Large vesicle extrusion from neurons may contribute to spreading pathogenic protein aggregates and promoting inflammatory responses, two mechanisms leading to neurodegenerative disease. Factors that regulate the extrusion of large vesicles, such as exophers produced by proteostressed C. elegans touch neurons, are poorly understood. Here, we document that mechanical force can significantly potentiate exopher extrusion from proteostressed neurons. Exopher production from the C. elegans ALMR neuron peaks at adult day 2 or 3, coinciding with the C. elegans reproductive peak. Genetic disruption of C. elegans germline, sperm, oocytes, or egg/early embryo production can strongly suppress exopher extrusion from the ALMR neurons during the peak period. Conversely, restoring egg production at the late reproductive phase through mating with males or inducing egg retention via genetic interventions that block egg-laying can strongly increase ALMR exopher production. Overall, genetic interventions that promote ALMR exopher production are associated with expanded uterus lengths and genetic interventions that suppress ALMR exopher production are associated with shorter uterus lengths. In addition to the impact of fertilized eggs, ALMR exopher production can be enhanced by filling the uterus with oocytes, dead eggs, or even fluid, supporting that distention consequences, rather than the presence of fertilized eggs, constitute the exopher-inducing stimulus. We conclude that the mechanical force of uterine occupation potentiates exopher extrusion from proximal proteostressed maternal neurons. Our observations draw attention to the potential importance of mechanical signaling in extracellular vesicle production and in aggregate spreading mechanisms, making a case for enhanced attention to mechanobiology in neurodegenerative disease.
Neurons are specialized cells in the brain and nervous system that transmit signals between the brain and the rest of the body, enabling humans and animals to react to internal and external stimuli. For this communication system to function effectively, neurons must remain healthy. Neurons maintain their function in a variety of ways, including by removing excess or damaged cellular components (such as organelles and protein aggregates) that could compromise neuron function. One way to do this is by extruding organelles and aggregates. During ‘extrusion events’, the material to be removed is gathered within a budding portion of the plasma membrane, which forms a vesicle that ejects the material from the neuron. However, the factors driving the extrusion process remained unknown. To investigate, Wang, Guasp, Salam et al. conducted experiments in the roundworm Caenorhabditis elegans, finding that the number of extrusion events in a certain type of neuron increases at the peak of reproduction. More specifically, a greater number of extrusion events were associated with the presence of fertilized eggs, which accumulate in the uterus before they are laid. Disrupting eggs, sperm or the fertilization process suppressed the increase in extrusion events, suggesting the presence of fertilized eggs is responsible. To determine how the eggs might trigger extrusion events, Wang et al. stretched the uterus using dead eggs, unfertilized eggs or by injecting fluid, finding that each of these approaches increased the number of extrusion events. Further analysis suggests that this mechanical stretching of the uterus signals to the neurons that reproduction has started, encouraging the neurons to remove old components and optimize their function. Wang et al. hypothesize that this stretch response could support neuronal behaviors that aid in successful reproduction, such as sensing food and selecting where to lay eggs. The findings increase our understanding of the factors that trigger vesicle extrusion in living organisms. These observations could have implications for human neurodegenerative diseases such as Alzheimer’s disease, in which protein aggregates accumulate in neurons. It is possible that mechanical signals generated by factors associated with Alzheimer’s disease, such as high blood pressure, could influence neuronal extrusion and contribute to some of the mechanisms underlying aggregate transfer in neurodegenerative diseases.

Mechanical stress within organoids is a pivotal indicator in disease modeling and pharmacokinetics, yet current tools lack the ability to rapidly and dynamically screen these mechanics. Here, we introduce biocompatible and compressible hollow microlasers that realize all-optical assessment of cellular stress within organoids. The laser spectroscopy yields identification of cellular deformation at the nanometer scale, corresponding to tens of pascals stress sensitivity. The compressibility enables the investigation of the isotropic component, which is the fundamental mechanics of multicellular models. By integrating with a microwell array, we demonstrate the high-throughput screening of mechanical cues in tumoroids, establishing a platform for mechano-responsive drug screening. Furthermore, we showcase the monitoring and mapping of dynamic contractile stress within human embryonic stem cell-derived cardiac organoids, revealing the internal mechanical inhomogeneity within a single organoid. This method eliminates time-consuming scanning and sample damage, providing insights into organoid mechanobiology.

OBJECTIVE: To assess the impact of elastic gradients formed among restorative material, cement, and substrate on the fracture resistance of tri-layer restorative systems.
METHODS: Four CAD/CAM materials were utilized, two glass-ceramics (IPS e.max CAD, Vita Suprinity) and two resin-ceramic hybrids (Vita Enamic, Lava Ultimate). Their fracture resistance was examined by biaxial flexure (n = 8) and Hertzian indentation (n = 10) tests. Statistical analysis was conducted using ANOVA and Tukey tests (p = 5 %). Finite element analysis (FEA) was employed to simulate the Hertzian indentation test and elucidate the stress-fields formed on the intaglio surface below the loading area.
RESULTS: The biaxial flexural strength (MPa) of glass-ceramics exceeded the hybrid materials (e.max 417a, Suprinity 230b, Enamic 138c, and Lava Ultimate 183bc). Conversely, the load-bearing capacity (N) of the materials bonded to dentin analog demonstrated the opposite trend, with the hybrid materials achieving superior results (e.max 830 C, Suprinity 660D, Enamic 1822B, and Lava Ultimate 2593 A). The stress-fields observed by FEA were coherent with the experimental results for Hertzian flexural stresses (MPa): e.max 501 A, Suprinity 342 C, Enamic 406B, whereas no tensile stress was observed at the intaglio surface of Lava Ultimate.
CONCLUSIONS: Detailed analysis of the fracture resistance of the tri-layer restorative systems showed that the elastic gradients play a more significant role than the flexural strength of the restorative materials. The coherence of the elastic moduli between the restorative material and supporting structures results in reduced tensile stress concentration at the intaglio surface beneath the loading area and enhances the ability to withstand load.

In vitro models are invaluable tools for deconstructing the biological complexity of the periodontal ligament (PDL). Model systems that closely reproduce the 3-dimensional (3D) configuration of cell-cell and cell-matrix interactions in native tissue can deliver physiologically relevant insights. However, 3D models of the PDL that incorporate mechanical loading are currently lacking. Hence, we developed a model where periodontal tissue constructs (PTCs) are made by casting PDL cells in a collagen gel suspended between a pair of slender, silicone posts for magnetic tensile loading. Specifically, one of the posts was rigid and the other was flexible with a magnet embedded in its tip so that PTCs could be subjected to tensile loading with an external magnet. Additionally, the deflection of the flexible post could be used to measure the contractile force of PDL cells in the PTCs. Prior to tensile loading, second harmonics generation analysis of collagen fibers in PTCs revealed that incorporation of PDL cells resulted in collagen remodeling. Biomechanical testing of PTCs by tensile loading revealed an elastic response at 4 h, permanent deformation by 1 d, and creep elongation by 1 wk. Subsequently, contractile forces of PDL cells were substantially lower for PTCs under tensile loading. Immunofluorescence analysis revealed that tensile loading caused PDL cells to increase in number, express higher levels of F-actin and α-smooth muscle actin, and become aligned to the tensile axis. Second harmonics generation analysis indicated that collagen fibers in PTCs progressively remodeled over time with tensile loading. Gene expression analysis also confirmed tension-mediated upregulation of the F-actin/Rho pathway and osteogenic genes. Our model is novel in demonstrating the mechanobiological behavior that results in cell-mediated remodeling of the PDL tissue in a 3D context. Hence, it can be a valuable tool to develop therapeutics for periodontitis, periodontal regeneration, and orthodontics.

Muscle wasting is a universal hallmark of aging which is displayed by a wide range of organisms, although the causes and mechanisms of this phenomenon are not fully understood. We used Drosophila to characterize the phenomenon of spontaneous muscle fiber degeneration (SMFD) during aging. We found that SMFD occurs across diverse types of somatic muscles, progresses with chronological age, and positively correlates with functional muscle decline. Data from vital dyes and morphological markers imply that degenerative fibers most likely die by necrosis. Mechanistically, SMFD is driven by the damage resulting from muscle contractions, and the nervous system may play a significant role in this process. Our quantitative model of SMFD assessment can be useful in identifying and validating novel genetic factors that influence aging-related muscle wasting.

Stiffening of the brain extracellular matrix (ECM) in glioblastoma promotes tumor progression. Previously, we discovered that protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK) plays a role in glioblastoma stem cell (GSC) adaptation to matrix stiffness through PERK/FLNA-dependent F-actin remodeling. Here, we examined the involvement of PERK in detecting stiffness changes via focal adhesion complex (FAC) formation. Compared to control GSCs, PERK-deficient GSCs show decreased vinculin and tensin expression, while talin and integrin-β1 remain constant. Furthermore, vimentin was also reduced while tubulin increased, and a stiffness-dependent increase of the differentiation marker GFAP expression was absent in PERK-deficient GSCs. In conclusion, our study reveals a novel role for PERK in FAC formation during matrix stiffening, which is likely linked to its regulation of F-actin remodeling.

VIP1, an Arabidopsis thaliana basic leucine zipper transcription factor, and its close homologs are imported from the cytoplasm to the nucleus when cells are exposed to mechanical stress. They bind to AGCTG (G/T) and regulate mechanical stress responses in roots. However, their role in leaves is unclear. To clarify this, mutant lines (QM1 and QM2) that lack the functions of VIP1 and its close homologs (bZIP29, bZIP30 and PosF21) were generated. Brushing more severely damaged QM1 and QM2 leaves than wild-type leaves. Genes regulating stress responses and cell wall properties were downregulated in brushed QM2 leaves and upregulated in brushed VIP1-GFP-overexpressing (VIP1-GFPox) leaves compared to wild-type leaves in a transcriptome analysis. The VIP1-binding sequence AGCTG (G/T) was enriched in the promoters of genes downregulated in brushed QM2 leaves compared to wild-type leaves and in those upregulated in brushed VIP1-GFPox leaves. Calmodulin-binding transcription activators (CAMTAs) are known regulators of mechanical stress responses, and the CAMTA-binding sequence CGCGT was enriched in the promoters of genes upregulated in the brushed QM2 leaves and in those downregulated in the brushed VIP1-GFPox leaves. These findings suggest that VIP1 and its homologs upregulate genes via AGCTG (G/T) and influence CAMTA-dependent gene expression to enhance mechanical stress tolerance in leaves.

BACKGROUND: This study investigates the potential of novel meniscal parameters as predictive factors for incident radiographic knee osteoarthritis (ROA) over a span of four years, as part of the Osteoarthritis Initiative (OAI) study.
OBJECTIVE: Quantitative measurements of meniscal parameters alteration could serve as predictors of OA\'s occurrence and progression.
METHODS: A nested matched case-control study design was used to select participants from OAI study. Case knees (n = 178) were defined as those with incident ROA (Kellgren Lawrence Grade (KLG) 0 or 1 at baseline (BL), evolving into KLG 2 or above by year 4). Control knees were matched one-to-one by sex, age and radiographic status with case knees. The mean distance from medial-to-lateral meniscal lesions [Mean(MLD)], mean value of tibial plateau width [Mean(TPW)] and the mean of the relative percentage of the medial-to-lateral meniscal lesions distance [Mean(RMLD)] were evaluated through coronal T2-weighted turbo spin echo (TSE) MRI at P-0 (visit when incident ROA was found on radiograph), P-1(one year prior to P-0) and baseline, respectively. Using the imaging data of one patient, the mechanism was investigated by finite element analysis.
RESULTS: Participants were on average 60.22 years old, predominantly female (66.7%) and overweight (mean BMI: 28.15). Mean(MLD) and Mean(RMLD) were significantly greater for incident knees compared to no incident knees at baseline, P-1 and P-0. [Mean(MLD), Mean(RMLD); (42.56-49.73) mean ± (7.70-9.52) mm SD vs. (38.14-40.78) mean ± (5.51-7.05)mm SD; (58.61-68.95) mean ± (8.52-11.40) mm SD vs. (52.52-56.35) mean ± (6.53-7.85)mm SD, respectively]. Baseline Mean(MLD) and Mean(RMLD), [Adjusted OR, 95%CI: 1.11(1.07 to 1.16) and 1.13(1.09 to 1.17), respectively], were associated with incident ROA during 4 years, However, Mean(TPW) [Adjusted OR, 95%CI: 0.98(0.94 to 1.02)] was not associated with incident ROA during 4 years. While Mean(TPW) at P-1 and P-0 was not associated with the risk of incident ROA, Mean(MLD) and Mean(RMLD) at P-1 and P-0 were significantly positively associated with the risk of incident ROA.
CONCLUSIONS: The meniscal parameters alteration could be an important imaging biomarker to predict the occurrence of ROA.

OBJECTIVE: To evaluate the fracture behavior of human molars with extensive MOD restorations using short-fiber-reinforced resin composite of varying viscosities.
METHODS: Human molars were randomly divided into seven groups (n = 12): intact teeth (control); restoration using conventional high-viscosity resin composite without (Filtek Z350XT, 3M) or with fibers (everX Posterior, GC); conventional low-viscosity resin composite without (Filtek Supreme Flowable, 3M) or with fibers (everX Flow Dentin Shade, GC); bulk-fill low-viscosity resin composite (Filtek Bulk Fill Flow, 3M) or with fibers (everX Flow Bulk Shade, GC). Restorations were performed on extensive MOD preparations, following the manufacturers\' recommendations for each material. Specimens underwent fracture strength testing (N) and fracture pattern (%) categorized as repairable, possibly repairable, or non-repairable. Results were analyzed using a generalized linear model (N) and Fisher\'s exact test (%), with α = 0.05.
RESULTS: Restorations performed with high-viscosity materials showed fracture strength values similar to the control and higher than those of restorations using low-viscosity resin composites (p < 0.0001), except for the bulk-fill low-viscosity resin composite with fibers (p > 0.05). Teeth restored using low-viscosity resin composite with fibers showed a higher % of repairable and possibly repairable fractures than the control (p = 0.0091).
CONCLUSIONS: The viscosity of materials mediated the fracture strength, with restorations using high-viscosity resin composites promoting values similar to the intact tooth; however, the presence of fibers influenced the fracture pattern.
CONCLUSIONS: Teeth with MOD cavities restored with high-viscosity resin composites showed similar fracture strength to intact teeth. Fiber-reinforced low-viscosity resin composite for the base of restoration resulted in a more repairable/possibly repairable fracture pattern.

The mammalian skull is very malleable and has notably radiated into highly diverse morphologies, fulfilling a broad range of functional needs. Although gnawing is relatively common in mammals, this behavior and its associated morphology are diagnostic features for rodents. These animals possess a very versatile and highly mechanically advantageous masticatory apparatus, which, for instance, allowed caviomorph rodents to colonize South America during the Mid-Eocene and successfully radiate in over 200 extant species throughout most continental niches. Previous work has shown that differences in bite force within caviomorphs could be better explained by changes in muscle development than in mechanical advantages (i.e., in cranial overall morphology). Considering the strong bites they apply, it is interesting to assess how the reaction forces upon the incisors (compression) and the powerful adductor musculature pulling (tension) mechanically affect the cranium, especially between species with different ecologies (e.g., chisel-tooth digging). Thus, we ran finite element analyses upon crania of the subterranean Talas\' tuco-tuco Ctenomys talarum, the semi-fossorial common degu Octodon degus, and the saxicolous long-tailed chinchilla Chinchilla lanigera to simulate: (A) in vivo biting in all species, and (B) rescaled muscle forces in non-ctenomyid rodents to match those of the tuco-tuco. Results show that the stress patterns correlate with the mechanical demands of distinctive ecologies, on in vivo-based simulations, with the subterranean tuco-tuco being the most stressed species. In contrast, when standardizing all three species (rescaled models), non-ctenomyid models exhibited a several-fold increase in stress, in both magnitude and affected areas. Detailed observations evidenced that this increase in stress was higher in lateral sections of the snout and, mainly, the zygomatic arch; between approximately 2.5-3.5 times in the common degu and 4.0-5.0 times in the long-tailed chinchilla. Yet, neither species, module, nor simulation condition presented load factor levels that would imply structural failure by strong, incidental biting. Our results let us conclude that caviomorphs have a high baseline for mechanical strength of the cranium because of the inheritance of a very robust \"rodent\" model, while interspecific differences are associated with particular masticatory habits and the concomitant level of development of the adductor musculature. Especially, the masseteric and zygomaticomandibular muscles contribute to >80% of the bite force, and therefore, their contraction is responsible for the highest strains upon their origin sites, that is, the zygomatic arch and the snout. Thus, the robust crania of the subterranean and highly aggressive tuco-tucos allow them to withstand much stronger forces than degus or chinchillas, such as the ones produced by their hypertrophied jaw adductor muscles or imparted by the soil reaction.