Mouse Lung Organoid Responses to Reduced, Increased, and Cyclic Stretch

Most lung development occurs in the context of cyclic stretch. Alteration of the mechanical microenvironment is a common feature of many pulmonary diseases with congenital diaphragmatic hernia (CDH) and fetal tracheal occlusion (FETO, a therapy for CDH) being extreme examples with changes in lung structure, cell differentiation and function. To address limitations in cell culture and in vivo mechanotransductive models we developed two mouse lung organoid (mLO) mechanotransductive models using postnatal day 5 (PND5) mouse lung CD326-positive cells and fibroblasts subjected to increased, decreased, and cyclic strain. In the first model, mLOs were exposed to forskolin (FSK) and/or disrupted (DIS) and evaluated at 20 hours. mLO cross-sectional area changed by +59%, +24% and -68% in FSK, control, and DIS mLOs respectively. FSK-treated organoids had twice as many proliferating cells as other organoids. In the second model, 20 hours of 10.25% biaxial cyclic strain increased the mRNAs of lung mesenchymal cell lineages compared to static stretch and no stretch. Cyclic stretch increased TGF-β and integrin-mediated signaling with upstream analysis indicating roles for histone deacetylases, microRNAs, and long non-coding RNAs. Cyclic stretch mLOs increased αSMA- and αSMA-PDGFRα-double positive cells compared to no stretch and static stretch mLOs. In this PND5 mLO mechanotransductive model, cell proliferation is increased by static stretch, and cyclic stretch induces mesenchymal gene expression changes important in postnatal lung development.

The Effect of Static Stretch and Dynamic Range of Motion Training on the Flexibility of the Hamstring Muscle

Introduction: A normal flexibility is required for an appropriate posture and entire activity in daily life. Hamstring is the muscle that is more prone for tightness. It present in all age groups and it increases with age and more over it is the muscle that is most prone to injuries during sporting activities. With regard to various methods that contribute to increasing flexibility of hamstring muscles, the current study aimed at investigating immediate effects of static stretching and dynamic range of motion training on hamstring flexibility increasing muscle flexibility. Methodology: 30 patients, age range 21-50 years, diagnosed with the Population who had hamstring tightness and divided equally into two groups-Group A and Group B .In Group B-Static Stretch exercise and in Group A-Dynamic Range of Motion Training exercise. Result: Pre and post evaluation of active knee extension test was taken. Result shows that there was significant improvement noted in both the groups (GROUP A: P value .000, t value:-17.9) and (GROUP B: P value: .000, t value:-17.6) Conclusion: Both the static stretch and dynamic exercise can be considered as an effective method for increasing flexibility and improving functions with hamstring tightness patients. Key words: Hamstring flexibility, Dynamic range of motion, and static stretch.

Modeling and drafting process simulation of cotton slivers based on an octahedron structure

In order to study the variation of the drafting force of cotton slivers, a three-dimensional model of cotton slivers was proposed. The model is based on the three-dimensional network structure of the fibers in the cotton sliver. The three-dimensional network structure is simulated by an octahedron. Based on the similarity between dynamic drawing and static drawing, the static drawing simulation of the model is carried out by using ANSYS software, and the static drawing force of different quantitative cotton slivers is simulated. The results show that the average relative error of the static stretch force and dynamic drafting force is 8.09%, and the maximum relative error is less than 15%. Then, the equations of the dynamic drafting force and static stretch force are obtained by linear regression, and the drawing force under other quantitative conditions is successfully predicted. Finally, static stretching is used to simulate the influence curve of different roller spacings on the dynamic drafting force, and the results show that the simulation results are consistent with the actual situation. Therefore, the octahedral cotton sliver model is effective, and the simulation results also provide a reference for the approximate prediction of dynamic drafting force.

Crosstalk Between CD11b and Piezo1 Mediates Macrophage Responses to Mechanical Cues

Macrophages are versatile cells of the innate immune system that perform diverse functions by responding to dynamic changes in their microenvironment. While the effects of soluble cues, including cytokines and chemokines, have been widely studied, the effects of physical cues, including mechanical stimuli, in regulating macrophage form and function are less well understood. In this study, we examined the effects of static and cyclic uniaxial stretch on macrophage inflammatory and healing activation. We found that cyclic stretch altered macrophage morphology and responses to IFNγ/LPS and IL4/IL13. Interestingly, we found that both static and cyclic stretch suppressed IFNγ/LPS induced inflammation. In contrast, IL4/IL13 mediated healing responses were suppressed with cyclic but enhanced with static stretch conditions. Mechanistically, both static and cyclic stretch increased expression of the integrin CD11b (αM integrin), decreased expression of the mechanosensitive ion channel Piezo1, and knock down of either CD11b or Piezo1 through siRNA abrogated stretch-mediated changes in inflammatory responses. Moreover, we found that knock down of CD11b enhanced the expression of Piezo1, and conversely knock down of Piezo1 enhanced CD11b expression, suggesting the potential for crosstalk between integrins and ion channels. Finally, stretch-mediated differences in macrophage activation were also dependent on actin, since pharmacological inhibition of actin polymerization abrogated the changes in activation with stretch. Together, this study demonstrates that the physical environment synergizes with biochemical cues to regulate macrophage morphology and function, and suggests a role for CD11b and Piezo1 crosstalk in mechanotransduction in macrophages.

Spontaneous myogenic fasciculation associated with the lengthening of cardiac muscle in response to static preloading

Force enhancement is one kind of myogenic spontaneous fasciculation in lengthening preload striated muscles. In cardiac muscle, the role of this biomechanical event is not well established. The physiological passive property is an essential part for maintaining normal diastole in the heart. In excessive preload heart, force enhancement relative erratic passive properties may cause muscle decompensating, implicate in the development of diastolic dysfunction. In this study, the force enhancement occurrence in mouse cardiac papillary muscle was evaluated by a microstepping stretch method. The intracellular Ca2+ redistribution during occurrence of force enhancement was monitored in real-time by a Flou-3 (2 mM) indicator. The force enhancement amplitude, the enhancement of the prolongation time, and the tension–time integral were analyzed by myography. The results indicated that the force enhancement occurred immediately after active stretching and was rapidly enhanced during sustained static stretch. The presence of the force and the increase in the amplitude synchronized with the acquisition and immediate transfer of Ca2+ to adjacent fibres. In highly preloaded fibres, the enhancement exceeded the maximum passive tension (from 4.49 ± 0.43 N/mm2 to 6.20 ± 0.51 N/mm2). The occurrence of force enhancement were unstable in each static stretch. The increased enhancement amplitude combined with the reduced prolongation time to induce a reduction in the tension–time integral. We concluded that intracellular Ca2+-synchronized force enhancement is one kind of interruption event in excessive preload cardiac muscle. During the cardiac muscle in its passive relaxation period, the occurrence of this interruption affected the rhythmic stability of the cardiac relaxation cycle.

Changes in Spinal-Reflex Excitability during Static Stretch and/or Explosive Contraction

To examine individual or combined effects of static stretch and explosive contraction on quadriceps spinal-reflex excitability (the peak Hoffmann’s reflex normalized by the peak motor-response) and the latency times of the Hoffmann’s reflex and motor-response. Fourteen healthy young males randomly experienced four conditions (stretch, contraction, stretch + contraction, and control—no intervention). For the stretch condition, three sets of a 30 s hold using the modified Thomas test on each leg were performed. For the contraction condition, three trials of maximal countermovement vertical jump were performed. Quadriceps spinal-reflex excitability and the latent period of each value on the right leg were compared at pre- and post-condition. All measurement values across conditions were not changed at any time point (condition × time) in spinal-reflex excitability (F6,143 = 1.10, p = 0.36), Hoffmann’s reflex latency (F6,143 = 0.45, p = 0.84), motor-response latency (F6,143 = 0.37, p = 0.90), and vertical jump heights (F2,65 = 1.82, p = 0.17). A statistical trend was observed in the contraction condition that spinal-reflex excitability was increased by 42% (effect size: 0.63). Neither static stretch nor explosive contraction changed the quadriceps spinal-reflex excitability, latency of Hoffmann’s reflex, and motor-response. Since our stretch protocol did not affect jumping performance and our contraction protocol induced the post-activation potentiation effect, either protocol could be used as pre-exercise activity.

The static preload associated myogenic spontaneous fasciculation response in lengthening cardiac muscle

The passive tension force enhancement is one kind of myogenic spontaneous fasciculation in muscles. However, its physiological properties in cardiac fibres are not well known. In this study, mice cardiac papillary muscle spontaneous force enhancement was evaluated by micro stepping stretch method. The occurrence of spontaneous force and real time cardiac fibre Ca2+ redistribution was tranced by Flou-3 (2mM) indicator. Force enhancement amplitude, enhancement prolonging time, and tension–time integral were analysis by myograph analyser. The results indicated that the spontaneous force occurred immediately after the active stretch, rapidly enhanced during tolerating the sustained static stretch. The force occurrence and amplitude enhance synchronized with the Ca2+ recruitment and lightning transmitted to adjacent fibres. In high preload fibres, the enhancement was forceful to over its maximum passive tension (6.20 ± 0.51 N/mm2 to 4.49 ± 0.43 N/mm2). The force occurrences were unsteadiness in each stretch. The increased enhancement amplitude combining with the shortening prolonging time induced reduction of tension–time integral. We concluded that the intracellular Ca2+ synchronized force enhancement is one kind of interruption event in overloading cardiac fibres. This interruption occurred during the relaxation processing in cardiac muscle, therefore affect the rhythmic stability of cardiac relaxation-contraction cycle.

Design of a 3D printed, motorized, uniaxial cell stretcher for microscopic and biochemical analysis of mechanotransduction

ABSTRACTCells respond to mechanical cues from their environment through a process of mechanosensing and mechanotransduction. Cell stretching devices are important tools to study the molecular pathways responsible for cellular responses to mechanobiological processes. We describe the development and testing of a uniaxial cell stretcher that has applications for microscopic as well as biochemical analyses. By combining simple fabrication techniques with adjustable control parameters, the stretcher is designed to fit a variety of experimental needs. The stretcher can be used for static and cyclic stretching. As a proof of principle, we visualize stretch induced deformation of cell nuclei via incremental static stretch, and changes in IEX1 expression via cyclic stretching. This stretcher is easily modified to meet experimental needs, inexpensive to build, and should be readily accessible for most laboratories with access to 3D printing.

The tolerance to stretch is linked with endogenous modulation of pain

Objectives The effect of stretching on joint range of motion is well documented, and although sensory perception has significance for changes in the tolerance to stretch following stretching the underlining mechanisms responsible for these changes is insufficiently understood. The aim of this study was to examine the influence of endogenous pain inhibitory mechanisms on stretch tolerance and to investigate the relationship between range of motion and changes in pain sensitivity. Methods Nineteen healthy males participated in this randomized, repeated-measures crossover study, conducted on 2 separate days. Knee extension range of motion, passive resistive torque, and pressure pain thresholds were recorded before, after, and 10 min after each of four experimental conditions; (i) Exercise-induced hypoalgesia, (ii) two bouts of static stretching, (iii) resting, and (iv) a remote, painful stimulus induced by the cold pressor test. Results Exercise-induced hypoalgesia and cold pressor test caused an increase in range of motion (p<0.034) and pressure pain thresholds (p<0.027). Moderate correlations in pressure pain thresholds were found between exercise-induced hypoalgesia and static stretch (Rho>0.507, p=0.01) and exercise-induced hypoalgesia and the cold pressor test (Rho=0.562, p=0.01). A weak correlation in pressure pain thresholds and changes in range of motion were found following the cold pressor test (Rho=0.460, p=0.047). However, a potential carryover hypoalgesic effect may have affected the results of the static stretch. Conclusions These results suggest that stretch tolerance may be linked with endogenous modulation of pain. Present results suggest, that stretch tolerance may merely be a marker for pain sensitivity which may have clinical significance given that stretching is often prescribed in the rehabilitation of different musculoskeletal pain conditions where reduced endogenous pain inhibition is frequently seen.