Indiana University-Purdue University Indianapolis (IUPUI) / The effect of weightlessness on bone and osteoblast precursor cells has previously been studied. A marked decrease in bone formation, an increase in less differentiated committed osteogenic cells (A+A'), and a decrease in preosteoblast cells (C+D) was noted. To date, the effect of hypergravity (2g) on osteoblast histogenesis has not been studied in vivo. In vitro studies using nonphysiologic high levels of gravity (20,40g) have shown an increased proliferation of cloned osteoblast-like cells. The purpose of the present study was to evaluate the effect of hypergravity (2g) on osteoblast histogenesis in the rat periodontal ligament (PDL), on the width of the mesial PDL, and on the percentage of forming bone surface on the mesial side of the tooth. Twenty male Wistar rats (SPF: Harlan Sprague Dawley) were randomly assigned to the centrifuge (experimental) or to the stationary (control) group. The experimental group was centrifuged for 14 days at 2g and the stationary group was housed in identical cages in the centrifuge room. The PDL of the mesial and distal surface of the mesial root of the first maxillary molar was analyzed microscopically 100 μm above and below the midroot area. Nuclear volume morphometry was used to classify periodontal ligament cells as: L cells (<40 μm3), A+A' cells (40-79 μm3), B cells (80-119 μm3), C cells (120-169 μm3), and D cells (>170 μm3). The percent of forming bone surface on the mesial side and the width of the PDL were also measured. A 2x2 factorial ANOVA with repeat measures revealed a significant (p < 0.05) decrease in the C cell population and a nearly significant (p < 0.06) increase in the A+A' cell population in the centrifuge group. Comparing bone surfaces, the forming surface had a significant (p < 0.01) increase in the C and D cell populations, a significant (p < 0.01) decrease in the L and A+A' cell populations, and a significant (p < 0.05) decrease in the B cell population. The stationary group weighed significantly (p < 0.01) more than the centrifuge group post-experiment. And an unpaired t-test revealed a nearly significant (p < 0.06) increase in the percent forming bone surface on the mesial side of the maxillary first molar and no significant difference in mesial PDL width. The results showed that the centrifuge group had a trend toward a block in preosteoblast formation. This is similar to that seen with hypogravity. However, it cannot be concluded at this time if this is a direct gravitational effect or related to other factors such as physiological response to stress. Physical stress has been suggested as a potential mechanism for the observed decrease in weight seen in centrifuged animal, while the PDL width does not seem to be affected by gravitational forces, and thus may not be a sensitive marker to osteoblast differentiation inhibition. Finally, the reason for the increase in forming bone surface in the centrifuged group is unclear.

京都大学 / 0048 / 新制・課程博士 / 博士(人間健康科学) / 甲第19278号 / 人健博第30号 / 新制||人健||3(附属図書館) / 32280 / 京都大学大学院医学研究科人間健康科学系専攻 / (主査)教授 市橋 則明, 教授 三谷 章, 教授 松田 秀一 / 学位規則第4条第1項該当 / Doctor of Human Health Sciences / Kyoto University / DFAM

L'obésité est associée à un risque de fragilité musculo-squelettique, en particulier d’arthrose (OA). Notre but est d’explorer leurs contributions des conditions métaboliques et du surpoids. L’obésité MM (mécanique et métabolique) est obtenue par un régime alimentaire. L’obésité M (mécanique) est mimée par hypergravité à 2g. L’OA est induite par acte chirurgicale. Nous avons exploré les effets des obésités MM et M sur le système musculo-squelettique de souris non OA. Les MM ont un os trabéculaire préservé, un os cortical détérioré et des muscles fragilisés. Chez les M, l’os est préservé et les muscles sont renforcés. Les troubles métaboliques sont responsables de la fragilisation de l’os cortical et du muscle. Dans une 2ème partie, les conséquences de l’OA sont évaluées chez des souris non obèses, MM ou M. L’OA chez les non obèses fragilise uniquement l’os trabéculaire. L’OA chez les MM accentue la diminution de l’épaisseur corticale. L’OA chez les souris M fragilise encore plus l’os cortical et le muscle que chez les souris MM. Cependant si on ne considère que les souris OA, la composante MM est toujours plus délétère que la composante M. Finalement, nous avons testé les vibrations corps entiers pendant les 4 dernières semaines comme traitement potentiel des détériorations musculo-squelettiques des MM couplée ou non à l’OA. Les vibrations n’impactent pas l’obésité et l’OA. Un effet musculaire est observé au niveau moléculaire, ces diminutions étant plus importantes chez les OA. Aucun changement de masse musculaire n’est observé. Le tissu osseux n’est pas influencé. / Obesity is associated with a risk of musculoskeletal fragility, especially osteoarthritis (OA). Our goal is to explore their contributions of metabolic and overweight conditions. MM obesity (mechanical and metabolic) is obtained through a diet. Obesity M (mechanical) is mimed by hypergravity at 2g. OA is induced by surgery. We explored the effects of MM and M obesity on the non-OA mouse musculoskeletal system. MMs have preserved trabecular bone, deteriorated cortical bone and weakened muscles. In M, bone is preserved and muscles are strengthened. Metabolic disorders are responsible for the weakening of cortical bone and muscle. In a second part, the consequences of OA are evaluated in non-obese mice, MM or M. OA in non-obese only weakens the trabecular bone. OA in MM accentuates the decrease in cortical thickness. OA in M mice further weakens cortical bone and muscle than in MM mice. However, if we consider only the OA mice, the MM component is always more deleterious than the M component. Finally, we tested entire body vibrations during the last 4 weeks as a potential treatment for musculoskeletal deterioration of MM, whether or not coupled to OA. Vibrations do not affect obesity and OA. A muscular effect is observed at the molecular level, these decreases being greater in OA. No change in muscle mass is observed. The bone tissue is not influenced.

Annular flow is an important flow regime in many industrial applications. The need for a better understanding of this flow regime is driven by the desire to improve the design of many terrestrial and space-based systems. Annular two-phase flow is frequently present in the drilling, production and transportation of oil and natural gas, boilers and condensers, and in heating and refrigeration systems. The flow regime is also important for the refueling of space vehicles, and heating and refrigeration systems for space use. Past studies on annular flow have dealt with varying the gas or liquid Reynolds numbers and studying the effect of such changes on the flow regimes and pressure drops. The effect of two other relevant dimensionless groups, namely the gas-to-liquid density ratio and the gas-to-liquid viscosity ratio, on the film characteristics are noticeably absent. As well, with the increased interest in the space environment, studies on the effect of the gravitational acceleration on two-phase flow would be beneficial. The effect of the gas density and the gravitational acceleration on the annular flow average film thickness and frictional pressure drop are examined. The film thickness was measured using two-wire conductance probes. Experimental data were collected in microgravity and hypergravity aboard the Novespace Zero-G Airbus microgravity simulator and normal gravity data were collected at the University of Saskatchewan. Data were collected for a range of annular flow set points by changing the liquid and gas mass flow rates. The liquid-to-gas density ratio was examined by collecting annular flow data using helium-water and air-water. The gravitational effect on the film thickness characteristics was examined by collecting the data during the microgravity and pull-up (hypergravity) portions of each parabolic flight. A direct comparison is possible between the normal gravity data and the microgravity data, due to the matching of the liquid and gas mass flow rates and the flow regime. The reduction in gravity causes the average film thickness to increase between two and four times from the normal gravity values. The microgravity average frictional pressure drop is within approximately 20% of the normal gravity pressure drop for the same flow conditions. For all gravity levels, the air-water and the helium-water flows give similar results, for both average film thickness and frictional pressure drop, when based on the specific energy of the gas. The hypergravity average film thickness results are larger than at normal gravity for the same flow conditions. However, no flow regime map exists for the hypergravity condition, so the similarity of the flow regime cannot be confirmed. The hypergravity flow appears more chaotic, and may be in the transition from a churn type flow. The average frictional pressure drop is increased by approximately 20% due to the increase in the gravitational acceleration. New non-dimensional equations, which include the effect of the gas density, are presented for each gravity level to predict the average film thickness and the average frictional pressure drop.

Annular flow is an important flow regime in many industrial applications. The need for a better understanding of this flow regime is driven by the desire to improve the design of many terrestrial and space-based systems. Annular two-phase flow is frequently present in the drilling, production and transportation of oil and natural gas, boilers and condensers, and in heating and refrigeration systems. The flow regime is also important for the refueling of space vehicles, and heating and refrigeration systems for space use. Past studies on annular flow have dealt with varying the gas or liquid Reynolds numbers and studying the effect of such changes on the flow regimes and pressure drops. The effect of two other relevant dimensionless groups, namely the gas-to-liquid density ratio and the gas-to-liquid viscosity ratio, on the film characteristics are noticeably absent. As well, with the increased interest in the space environment, studies on the effect of the gravitational acceleration on two-phase flow would be beneficial. The effect of the gas density and the gravitational acceleration on the annular flow average film thickness and frictional pressure drop are examined. The film thickness was measured using two-wire conductance probes. Experimental data were collected in microgravity and hypergravity aboard the Novespace Zero-G Airbus microgravity simulator and normal gravity data were collected at the University of Saskatchewan. Data were collected for a range of annular flow set points by changing the liquid and gas mass flow rates. The liquid-to-gas density ratio was examined by collecting annular flow data using helium-water and air-water. The gravitational effect on the film thickness characteristics was examined by collecting the data during the microgravity and pull-up (hypergravity) portions of each parabolic flight. A direct comparison is possible between the normal gravity data and the microgravity data, due to the matching of the liquid and gas mass flow rates and the flow regime. The reduction in gravity causes the average film thickness to increase between two and four times from the normal gravity values. The microgravity average frictional pressure drop is within approximately 20% of the normal gravity pressure drop for the same flow conditions. For all gravity levels, the air-water and the helium-water flows give similar results, for both average film thickness and frictional pressure drop, when based on the specific energy of the gas. The hypergravity average film thickness results are larger than at normal gravity for the same flow conditions. However, no flow regime map exists for the hypergravity condition, so the similarity of the flow regime cannot be confirmed. The hypergravity flow appears more chaotic, and may be in the transition from a churn type flow. The average frictional pressure drop is increased by approximately 20% due to the increase in the gravitational acceleration. New non-dimensional equations, which include the effect of the gas density, are presented for each gravity level to predict the average film thickness and the average frictional pressure drop.

Diss. (sammanfattning) Linköping : Univ., 2004. / Härtill 4 uppsatser.

Les vols spatiaux affaiblissent le système immunitaire. Les objectifs de cette thèse étaient de déterminer, à l’aide de modèles terrestres, les conséquences d’une exposition à la micropesanteur simulée, l’hypergravité ou des stress chroniques ultra légers sur l’immunité humorale murine. Nous avons ainsi montré que la suspension anti-Orthostatique, qui mime certains effets de la micropesanteur, induit une diminution de 59% du nombre de lymphocytes B spléniques et une inversion du rapport entre lymphocytes B et T. L’affaiblissement de la lymphopoïèse B mis en évidence par la diminution du nombre de progéniteurs lymphoïdes et des cellules pré-B en est probablement la cause. De plus, la position anti-Orthostatique entraîne, en l’absence de stress, une diminution de la réponse in vitro des lymphocytes B au LPS, plus importante que celle des lymphocytes T à la ConA, comme en hypergravité. Afin de déterminer les mécanismes responsables de la diminution de la réponse des lymphocytes B chez les souris hypergravitaires, les transcrits de gènes impliqués dans la voie TLR-4, le récepteur du LPS, ont été quantifiés. L’expression de plusieurs gènes de la voie MyD88-Dépendante est augmentée après 21 jours d’hypergravité, mais aucune modification n’a été observée dans les lymphocytes B stimulés avec du LPS. Enfin, la contribution de stress chroniques ultra légers a été évaluée. Les proportions de lymphocytes spléniques ne sont pas affectées par ces stress. Par contre, le taux d’IgA sériques s’est révélé augmenté et une baisse des cytokines pro-Inflammatoires de type Th1 a été observée comme chez les astronautes. Ainsi, ces stress n’expliquent pas complètement l’impact négatif des vols spatiaux sur l’immunité humorale. Ces recherches sont importantes pour identifier la/les cause(s) de l’altération de l’immunité humorale en vol, afin de tester/développer des contremesures efficaces pour renforcer le système immunitaire des astronautes, mais également de personnes stressées ou âgées / Spaceflight weaken the immune system. The aims of this thesis were to determine the effects of exposure to simulated microgravity, hypergravity or chronic ultra-Mild stress on murine humoral immunity, using ground-Based models. We were able to show that the anti-Orthostatic suspension, that mimics some of the effects of microgravity, caused a 59% decrease of the number of splenic B cells and an inversion of the ratio between B and T lymphocytes. A decrease in B lymphopoiesis, as evidenced by the decrease of lymphoid progenitors and pre-B cells is likely the cause. Furthermore, we showed that the anti-Orthostatic position, in the absence of stress, leads to a decreased in vitro response of B cells to LPS, more important than the one of T cells to ConA, as did hypergravity exposure. To determine the mechanisms responsible of the decreased response of B cells from hypergravity mice, transcripts encoding genes involved in the TLR-4 signaling pathway, the LPS receptor on B cells, have been quantified. The expression of several genes of the MyD88-Dependent pathway was increased after 21 days of hypergravity, but no change was observed in B cells stimulated with LPS. Finally, the contribution of chronic ultra-Mild stresses was evaluated. The proportions of splenic lymphocytes were not affected by these stresses. However, the levels of serum IgA were increased and those of Th1 pro-Inflammatory cytokines were decreased, as in astronauts. Thus, these stresses do not fully explain the negative impact of spaceflight conditions on humoral immunity. These researches are important to identify the cause(s) of spaceflight-Associated impaired immunity in order to test/develop effective countermeasures to strengthen the immune system of astronauts, but also of stressed or elderly people

Experiments with microbial biofilms in microgravity and simulated microgravity have revealed altered growth kinetics, but geomicrobial biofilms have not yet been studied in low gravity environments. No characterisation of biofilms, geomicrobial or otherwise, have been conducted at hypergravity. This thesis explores factors affecting microbe-mineral interactions under terrestrial conditions, lays the groundwork for a scheduled microgravity experiment, and provides the first data on biofilms grown at hypergravity. As a first step in understanding microbe-mineral interactions in altered gravity environments, experiments were undertaken to identify factors that constrain attachment in a terrestrial environment. The model organism Sphingomonas desiccabilis and basaltic rock from Iceland were selected, and the minerals that make up the basalt were identified and procured in their pure form. The relative significance of physical factors such as hydrophobicity, surface charge, porosity and nutritional value were examined in relationship to the success with which biofilms colonised the mineral surfaces. Growth was measured by the quantity of biofilm biomass after a ifxed time period, using Crystal Violet stain, in order to draw conclusions about the most influential physical conditions on biofilm attachment to a substrate. It was found that mineral attachment is influenced more by porosity and nutritional value than by hydrophobicity or surface charge. To explore how reduced gravity affects biofilm formation and weathering rates, a European Space Agency experiment, BioRock, is underway. Samples of basalt, with monocultures of three different organisms, will be sent to the International Space Station in 2019 for long-term exposure to Martian and micro-gravity. Research testing proof of concepts, material compatibility, and experimental procedure and equipment is described. Confocal laser scanning microscopy (CLSM) was used to image the biofilms, and inductively coupled plasma mass spectroscopy (ICP-MS) experiments were conducted to compare biotic and abiotic elemental release rates from basalt. Both of these methods will be employed for post-flight analysis of BioRock. Preliminary terrestrial ICP-MS experiments indicated that rare Earth elements (REEs) showed the most reliable reflection of leaching patterns overall, as a consequence of their high molecular weight and low volatility during the ashing procedure. To fully understand gravity's effect on microbiological processes it is important to investigate what occurs when its influences are removed, but also to establish what occurs when extra gravitational force is applied. Using simulated hypergravity, achieved through hyper-acceleration on a geotechnical centrifuge, the effects of 10 x g on biofilm development and the leaching of basalt were investigated. As this was the first time that biofilms had been studied under hypergravity, additional substrates were included with the basalt, to enable characterisation of the more general response of biofilms to hypergravity. In contrast to previous experiments conducted on planktonic bacteria, which found decreased population sizes, the biofilms grown at 10 x g showed greater biomass than the 1 x g samples. ICP-MS showed no difference in the average weathering rates, but greater variability in the higher gravity samples. The data collected here advances our understanding of microbial interactions with geologically important substrates, with implications for an ISS microgravity experiment and future human space exploration. It also presents new intelligence on the previously unstudied effects of hypergravity on biofilms and rock weathering.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2006. / Härtill 5 uppsatser.

Le principal objectif de notre projet de recherche était l’étude approfondie des mécanismes fondamentaux qui sous-tendent l’adaptation de l’os à la contrainte mécanique liée à la gravité, en évaluant les effets de deux modèles d’hypergravité sur le squelette de jeune souris saines. Des expériences de même durée, portant sur des animaux de même souche et de même âge, nous ont permis d’établir des comparaisons entre les différents types d’adaptation du squelette à une hypergravité continue, générée par centrifugation, et une hypergravité intermittente, obtenue par vibrations du corps entier (WBV). Nous avons observé que la centrifugation réduit la résorption et augmente la formation dans l’os trabéculaire alors que les WBV ne découplent pas les activités de résorption et formation qui sont, l’une et l’autre, stimulées. La centrifugation induit une réorganisation de la microarchitecture trabéculaire au niveau du fémur et des vertèbres mais n’a pas d’effet sur les paramètres de masse osseuse corticale. En revanche, les WBV stimulent l’expansion de l’os cortical et augmente sa densité minérale osseuse. Les deux modèles d’hypergravité induisent une diminution de l’expression de la sclérostine (inhibitrice de la formation) et une augmentation de celle de DMP1 (responsable de la minéralisation) dans le cortex fémoral. De plus, les deux modèles augmentent le nombre de vaisseaux sanguins dans la diaphyse fémorale. Sur le plan technique, nous avons développé avec succès une méthode d’IHC quantitative qui nous a permis de détecter et de valider statistiquement de faibles variations, induites par nos expérimentations, dans l’expression de protéines ostéocytaires. Nous pensons que les résultats obtenus en IHC devraient faire l’objet d’une analyse quantitative systématique et fournissons, à cet effet, un outil adapté aux échantillons murins ou humains inclus en paraffine ou MMA / The main focus of the research project was to further study fundamental mechanisms underlying bone adaptation to gravity-induced mechanical loading and to assess effects of two different hypergravity models on skeleton of young healthy mice. Same duration of the experiments and the use of animals of the same type and age allowed us to make comparisons between different skeleton adaptations to continuous hypergravity generated by centrifugation and to intermittent one generated by WBV. We observed that centrifugation reduced resorption and increased formation in trabecular bone, whereas WBV did not uncoupled resorption and formation activities and stimulated both of them simultaneously. Centrifugation resulted in reorganized trabecular microarchitecture in femur and vertebra but had no effect on cortical bone mass-structural parameters. In contrast, WBV stimulated cortical bone geometrical expansion in 3-week experiment and increased cortical mineral density in 9 weeks. Both hypergravity models resulted in lower Sclerostin and higher DMP1 expressions in femoral cortex. Also, both models resulted in higher number of blood vessels in femoral metaphysis, however only centrifugation increased vessels volume. In relation to technical objects of the research project, we successfully developed a method of quantitative IHC, which allowed us to detect and verify statistically even modest alterations of osteocyte protein expressions in our experiments. We believe that IHC results should always be quantitatively analyzed and we provide a tool for both mice and human bone samples embedded in paraffin or MMA