Linking Metastatic Potential and Viscoelastic Properties of Breast Cancer Spheroids via Dynamic Compression and Relaxation in Microfluidics.

The growth and invasion of solid tumors are associated with changes in their viscoelastic properties, influenced by both internal cellular factors and physical forces in the tumor microenvironment. Due to the lack of a comprehensive investigation of tumor tissue viscoelasticity, the relationship between such physical properties and cancer malignancy remains poorly understood. Here, the viscoelastic properties of breast cancer spheroids, 3D (in vitro) tumor models, are studied in relation to their metastatic potentials by imposing controlled, dynamic compression within a microfluidic constriction, and subsequently monitoring the relaxation of the imposed deformation. By adopting a modified Maxwell model to extract viscoelastic properties from the compression data, the benign (MCF-10A) spheroids are found to have higher bulk elastic modulus and viscosity compared to malignant spheroids (MCF-7 and MDA-MB-231). The relaxation is characterized by two timescales, captured by a double exponential fitting function, which reveals a similar fast rebound for MCF-7 and MCF-10A. Both the malignant spheroids exhibit similar long-term relaxation and display residual deformation. However, they differ significantly in morphology, particularly in intercellular movements. These differences between malignant spheroids are demonstrated to be linked to their cytoskeletal organization, by microscopic imaging of F-actin within the spheroids, together with cell-cell adhesion strength.

Ex vivo SIM-AFM measurements reveal the spatial correlation of stiffness and molecular distributions in 3D living tissue.

Living tissues each exhibit a distinct stiffness, which provides cells with key environmental cues that regulate their behaviors. Despite this significance, our understanding of the spatiotemporal dynamics and the biological roles of stiffness in three-dimensional tissues is currently limited due to a lack of appropriate measurement techniques. To address this issue, we propose a new method combining upright structured illumination microscopy (USIM) and atomic force microscopy (AFM) to obtain precisely coordinated stiffness maps and biomolecular fluorescence images of thick living tissue slices. Using mouse embryonic and adult skin as a representative tissue with mechanically heterogeneous structures inside, we validate the measurement principle of USIM-AFM. Live measurement of tissue stiffness distributions revealed the highly heterogeneous mechanical nature of skin, including nucleated/enucleated epithelium, mesenchyme, and hair follicle, as well as the role of collagens in maintaining its integrity. Furthermore, quantitative analysis comparing stiffness distributions in live tissue samples with those in preserved tissues, including formalin-fixed and cryopreserved tissue samples, unveiled the distinct impacts of preservation processes on tissue stiffness patterns. This series of experiments highlights the importance of live mechanical testing of tissue-scale samples to accurately capture the true spatiotemporal variations in mechanical properties. Our USIM-AFM technique provides a new methodology to reveal the dynamic nature of tissue stiffness and its correlation with biomolecular distributions in live tissues and thus could serve as a technical basis for exploring tissue-scale mechanobiology. STATEMENT OF SIGNIFICANCE: Stiffness, a simple mechanical parameter, has drawn attention in understanding the mechanobiological principles underlying the homeostasis and pathology of living tissues. To explore tissue-scale mechanobiology, we propose a technique integrating an upright structured illumination microscope and an atomic force microscope. This technique enables live measurements of stiffness distribution and fluorescent observation of thick living tissue slices. Experiments revealed the highly heterogeneous mechanical nature of mouse embryonic and adult skin in three dimensions and the previously unnoticed influences of preservation techniques on the mechanical properties of tissue at microscopic resolution. This study provides a new technical platform for live stiffness measurement and biomolecular observation of tissue-scale samples with micron-scale resolution, thus contributing to future studies of tissue- and organ-scale mechanobiology.

Data-driven continuum damage mechanics with built-in physics.

Soft materials such as rubbers and soft tissues often undergo large deformations and experience damage degradation that impairs their function. This energy dissipation mechanism can be described in a thermodynamically consistent framework known as continuum damage mechanics. Recently, data-driven methods have been developed to capture complex material behaviors with unmatched accuracy due to the high flexibility of deep learning architectures. Initial efforts focused on hyperelastic materials, and recent advances now offer the ability to satisfy physics constraints such as polyconvexity of the strain energy density function by default. However, modeling inelastic behavior with deep learning architectures and built-in physics has remained challenging. Here we show that neural ordinary differential equations (NODEs), which we used previously to model arbitrary hyperelastic materials with automatic polyconvexity, can be extended to model energy dissipation in a thermodynamically consistent way by introducing an inelastic potential: a monotonic yield function. We demonstrate the inherent flexibility of our network architecture in terms of different damage models proposed in the literature. Our results suggest that our NODEs re-discover the true damage function from synthetic stress-deformation history data. In addition, they can accurately characterize experimental skin and subcutaneous tissue data.

Characterization of pediatric porcine pulmonary valves as a model for tissue engineered heart valves.

Heart valve tissue engineering holds the potential to transform the surgical management of congenital heart defects affecting the pediatric pulmonary valve (PV) by offering a viable valve replacement. While aiming to recapitulate the native valve, the minimum requirement for tissue engineered heart valves (TEHVs) has historically been adequate mechanical function at implantation. However, long-term in situ functionality of TEHVs remains elusive, suggesting that a closer approximation of the native valve is required. The realization of biomimetic engineered pediatric PV is impeded by insufficient characterization of healthy pediatric tissue. In this study, we comprehensively characterized the planar biaxial tensile behaviour, extracellular matrix (ECM) composition and organization, and valvular interstitial cell (VIC) phenotypes of PVs from piglets to provide benchmarks for TEHVs. The piglet PV possessed an anisotropic and non-linear tension-strain profile from which material constants for a predictive constitutive model were derived. The ECM of the piglet PV possessed a trilayer organization populated by collagen, glycosaminoglycans, and elastin. Biochemical quantification of ECM content normalized to wet weight and DNA content of PV tissue revealed homogeneous distribution across sampled regions of the leaflet. Finally, VICs in the piglet PV were primarily quiescent vimentin-expressing fibroblasts, with a small proportion of activated α-smooth muscle actin-expressing myofibroblasts. Overall, piglet PV properties were consistent with those reported anecdotally for pediatric human PVs and distinct from those of adult porcine and human PVs, supporting the utility of the properties determined here to inform the design of tissue engineered pediatric PVs. STATEMENT OF SIGNIFICANCE: Heart valve tissue engineering has the potential to transform treatment for children born with defective pulmonary valves by providing living replacement tissue that can grow with the child. The design of tissue engineered heart valves is best informed by native valve properties, but native pediatric pulmonary valves have not been fully described to date. Here, we provide comprehensive characterization of the planar biaxial tensile behaviour, extracellular matrix composition and organization, and valvular interstitial cell phenotypes of pulmonary valves from piglets as a model for the native human pediatric valve. Together, these findings provide standards that inform engineered heart valve design towards generation of biomimetic pediatric pulmonary valves.

A viscoelastic constitutive framework for aging muscular and elastic arteries.

The evolution of arterial biomechanics and microstructure with age and disease plays a critical role in understanding the health and function of the cardiovascular system. Accurately capturing these adaptative processes and their effects on the mechanical environment is critical for predicting arterial responses. This challenge is exacerbated by the significant differences between elastic and muscular arteries, which have different structural organizations and functional demands. In this study, we aim to shed light to these adaptive processes by comparing the viscoelastic mechanics of autologous thoracic aortas (TA) and femoropopliteal arteries (FPA) in different age groups. We have extended our fractional viscoelastic framework, originally developed for FPA, to both types of arteries. To evaluate this framework, we analyzed experimental mechanical data from TA and FPA specimens from 21 individuals aged 13 to 73 years. Each specimen was subjected to a multi-ratio biaxial mechanical extension and relaxation test complemented by bidirectional histology to quantify the structural density and microstructural orientations. Our new constitutive model accurately captured the mechanical responses and microstructural differences of the tissues and closely matched the experimentally measured densities. It was found that the viscoelastic properties of collagen and smooth muscle cells (SMCs) in both the FPA and TA remained consistent with age, but the viscoelasticity of the SMCs in the FPA was twice that of the TA. Additionally, changes in collagen nonlinearity with age were similar in both TA and FPA. This model provides valuable insights into arterial mechanophysiology and the effects of pathological conditions on vascular biomechanics. STATEMENT OF SIGNIFICANCE: Developing durable treatments for arterial diseases necessitates a deeper understanding of how mechanical properties evolve with age in response to mechanical environments. In this work, we developed a generalized viscoelastic constitutive model for both elastic and muscular arteries and analyzed both the thoracic aorta (TA) and the femoropopliteal artery (FPA) from 21 donors aged 13 to 73. The derived parameters correlate well with histology, allowing further examination of how viscoelasticity evolves with age. Correlation between the TA and FPA of the same donors suggest that the viscoelasticity of the FPA may be influenced by the TA, necessitating more detailed analysis. In summary, our new model proves to be a valuable tool for studying arterial mechanophysiology and exploring pathological impacts.

Orchestrated Movement Sequences and Shape-Memory-like Effects in Pine Cones.

Hygroscopic seed-scale movement is responsible for the weather-adaptive opening and closing of pine cones and for facilitating seed dispersal under favorable environmental conditions. Although this phenomenon has long been investigated, many involved processes are still not fully understood. To gain a deeper mechanical and structural understanding of the cone and its functional units, namely the individual seed scales, we have investigated their desiccation- and wetting-induced movement processes in a series of analyses and manipulative experiments. We found, for example, that the abaxial scale surface is responsible for the evaporation of water from the closed cone and subsequent cone opening. Furthermore, we tested the capability of dry and deformed scales to restore their original shape and biomechanical properties by wetting. These results shed new light on the orchestration of scale movement in cones and the involved forces and provide information about the functional robustness and resilience of cones, leading to a better understanding of the mechanisms behind hygroscopic pine cone opening, the respective ecological framework, and, possibly, to the development of smart biomimetic actuators.

Analysis of image formation in optical palpation.

Optical palpation is an emerging elastography technique that generates two-dimensional images of mechanical stress at the tissue surface, with clinical applications such as intraoperative cancer detection and scar assessment. It has been implemented using various imaging systems, however, an analysis of how deformation of the sample and layer influences image formation has not been performed. Here, an analysis framework is presented, which assesses performance independently of the imaging system used. Optical palpation of varying samples and layers is simulated using finite element analysis and validated with experiments on silicone phantoms, providing a characterization of detectability, feature resolution, and contrast ratio. Using our framework, we demonstrate that computational optical palpation, which incorporates realistic assumptions of layer deformation, improves the feature resolution up to a factor of four. This framework can guide the development of optical palpation and aid in the selection of appropriate imaging system and layer properties for a given application.

Heterogeneous Mechanical Stress and Interstitial Fluid Flow Predictions Derived from DCE-MRI for Rat U251N Orthotopic Gliomas.

Mechanical stress and fluid flow influence glioma cell phenotype in vitro, but measuring these quantities in vivo continues to be challenging. The purpose of this study was to predict these quantities in vivo, thus providing insight into glioma physiology and potential mechanical biomarkers that may improve glioma detection, diagnosis, and treatment. Image-based finite element models of human U251N orthotopic glioma in athymic rats were developed to predict structural stress and interstitial flow in and around each animal's tumor. In addition to accounting for structural stress caused by tumor growth, our approach has the advantage of capturing fluid pressure-induced structural stress, which was informed by in vivo interstitial fluid pressure (IFP) measurements. Because gliomas and the brain are soft, elevated IFP contributed substantially to tumor structural stress, even inverting this stress from compressive to tensile in the most compliant cases. The combination of tumor growth and elevated IFP resulted in a concentration of structural stress near the tumor boundary where it has the greatest potential to influence cell proliferation and invasion. MRI-derived anatomical geometries and tissue property distributions resulted in heterogeneous interstitial fluid flow with local maxima near cerebrospinal fluid spaces, which may promote tumor invasion and hinder drug delivery. In addition, predicted structural stress and interstitial flow varied markedly between irradiated and radiation-naïve animals. Our modeling suggests that relative to tumors in stiffer tissues, gliomas experience unusual mechanical conditions with potentially important biological (e.g., proliferation and invasion) and clinical consequences (e.g., drug delivery and treatment monitoring).

Sensitivity of knee cartilage biomechanics in finite element analysis to selected Musculoskeletal models.

Knee joint kinematics and kinetics analyzed by musculoskeletal (MS) modeling are often utilized in finite element (FE) models, estimating tissue-level mechanical responses. We compared knee cartilage stresses, strains, and centers of pressure of FE models driven by two widely used MS models, implemented in AnyBody and OpenSim. Minor discrepancies in the results were observed between the models. AnyBody-driven FE models showed slightly higher stresses in the medial tibial cartilage, while OpenSim-driven FE models estimated more anterior and lateral center of pressure. Recognizing these differences in the MS-FE models is important to ensure reliable analysis of cartilage mechanics and failure and simulation of rehabilitation.

Differential proliferation regulates multi-tissue morphogenesis during embryonic axial extension: integrating viscous modeling and experimental approaches.

A major challenge in biology is to understand how mechanical interactions and cellular behavior affect the shapes of tissues and embryo morphology. The extension of the neural tube and paraxial mesoderm, which form the spinal cord and musculoskeletal system, respectively, results in the elongated shape of the vertebrate embryonic body. Despite our understanding of how each of these tissues elongates independently of the others, the morphogenetic consequences of their simultaneous growth and mechanical interactions are still unclear. Our study investigates how differential growth, tissue biophysical properties and mechanical interactions affect embryonic morphogenesis during axial extension using a 2D multi-tissue continuum-based mathematical model. Our model captures the dynamics observed in vivo by time-lapse imaging of bird embryos, and reveals the underestimated influence of differential tissue proliferation rates. We confirmed this prediction in quail embryos by showing that decreasing the rate of cell proliferation in the paraxial mesoderm affects long-term tissue dynamics, and shaping of both the paraxial mesoderm and the neighboring neural tube. Overall, our work provides a new theoretical platform upon which to consider the long-term consequences of tissue differential growth and mechanical interactions on morphogenesis.

The Biomechanical, Biochemical, and Morphological Properties of 19 Human Cadaveric Lower Limb Tendons and Ligaments: An Open-Access Data Set.

BACKGROUND: Methodological heterogeneity hinders data comparisons across isolated studies of tendon and ligament properties, limiting clinical understanding and affecting the development and evaluation of replacement materials. PURPOSE: To create an open-access data set on the morphological, biomechanical, and biochemical properties of clinically important tendons and ligaments of the lower limb, using consistent methodologies, to enable direct tendon/ligament comparisons. STUDY DESIGN: Descriptive laboratory study. METHODS: Nineteen distinct lower limb tendons and ligaments were retrieved from 8 fresh-frozen human cadavers (5 male, 3 female; aged 49-65 years) including Achilles, tibialis posterior, tibialis anterior, fibularis (peroneus) longus, fibularis (peroneus) brevis, flexor hallucis longus, extensor hallucis longus, plantaris, flexor digitorum longus, quadriceps, patellar, semitendinosus, and gracilis tendons; anterior cruciate, posterior cruciate, medial collateral, and lateral collateral ligaments; and 10 mm-wide grafts from the contralateral quadriceps and patellar tendons. Outcomes included morphology (tissue length, ultrasound-quantified cross-sectional area [CSAUS], and major and minor axes), biomechanics (failure load, ultimate tensile strength [UTS], failure strain, and elastic modulus), and biochemistry (sulfated glycosaminoglycan [sGAG] and hydroxyproline contents). Tissue differences were analyzed using mixed-model regression. RESULTS: There was a range of similarities and differences between tendons and ligaments across outcomes. A key finding relating to potential graft tissue suitability was the comparable failure loads, UTS, CSAUS, sGAG, and hydroxyproline present between hamstring tendons (a standard graft source) and 5 tendons not typically used for grafting: fibularis (peroneus) longus and brevis, flexor and extensor hallucis longus, and flexor digitorum longus tendons. CONCLUSION: This study of lower limb tendons and ligaments has enabled direct comparison of morphological, biomechanical, and biochemical human tissue properties-key factors in the selection of suitable graft tissues. This analysis has identified 6 potential new donor tissues with properties comparable to currently used grafts. CLINICAL RELEVANCE: This extensive data set reduces the need to utilize data from incompatible sources, which may aid surgical decisions (eg, evidence to expand the range of tendons considered suitable for use as grafts) and may provide congruent design inputs for new biomaterials and computational models. The complete data set has been provided to facilitate further investigations, with the capacity to expand the resource to include additional outcomes and tissues.

Mechanical and morphological characterization of the emphysematous lung tissue.

Irreversible alveolar airspace enlargement is the main characteristic of pulmonary emphysema, which has been extensively studied using animal models. While the alterations in lung mechanics associated with these morphological changes have been documented in the literature, the study of the mechanical behavior of parenchymal tissue from emphysematous lungs has been poorly investigated. In this work, we characterize the mechanical and morphological properties of lung tissue in elastase-induced emphysema rat models under varying severity conditions. We analyze the non-linear tissue behavior using suitable hyperelastic constitutive models that enable to compare different non-linear responses in terms of hyperelastic material parameters. We further analyze the effect of the elastase dose on alveolar morphology and tissue material parameters and study their connection with respiratory-system mechanical parameters. Our results show that while the lung mechanical function is not significantly influenced by the elastase treatment, the tissue mechanical behavior and alveolar morphology are markedly affected by it. We further show a strong association between alveolar enlargement and tissue softening, not evidenced by respiratory-system compliance. Our findings highlight the importance of understanding tissue mechanics in emphysematous lungs, as changes in tissue properties could detect the early stages of emphysema remodeling. STATEMENT OF SIGNIFICANCE: Gas exchange is vital for life and strongly relies on the mechanical function of the lungs. Pulmonary emphysema is a prevalent respiratory disease where alveolar walls are damaged, causing alveolar enlargement that induces harmful changes in the mechanical response of the lungs. In this work, we study how the mechanical properties of lung tissue change during emphysema. Our results from animal models show that tissue properties are more sensitive to alveolar enlargement due to emphysema than other mechanical properties that describe the function of the whole respiratory system.

Notochord segmentation in zebrafish controlled by iterative mechanical signaling.

In bony fishes, patterning of the vertebral column, or spine, is guided by a metameric blueprint established in the notochord sheath. Notochord segmentation begins days after somitogenesis concludes and can occur in its absence. However, somite patterning defects lead to imprecise notochord segmentation, suggesting that these processes are linked. Here, we identify that interactions between the notochord and the axial musculature ensure precise spatiotemporal segmentation of the zebrafish spine. We demonstrate that myoseptum-notochord linkages drive notochord segment initiation by locally deforming the notochord extracellular matrix and recruiting focal adhesion machinery at these contact points. Irregular somite patterning alters this mechanical signaling, causing non-sequential and dysmorphic notochord segmentation, leading to altered spine development. Using a model that captures myoseptum-notochord interactions, we find that a fixed spatial interval is critical for driving sequential segment initiation. Thus, mechanical coupling of axial tissues facilitates spatiotemporal spine patterning.

Active viscoelastic models for cell and tissue mechanics.

Living cells are out of equilibrium active materials. Cell-generated forces are transmitted across the cytoskeleton network and to the extracellular environment. These active force interactions shape cellular mechanical behaviour, trigger mechano-sensing, regulate cell adaptation to the microenvironment and can affect disease outcomes. In recent years, the mechanobiology community has witnessed the emergence of many experimental and theoretical approaches to study cells as mechanically active materials. In this review, we highlight recent advancements in incorporating active characteristics of cellular behaviour at different length scales into classic viscoelastic models by either adding an active tension-generating element or adjusting the resting length of an elastic element in the model. Summarizing the two groups of approaches, we will review the formulation and application of these models to understand cellular adaptation mechanisms in response to various types of mechanical stimuli, such as the effect of extracellular matrix properties and external loadings or deformations.

Organoids and organ-on-chip technology for investigating host-microorganism interactions.

Recent advances in organoid and organ-on-chip (OoC) technologies offer an unprecedented level of tissue mimicry. These models can recapitulate the diversity of cellular composition, 3D organization, and mechanical stimulation. These approaches are intensively used to understand complex diseases. This review focuses on the latest advances in this field to study host-microorganism interactions.

A scientometric analysis of bone cutting tools & methodologies: Mapping the research landscape.

This study undertakes a Scientometric analysis of bone-cutting tools, investigating a corpus of 735 papers from the Scopus database between 1941 and 2023. It employs bibliometric methodologies such as keyword coupling, co-citation, and co-authorship analysis to map the intellectual landscape and collaborative networks within this research domain. The analysis highlights a growing interest and significant advancements in bone-cutting tools, focusing on their design, the materials used, and the cutting processes involved. It identifies key research fronts and trends, such as the emphasis on surgical precision, material innovation, and the optimization of tool performance. Further, the study reveals a broad collaboration among researchers from various disciplines, including engineering, materials science, and medical sciences, reflecting the field's interdisciplinary nature. Despite the progress, the analysis points out several gaps, notably in tool design optimization and the impact of materials on bone health. This comprehensive review not only charts the evolution of bone-cutting tool research but also calls attention to areas requiring further investigation, aiming to inspire future studies that address these identified gaps and enhance surgical outcomes.

The interplay of collagen, macrophages, and microcalcification in atherosclerotic plaque cap rupture mechanics.

The rupture of an atherosclerotic plaque cap overlying a lipid pool and/or necrotic core can lead to thrombotic cardiovascular events. In essence, the rupture of the plaque cap is a mechanical event, which occurs when the local stress exceeds the local tissue strength. However, due to inter- and intra-cap heterogeneity, the resulting ultimate cap strength varies, causing proper assessment of the plaque at risk of rupture to be lacking. Important players involved in tissue strength include the load-bearing collagenous matrix, macrophages, as major promoters of extracellular matrix degradation, and microcalcifications, deposits that can exacerbate local stress, increasing tissue propensity for rupture. This review summarizes the role of these components individually in tissue mechanics, along with the interplay between them. We argue that to be able to improve risk assessment, a better understanding of the effect of these individual components, as well as their reciprocal relationships on cap mechanics, is required. Finally, we discuss potential future steps, including a holistic multidisciplinary approach, multifactorial 3D in vitro model systems, and advancements in imaging techniques. The obtained knowledge will ultimately serve as input to help diagnose, prevent, and treat atherosclerotic cap rupture.

Cell division angle predicts the level of tissue mechanics that tune the amount of cerebellar folding.

Modeling has led to proposals that the amount of neural tissue folding is set by the level of differential expansion between tissue layers and that the wavelength is set by the thickness of the outer layer. Here, we used inbred mouse strains with distinct amounts of cerebellar folding to investigate these predictions. We identified a distinct critical period during which the folding amount diverges between the two strains. In this period, regional changes in the level of differential expansion between the external granule layer (EGL) and underlying core correlate with the folding amount in each strain. Additionally, the thickness of the EGL varies regionally during the critical period alongside corresponding changes in wavelength. The number of SHH-expressing Purkinje cells predicts the folding amount, but the proliferation rate in the EGL is the same between the strains. However, regional changes in the cell division angle within the EGL predicts both the tangential expansion and the thickness of the EGL. Cell division angle is likely a tunable mechanism whereby both the level of differential expansion along the perimeter and the thickness of the EGL are regionally tuned to set the amount and wavelength of folding.

On the soft tissue ultrasound elastography using FEM based inversion approach.

Elastography is a medical imaging modality that enables visualization of tissue stiffness. It involves quasi-static or harmonic mechanical stimulation of the tissue to generate a displacement field which is used as input in an inversion algorithm to reconstruct tissue elastic modulus. This paper considers quasi-static stimulation and presents a novel inversion technique for elastic modulus reconstruction. The technique follows an inverse finite element framework. Reconstructed elastic modulus maps produced in this technique do not depend on the initial guess, while it is computationally less involved than iterative reconstruction approaches. The method was first evaluated using simulated data (in-silico) where modulus reconstruction's sensitivity to displacement noise and elastic modulus was assessed. To demonstrate the method's performance, displacement fields of two tissue mimicking phantoms determined using three different motion tracking techniques were used as input to the developed elastography method to reconstruct the distribution of relative elastic modulus of the inclusion to background tissue. In the next stage, the relative elastic modulus of three clinical cases pertaining to liver cancer patient were determined. The obtained results demonstrate reasonably high elastic modulus reconstruction accuracy in comparison with similar direct methods. Also it is associated with reduced computational cost in comparison with iterative techniques, which suffer from convergence and uniqueness issues, following the same formulation concept. Moreover, in comparison with other methods which need initial guess, the presented method does not require initial guess while it is easy to understand and implement.

The extracellular matrix in tissue morphogenesis: No longer a backseat driver.

The forces driving tissue morphogenesis are thought to originate from cellular activities. While it is appreciated that extracellular matrix (ECM) may also be involved, ECM function is assumed to be simply instructive in modulating the cellular behaviors that drive changes to tissue shape. However, there is increasing evidence that the ECM may not be the passive player portrayed in developmental biology textbooks. In this review we highlight examples of embryonic ECM dynamics that suggest cell-independent activity, along with developmental processes during which localized ECM alterations and ECM-autonomous forces are directing changes to tissue shape. Additionally, we discuss experimental approaches to unveil active ECM roles during tissue morphogenesis. We propose that it may be time to rethink our general definition of morphogenesis as a cellular-driven phenomenon and incorporate an underappreciated, and surprisingly dynamic ECM.