First phase microcirculatory reaction to mechanical skin irritation: a three layer model of a compliant vascular tree

Mechanical skin irritation creates vasodilatation in the line of a stroke and in the surrounding tissue. To obtain further insight on underlying physiological mechanisms we developed a model of the vascular network comprised of three layers, where the first and the last one have a tree structure. They represent the arterial and the venous system, respectively. Both are connected via an intermediate zone representing the core of the microcirculation, which is described by means of a compliant compartment model. Irritation induces change in compliance of vessels situated at the entrance of the intermediate zone. Thus the model describes flow and pressure behavior due to mechanical skin irritation.     

Enhanced film forming and film properties of amylopectin using micro-fibrillated cellulose

This work describes a novel approach to produce amylopectin films with enhanced properties by the addition of microfibrillated cellulose (MFC). Aqueous dispersions of gelatinized amylopectin, glycerol (0–38 wt%) and MFC (0–10 wt%) were cast at ambient temperature and 50% relative humidity and, after 10 days of storage, the tensile properties were investigated. The structure of the composite films was revealed by optical, atomic force and transmission electron microscopy. The moisture content was determined by thermogravimetry and the temperature-dependent film rigidity was measured by thermal mechanical analysis. Synchrotron simultaneous small- and wide-angle X-ray measurements revealed that the solutions had to be heated to above 85 °C in order to achieve complete gelatinization. Optical microscopy and atomic force microscopy revealed uniformly distributed MFC aggregates in the films, with a length of 10–90 μm and a width spanning from a few hundred nanometers to several microns. Transmission electron microscopy showed that, in addition to aggregates, single MFC microfibrils were also embedded in the amylopectin matrix. It was impossible to cast amylopectin films of sufficient quality with less than 38 wt% glycerol. However, when MFC was added it was possible to produce high quality films even without glycerol. The film without glycerol was stiff and strong but not brittle. It was suggested that this remarkable effect was due to its comparatively high moisture content. Consequently MFC acted both as a “conventional” reinforcement because of its fibrous structure and also indirectly as a plasticiser because its presence led to an increase in film moisture content.     

The effects of Achilles tendon compliance on triceps surae mechanics and energetics in walking

Achilles tendon (AT) compliance can affect the generation and transmission of triceps surae muscle forces, and thus has important biomechanical consequences for walking performance. However, the uniarticular soleus (SOL) and the biarticular (GAS) function differently during walking, with in vivo evidence suggesting that their associated fascicles and tendinous structures exhibit unique kinematics during walking. Given the strong association between muscle fiber length, velocity and force production, we conjectured that SOL and GAS mechanics and energetic behavior would respond differently to altered AT compliance. To test this, we characterized GAS and SOL muscle and tendon mechanics and energetics due to systematic changes in tendon compliance using musculoskeletal simulations of walking. Increased tendon compliance enlarged GAS and SOL tendon excursions, shortened fiber operation lengths and affected muscle excitation patterns. For both muscles, an optimal tendon compliance (tendon strains of approximately 5% with maximum isometric force) existed that minimized metabolic energy consumption. However, GAS muscle-tendon mechanics and energetics were significantly more sensitive to changes in tendon compliance than were those for SOL. In addition, GAS was not able to return stored tendon energy during push-off as effectively as SOL, particularly for larger values of tendon compliance. These fundamental differences between GAS and SOL sensitivity to altered tendon compliance seem to arise from the biarticular nature of GAS. These insights are potentially important for understanding the functional consequences of altered Achilles tendon compliance due to aging, injury, or disease.     

Spatial development of gingival fibroblasts and dental pulp cells: Effect of extracellular matrix

Cells sensing changes in their microenvironmental stiffness and composition alter their responses, accordingly. This study determines whether gingival fibroblasts (GFs) and dental pulp mesenchymal stem cells (DPMSCs) support the formation of continuous layers in vitro by mimicking the stiffness and protein composition of their native extracellular matrix (ECM). Immortalized cells were incubated with (i) 0–100% Matrigel-ECM (M-ECM) for 7-28d, and with (ii) collagen and fibrin matrices for 14d. Cultures were analyzed by phase-contrast, fluorescence and confocal microscopies. The diameters and surface areas were measured via ImageJ. Self-renewal markers were detected by RT-PCR and immunocytochemistry assays. GFs and DPMSCs developed spheroids interconnected by elongated cell bundles or layers, respectively, expressing the self-renewal markers. Increased matrix stiffness resulted in spheroids replacement by the interconnecting cells/layers. Both cells required 100% M-ECM to reduce their spheroid diameter. However, it reduced the surface area of the interconnecting layers. Those differences led to extended, spindle-shaped GFs vs. compact, ring-shaped DPMSCs constructs. Collagen and fibrin matrices developed continuous layers of tightly connected cells vs. distinctive scattered cell aggregates, respectively. The ability of GFs and DPMSCs to create tissue-like multicellular layers at various matrix conditions may be imprinted by cells’ adaptation to mechanical forces and composition in vivo.     

Esterification of cellulose-enriched agricultural by-products and characterization of mechanical properties of cellulosic films

Two agricultural by-products, wheat bran and maize bran have been examined for their suitability to be transformed into biomaterials by esterification by lauroyl chloride. Influence of biochemical characteristic of cellulose (cellulose content, viscosity-average degree of polymerization, crystallinity) was studied on eight samples enriched in cellulose after chemical removal of heteroxylans and lignin. After an acidic pre-treatment, esterification was carried out with lauroyl chloride and an optimized reaction time of 8 h was used.

Chemical compositions were similar for all cellulose esters obtained, but cellulose content of initial material had a marked influence on the amount of esterified product. A film was easily obtained by casting and the mechanical (tensile strength and elongation), thermomechanical and calorimetric properties were determined. The possible role of grafted fatty acid as internal plasticizer was finally discussed.     

Current strategies of mechanical stimulation for maturation of cardiac microtissues

The most advanced in vitro cardiac models are today based on the use of induced pluripotent stem cells (iPSCs); however, the maturation of cardiomyocytes (CMs) has not yet been fully achieved. Therefore, there is a rising need to move towards models capable of promoting an adult-like cardiomyocytes phenotype. Many strategies have been applied such as co-culture of cardiomyocytes, with fibroblasts and endothelial cells, or conditioning them through biochemical factors and physical stimulations. Here, we focus on mechanical stimulation as it aims to mimic the different mechanical forces that heart receives during its development and the post-natal period. We describe the current strategies and the mechanical properties necessary to promote a positive response in cardiac tissues from different cell sources, distinguishing between passive stimulation, which includes stiffness, topography and static stress and active stimulation, encompassing cyclic strain, compression or perfusion. We also highlight how mechanical stimulation is applied in disease modelling.     

Role of Ca2+ signaling in initiation of stretch-induced apoptosis in neonatal heart cells

Abnormal mechanical load, as seen in hypertension, is found to induce heart cell apoptosis, yet the signaling link between cell stretch and apoptotic pathways is not known. Using an in vitro stretch model mimicking diastolic pressure stress, here we show that Ca(2+) signaling participates essentially in the early stage of stretch-induced apoptosis. In neonatal rat cardiomyocytes, the moderate 20% stretch resulted in tonic elevation of intracellular free Ca(2+) ([Ca(2+)](i)). Buffering [Ca(2+)](i) by EGTA-AM, suppressing ryanodine-sensitive Ca(2+) release, and blocking L-type Ca(2+) channels all prevented the stretch-induced apoptosis as assessed by phosphatidylserine exposure and nuclear fragmentation. Notably, Ca(2+) suppression also prevented known stretch-activated apoptotic events, including caspase-3/-9 activation, mitochondrial membrane potential corruption, and reactive oxygen species production, suggesting that Ca(2+) signaling is the upstream of these events. Since [Ca(2+)](i) did not change without activating mechanosensitive Ca(2+) entry, we conclude that stretch-induced Ca(2+) entry, via the Ca(2+)-induced Ca(2+) release mechanism, plays an important role in initiating apoptotic signaling during mechanical stress.     

提高人胚胎干细胞建系率的优选培养体系与方法

目的:建立一种从废弃胚胎中提高囊胚形成率和质量的培养体系,寻找多种促进内细胞团(ICM)数目增多、贴壁、增值的方法,提高人胚胎干细胞(human embryonic stem cell,hESC)建系效率,建立人胚胎干细胞库。方法:将179枚IVFDay3废弃的胚胎放入优选培养体系中培养(G2.5培养液中添加10%人血清蛋白,人白细胞抑制生长因子(hLIF),碱性成纤维细胞生长因子(bFGF))。到Day7将形成的囊胚全部用机械法分离ICM,接种于丝裂霉素C灭活处理的原代小鼠胚胎成纤维细胞(MEF)上,培养8-9天,每4-5天传代1次。结果:优选培养体系的囊胚形成率为29.1%(52/179),其中A级囊胚形成率为11.2%(20/179),50个ICM贴壁生长,20个出现克隆形态,成功建立11株hESC(FY-hES-11至FY-hES-21)。11株hESC均具有共同的多能性生物学特性。结论:优选培养体系可以明显提高囊胚形成的质量,促进ICM的增值,纯熟的机械切割法可以避免损伤ICM并提高其贴壁率,原代灭活的MEF饲养层可以明显促进细胞增殖。     

Investigation of Early Plasma Evolution Induced by Ultrashort Laser Pulses

Early plasma is generated owing to high intensity laser irradiation of target and the subsequent target material ionization. Its dynamics plays a significant role in laser-material interaction, especially in the air environment^1-11.Early plasma evolution has been captured through pump-probe shadowgraphy^1-3 and interferometry^1,4-7. However, the studied time frames and applied laser parameter ranges are limited. For example, direct examinations of plasma front locations and electron number densities within a delay time of 100 picosecond (ps) with respect to the laser pulse peak are still very few, especially for the ultrashort pulse of a duration around 100 femtosecond (fs) and a low power density around 10¹⁴ W/cm². Early plasma generated under these conditions has only been captured recently with high temporal and spatial resolutions¹². The detailed setup strategy and procedures of this high precision measurement will be illustrated in this paper. The rationale of the measurement is optical pump-probe shadowgraphy: one ultrashort laser pulse is split to a pump pulse and a probe pulse, while the delay time between them can be adjusted by changing their beam path lengths. The pump pulse ablates the target and generates the early plasma, and the probe pulse propagates through the plasma region and detects the non-uniformity of electron number density. In addition, animations are generated using the calculated results from the simulation model of Ref. ¹² to illustrate the plasma formation and evolution with a very high resolution (0.04 ~ 1 ps).Both the experimental method and the simulation method can be applied to a broad range of time frames and laser parameters. These methods can be used to examine the early plasma generated not only from metals, but also from semiconductors and insulators.     

Role of vesicular nucleotide transporter VNUT (SLC17A9) in release of ATP from AR42J cells and mouse pancreatic acinar cells

ATP is released from cells in response to various stimuli. Our previous studies on pancreas indicated that pancreatic acini could be major stores of secreted ATP. In the present study, our aim was to establish the role of the vesicular nucleotide transporter (VNUT), SLC17A9, in storage and release of ATP. Freshly prepared acini from mice and AR42J rat acinar cells were used in this study. We illustrate that in AR42J cells, quinacrine (an ATP store marker) and Bodipy ATP (a fluorescent ATP analog) co-localized with VNUT-mCherry to vesicles/granules. Furthermore, in acini and AR42J cells, a marker of the zymogen granule membranes, Rab3D, and VNUT co-localized. Dexamethasone treatment of AR42J cells promoted formation of acinar structures, paralleled by increased amylase and VNUT expression, and increased ATP release in response to cholinergic stimulation. Mechanical stimulus (pressure) and cell swelling also induced ATP release, but this was not influenced by dexamethasone, most likely indicating different non-zymogen-related release mechanism. In conclusion, we propose that VNUT-dependent ATP release pathway is associated with agonist-induced secretion process and downstream purinergic signalling in pancreatic ducts.