In vivo measurement of human brain elasticity using a light aspiration device

The brain deformation that occurs during neurosurgery is a serious issue impacting the patient "safety" as well as the invasiveness of the brain surgery. Model-driven compensation is a realistic and efficient solution to solve this problem. However, a vital issue is the lack of reliable and easily obtainable patient-specific mechanical characteristics of the brain which, according to clinicians' experience, can vary considerably. We designed an aspiration device that is able to meet the very rigorous sterilization and handling process imposed during surgery, and especially neurosurgery. The device, which has no electronic component, is simple, light and can be considered as an ancillary instrument. The deformation of the aspirated tissue is imaged via a mirror using an external camera. This paper describes the experimental setup as well as its use during a specific neurosurgery. The experimental data was used to calibrate a continuous model. We show that we were able to extract an in vivo constitutive law of the brain elasticity: thus for the first time, measurements are carried out per-operatively on the patient, just before the resection of the brain parenchyma. This paper discloses the results of a difficult experiment and provide for the first time in-vivo data on human brain elasticity. The results point out the softness as well as the highly non-linear behavior of the brain tissue.Comment: Medical Image Analysis (2009) accept\'

Formation and optogenetic control of engineered 3D skeletal muscle bioactuators

Densely arrayed skeletal myotubes are activated individually and as a group using precise optical stimulation with high spatiotemporal resolution. Skeletal muscle myoblasts are genetically encoded to express a light-activated cation channel, Channelrhodopsin-2, which allows for spatiotemporal coordination of a multitude of skeletal myotubes that contract in response to pulsed blue light. Furthermore, ensembles of mature, functional 3D muscle microtissues have been formed from the optogenetically encoded myoblasts using a high-throughput device. The device, called “skeletal muscle on a chip”, not only provides the myoblasts with controlled stress and constraints necessary for muscle alignment, fusion and maturation, but also facilitates the measurement of forces and characterization of the muscle tissue. We measured the specific static and dynamic stresses generated by the microtissues and characterized the morphology and alignment of the myotubes within the constructs. The device allows testing of the effect of a wide range of parameters (cell source, matrix composition, microtissue geometry, auxotonic load, growth factors and exercise) on the maturation, structure and function of the engineered muscle tissues in a combinatorial manner. Our studies integrate tools from optogenetics and microelectromechanical systems (MEMS) technology with skeletal muscle tissue engineering to open up opportunities to generate soft robots actuated by a multitude of spatiotemporally coordinated 3D skeletal muscle microtissues.National Science Foundation (U.S.) (Science and Technology Center—Emergent Behaviors of Integrated Cellular Systems (EBICS) grant No. CBET-0939511)National Institutes of Health (U.S.) (EB00262)National Science Foundation (U.S.) (GM74048)National Science Foundation (U.S.) (HL90747)National Institute for Biomedical Imaging and Bioengineering (U.S.) (RESBIO, Integrapted Technologies for Polymeric Biomaterial)University of Pennsylvania. Center for Engineering Cells and RegenerationSingapore-MIT Alliance for Research and Technolog

Confinement-Induced Transition between Wavelike Collective Cell Migration Modes

International audienceThe structural and functional organization of biological tissues relies on the intricate interplay between chemical and mechanical signaling. Whereas the role of constant and transient mechanical perturbations is generally accepted, several studies recently highlighted the existence of longrange mechanical excitations (i.e., waves) at the supracellular level. Here, we confine epithelial cell mono-layers to quasi-one dimensional geometries, to force the establishment of tissue-level waves of well-defined wavelength and period. Numerical simulations based on a self-propelled Voronoi model reproduce the observed waves and exhibit a phase transition between a global and a multi-nodal wave, controlled by the confinement size. We conrm experimentally the existence of such a phasetransition, and show that wavelength and period are independent of the confinement length. Together, these results demonstrate the intrinsic origin of tissue oscillations, which could provide cells with a mechanism to accurately measure distances at the supracellular level

Free-standing polyelectrolyte membranes made of chitosan and alginate

Free-standing films have increasing applications in the biomedical field as drug delivery systems for wound healing and tissue engineering. Here, we prepared free-standing membranes by the layer-by-layer assembly of chitosan and alginate, two widely used biomaterials. Our aim was to produce a thick membrane and to study the permeation of model drugs and the adhesion of muscle cells. We first defined the optimal growth conditions in terms of pH and alginate concentration. The membranes could be easily detached from polystyrene or polypropylene substrate without any postprocessing step. The dry thickness was varied over a large range from 4 to 35 μm. A 2-fold swelling was observed by confocal microscopy when they were immersed in PBS. In addition, we quantified the permeation of model drugs (fluorescent dextrans) through the free-standing membrane, which depended on the dextran molecular weight. Finally, we showed that myoblast cells exhibited a preferential adhesion on the alginate-ending membrane as compared to the chitosan-ending membrane or to the substrate side.This work was financially supported by Foundation for Science and Technology (FCT) through the Scholarship SFRH/BD/64601/2009 granted to S.G.C. C.M. is indebted to Grenoble INP for financial support via a postdoctoral fellowship. This work was supported by the European Commission (FP7 Program) via a European Research Council starting grant (BIOMIM, GA 259370 to C.P.). C.P. is also grateful to Institut Universitaire de France and to Grenoble Institute of Technology for financial support. We thank Isabelle Paintrand for her technical help with the confocal apparatus and Patrick Chaudouet for his help with SEM imaging

Bone Tumor Environment as a Potential Therapeutic Target in Ewing Sarcoma

Ewing sarcoma is the second most common pediatric bone tumor, with three cases per million worldwide. In clinical terms, Ewing sarcoma is an aggressive, rapidly fatal malignancy that mainly develops not only in osseous sites (85%) but also in extra-skeletal soft tissue. It spreads naturally to the lungs, bones, and bone marrow with poor prognosis in the two latter cases. Bone lesions from primary or secondary (metastases) tumors are characterized by extensive bone remodeling, more often due to osteolysis. Osteoclast activation and subsequent bone resorption are responsible for the clinical features of bone tumors, including pain, vertebral collapse, and spinal cord compression. Based on the "vicious cycle" concept of tumor cells and bone resorbing cells, drugs, which target osteoclasts, may be promising agents as adjuvant setting for treating bone tumors, including Ewing sarcoma. There is also increasing evidence that cellular and molecular protagonists present in the bone microenvironment play a part in establishing a favorable "niche" for tumor initiation and progression. The purpose of this review is to discuss the potential therapeutic value of drugs targeting the bone tumor microenvironment in Ewing sarcoma. The first part of the review will focus on targeting the bone resorbing function of osteoclasts by means of bisphosphonates or drugs blocking the pro-resorbing cytokine receptor activator of NF-kappa B ligand. Second, the role of this peculiar hypoxic microenvironment will be discussed in the context of resistance to chemotherapy, escape from the immune system, or neo-angiogenesis. Therapeutic interventions based on these specificities could be then proposed in the context of Ewing sarcoma

Cardiomyocytes from human pluripotent stem cells: from laboratory curiosity to industrial biomedical platform

Cardiomyocytes from human pluripotent stem cells (hPSCs-CMs) could revolutionise biomedicine. Global burden of heart failure will soon reach USD $90bn, while unexpected cardiotoxicity underlies 28% of drug withdrawals. Advances in hPSC isolation, Cas9/CRISPR genome engineering and hPSC-CM differentiation have improved patient care, progressed drugs to clinic and opened a new era in safety pharmacology. Nevertheless, predictive cardiotoxicity using hPSC-CMs contrasts from failure to almost total success. Since this likely relates to cell immaturity, efforts are underway to use biochemical and biophysical cues to improve many of the ~ 30 structural and functional properties of hPSC-CMs towards those seen in adult CMs. Other developments needed for widespread hPSC-CM utility include subtype specification, cost reduction of large scale differentiation and elimination of the phenotyping bottleneck. This review will consider these factors in the evolution of hPSC-CM technologies, as well as their integration into high content industrial platforms that assess structure, mitochondrial function, electrophysiology, calcium transients and contractility. This article is part of a Special Issue entitled: Cardiomyocyte Biology: Integration of Developmental and Environmental Cues in the Heart edited by Marcus Schaub and Hughes Abriel

The impact of natural and anthropogenic Dissolved Organic Carbon (DOC), and pH on the toxicity of triclosan to the crustacean Gammarus pulex (L.).

Regulatory ecotoxicology testing rarely accounts for the influence of natural water chemistry on the bioavailability and toxicity of a chemical. Therefore, this study identifies whether key omissions in relation to Dissolved Organic Carbon (DOC) and pH have an impact on measured effect concentrations (EC). Laboratory ecotoxicology tests were undertaken for the widely used antimicrobial compound triclosan, using adult Gammarus pulex (L.), a wild-type amphipod using synthetic fresh water, humic acid solutions and wastewater treatment works effluent. The toxicity of triclosan was tested at two different pHs of 7.3 and 8.4, with and without the addition of DOC and 24 and 48hour EC values with calculated 95% confidence intervals calculated. Toxicity tests undertaken at a pH above triclosan's pKa and in the presents of humic acid and effluent, containing 11 and 16mgL(-1) mean DOC concentrations respectively, resulted in significantly decreased triclosan toxicity. This was most likely a result of varying triclosan speciation and complexation due to triclosan's pKa and high hydrophobicity controlling its bioavailability. The mean 48hour EC50 values varied between 0.75±0.45 and 1.93±0.12mgL(-1) depending on conditions. These results suggest that standard ecotoxicology tests can cause inaccurate estimations of triclosan's bioavailability and subsequent toxicity in natural aquatic environments. These results highlight the need for further consideration regarding the role that water chemistry has on the toxicity of organic contaminants and how ambient environmental conditions are incorporated into the standard setting and consenting processes in the future

Une méthode innovante pour l identification des propriétés mécaniques d échantillons isotropes compressibles basée sur l aspiration par micropipette (Application à l étude des gels de polyacrylamide dans des domaines linéaire et non-linéaire de déformations )

Nous avons cherché ici à développer une technique permettant de caractériser la rhéologie d'échantillons biologiques: l'aspiration par micropipette. Parallèlement à la mise en œuvre de l'expérience, nous avons développé un modèle théorique de l'enpérience. Ainsi, et grâce à la mise en place d'une modélisation aux éléments finis, nous avons réussi à proposer une méthode originale permettant de quantifier les propriétés mécaniques d'échantillons biologiques isotropes. Afin de valider cette méthode, nous nous sommes intéressé à des substrats cellulaires bien connus en biologie, les gels de polyacrylamide. Ces polymères sont en effet régulièrement utilisés afin d'étudier l'influence de la rhéologie du substrat sur les processus cellulaires ou pour mesurer les forces de traction cellulaires. Il existait donc le besoin fort d'une quantification précise des propriétés mécaniques de ces gels auquel nous avons répondu en établissant leur quasi-incompressibilité et en proposant une relation entre rigidité et concentrations initiales en monomères.Ayant observé que ces gels pouvaient présenter un comportement non-linéaire lorsqu'ils étaient soumis à de fortes aspirations, nous avons étendu notre modèle numérique et nous avons réussi à développer une méthode de caractérisation des propriétés hyperélastiques non-linéaires grâce à laquelle nous avons caractérisé le comportement rhéologique de ces gels soumis à de grandes déformations.En étudiant l'importance de ce comportement sur des mesures de forces de traction cellulaires précédemment publiées, nous avons observé des écarts de près de 50% lorsque le comportement non-linéaire était ou non pris en compte.The quantification of the mechanical properties of biological samples is a key issue for the modern biology and we focused here on extending the micropipette aspiration technique to characterize the rheology of biological samples. Using a finite element analysis of the sample aspiration into the micropipette, we developed an original method for simultaneously quantifying rigidity and compressibility of isotropic biological samples. ln order to validate this mehod, experiments were conducted on adherent polyacrylamide gels, polymers whieh are frequently used in biology for studying the influence of the substrate rheology on cell processes or for measuring cell traction forces. There was thus a strong need for an accurate characterization of their mechanieal properties. We established here the quasi-incompressibility of these gels and we proposed a relationship between their rigidity and the initial concentrations of monomers.Moreover, we observed a non-linear behaviour of these polymers when they were submitted to large pressures. We extended our previous numerical model and we succeed at developing a new method of quantification of the hyperelastic non-linear properties, which was used to fully characterize the rheological behaviour of these polyacrylamide gels submitted to large deformations. As an illustration of the implications of our results, we considered the quantification of cell traction stresses obtained b traction force microscopy methods. We thus observed significant differences when considering polyacrylamide gels either as linear elastic or nonlinear hyperelastic media.GRENOBLE1-BU Sciences (384212103) / SudocSudocFranceF

Polyelectrolyte Multilayer Films Based on Polysaccharides: From Physical Chemistry to the Control of Cell Differentiation

International audienc