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Background: Simulated microgravity (SMG) has not been well characterized in terms of its impact on cell division structures. This research aimed to assess the changes in cell division in Chang liver cells (CCL-13 cells) under SMG conditions. Methods: CCL-13 cells were exposed to SMG conditions via a 3D clinostat for 72 h. The cells from the control group were kept under the same conditions, without exposure to SMG. The changes in cell division were assessed via cell cycle progression analysis, the transcript expression of the genes associated with the cell cycle, and the appearance of the contractile ring, microvilli, and spindle in CCL-13 cells. Results: The CCL-13 cells from both the control group and the SMG group exhibited a typical epithelial-like shape. The CCL-13 cells of both groups displayed normal nuclear morphologies and were devoid of fragmentation and condensation, which are signs of apoptosis. There were changes in the cell cycle of CCL-13 cells in the SMG condition, which were shown via an increase in the cell percentage in the G0/G1 phase and a decrease in the S phase and G2/M phase. The cell area of the SMG-exposed CCl-13 cells increased, while their nuclear area decreased, which led to a reduction in the nuclear/cytoplasmic ratio. Moreover, the transcript expression of cyclin b1, cyclin d1, cdk2, and cdk6 was downregulated in CCL-13 cells under SMG conditions compared to the control group. Interestingly, SMG-exposed CCL-13 cells exhibited a decreased appearance of microvilli, changes in the formation of the contractile ring, and polar spindle microtubules during cytokinesis. Conclusions: SMG attenuated the cell division of CCL-13 cells by driving cells into the arrest phase and altering the cell division structures.

Life on Earth has evolved under the influence of gravity. This force has played an important role in shaping development and morphology from the molecular level to the whole organism. Although aquatic life experiences reduced gravity effects, land plants have evolved under a 1- environment. Understanding gravitational effects requires changing the magnitude of this force. One method of eliminating gravity''s influence is to enter into a free-fall orbit around the planet, thereby achieving a balance between centripetal force of gravity and the centrifugal force of the moving object. This balance is often mistakenly referred to as microgravity, but is best described as weightlessness. In addition to actually compensating gravity, instruments such as clinostats, random-positioning machines (RPM), and magnetic levitation devices have been used to eliminate effects of constant gravity on plant growth and development. However, these platforms do not reduce gravity but constantly change its direction. Despite these fundamental differences, there are few studies that have investigated the comparability between these platforms and weightlessness. Here, we provide a review of the strengths and weaknesses of these analogs for the study of plant growth and development compared to spaceflight experiments. We also consider reduced or partial gravity effects spaceflight and analog methods. While these analogs are useful, the fidelity of the results relative to spaceflight depends on biological parameters and environmental conditions that cannot be simulated in ground-based studies.

An organoid is a 3D organization of cells that can recapitulate some of the structure and function of native tissue. Recent work has seen organoids gain prominence as a valuable model for studying tissue development, drug discovery, and potential clinical applications. The requirements for the successful culture of organoids in vitro differ significantly from those of traditional monolayer cell cultures. The generation and maturation of high-fidelity organoids entails developing and optimizing environmental conditions to provide the optimal cues for growth and 3D maturation, such as oxygenation, mechanical and fluidic activation, nutrition gradients, etc. To this end, we discuss the four main categories of bioreactors used for organoid culture: stirred bioreactors (SBR), microfluidic bioreactors (MFB), rotating wall vessels (RWV), and electrically stimulating (ES) bioreactors. We aim to lay out the state-of-the-art of both commercial and in-house developed bioreactor systems, their benefits to the culture of organoids derived from various cells and tissues, and the limitations of bioreactor technology, including sterilization, accessibility, and suitability and ease of use for long-term culture. Finally, we discuss future directions for improvements to existing bioreactor technology and how they may be used to enhance organoid culture for specific applications.

Background: Irisin, a novel exercise-induced myokine, was shown to mediate beneficial effects of exercise in osteoporosis. Microgravity is a major threat to bone homeostasis of astronauts during long-term spaceflight, which results in decreased bone formation. Methods: The hind-limb unloading mice model and a random position machine are respectively used to simulate microgravity in vivo and in vitro. Results: We demonstrate that not only are bone formation and osteoblast differentiation decreased, but the expression of fibronectin type III domain-containing 5 (Fdnc5; irisin precursor) is also downregulated under simulated microgravity. Moreover, a lower dose of recombinant irisin (r-irisin) (1 nM) promotes osteogenic marker gene (alkaline phosphatase (Alp), collagen type 1 alpha-1(ColIα1)) expressions, ALP activity, and calcium deposition in primary osteoblasts, with no significant effect on osteoblast proliferation. Furthermore, r-irisin could recover the decrease in osteoblast differentiation induced by simulated microgravity. We also find that r-irisin increases β-catenin expression and partly neutralizes the decrease in β-catenin expression induced by simulated microgravity. In addition, β-catenin overexpression could also in part attenuate osteoblast differentiation reduction induced by simulated microgravity. Conclusions: The present study is the first to show that r-irisin positively regulates osteoblast differentiation under simulated microgravity through increasing β-catenin expression, which may reveal a novel mechanism, and it provides a prevention strategy for bone loss and muscle atrophy induced by microgravity.

Purpose: The current study investigates the adaptive responses of four bacterial strains in terms of physiology, morphology, and genetic behaviour under simulated microgravity (SMG) compared to their real exposure to the outer space stratosphere. Design/methodology/approach: Four nonpathogenic bacterial strains: Escherichia coli, Bacillus subtilis, Salmonella typhimurium, and Staphylococcus aureus, were cultured under SMG using a 3D clinostat and their growth, viability, morphology, antibiotic susceptibility, and gene stability were tested. Findings: The bacterial strains under study survived SMG, with certain species showing phenotypic changes, shifts in antibiotic susceptibility, and a few genetic variations. Originality/value: The study showed gravitational effects and shed light about the importance for control of space contamination and microbial risk, which contribute to SDG 3, SDG 9, and SDG 13. Research limitations: Longer- duration exposure, and multi‑omics analyses are recommended. Practical implications: The findings point to health management of space missions and the design of space relevant microbial bioprocesses. Keywords: Simulated Microgravity; Bacterial Adaptation; Clinostat; Antibiotic Sensitivity; Genomics; Sustainable Development. Citation: Malkawi (2025): Bacterial Growth Responses to Simulated Microgravity: Implications for Space Biotechnology and Sustainable Development. World Journal of Science, Technology and Sustainable Development (WJSTSD), Vol. 20, No. 4, pp. 367-382.

A variety of physiological changes in the human body have been observed to occur under the microgravity conditions of space. 3D clinostat devices capable of implementing time-averaged simulated microgravity (taSMG) have been widely used to predict these changes on Earth and to identify their underlying mechanisms. Recently, the concept of time-averaged simulated partial gravity (taSPG), which mimics the gravitational environments of the Moon (0.17 g) and Mars (0.38 g), has been proposed as an extension of taSMG, and clinostat control algorithms capable of implementing it have been developed. However, existing taSPG algorithms are dependent on specific hardware, limiting their versatility. Further, they are unable to generate taSPG levels exceeding 0.44 g. To address this limitation, we propose an improved control algorithm and validate it through both simulation and experiments. By applying an algorithm that does not depend on the characteristics of individual clinostat hardware, we confirmed the accurate implementation of taSPG up to 0.809 g. By adjusting the parameters, taSPG levels approaching 1 g can also be achieved. Notably, for taSPG in the range of 0.265 g to 0.635 g, the experimental values demonstrated refined accuracy with approximately 1% or less deviation from the simulation results.

A comprehensive understanding of spaceflight factors involved in immune dysfunction and the evaluation of biomarkers to assess in-flight astronaut health are essential goals for NASA. An elevated neutrophil-to-lymphocyte ratio (NLR) is a potential biomarker candidate, as leukocyte differentials are altered during spaceflight. In the reduced gravity environment of space, rodents and astronauts displayed elevated NLR and granulocyte-to-lymphocyte ratios (GLR), respectively. To simulate microgravity using two well-established ground-based models, we cultured human whole blood-leukocytes in high-aspect rotating wall vessels (HARV-RWV) and used hindlimb unloaded (HU) mice. Both HARV-RWV simulation of leukocytes and HU-exposed mice showed elevated NLR profiles comparable to spaceflight exposed samples. To assess mechanisms involved, we found the simulated microgravity HARV-RWV model resulted in an imbalance of redox processes and activation of myeloperoxidase-producing inflammatory neutrophils, while antioxidant treatment reversed these effects. In the simulated microgravity HU model, mitochondrial catalase-transgenic mice that have reduced oxidative stress responses showed reduced neutrophil counts, NLR, and a dampened release of selective inflammatory cytokines compared to wildtype HU mice, suggesting simulated microgravity induced oxidative stress responses that triggered inflammation. In brief, both spaceflight and simulated microgravity models caused elevated NLR, indicating this as a potential biomarker for future in-flight immune health monitoring.

Space agriculture, pivotal for sustainable extraterrestrial missions, requires plants that can adapt to altered gravitational conditions. This study delves into the adaptive responses to altered gravity of Wolffia globosa , an aquatic plant known for its rapid growth and high nutritional value. The research aimed to analyse the effect of simulated microgravity and hypergravity on relative growth rate (RGR), morphological characteristics, protein content, and the correlation between plant size and growth rate of Wolffia globosa . The study highlighted the responses of the species to altered gravity, uncovering inherent variability among seven different clones of W. globosa . Results show a base variability among clones in terms of RGR, size and protein content. Furthermore, some clones are affected by simulated microgravity, showing a decrease in RGR. Differently, under hypergravity, clones showed RGR higher than in 1 g control, therefore revealing a novel plant response to hypergravity. Morphological adaptations to gravity alterations were also evident. Among the studied clones, significant morphological changes were observed, further underlining the peculiar adaptation to the hypergravity environment. Differently, under simulated microgravity, morphology was generally stable across clones. A key finding of the study was the significant negative correlation between RGR and the physical dimensions of the plants: the fastest growth was associated with the smallest dimensions of the plants. This correlation might have practical implications in selecting clones for space cultivation, that leads to compact yet highly productive clones. The analysis of the protein content of all the clones revealed mostly no significant changes under hypergravity. Otherwise, a general decrease in protein content was observed under simulated microgravity. Overall, the study confirms the suitability of W. globosa for space agriculture and provides new insights into the perspective of using W. globosa as an alternative crop species for protein production for manned Space missions. Furthermore, it underscores the need for focusing on the clones and the selection of the W. globosa plants that are best adapted to the environmental conditions of space; therefore, selecting those with the best combination of biomass production (by means of growth rate, size), and protein content.

Human space travel and exploration are of interest to both the industrial and scientific community. However, there are many adverse effects of spaceflight on human physiology. In particular, there is a lack of understanding of the extent to which microgravity affects the immune system. T cells, key players of the adaptive immune system and long-term immunity, are present not only in blood circulation but also reside within the tissue. As of yet, studies investigating the effects of microgravity on T cells are limited to peripheral blood or traditional 2D cell culture that recapitulates circulating blood. To better mimic interstitial tissue, 3D cell culture has been well established for physiologically and pathologically relevant models. In this work, we utilize 2D cell culture and 3D collagen matrices to gain an understanding of how simulated microgravity, using a random positioning machine, affects both circulating and tissue-resident T cells. T cells were studied in both resting and activated stages. We found that 3D cell culture attenuates the effects of simulated microgravity on the T cells transcriptome and nuclear irregularities compared to 2D cell culture. Interestingly, simulated microgravity appears to have less effect on activated T cells compared to those in the resting stage. Overall, our work provides novel insights into the effects of simulated microgravity on circulating and tissue-resident T cells which could provide benefits for the health of space travellers.

Disuse osteoporosis is characterized by decreased bone mass caused by abnormal mechanical stimulation of bone. Piezo1 is a major mechanosensitive ion channel in bone homeostasis. However, whether intervening in the action of Piezo1 can rescue disuse osteoporosis remains unresolved. In this study, a commonly‐used hindlimb‐unloading model is employed to simulate microgravity. By single‐cell RNA sequencing, bone marrow‐derived mesenchymal stem cells (BMSCs) are the most downregulated cell cluster, and coincidentally, Piezo1 expression is mostly enriched in those cells, and is substantially downregulated by unloading. Importantly, activation of Piezo1 by systemically‐introducing yoda1 mimics the effects of mechanical stimulation and thus ameliorates bone loss under simulated microgravity. Mechanistically, Piezo1 activation promotes the proliferation and osteogenic differentiation of Gli1 + BMSCs by activating the β‐catenin and its target gene activating transcription factor 4 (ATF4). Inhibiting β‐catenin expression substantially attenuates the effect of yoda1 on bone loss, possibly due to inhibited proliferation and osteogenic differentiation capability of Gli1 + BMSCs mediated by ATF4. Lastly, Piezo1 activation also slightly alleviates the osteoporosis of OVX and aged mice. In conclusion, impaired function of Piezo1 in BMSCs leads to insufficient bone formation especially caused by abnormal mechanical stimuli, and is thus a potential therapeutic target for osteoporosis.