The impact of cell density variations on nanoparticle uptake across bioprinted A549 gradients

Frontiers in Bioengineering and Biotechnology, Год журнала: 2025, Номер 13

Опубликована: Апрель 30, 2025

The safe-by-design of engineered nanoparticles (NPs) for any application requires a detailed understanding how the particles interact with single cells. Most studies are based on two-dimensional, uniformly dense cell cultures, which do not represent diverse and inhomogeneous environments found in situ. In-vitro models that accurately tissue complexity, including realistic densities, essential to increase predictive accuracy cell-NP interactions. This study uses bioprinted gradient model examine relation between density NP uptake one dish. A549 lung epithelial gradients within inserts were produced bioprinter by modulating inter-droplet distances. After two days culture, cells exposed Cy5-labeled silica NPs (SiO2 NPs, ∼112 nm, 20 μg/mL) up 48 h. Confocal fluorescence microscopy 3D image analysis used quantify uptake, surface area, volume. relationship other parameters was then investigated statistically. Bioprinting enabled creation reproducible linear gradients, allowing controlled modeling variations while preserving viability throughout experiment. Increasing distances, from 0.1 mm 0.6 mm, achieve decreasing densities. SiO2 per around 50% higher low-density regions compared high-density areas across all time points, i.e., 6, 24, h post-exposure. inverse correlated greater average area lower-density regions, differences proliferation rates at varying densities did significantly impact uptake. is enhanced lower mainly due increased available revealing potential interaction tissues present variability. Our drop-on-demand bioprinting successfully supports implementation in-vitro their relevance as new approach methodologies (NAMs) next-generation risk assessment strategies.

Язык: Английский

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Colony context and size-dependent compensation mechanisms give rise to variations in nuclear growth trajectories

Cell Systems, Год журнала: 2025, Номер unknown, С. 101265 - 101265

Опубликована: Май 1, 2025

To investigate how cellular variations arise across spatiotemporal scales in a population of identical healthy cells, we performed data-driven analysis nuclear growth hiPS cell colonies as model system. We generated 3D timelapse dataset thousands nuclei over multiple days and developed open-source tools for image data feature-based exploration. Together, these data, tools, workflows comprise framework systematic quantitative dynamics at individual levels, the further highlights important aspects to consider when interpreting data. found that volume trajectories from short-timescale attributable their context within colony. identified time-invariant compensation relationship between duration starting population. Notably, discovered inheritance plays crucial role determining two key features while other are determined by not inherited.

Язык: Английский

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A novel tool for the unbiased characterization of epithelial monolayer development in culture

Molecular Biology of the Cell, Год журнала: 2023, Номер 34(4)

Опубликована: Янв. 25, 2023

The function of an epithelial tissue is intertwined with its architecture. Epithelial tissues are often described as pseudo-two-dimensional, but this view may be partly attributed to experimental bias: many model epithelia, including cultured cell lines, easiest image from the "top-down." We measured three-dimensional architecture cells in culture and found that it varies dramatically across regions, presenting a challenge for reproducibility cross-study comparisons. therefore developed novel tool (Automated Layer Analysis, "ALAn") characterize unbiased manner. Using ALAn, we find can organize into four distinct architectures correlates density. Cells exhibit biological properties each Organization apical-basal axis determined early monolayer development by substrate availability, while disorganization arises inability form connections. Our work highlights need carefully control when using system biology introduces tool, built on set rules widely applied culture.

Язык: Английский

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Unveiling the Intricate Connection: Cell Volume as a Key Regulator of Mechanotransduction

Annual Review of Biophysics, Год журнала: 2024, Номер 53(1), С. 299 - 317

Опубликована: Март 1, 2024

The volumes of living cells undergo dynamic changes to maintain the cells' structural and functional integrity in many physiological processes. Minor fluctuations cell volume can serve as intrinsic signals that play a crucial role fate determination during mechanotransduction. In this review, we discuss variability its vivo, along with an overview mechanisms governing regulation. Additionally, provide insights into current approaches used control vitro. Furthermore, summarize biological implications regulation recent advances understanding fundamental relationship between Finally, delve potential underlying mechanisms, including intracellular macromolecular crowding cellular mechanics, govern global response volume. By exploring intricate interplay mechanotransduction, underscore importance considering signaling cue unravel basic principles propose future research directions extend our Overall, review highlights significance signal mechanotransduction points out possibility controlling fate, mitigate disease-related damage, facilitate healing damaged tissues.

Язык: Английский

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Proliferation symmetry breaking in growing tissues

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Сен. 6, 2024

Morphogenesis of developing tissues results from anisotropic growth, typically driven by polarized patterns gene expression. Here we propose an alternative model growth self-organized feedback between cell polarity, mechanical pressure, and division rates. Specifically, polarity alignment can induce spontaneous symmetry breaking in proliferation, resulting the distribution pressure tissue. We show that proliferation anisotropy be controlled cellular elasticity, motility contact inhibition, thereby elucidating design principles for morphogenesis.

Язык: Английский

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Regulation of epithelial cell jamming transition by cytoskeleton and cell–cell interactions

Biophysics Reviews, Год журнала: 2024, Номер 5(4)

Опубликована: Окт. 14, 2024

Multicellular systems, such as epithelial cell collectives, undergo transitions similar to those in inert physical systems like sand piles and foams. To remodel or maintain tissue organization during development disease, these collectives transition between fluid-like solid-like states, undergoing jamming unjamming transitions. While share principles with understanding their regulation implications biology is challenging. Although follow physics described by the diagram, they are fundamentally biological processes. In this review, we explore how cellular processes interactions regulate We begin an overview of control remodeling model describe recent findings governing solidification fluidization. then mechanistic pathways that modulate phase diagram axes, focusing on fluctuations geometric compatibility. Drawing upon seminal works biology, discuss roles cytoskeleton cell-cell adhesion controlling motility geometry. This comprehensive view illustrates molecular unjamming, crucial for various contexts.

Язык: Английский

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Mechanochemical control systems regulating animal cell size

Current Opinion in Cell Biology, Год журнала: 2024, Номер 91, С. 102443 - 102443

Опубликована: Ноя. 6, 2024

Язык: Английский

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Membrane potential as master regulator of cellular mechano-transduction

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2023, Номер unknown

Опубликована: Ноя. 3, 2023

Mechanical forces have been shown to influence cellular decisions grow, die, or differentiate, through largely mysterious mechanisms. Separately, changes in resting membrane potential observed development, differentiation, regeneration, and cancer. We now demonstrate that is the central mediator of response mechanical pressure. show acting on cell change biomass density, which turn alters potential. Membrane then regulates number density epithelia by controlling growth, proliferation, elimination. Mechanistically, we control signaling Hippo MAPK pathways, potentially other pathways originate at membrane. While many molecular interactions are known affect signaling, upstream signal activates canonical pathway has previously elusive. Our results establish as a regulator growth tissue homeostasis.

Язык: Английский

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Contact Inhibition and Mechanotransduction

Опубликована: Дек. 19, 2023

Contact inhibition (CI) represents a crucial tumor-suppressive mechanism responsible for controlling the unbridled growth of cells, thus preventing formation cancerous tissues. CI can be further categorized into two distinct yet interrelated components: locomotion (CIL) and proliferation (CIP). These components have historically been viewed as separate processes, but emerging research indicates that they share both distinctive common pathways. Moreover, exhibit intriguing intersections with mechanotransduction, process involves cells sensing responding to mechanical forces. This review article describes intricate interplay between forces CI, shedding light on how these regulate CIL CIP. Emphasis is placed filamin A (FLNA)-mediated elucidating FLNA senses translates them biochemical signals cell proliferation. In addition FLNA, trans-acting factors (TAFs) emerge sensitive players in mechanotransduction process. presents methods identifying TAFs profiling associated changes chromatin structure, offering valuable insights regulation. Finally, it addresses unanswered questions fields delineates their possible future directions.

Язык: Английский

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Confinement in fibrous environments positions and orients mitotic spindles

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Апрель 15, 2024

Abstract Accurate positioning of the mitotic spindle within rounded cell body is critical to physiological maintenance. Adherent cells encounter confinement from neighboring or extracellular matrix (ECM), which can cause rotation spindles and, consequently, titling metaphase plate (MP). To understand and orientation under by fibers (ECM-confinement), we use flexible ECM-mimicking nanofibers that allow natural rounding while confining it differing levels. Rounded bodies are anchored in place actin retraction (RFs) originating adhesion clusters on fibers. We discover extent ECM-confinement patterns RFs 3D: triangular band-like at low high confinement, respectively. A stochastic Monte-Carlo simulation centrosome (CS), chromosome (CH), membrane interactions, 3D arrangement recovers MP tilting trends observed experimentally. Our mechanistic analysis reveals shape primary driver rotation. Under ECM-confinement, mechanically pinch cortex, causing have localized deformations contact sites with Interestingly, leads tilts, mechanistically depend upon cortical deformation, RF patterning, position. identify deformation work tandem limit tilt, asymmetric tilts. Overall, provide fundamental insights into how mitosis may proceed fibrous ECM-confining microenvironments vivo.

Язык: Английский

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