop growth and consuming the plants, meaningfulness, performance, connections with others and the Earth, desire to work with and eat plants, and experiences with struggling or dying plants. Higher ends of the 7-point Likert scale indicated more positive outcomes, lower ends of the scale were anchored with more negative outcomes, and the scale midpoints indicated neutral positions. Participants selected a specific survey version based on whether they interacted with the plant system(s) or not, and we included only the scores from the crewmembers that participated in any given activity. RESULTS & DISCUSSION Results indicated that participants generally rated tasks related to the plants and crop growth as enjoyable. Consuming the harvested plants and voluntary viewing were the most enjoyable activities. Tending to the plants was reported as moderately enjoyable, indicating BHP benefits from this work task. The BHP impacts of whether the task was engaging, meaningful, supported well-being, or was demanding for a crewmember were generally positive and increased over time. As shown in the Figure, working with plants was reported as more engaging, meaningful, and beneficial to well-being over time, while the demand from the task was moderate to low and consistent over time. Engaging, meaningful, and supporting well-being were strongly, positively correlated with each other, which is consistent with research that suggests engaging and meaningful work can support well-being. Plant activities such as voluntary viewing, watering, harvesting, and consuming, were positively related to task engagement and meaning, and enjoyment and well-being. Findings suggest that working with plants and consuming them can be a behavioral health countermeasure for long-duration spaceflight. REFERENCES [1] Schlacht I, et al. (2020) Impact of plants in isolation: The EDEN-ISS…in Antarctica. In Advances in Human Factors of Transportation, Washington DC. [2] Vessel E, Russo S (2015) Effects of reduced sensory stimulation and…countermeasures…. NASA/TM-2015-218576. [3] Odeh R, Guy C (2017) Gardening for therapeutic people-plant interactions during LDSM. Open Agriculture, 2(1), 1-13. ACKNOWLEDGEMENTS This research was supported by the Human Health and Performance Contract NNJ15HK11B, and NASA grant MTL #1075 (PI G.D. Massa).

behavioral health↗

Behavioral Health Benefits of the ISS Pick-and-Eat Crop Growth System

Summary: We summarize findings related to assessing the pick-and-eat salad-crop productivity and acceptability of the ISS food system as a countermeasure to maintain behavioral health in long-duration spaceflight. BACKGROUND Plants are a potential countermeasure for the stresses of living in space. Caring for plants and eating fresh food have been shown to serve as a psychological benefit for previous astronauts and for others in analogous environments such as Antarctica [1,2]. Gardening can be therapeutic in terrestrial settings [3]. These effects may carry over or be more pronounced in austere environments such as spaceflight. It can reduce stress and increase sensory stimulation and enjoyment [2]. Growing crops may offer astronauts meaningful and engaging work as they care for living things in an austere environment away from Earth’s nature and as they provide sustenance for their crew. We examined the extent to which interacting with plants (e.g., tending to, consuming) was related to behavioral health outcomes in long-duration space missions. METHOD Participants were 27 astronauts that interacted with the plant system VEG-04, VEG-05, and HRF-VEG (VEG-03 I-L, PH-04). In total, there were 106 in-flight observations. Participants completed monthly measures of mood and well-being, enjoyment and time spent performing specific crop growth and consuming the plants, meaningfulness, performance, connections with others and the Earth, desire to work with and eat plants, and experiences with struggling or dying plants. Higher ends of the 7-point Likert scale indicated more positive outcomes, lower ends of the scale were anchored with more negative outcomes, and the scale midpoints indicated neutral positions. Participants selected a specific survey version based on whether they interacted with the plant system(s) or not, and we included only the scores from the crewmembers that participated in any given activity. RESULTS & DISCUSSION Results indicated that participants generally rated tasks related to the plants and crop growth as enjoyable. Consuming the harvested plants and voluntary viewing were the most enjoyable activities. Tending to the plants was reported as moderately enjoyable, indicating BHP benefits from this work task. The BHP impacts of whether the task was engaging, meaningful, supported well-being, or was demanding for a crewmember were generally positive and increased over time. As shown in the Figure, working with plants was reported as more engaging, meaningful, and beneficial to well-being over time, while the demand from the task was moderate to low and consistent over time. Engaging, meaningful, and supporting well-being were strongly, positively correlated with each other, which is consistent with research that suggests engaging and meaningful work can support well-being. Plant activities such as voluntary viewing, watering, harvesting, and consuming, were positively related to task engagement and meaning, and enjoyment and well-being. Findings suggest that working with plants and consuming them can be a behavioral health countermeasure for long-duration spaceflight. REFERENCES [1] Schlacht I, et al. (2020) Impact of plants in isolation: The EDEN-ISS…in Antarctica. In Advances in Human Factors of Transportation, Washington DC. [2] Vessel E, Russo S (2015) Effects of reduced sensory stimulation and…countermeasures…. NASA/TM-2015-218576. [3] Odeh R, Guy C (2017) Gardening for therapeutic people-plant interactions during LDSM. Open Agriculture, 2(1), 1-13. ACKNOWLEDGEMENTS This research was supported by the Human Health and Performance Contract NNJ15HK11B, and NASA grant MTL #1075 (PI G.D. Massa).

countermeasure↗

A novel calmodulin-regulated Ca2+-ATPase (ACA2) from Arabidopsis with an N-terminal autoinhibitory domain

To study transporters involved in regulating intracellular Ca2+, we isolated a full-length cDNA encoding a Ca2+-ATPase from a model plant, Arabidopsis, and named it ACA2 (Arabidopsis Ca2+-ATPase, isoform 2). ACA2p is most similar to a "plasma membrane-type" Ca2+-ATPase, but is smaller (110 kDa), contains a unique N-terminal domain, and is missing a long C-terminal calmodulin-binding regulatory domain. In addition, ACA2p is localized to an endomembrane system and not the plasma membrane, as shown by aqueous-two phase fractionation of microsomal membranes. ACA2p was expressed in yeast as both a full-length protein (ACA2-1p) and an N-terminal truncation mutant (ACA2-2p; Delta residues 2-80). Only the truncation mutant restored the growth on Ca2+-depleted medium of a yeast mutant defective in both endogenous Ca2+ pumps, PMR1 and PMC1. Although basal Ca2+-ATPase activity of the full-length protein was low, it was stimulated 5-fold by calmodulin (50% activation around 30 nM). In contrast, the truncated pump was fully active and insensitive to calmodulin. A calmodulin-binding sequence was identified within the first 36 residues of the N-terminal domain, as shown by calmodulin gel overlays on fusion proteins. Thus, ACA2 encodes a novel calmodulin-regulated Ca2+-ATPase distinguished by a unique N-terminal regulatory domain and a non-plasma membrane localization.

Non-NASA Center↗

Rapid systemic up-regulation of genes after heat-wounding and electrical stimulation

When one leaf of a tomato plant is electrically-stimulated or heat-wounded, proteinase inhibitor genes are rapidly up-regulated in distant leaves. The identity of the systemic wound signal(s) is not yet known, but major candidates include hormones transmitted via the phloem or the xylem, the electrically-stimulated self-propagating electrical signal in the phloem (the action potential, AP), or the heat-wound-induced surge in hydraulic pressure in the xylem evoking a local change in membrane potential in adjacent living cells (the variation potential, VP). In order to discriminate between these signals we have adopted two approaches. The first approach involves applying stimuli that evoke known signals and determining whether these signals have similar effects on the "model" transcripts for proteinase inhibitors (pin) and calmodulin (cal). Here we show that a heat wound almost invariably evokes a VP, while an electrical stimulation occasionally evokes an AP, and both of these signals induce accumulation of transcripts encoding proteinase inhibitors. The second approach involves identifying the array of genes turned on by heat-wounding. To this end, we have constructed a subtractive library for heat-wounded tissue, isolated over 800 putatively up-regulated clones, and shown that all but two of the fifty that we have analyzed by Northern hybridization are, indeed, up-regulated. Here we show the early kinetics of up-regulation of three of these transcripts in the terminal (4th) leaf in response to heat-wounding the 3rd leaf, about 5 cm away. Even though these transcripts show somewhat different time courses of induction, with one peaking at 30 min, another at 15 min, and another at 5 min after flaming of a distant leaf, they all exhibit a similar pattern, i.e., a transient period of transcript accumulation preceding a period of transcript decrease, followed by a second period of transcript accumulation.

NASA Discipline Plant Biology↗

CHOICE - Directed Study: Consequences of longterm- Confinement and Hypobaric HypOxia on Immunity in the Antarctic Concordia Environment

Concerning ground-based space physiological research, the choice of analog must carefully match the system of interest. For spaceflight-associated immune dysregulation (SAID), Antarctica winter-over has emerged as potentially the best terrestrial analog. The prolonged mission durations, extreme/dangerous environment, station-based lifestyle, isolation from outside world, disrupted circadian rhythms, and other psychological aspects make this analog extremely high fidelity for exploration-class space missions (long duration lunar, Mars). NASA, ESA and RSA are currently investigating SAID, with NASA currently operating the Integrated Immune flight study. It is desirable to have a ground analog for SAID validated, so that potential countermeasures might be validated terrestrially prior to during flight. For this presentation, NASA data collected on the winterover 2009 crewmembers, baseline through early deployment will be presented. Through early deployment (approximately 2-3 weeks at Concordia), phenotypic alterations included increased levels of memory T cells, shifts among the CD8+ T cell compartment to a more mature phenotype, and increases in constitutively activated T cells. CD8+/IFNg+ T cell percentages, and T cell blastogenesis functional responses were depressed early deployment as compared to healthy controls. In four compatible subjects, secreted T cell Th1/Th2 cytokines were measured following culture stimulation, and a Th2 shift was observed as compared to controls. Post-winter over frozen sample return will be required to determine if this shift persisted during the winter over period. Additionally, circadian rhythms remained altered compared to baseline, as determined through 5x daily cortisol measurements. Latent viral reactivation will not be determined until frozen sample return occurs.

Sams, Clarence↗

Anaerobic Psychrophiles from Lake Zub and Lake Untersee, Antarctica

The study of samples from Antarctica 2008 and 2009 expeditions organized and successfully conducted by Richard Hoover led to the isolation of diverse anaerobic strains with psychrotolerant and psychrophilic physiology. Due to the fact that Lake Untersee has never been subject to microbiological study, this work with the samples has significant and pioneering impact to the knowledge about the biology of this unique ecosystem. Also, the astrobiological significance for the study of these ecosystems is based on new findings of ice covered water systems on other bodies of our solar system. Anaerobic psychrotolerant strain LZ-22 was isolated from a frozen sample of green moss with soils around the rhizosphere collected near Lake Zub in Antarctica. Morphology of strain LZ-22 was observed to be motile, rod shaped and spore-forming cells with sizes 1 x 5-10 micron. This new isolate is a mesophile with the maximum temperature of growth at 40C. Strain LZ-22 is able to live on media without NaCl and in media with up to 7% (w/v) NaCl. It is catalase negative and grows only on sugars with the best growth rate being on lactose. The strain is a neutrophile and grows between pH 5 and 9.0 with the optimum at 7.8. Another two strains UL7-96mG and LU-96m7P were isolated from deep water samples of Lake Untersee. Proteolytic strain LU-96m7P had a truly psychrophilic nature and refused to grow at room temperature. Sugarlytic strain UL7-96mG was found to be psychrotolerant, but its rate of growth at 3C was very high compared with other mesophiles. Two homoacetogenic psychrophilic strains A7AC-96m and AC-DS7 were isolated and purified from samples of Lake Untersee; both of them are able to grow chemolithotrophically on H2+CO2. In the presence of lactate, these strains are able to grow only at 0-18C, and growth at 22C was observed only with yeast extract stimulation. In this paper, physiological and morphological characteristics of novel psychrophilic and psychrotolerant isolates from Antarctica 2008 and 2009 expeditions will be discussed.

Townsend, Alisa↗

Improved Whole-Blood-Staining Device

Dramatic improvements have been made in NASA s Whole Blood Staining Device (WBSD) since it was last described in "Whole-Blood-Staining Device," NASA Tech Briefs, Vol. 23, No. 10 (October 1999), page 64. The new system has a longer shelf life, a simpler and more effective operational procedure, improved interface with instrumentation, and shorter processing time. More specifically, the improvements have targeted bag and locking clip materials, sampling ports, and air pocket prevention. The WBSD stains whole blood collected during spaceflight for subsequent flow cytometric analysis. In short, the main device stains white blood cells by use of monoclonal antibodies conjugated to various fluorochromes, followed by lysing and fixing of the cells by use of a commercial reagent that has been diluted according to NASA safety standards. This system is compact, robust, and does not require electric power, precise mixing, or precise incubation times. Figure 1 depicts the present improved version for staining applications, which is a poly(tetrafluoroethylene) bag with a Luer-lock port and plastic locking clips. An InterLink (or equivalent) intravenous- injection port screws into the Luer-lock port. The inflatable/collapsible nature of the bag facilitates loading and helps to minimize the amount of air trapped in the fully loaded bag. Some additional uses have been identified for the device beyond whole blood staining. The WBSD has been configured for functional assays that require culture of live cells by housing sterile culture media, mitogens, and fixatives prior to use [Figure 2(a)]. Simple injection of whole blood allows cell-stimulation culture to be performed in reduced gravity conditions, and product stabilization prior to storage, while protecting astronauts from liquid biohazardous materials. Also, the improved WBSD has reconstituted powdered injectable antibiotics by mixing them with diluent liquids [Figure 2(b)]. Although such mixing can readily be performed on Earth by shaking in glass vials, it cannot readily be performed this way in outer space without entraining air bubbles. The present device can be preloaded with the powder and diluent(s) in separate compartments. The powder and diluent( s) can be mixed, without introducing air bubbles, by removing the clip(s), then shaking. This use of the device could also be advantageous in terrestrial applications because it maintains the isolation of the constituents until the time of use.

Sams, Clarence F.↗

Inhibition of nitrate transport by anti-nitrate reductase IgG fragments and the identification of plasma membrane associated nitrate reductase in roots of barley seedlings

Membrane associated nitrate reductase (NR) was detected in plasma membrane (PM) fractions isolated by aqueous two-phase partitioning from barley (Hordeum vulgare L. var CM 72) roots. The PM associated NR was not removed by washing vesicles with 500 millimolar NaCl and 1 millimolar EDTA and represented up to 4% of the total root NR activity. PM associated NR was stimulated up to 20-fold by Triton X-100 whereas soluble NR was only increased 1.7-fold. The latency was a function of the solubilization of NR from the membrane. NR, solubilized from the PM fraction by Triton X-100 was inactivated by antiserum to Chlorella sorokiniana NR. Anti-NR immunoglobulin G fragments purified from the anti-NR serum inhibited NO3- uptake by more than 90% but had no effect on NO2- uptake. The inhibitory effect was only partially reversible; uptake recovered to 50% of the control after thorough rinsing of roots. Preimmune serum immunoglobulin G fragments inhibited NO3- uptake 36% but the effect was completely reversible by rinsing. Intact NR antiserum had no effect on NO3- uptake. The results present the possibility that NO3- uptake and NO3- reduction in the PM of barley roots may be related.

NASA Discipline Number 61-10↗

Conditions of the Toll-Like Receptors System of the Human Innate Immunity Cells During the Long-Term Isolation

During long duration missions onboard the International Space Station (ISS), alterations in T cell function and plasma cytokine profiles have been observed. Recent studies involving pattern recognition receptors (PRRs) have established that adaptation of innate immunity to long duration space flight occurs through changes in signal PRRs, Toll-like receptors (TLRs). Despite the fact that fundamental research of PRR molecular biology is far from complete, there are an increasing number of arguments confirming the important role of this system in regulating the initial stages of the adaptive immune response, as well as in the formation of autoimmune and allergic diseases. Long duration isolation is widely used by NASA as an analog for space flight with previous missions including several Antarctic bases, an undersea research station (NEEMO), and other isolation chambers. ‘SIRIUS’ is a multi-compartment pressurized facility at the IMBP in Moscow, Russia, in which subjects will spend 8 months executing tasks similar to those performed by astronauts in space. Prolonged isolation studies like SIRIUS replicate many of the influences expected to occur during long duration spaceflight missions including isolation, stress, confinement, and living in remote conditions. The immune study to be performed on the upcoming SIRIUS mission is being led by Dr. Sergey Ponomarev at the Institute of Biomedical Problems in Moscow, Russia. The NASA JSC Immunology lab will provide analysis of immune cell function, serum cytokines and stress hormones, and salivary cortisol, cytokines, and viruses. This study will be conducted with SIRIUS subjects under the auspices of the parent program. The goal of the study is to evaluate the effect of the 8-month isolation mission on components of human innate immunity at the molecular and cellular level. Successful implementation and completion of this project will provide a unique assortment of new data, significantly expanding the current understanding of the molecular and cellular changes in TLRs during long-term isolation. We hypothesize that mission-like isolation in the Russian NEK facility, consisting of circadian misalignment, isolation, stress, and ‘station lifestyle’, will result in a pattern of immune dysregulation similar to that observed in astronauts onboard ISS. This pattern of alterations can be summarized as phenotypic changes (altered distribution of peripheral cytotoxic, central memory, CD8+ T cell subsets), reductions in T cell function, alterations in plasma cytokine profiles (increased inflammatory and certain chemokines), and reductions in mitogen stimulated cytokine profiles (spans Th1, Th2, Th17). We further hypothesize that these immune changes will correlate with the reactivation of latent herpesviruses (which persists during orbital flight), to be used as measurable adverse clinical manifestation or with other observed symptomology. The SIRIUS-21 mission began in November of 2021 and successfully concluded with the crew emerging in July of 2022. Currently, the logistics of shipping the samples from Russia to the United States is being organized.

Cody L Gutierrez↗

Alteration of mineral crystallinity and collagen cross-linking of bones in osteopetrotic toothless (tl/tl) rats and their improvement after treatment with colony stimulating factor-1

A common feature of various types of mammalian osteopetroses is a marked increase in bone mass accompanied by spontaneous bone fractures. The toothless (tl/tl) rat osteopetrotic mutation is characterized by drastically reduced bone resorption due to a profound deficiency of osteoclasts and their precursors. An altered bone morphology has also been observed. The mutants cannot be cured by bone marrow transplantation, but skeletal defects are greatly reduced after treatment with colony stimulating factor 1 (CSF-1). The objectives of this study were to characterize mineral and collagen matrices in cancellous and compact bone isolated from long bones of 6-week-old normal littermates, tl/tl osteopetrotic mutants and mutants (tl/tl) treated with CSF-1. There were no differences in bone mineral content, but a significant decrease in the crystallinity of mineral evaluated by the method based on electron paramagnetic resonance spectrometry was observed in all bones of tl/tl mutants as compared to that of controls. Within the collagen matrix, slight decreases in the labile cross-links, but significant increases in the content of the stable cross-links, pyridinoline, and deoxypyridinoline, were observed in both cancellous and compact bone of osteopetrotic mutants. In tl/tl mutants treated with human recombinant CSF-1, the normalization of the crystallinity of bone mineral as well as collagen cross-links was found. Our results indicate that remodeling of bone matrix in tl/tl mutants is highly suppressed, but that after treatment with CSF-1, this activity recovers significantly. Taken together, these data provide further support for the hypothesis that CSF-1 is an essential factor for normal osteoclast differentiation and bone remodelling.

NASA Discipline Musculoskeletal↗