Search bioRxiv⌕ Search

Biology subjects

Tassieri, M.

Publications and source records attributed to Tassieri, M..

6 recordsLinked to original sources

Broadband Rheometry of Native Soft Tissues via i-Rheo-Indent

Characterising the frequency-dependent viscoelasticity of soft biological tissues is important for mechanobiology, disease phenotyping, and the design of biomimetic materials. However, conventional oscillatory rheometry becomes increasingly compromised by instrumental inertia at high frequencies and often requires specimens to be cut, reshaped, or compressed to conform to predefined measuring geometries, while local indentation methods probe only limited tissue volumes. Here, we apply i-Rheo-Indent, a macroscale indentation approach that transforms the measured force-relaxation and indentation histories from a single experiment directly into the broadband viscoelastic spectrum, without fitting to a predefined constitutive model. Measurements on a 3D-printed porcine liver analogue and murine quadriceps and brain showed good agreement with conventional oscillatory rheometry over the frequency range in which the latter remained reliable, while extending the accessible response up to 500 rad/s, beyond the regime increasingly dominated by instrument inertia. The method also enabled direct characterisation of intact tubular murine colon, yielding an elastic response consistent with published indentation measurements while additionally resolving the frequency-dependent viscous contribution. Intact murine livers exhibited substantially larger viscoelastic moduli than biopsy-punched specimens prepared for parallel-plate rheometry, indicating that specimen preparation can materially influence the measured mechanical response. By combining rapid broadband acquisition, macroscale probing, minimal specimen manipulation, and comparatively simple instrumentation, i-Rheo-Indent provides a practical route to characterising soft tissues closer to their native mechanical state, with potential applications in mechanobiology, disease phenotyping, and the development of tissue-engineered and biomimetic materials.

bioengineering↗

The mitotic chromosome periphery: a fluid coat that mediates chromosome mechanics

Mitotic chromosomes are specialised packets of condensed genetic material with dynamic mechanical properties. Each chromosome is coated by a sheath of proteins and RNA, called the mitotic chromosome periphery (MCP). The MCP is widely considered as an essential chromosome compartment where its multiple functions bestow material properties important for successful cell division. However, the details of the micromechanical properties of mitotic chromosomes, and specifically if and how the MCP contributes to these features, remain poorly understood. In this study, we present the most comprehensive characterisation of single-chromosome mechanics to date spanning a broadband frequency range, using optical tweezers and a novel microrheology technique. We extend this analysis to the first direct measurements of MCP micromechanics by manipulating levels of Ki-67, the chief organiser of this compartment, and apply a rheological model to isolate its contribution to chromosome dynamics. We report that the MCP governs high-frequency self-reorganisation dynamics and acts as a structural constraint, providing force-damping properties that mitigate mitotic stress. This work significantly advances our understanding of chromosome micromechanics and how the MCP contributes to the fundamental properties of chromosomes.

biophysics↗

Mind the viscous modulus: The mechanotransductive response to the viscous nature of isoelastic matrices regulates stem cell chondrogenesis

The design of hydrogels as mimetics of tissues matrices typically disregards the viscous nature of native tissues and focuses only on their elastic properties. In the case of stem cell chondrogenesis, this has led to contradictory results, likely due to unreported changes of the matrices viscous modulus. Here, by employing isoelastic matrices with a Youngs modulus of ~12 kPa, we demonstrate that variations in viscous properties alone (i.e., loss tangent between 0.1-0.25) are sufficient to drive efficient growth factor-free chondrogenesis of human mesenchymal stem cells, both in 2D and 3D cultures. The increase of the viscous component of RGD-functionalised polyacrylamide or polyethylene glycol maleimide hydrogels promotes a phenotype with reduced adhesion, alters mechanosensitive signalling, and boosts cell-cell contacts. In turn, this upregulates the chondrogenic transcription factor SOX9 and supports neocartilage formation, demonstrating that the mechanotransductive response to the viscous nature of the matrix can be harnessed to direct cell fate.

bioengineering↗

OptoRheo: Simultaneous in situ micro-mechanical sensing and 3D imaging of live cell cultures

Biomechanical cues from the extracellular matrix (ECM) are essential for directing many cellular processes, from normal development and repair, to disease progression. To better understand cell-matrix interactions, we have developed a new instrument named OptoRheo that combines light sheet fluorescence microscopy with particle tracking microrheology. OptoRheo lets us image cells in 3D as they proliferate over several days while simultaneously sensing the mechanical properties of the surrounding extracellular and pericellular matrix at a sub-cellular length scale. OptoRheo can be used in two operational modalities (with and without an optical trap) to extend the dynamic range of microrheology measurements. We corroborated this by characterising the ECM surrounding live breast cancer cells in two distinct culture systems, cell clusters in 3D hydrogels and spheroids in suspension culture. This cutting-edge instrument will transform the exploration of drug transport through complex cell culture matrices and optimise the design of the next-generation of disease models.

bioengineering↗

Nanovibrational stimulation of mesenchymal stem cells induces therapeutic reactive oxygen species and inflammation for 3D bone tissue engineering

There is a pressing clinical need to develop cell-based bone therapies due to a lack of viable, autologous bone grafts and a growing demand for bone grafts in musculoskeletal surgery. Such therapies can be tissue engineered and cellular, such as osteoblasts combined with a material scaffold. Because mesenchymal stem cells (MSCs) are both available and fast growing compared to mature osteoblasts, therapies that utilise these progenitor cells are particularly promising. We have developed a nanovibrational bioreactor that can convert MSCs into bone-forming osteoblasts in 2D and 3D but the mechanisms involved in this osteoinduction process remain unclear. Here, to elucidate this mechanism, we use increasing vibrational amplitude, from 30 nm (N30) to 90 nm (N90) amplitudes at 1000 Hz, and assess MSC metabolite, gene and protein changes. These approaches reveal that dose-dependent changes occur in MSCs responses to increased vibrational amplitude, particularly in adhesion and mechanosensitive ion channel expression, and that energetic metabolic pathways are activated, leading to low-level reactive oxygen species (ROS) production and to low-level inflammation, as well as to ROS- and inflammation-balancing pathways. These events are analogous to those that occur in the natural bone-healing processes. We have also developed a tissue engineered MSC-laden scaffold designed using cells mechanical memory, driven by the stronger N90 stimulation. These new mechanistic insights and cell-scaffold design are underpinned by a process that is free of inductive chemicals.

bioengineering↗

Microrheology reveals microscale viscosity gradients in planktonic systems

Microbial activity in planktonic systems creates a dynamic and heterogeneous microscale seascape that harbours a diverse community of microorganisms and ecological interactions of global significance. In recent decades a great effort has been put into understanding this complex system, particularly focusing on the role of chemical patchiness, while overlooking a physical parameter that governs microbial life and is affected by biological activity: viscosity. Here we use microrheological techniques to measure viscosity at length scales relevant to microorganisms. Our results reveal the viscous nature and the spatial extent of the phycosphere, the microenvironment surrounding phytoplankton cells, and show heterogeneity in viscosity at the microscale. Such heterogeneity affects the distribution of chemicals and microorganisms, with pervasive and profound implications for the functioning of the planktonic ecosystem. One Sentence SummaryMicrorheology measurements unveil the existence of layers of increased viscosity surrounding phytoplankton cells and within aggregates.

ecology↗