Effects Of Simulated Microgravity Research Articles

Comparative Studies on Effects of Simulated Microgravity on Growth and Photosynthetic Parameters in Rice and Mungbean

Microgravity or low gravity environment of space can be simulated on Earth by using clinorotation with appropriate speed of rotation. Previous reports show that microgravity environment (real or simulated) affects growth, chlorophyll content and photosynthetic performance in different plants. However, most of the reports showed variation in results. Earlier studies carried out on simulated microgravity (SMG) using clinorotation in our laboratory showed increase in growth, chlorophyll content and various yield attributes in case of laboratory and field grown rice. Subsequent studies were carried out on mungbean seedlings. In the present work, comparative aspects of these studies are reported. Results of the present study showed increase in growth, chlorophyll content and chlorophyll fluorescence parameters in SMG samples as compared to control samples in rice and mungbean seedlings. Interestingly, percentage of enhancement in growth, chlorophyll content and chlorophyll fluorescence parameters of SMG samples is significantly higher in rice (monocotyledonous) seedlings as compared to mungbean (dicotyledonous).

Effects of Simulated Microgravity on Rat Reproductive System: Potential Benefits of Vitamin D3 Intervention.

Gravity in space can have a negative impact on the reproductive system. Given that the reproductive system is one of vitamin D's objectives, this study will use a simulated microgravity model to evaluate its impact on the rat reproductive system.Twenty-two male Wistar rats were allocated into four groups at random. Under microgravity circumstances, the rats were housed in both special and standard cages. Each group was then separated into two subgroups, one of which received vitamin D3 and the other did not. Blood was drawn twice to determine blood levels of vitamin D3, LH, FSH, and testosterone. Rat testes were isolated for histological analysis, as well as a piece of epididymis for sperm count and morphological examination.Microgravity had a detrimental effect on testicular tissue, resulting in lower serum levels of LH and testosterone (p-value < 0.001). Spermatogenesis was largely inhibited under microgravity. During microgravity conditions, however, vitamin D3 had a good effect on testicular structure, and the total number of sperm. Simulated microgravity affects the male reproductive system, compromising testicular morphology, sperm parameters, and hormonal balance. However, this study shows that vitamin D3 supplementation can act as a preventative strategy, minimizing the negative consequences of microgravity. The beneficial effect of vitamin D3 on testicular health and sperm quality implies that it may be useful in protecting male reproductive function in space-related situations.

Combined effects of radiation and simulated microgravity on intestinal tumorigenesis in C3B6F1 ApcMin/+ mice

Explorations of the Moon and Mars are planned as future manned space missions, during which humans will be exposed to both radiation and microgravity. We do not, however, know the health effects for such combined exposures. In a ground-based experiment, we evaluated the combined effects of radiation and simulated microgravity on tumorigenesis by performing X-irradiation and tail suspension in C3B6F1 ApcMin/+ mice, a well-established model for intestinal tumorigenesis. Mice were irradiated at 2 weeks of age and underwent tail suspension for 3 or 11 weeks using a special device that avoids damage to the tail. The tail suspension treatment significantly reduced the thymus weight after 3 weeks but not 11 weeks, suggesting a transient stress response. The combination of irradiation and tail suspension significantly increased the number of small intestinal tumors less than 2 mm in diameter as compared with either treatment alone. The combined treatment also increased the fraction of malignant tumors among all small intestinal tumors as compared with the radiation-only treatment. Thus, the C3B6F1 ApcMin/+ mouse is a useful model for assessing cancer risk in a simulated space environment, in which simulated microgravity accelerates tumor progression when combined with radiation exposure.

Effects of simulated microgravity on current generation of electrochemically active bacteria: Insights from case-control study using random positioning machine

Electrochemically active bacteria (EAB) exhibit promising prospects for space exploration and life support systems. However, the effects of the space environment on EAB are unclear. In this study, the effects of simulated microgravity on the current generation of mixed-culture EAB were illustrated, and the underlying mechanism was elucidated. The results demonstrated that the electrochemical activity of mixed-culture EAB was enhanced, which was mainly due to the enrichment of Geobacter and the increase in EAB biomass. Additionally, the genes and proteins of the biofilm changed obviously under simulated microgravity conditions, including: I) genes related to signal transfer, II) genes related to cell wall synthesis, and III) genes related to riboflavin synthesis. This study first revealed the enrichment in EAB abundance, the increase in EAB biomass, and the promotion of current generation under simulated microgravity.

The effects of microgravity on cellular elements of the central nervous system - astrocytes, microglia, neurons, oligodendrocytes, and oligodendrocyte precursor cells

The microgravity of space has a number of detrimental effects on the central nervous system of astronauts and cosmonauts. This includes the development of increased intracranial pressure and cerebrospinal fluid volume, thickening of the meninges, attrition of gray matter and the number of neuronal synapses, aberrant astrocyte structure and function, changes in the morphology, distribution and proinflammatory activity of microglia, and detrimental effects on the ontogeny and functioning of oligodendrocytes and their precursor cells. In this article, I review what is known about the ontogeny, physiology and anatomy of astrocytes, microglia, neurons, oligodendrocytes, and oligodendrocyte precursor cells and detail the effects of microgravity and/or simulated microgravity on these key cellular elements of the central nervous system.

Simulated microgravity attenuates myogenesis and contractile function of 3D engineered skeletal muscle tissues

While the effects of microgravity on inducing skeletal muscle atrophy have been extensively studied, the impacts of microgravity on myogenesis and its mechanisms remain unclear. In this study, we developed a microphysiological system of engineered muscle tissue (EMT) fabricated using a collagen / Matrigel composite hydrogel and murine skeletal myoblasts. This 3D EMT model allows non-invasive quantitative assessment of contractile function. After applying a 7-day differentiation protocol to induce myotube formation, the EMTs clearly exhibited sarcomerogenesis, myofilament formation, and synchronous twitch and tetanic contractions with electrical stimuli. Using this 3D EMT system, we investigated the effects of simulated microgravity at 10−3 G on myogenesis and contractile function utilizing a random positioning machine. EMTs cultured for 5 days in simulated microgravity exhibited significantly reduced contractile forces, myofiber size, and differential expression of muscle contractile, myogenesis regulatory, and mitochondrial biogenesis-related proteins. These results indicate simulated microgravity attenuates myogenesis, resulting in impaired muscle function.

Morphological Changes of 3T3 Cells under Simulated Microgravity.

Cells are sensitive to changes in gravity, especially the cytoskeletal structures that determine cell morphology. The aim of this study was to assess the effects of simulated microgravity (SMG) on 3T3 cell morphology, as demonstrated by a characterization of the morphology of cells and nuclei, alterations of microfilaments and microtubules, and changes in cycle progression. 3T3 cells underwent induced SMG for 72 h with Gravite®, while the control group was under 1G. Fluorescent staining was applied to estimate the morphology of cells and nuclei and the cytoskeleton distribution of 3T3 cells. Cell cycle progression was assessed by using the cell cycle app of the Cytell microscope, and Western blot was conducted to determine the expression of the major structural proteins and main cell cycle regulators. The results show that SMG led to decreased nuclear intensity, nuclear area, and nuclear shape and increased cell diameter in 3T3 cells. The 3T3 cells in the SMG group appeared to have a flat form and diminished microvillus formation, while cells in the control group displayed an apical shape and abundant microvilli. The 3T3 cells under SMG exhibited microtubule distribution surrounding the nucleus, compared to the perinuclear accumulation in control cells. Irregular forms of the contractile ring and polar spindle were observed in 3T3 cells under SMG. The changes in cytoskeleton structure were caused by alterations in the expression of major cytoskeletal proteins, including β-actin and α-tubulin 3. Moreover, SMG induced 3T3 cells into the arrest phase by reducing main cell cycle related genes, which also affected the formation of cytoskeleton structures such as microfilaments and microtubules. These results reveal that SMG generated morphological changes in 3T3 cells by remodeling the cytoskeleton structure and downregulating major structural proteins and cell cycle regulators.

Simulated microgravity improves maturation of cardiomyocytes derived from human induced pluripotent stem cells

Cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) possess tremendous potential for basic research and translational application. However, these cells structurally and functionally resemble fetal cardiomyocytes, which is a major limitation of these cells. Microgravity can significantly alter cell behavior and function. Here we investigated the effect of simulated microgravity on hiPSC-CM maturation. Following culture under simulated microgravity in a random positioning machine for 7 days, 3D hiPSC-CMs had increased mitochondrial content as detected by a mitochondrial protein and mitochondrial DNA to nuclear DNA ratio. The cells also had increased mitochondrial membrane potential. Consistently, simulated microgravity increased mitochondrial respiration in 3D hiPSC-CMs, as indicated by higher levels of maximal respiration and ATP content, suggesting improved metabolic maturation in simulated microgravity cultures compared with cultures under normal gravity. Cells from simulated microgravity cultures also had improved Ca2+ transient parameters, a functional characteristic of more mature cardiomyocytes. In addition, these cells had improved structural properties associated with more mature cardiomyocytes, including increased sarcomere length, z-disc length, nuclear diameter, and nuclear eccentricity. These findings indicate that microgravity enhances the maturation of hiPSC-CMs at the structural, metabolic, and functional levels.

Morphologic and genomic changes of thyroid cancer cell lines exposed to conditions of simulated microgravity

Microgravity in space impacts human health. In particular, thyroid cancer, which has a high incidence rate, has been the subject of numerous studies with respect to microgravity. However, most studies have focused on Western follicular thyroid cancer cell lines, while data regarding the effects of microgravity on Asian cell lines are lacking. Therefore, we aimed to investigate the effect of simulated ground-based microgravity on two Korean thyroid cancer cell lines, namely SNU-790 and SNU-80. We found that both cell lines formed multicellular spheroids under simulated microgravity. Gene expression analysis revealed that in SNU-790 cells, histone-related genes were upregulated and microRNA-related genes were downregulated. Meanwhile, in SNU-80 cells, genes related to the cellular response to hypoxia were downregulated. These findings contribute to a better understanding of the effects of microgravity on thyroid cancer cells. Further validation studies and clinical significance analyses are needed to fully understand the implications of these findings.

Whole transcriptome analysis highlights nutrient limitation of nitrogen cycle bacteria in simulated microgravity

Regenerative life support systems (RLSS) will play a vital role in achieving self-sufficiency during long-distance space travel. Urine conversion into a liquid nitrate-based fertilizer is a key process in most RLSS. This study describes the effects of simulated microgravity (SMG) on Comamonas testosteroni, Nitrosomonas europaea, Nitrobacter winogradskyi and a tripartite culture of the three, in the context of nitrogen recovery for the Micro-Ecological Life Support System Alternative (MELiSSA). Rotary cell culture systems (RCCS) and random positioning machines (RPM) were used as SMG analogues. The transcriptional responses of the cultures were elucidated. For CO2-producing C. testosteroni and the tripartite culture, a PermaLifeTM PL-70 cell culture bag mounted on an in-house 3D-printed holder was applied to eliminate air bubble formation during SMG cultivation. Gene expression changes indicated that the fluid dynamics in SMG caused nutrient and O2 limitation. Genes involved in urea hydrolysis and nitrification were minimally affected, while denitrification-related gene expression was increased. The findings highlight potential challenges for nitrogen recovery in space.

Effects of altered gravity on growth and morphology in Wolffia globosa implications for bioregenerative life support systems and space-based agriculture

Understanding the response of plants to varied gravitational conditions is vital for developing effective food production in space bioregenerative life support systems. This study examines the impact of altered gravity conditions on the growth and morphological responses of Wolffia globosa (commonly known as “water lentils” or “duckweed”), assessing its potential as a space crop. Although an experiment testing the effect of simulated microgravity on Wolffia globosa has been previously conducted, for the first time, we investigated the effect of multiple gravity levels on the growth and morphological traits of Wolffia globosa plants. The plant responses to simulated microgravity, simulated partial gravity (Moon), and hypergravity environments were evaluated using random positioning machines and the large-diameter centrifuge. As hypothesized, we observed a slight reaction to different gravitational levels in the growth and morphological traits of Wolffia globosa. The relative growth rates (RGR) of plants subjected to simulated microgravity and partial gravity were reduced when compared to those in other gravity levels. The morphological analysis revealed differences in plant dimensions and frond length-to-width ratios under diverse gravity conditions. Our findings showed that Wolffia globosa is responsive to gravitational changes, with its growth and morphological adaptations being slightly influenced by varying gravitational environments. As for other crop species, growth was reduced by the microgravity conditions; however, RGR remained substantial at 0.33 a day. In conclusion, this study underscores the potential of Wolffia globosa as a space crop and its adaptability to diverse gravitational conditions, contributing to the development of sustainable food production and bioregenerative life support systems for future space exploration missions.

The Effects of Simulated and Real Microgravity on Vascular Smooth Muscle Cells

Abstract As considerations are being made for the limitations and safety of long-term human spaceflight, the vasculature is important given its connection to and impact on numerous organ systems. As a major constituent of blood vessels, vascular smooth muscle cells are of interest due to their influence over vascular tone and function. Additionally, vascular smooth muscle cells are responsive to pressure and flow changes. Therefore, alterations in these parameters under conditions of microgravity can be functionally disruptive. As such, here we review and discuss the existing literature that assesses the effects of microgravity, both actual and simulated, on smooth muscle cells. This includes the various methods for achieving or simulating microgravity, the animal models or cells used, and the various durations of microgravity assessed. We also discuss the various reported findings in the field, which include changes to cell proliferation, gene expression and phenotypic shifts, and renin-angiotensin-aldosterone system (RAAS), nitric oxide synthase (NOS), and Ca2+ signaling. Additionally, we briefly summarize the literature on smooth muscle tissue engineering in microgravity as well as considerations of radiation as another key component of spaceflight to contextualize spaceflight experiments, which by their nature include radiation exposure. Finally, we provide general recommendations based on the existing literature's focus and limitations.

Effects of short-term simulated microgravity on changes in extracellular space structure and substance diffusion and clearance

Substance transportation and clearance in the brain extracellular space (ECS) is crucial to brain function, but simulated microgravity disrupts substance diffusion and clearance in the brain ECS and there is limited knowledge regarding these disruptions. This study employed a tracer-based magnetic resonance imaging (MRI) technique to examine the impact of simulated microgravity on hippocampal ECS structure and the drainage of brain interstitial fluid (ISF) in a tail-suspended hindlimb-unloading rat model. The results demonstrated that under simulated microgravity conditions for 3, 7, and 14 days, the volume fraction of the hippocampal ECS increased in hindlimb unloading rats, and short-term (3 days) simulated microgravity expedited the drainage of the hippocampal ISF, but the drainage slowed under longer (7 or 14 days) simulated microgravity. These findings suggested that simulated microgravity alters the structure of the brain ECS and the speed of brain ISF drainage, which may have implications for space-flight-related brain injuries and relevant neuroprotective approaches via the brain ECS.

Use of a microgravity analog to explore the effects of simulated microgravity on the development of Escherichia coli K12 biofilms.

The rapid development of space technologies and the increase of human presence in space has brought the discussion of the effects of microgravity on cells into the undergraduate classroom. This paper proposes an idea to simulate microgravity on a bacterial culture, suitable for an introductory microbiology laboratory. For this purpose, we show the use of a 2D clinostat designed for microbial studies, along with traditional microbiology techniques such as optical density, plate counts, and biofilm biomass measurement to test the effect of simulated microgravity on the growth of Escherichia coli K12. This exercise aims to facilitate further discussions on the effects of microgravity on bacteria growth and communication, as well as the use of technology to simulate space and predict physiological changes in cells.

Preparation for mice spaceflight: Indications for training C57BL/6J mice to adapt to microgravity effect with three-dimensional clinostat on the ground

Like astronauts, animals need to undergo training and screening before entering space. At present, pre-launch training for mice mainly focuses on adaptation to habitat system. Training for the weightless environment of space in mice has not received much attention. Three-dimensional (3D) clinostat is a method to simulate the effects of microgravity on Earth. However, few studies have used a 3D clinostat apparatus to simulate the effects of microgravity on animal models. Therefore, we conducted a study to evaluate the feasibility and effects of long-term treatment with three-dimensional clinostat in C57BL/6 J mice. Thirty 8-week-old male C57BL/6 J mice were randomly assigned to three groups: mice in individually ventilated cages (MC group, n = 6), mice in survival boxes (SB group, n = 12), and mice in survival boxes receiving 3D clinostat treatment (CS group, n = 12). The mice showed good tolerance after 12 weeks of alternate day training. To evaluate the biological effects of simulated microgravity, the changes in serum metabolites were monitored using untargeted metabolomics, whereas bone loss was assessed using microcomputed tomography of the left femur. Compared with the metabolome of the SB group, the metabolome of the CS group showed significant differences during the first three weeks and the last three weeks. The KEGG pathways in the late stages were mainly related to the nervous system, indicating the influence of long-term microgravity on the central nervous system. Besides, a marked reduction in the trabecular number (P < 0.05) and an increasing trend of trabecular spacing (P < 0.1) were observed to occur in a time-dependent manner in the CS group compared with the SB group. These results showed that mice tolerated well in a 3D clinostat and may provide a new strategy in pre-launch training for mice and conducting relevant ground-based modeling experiments.

Simulated Microgravity Exposure Induces Antioxidant Barrier Deregulation and Mitochondria Enlargement in TCam-2 Cell Spheroids.

One of the hallmarks of microgravity-induced effects in several cellular models is represented by the alteration of oxidative balance with the consequent accumulation of reactive oxygen species (ROS). It is well known that male germ cells are sensitive to oxidative stress and to changes in gravitational force, even though published data on germ cell models are scarce. We previously studied the effects of simulated microgravity (s-microgravity) on a 2D cultured TCam-2 seminoma-derived cell line, considered the only human cell line available to study in vitro mitotically active human male germ cells. In this study, we used a corresponding TCam-2 3D cell culture model that mimics cell-cell contacts in organ tissue to test the possible effects induced by s-microgravity exposure. TCam-2 cell spheroids were cultured for 24 h under unitary gravity (Ctr) or s-microgravity conditions, the latter obtained using a random positioning machine (RPM). A significant increase in intracellular ROS and mitochondria superoxide anion levels was observed after RPM exposure. In line with these results, a trend of protein and lipid oxidation increase and increased pCAMKII expression levels were observed after RPM exposure. The ultrastructural analysis via transmission electron microscopy revealed that RPM-exposed mitochondria appeared enlarged and, even if seldom, disrupted. Notably, even the expression of the main enzymes involved in the redox homeostasis appears modulated by RPM exposure in a compensatory way, with GPX1, NCF1, and CYBB being downregulated, whereas NOX4 and HMOX1 are upregulated. Interestingly, HMOX1 is involved in the heme catabolism of mitochondria cytochromes, and therefore the positive modulation of this marker can be associated with the observed mitochondria alteration. Altogether, these data demonstrate TCam-2 spheroid sensitivity to acute s-microgravity exposure and indicate the capability of these cells to trigger compensatory mechanisms that allow them to overcome the exposure to altered gravitational force.

Simulated Microgravity-Induced Changes to Drug Response in Cancer Cells Quantified Using Fluorescence Morphometry.

Unlike plants that have special gravity-sensing cells, such special cells in animals are yet to be discovered. However, microgravity, the condition of apparent weightlessness, causes bone, muscular and immune system dysfunctions in astronauts following spaceflights. Decades of investigations show correlations between these organ and system-level dysfunctions with changes induced at the cellular level both by simulated microgravity as well as microgravity conditions in outer space. Changes in single bone, muscle and immune cells include morphological abnormalities, altered gene expression, protein expression, metabolic pathways and signaling pathways. These suggest that human cells mount some response to microgravity. However, the implications of such adjustments on many cellular functions and responses are not clear. Here, we addressed the question whether microgravity induces alterations to drug response in cancer cells. We used both adherent cancer cells (T98G) and cancer cells in suspension (K562) to confirm the known effects of simulated microgravity and then treated the K562 cells with common cancer drugs (hydroxyurea and paclitaxel) following 48 h of exposure to simulated microgravity via a NASA-developed rotary cell culture system. Through fluorescence-guided morphometry, we found that microgravity abolished a significant reduction (p < 0.01) in the nuclear-to-cytoplasm ratio of cancer cells treated with hydroxyurea. Our results call for more studies on the impact of microgravity on cellular drug response, in light of the growing need for space medicine, as space exploration grows.

Effects of an aged tissue niche on the immune potency of dendritic cells using simulated microgravity

Microgravity accelerates the aging of various physiological systems, and it is well acknowledged that aged individuals and astronauts both have increased susceptibility to infections and poor response to vaccination. Immunologically, dendritic cells (DCs) are the key players in linking innate and adaptive immune responses. Their distinct and optimized differentiation and maturation phases play a critical role in presenting antigens and mounting effective lymphocyte responses for long-term immunity. Despite their importance, no studies to date have effectively investigated the effects of microgravity on DCs in their native microenvironment, which is primarily located within tissues. Here, we address a significantly outstanding research gap by examining the effects of simulated microgravity via a random positioning machine on both immature and mature DCs cultured in biomimetic collagen hydrogels, a surrogate for tissue matrices. Furthermore, we explored the effects of loose and dense tissues via differences in collagen concentration. Under these various environmental conditions, the DC phenotype was characterized using surface markers, cytokines, function, and transcriptomic profiles. Our data indicate that aged or loose tissue and exposure to RPM-induced simulated microgravity both independently alter the immunogenicity of immature and mature DCs. Interestingly, cells cultured in denser matrices experience fewer effects of simulated microgravity at the transcriptome level. Our findings are a step forward to better facilitate healthier future space travel and enhance our understanding of the aging immune system on Earth.

Microgravity and Musculoskeletal Health: What Strategies Should Be Used for a Great Challenge?

Space colonization represents the most insidious challenge for mankind, as numerous obstacles affect the success of space missions. Specifically, the absence of gravitational forces leads to systemic physiological alterations, with particular emphasis on the musculoskeletal system. Indeed, astronauts exposed to spaceflight are known to report a significant impairment of bone microarchitecture and muscle mass, conditions clinically defined as osteoporosis and sarcopenia. In this context, space medicine assumes a crucial position, as the development of strategies to prevent and/or counteract weightlessness-induced alterations appears to be necessary. Furthermore, the opportunity to study the biological effects induced by weightlessness could provide valuable information regarding adaptations to spaceflight and suggest potential treatments that can preserve musculoskeletal health under microgravity conditions. Noteworthy, improving knowledge about the latest scientific findings in this field of research is crucial, as is thoroughly investigating the mechanisms underlying biological adaptations to microgravity and searching for innovative solutions to counter spaceflight-induced damage. Therefore, this narrative study review, performed using the MEDLINE and Google Scholar databases, aims to summarize the most recent evidence regarding the effects of real and simulated microgravity on the musculoskeletal system and to discuss the effectiveness of the main defence strategies used in both real and experimental settings.

Effects of Simulated Microgravity on the Internalization of Cerium Oxide Nanoparticles by Proliferating Human Skeletal Myoblasts

Spaceflight typically exerts detrimental effects on living organisms by promoting/accelerating some degenerative processes associated with aging and pathology onset on Earth, like muscle degeneration. As possible countermeasures to spaceflight effects, a few recent studies successfully tested the administration of nanomaterials for tuning of cellular activities and for promotion of certain cell phenotypes in microgravity, but largely missed investigation of early interactions of these nanomaterials with their cellular targets under altered gravity conditions. This study aims at filling this gap by elucidation of early interactions of a selected typology of redox-active nanoparticles (cerium oxide nanoparticles, also termed nanoceria, NC) with proliferating human skeletal myoblasts (HSkM) by concomitant exposure to simulated microgravity (achieved by a random positioning machine, RPM). To this purpose, NC were synthesized by a direct precipitation method and were chemically, structurally, and functionally characterized by several independent techniques prior to administration to skeletal muscle cell cultures and loading on the RPM. Confocal and electron microscopy evidence of nanoparticle internalization by several mechanisms (mostly involving macropinocytosis) in HSkM is provided, along with evidence of transcriptional regulation of key antioxidant response markers (Nos1, Sod2, and Sod3), promising oxidative stress alleviation and muscle protection under mechanical unloading conditions.