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Technical feasibility of constant-load and high-intensity interval training for cardiopulmonary conditioning using a re-engineered dynamic leg press. 使用重新设计的动态压腿机进行恒定负荷和高强度间歇训练以促进心肺功能调节的技术可行性。
Pub Date : 2019-10-03 eCollection Date: 2019-01-01 DOI: 10.1186/s42490-019-0025-9
Farouk Chrif, Tobias Nef, Kenneth J Hunt

Background: Leg-press devices are one of the most widely used training tools for musculoskeletal strengthening of the lower-limbs, and have demonstrated important cardiopulmonary benefits for healthy and patient populations. Further engineering development was done on a dynamic leg-press for work-rate estimation by integrating force and motion sensors, power calculation and a visual feedback system for volitional work-rate control. This study aimed to assess the feasibility of the enhanced dynamic leg press for cardiopulmonary exercise training in constant-load training and high-intensity interval training. Five healthy participants aged 31.0±3.9 years (mean ± standard deviation) performed two cardiopulmonary training sessions: constant-load training and high-intensity interval training. Participants carried out the training sessions at a work rate that corresponds to their first ventilatory threshold for constant-load training, and their second ventilatory threshold for high-intensity interval training.

Results: All participants tolerated both training protocols, and could complete the training sessions with no complications. Substantial cardiopulmonary responses were observed. The difference between mean oxygen uptake and target oxygen uptake was 0.07±0.34 L/min (103 ±17%) during constant-load training, and 0.35±0.66 L/min (113 ±27%) during high-intensity interval training. The difference between mean heart rate and target heart rate was -7±19 bpm (94 ±15%) during constant-load training, and 4.2±16 bpm (103 ±12%) during high-intensity interval training.

Conclusions: The enhanced dynamic leg press was found to be feasible for cardiopulmonary exercise training, and for exercise prescription for different training programmes based on the ventilatory thresholds.

背景:压腿装置是最广泛使用的下肢肌肉骨骼强化训练工具之一,对健康人群和病人的心肺功能有重要益处。通过整合力和运动传感器、功率计算和视觉反馈系统,我们对动态压腿器进行了进一步的工程开发,以估算工作速率。本研究旨在评估增强型动态压腿器在恒定负荷训练和高强度间歇训练中用于心肺运动训练的可行性。五名年龄为 31.0±3.9 岁(平均值±标准差)的健康参与者进行了两次心肺训练:恒定负荷训练和高强度间歇训练。在进行恒定负荷训练时,参与者的工作速率与他们的第一通气阈值相对应;在进行高强度间歇训练时,参与者的工作速率与他们的第二通气阈值相对应:结果:所有参与者都能耐受两种训练方案,并能顺利完成训练。观察到了显著的心肺反应。在恒定负荷训练中,平均摄氧量与目标摄氧量之差为 0.07±0.34 升/分钟(103 ±17%);在高强度间歇训练中,平均摄氧量与目标摄氧量之差为 0.35±0.66 升/分钟(113 ±27%)。在恒定负荷训练中,平均心率与目标心率的差值为-7±19 bpm(94±15%);在高强度间歇训练中,平均心率与目标心率的差值为4.2±16 bpm(103±12%):结论:研究发现,增强型动态压腿对心肺运动训练和根据通气阈值为不同训练计划制定运动处方是可行的。
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引用次数: 0
Detection of stretch reflex onset based on empirical mode decomposition and modified sample entropy. 基于经验模式分解和修正样本熵的拉伸反射起始检测。
Pub Date : 2019-09-26 eCollection Date: 2019-01-01 DOI: 10.1186/s42490-019-0023-y
Mingjia Du, Baohua Hu, Feiyun Xiao, Ming Wu, Zongjun Zhu, Yong Wang

Background: Accurate spasticity assessment provides an objective evaluation index for the rehabilitation treatment of patients with spasticity, and the key is detecting stretch reflex onset. The surface electromyogram of patients with spasticity is prone to false peaks, and its data length is unstable. These conditions decrease signal differences before and after stretch reflex onset. Therefore, a method for detecting stretch reflex onset based on empirical mode decomposition denoising and modified sample entropy recognition is proposed in this study.

Results: The empirical mode decomposition algorithm is better than the wavelet threshold algorithm in denoising surface electromyogram signal. Without adding Gaussian white noise to the electromyogram signal, the stretch reflex onset recognition rate of the electromyogram signal before and after empirical mode decomposition denoising was increased by 56%. In particular, the recognition rate of stretch reflex onset under the optimal parameter of the modified sample entropy can reach up to 100% and the average recognition rate is 93%.

Conclusions: The empirical mode decomposition algorithm can eliminate the baseline activity of the surface electromyogram signal before stretch reflex onset and effectively remove noise from the signal. The identification of stretch reflex onset using combined empirical mode decomposition and modified sample entropy is better than that via modified sample entropy alone, and stretch reflex onset can be accurately determined.

背景:准确的痉挛评估为痉挛患者的康复治疗提供了客观的评价指标,而关键在于检测伸展反射的发生。痉挛患者的表面肌电图容易出现假峰值,而且数据长度不稳定。这些情况会降低拉伸反射开始前后的信号差异。因此,本研究提出了一种基于经验模式分解去噪和修正样本熵识别的拉伸反射起始检测方法:结果:在表面肌电信号去噪方面,经验模式分解算法优于小波阈值算法。在不添加高斯白噪声的情况下,经验模式分解去噪前后肌电信号的拉伸反射发病识别率提高了 56%。其中,在修正样本熵的最优参数下,拉伸反射起始点的识别率可达 100%,平均识别率为 93%:结论:经验模式分解算法可以消除拉伸反射开始前表面肌电信号的基线活动,有效去除信号中的噪声。结论:经验模式分解算法能消除拉伸反射开始前的表面肌电信号基线活动,有效去除信号中的噪声,使用经验模式分解和修正样本熵相结合的方法识别拉伸反射开始比单独使用修正样本熵的方法识别效果更好,能准确判断拉伸反射开始。
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引用次数: 0
Human motor decoding from neural signals: a review. 从神经信号解码人类运动:综述。
Pub Date : 2019-09-03 eCollection Date: 2019-01-01 DOI: 10.1186/s42490-019-0022-z
Wing-Kin Tam, Tong Wu, Qi Zhao, Edward Keefer, Zhi Yang

Many people suffer from movement disability due to amputation or neurological diseases. Fortunately, with modern neurotechnology now it is possible to intercept motor control signals at various points along the neural transduction pathway and use that to drive external devices for communication or control. Here we will review the latest developments in human motor decoding. We reviewed the various strategies to decode motor intention from human and their respective advantages and challenges. Neural control signals can be intercepted at various points in the neural signal transduction pathway, including the brain (electroencephalography, electrocorticography, intracortical recordings), the nerves (peripheral nerve recordings) and the muscles (electromyography). We systematically discussed the sites of signal acquisition, available neural features, signal processing techniques and decoding algorithms in each of these potential interception points. Examples of applications and the current state-of-the-art performance were also reviewed. Although great strides have been made in human motor decoding, we are still far away from achieving naturalistic and dexterous control like our native limbs. Concerted efforts from material scientists, electrical engineers, and healthcare professionals are needed to further advance the field and make the technology widely available in clinical use.

许多人由于截肢或神经系统疾病而患有运动障碍。幸运的是,随着现代神经技术的发展,现在有可能在神经转导通路的各个点拦截运动控制信号,并利用这些信号驱动外部设备进行通信或控制。在此,我们将回顾人类运动解码的最新进展。本文综述了人类运动意向解码的各种策略,以及各自的优势和挑战。神经控制信号可以在神经信号转导通路的各个点上被截获,包括大脑(脑电图、皮质电图、皮质内记录)、神经(周围神经记录)和肌肉(肌电图)。我们系统地讨论了每个潜在截获点的信号采集位置、可用的神经特征、信号处理技术和解码算法。还审查了应用实例和目前最先进的性能。尽管在人类运动解码方面已经取得了巨大的进步,但我们离实现像我们天生的四肢那样自然而灵巧的控制还很遥远。需要材料科学家、电气工程师和医疗保健专业人员共同努力,进一步推动该领域的发展,并使该技术广泛应用于临床。
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引用次数: 44
User perspective and higher cognitive task-loads influence movement and performance in immersive training environments. 用户视角和更高的认知任务负荷影响沉浸式训练环境中的运动和表现。
Pub Date : 2019-08-30 eCollection Date: 2019-01-01 DOI: 10.1186/s42490-019-0021-0
Juan Trelles Trabucco, Andrea Rottigni, Marco Cavallo, Daniel Bailey, James Patton, G Elisabeta Marai

Background: In virtual reality (VR) applications such as games, virtual training, and interactive neurorehabilitation, one can employ either the first-person user perspective or the third-person perspective to perceive the virtual environment; however, applications rarely offer both perspectives for the same task. We used a targeted-reaching task in a large-scale virtual reality environment (N=30 healthy volunteers) to evaluate the effects of user perspective on the head and upper extremity movements, and on user performance. We further evaluated how different cognitive challenges would modulate these effects. Finally, we obtained the user-reported engagement level under the different perspectives.

Results: We found that first-person perspective resulted in larger head movements (3.52±1.3m) than the third-person perspective (2.41±0.7m). First-person perspective also resulted in more upper-extremity movement (30.08±7.28m compared to 26.66±4.86m) and longer completion times (61.3±16.4s compared to 53±10.4s) for more challenging tasks such as the "flipped mode", in which moving one arm causes the opposite virtual arm to move. We observed no significant effect of user perspective alone on the success rate. Subjects reported experiencing roughly the same level of engagement in both first-person and third-person perspectives (F(1.58)=0.9,P=.445).

Conclusion: User perspective and its interaction with higher-cognitive load tasks influences the extent of movement and user performance in a virtual theater environment, and may influence the choice of the interface type (first or third person) in immersive training depending on the user conditions and exercise requirements.

背景:在虚拟现实(VR)应用中,如游戏、虚拟训练和交互式神经康复,人们可以采用第一人称视角或第三人称视角来感知虚拟环境;然而,应用程序很少为同一任务提供两种透视图。我们在一个大规模的虚拟现实环境中(N=30名健康志愿者)使用了一个目标到达任务来评估用户视角对头部和上肢运动以及用户表现的影响。我们进一步评估了不同的认知挑战如何调节这些影响。最后,我们获得了不同视角下的用户报告粘性水平。结果:我们发现第一人称视角的头部运动(3.52±1.3m)大于第三人称视角(2.41±0.7m)。第一人称视角还导致上肢运动更多(30.08±7.28米比26.66±4.86米),完成更具有挑战性的任务(如“翻转模式”,移动一只手臂会导致另一只虚拟手臂移动)所需的时间更长(61.3±16.4秒比53±10.4秒)。我们观察到用户视角对成功率没有显著影响。受试者报告在第一人称和第三人称视角下的参与度大致相同(F(1.58)=0.9,P=.445)。结论:用户视角及其与高认知负荷任务的交互影响虚拟剧场环境中的运动程度和用户表现,并可能根据用户条件和锻炼要求影响沉浸式训练中界面类型(第一人称或第三人称)的选择。
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引用次数: 2
Treadmill training augmented with real-time visualisation feedback and function electrical stimulation for gait rehabilitation after stroke: a feasibility study. 跑步机训练增强实时可视化反馈和功能电刺激中风后的步态康复:可行性研究。
Pub Date : 2019-08-22 eCollection Date: 2019-01-01 DOI: 10.1186/s42490-019-0020-1
Chanwit Phongamwong, Philip Rowe, Karen Chase, Andrew Kerr, Lindsay Millar

Background: Stroke rehabilitation often uses the motor relearning concept that require patients to perform active practice of skill-specific training and to receive feedback. Treadmill training augmented with real-time visualisation feedback and functional electrical stimulation may have a beneficial synergistic effect on motor recovery. This study aims to determine the feasibility of this kind of enhanced treadmill training for gait rehabilitation among patients after stroke. A system for dynamic visualisation of lower-limb movement based on 3-dimentional motion capture and a computer timed functional electrical stimulation system was developed. Participants received up to 20-min enhanced treadmill training instead of their over-ground gait training once or twice a week for 6 weeks at Coathill hospital, Lanarkshire, United Kingdom. Number of training sessions attended, and training duration were used to assess feasibility. Ankle kinematics in the sagittal plane of walking with and without functional electrical stimulation support of the pre-tibial muscles were also compared and used to confirm the functional electrical stimulation was triggered at the targeted time.

Results: Six patients after stroke participated in the study. The majority of participants were male (5/6) with a age range from 30 to 84 years and 4/6 had left hemiplegia. All participants suffered from brain infarction and were at least 3 months after stroke. Number of training sessions attended ranged from 5 to 12. The duration of training sessions ranged from 11 to 20 min. No serious adverse events were reported. The computerised functional electrical stimulation to the pre-tibial muscles was able to reduce plantarflexion angle during the swing phase with statistical significance (p = 0.015 at 80%; p = 0.008 at 90 and 100% of the gait cycle).

Conclusions: It is safe and feasible to use treadmill gait training augmented with real-time visual feedback and computer-controlled functional electrical stimulation with patients after stroke in routine clinical practice.

Trial registration: NCT03348215. Registered 20 November 2017.

背景:脑卒中康复通常采用运动再学习的概念,要求患者进行主动的技能专项训练并接受反馈。跑步机训练辅以实时视觉反馈和功能性电刺激可能对运动恢复有有益的协同作用。本研究旨在确定这种强化跑步机训练在脑卒中患者步态康复中的可行性。开发了一种基于三维运动捕捉和计算机定时功能电刺激的下肢运动动态可视化系统。参与者在英国拉纳克郡的Coathill医院接受了长达20分钟的强化跑步机训练,而不是他们的地面步态训练,每周一次或两次,持续6周。采用参加培训的次数和培训时间来评估可行性。在有和没有胫骨前肌肉功能性电刺激支持的情况下,我们还比较了行走时踝关节矢状面运动学,并用于确认功能性电刺激在目标时间触发。结果:6例脑卒中后患者参与研究。大多数参与者为男性(5/6),年龄在30至84岁之间,4/6患有左偏瘫。所有参与者都患有脑梗死,并且中风后至少3个月。参加的培训课程从5到12次不等。训练时间从11分钟到20分钟不等。无严重不良事件报告。计算机化的胫骨前肌功能性电刺激能够在摆动阶段降低跖屈曲角度,具有统计学意义(p = 0.015, 80%;在90和100%的步态周期P = 0.008)。结论:在常规临床实践中,对脑卒中后患者采用增强实时视觉反馈和计算机控制功能电刺激的跑步机步态训练是安全可行的。试验注册:NCT03348215。2017年11月20日注册。
{"title":"Treadmill training augmented with real-time visualisation feedback and function electrical stimulation for gait rehabilitation after stroke: a feasibility study.","authors":"Chanwit Phongamwong,&nbsp;Philip Rowe,&nbsp;Karen Chase,&nbsp;Andrew Kerr,&nbsp;Lindsay Millar","doi":"10.1186/s42490-019-0020-1","DOIUrl":"https://doi.org/10.1186/s42490-019-0020-1","url":null,"abstract":"<p><strong>Background: </strong>Stroke rehabilitation often uses the motor relearning concept that require patients to perform active practice of skill-specific training and to receive feedback. Treadmill training augmented with real-time visualisation feedback and functional electrical stimulation may have a beneficial synergistic effect on motor recovery. This study aims to determine the feasibility of this kind of enhanced treadmill training for gait rehabilitation among patients after stroke. A system for dynamic visualisation of lower-limb movement based on 3-dimentional motion capture and a computer timed functional electrical stimulation system was developed. Participants received up to 20-min enhanced treadmill training instead of their over-ground gait training once or twice a week for 6 weeks at Coathill hospital, Lanarkshire, United Kingdom. Number of training sessions attended, and training duration were used to assess feasibility. Ankle kinematics in the sagittal plane of walking with and without functional electrical stimulation support of the pre-tibial muscles were also compared and used to confirm the functional electrical stimulation was triggered at the targeted time.</p><p><strong>Results: </strong>Six patients after stroke participated in the study. The majority of participants were male (5/6) with a age range from 30 to 84 years and 4/6 had left hemiplegia. All participants suffered from brain infarction and were at least 3 months after stroke. Number of training sessions attended ranged from 5 to 12. The duration of training sessions ranged from 11 to 20 min. No serious adverse events were reported. The computerised functional electrical stimulation to the pre-tibial muscles was able to reduce plantarflexion angle during the swing phase with statistical significance (<i>p</i> = 0.015 at 80%; <i>p</i> = 0.008 at 90 and 100% of the gait cycle).</p><p><strong>Conclusions: </strong>It is safe and feasible to use treadmill gait training augmented with real-time visual feedback and computer-controlled functional electrical stimulation with patients after stroke in routine clinical practice.</p><p><strong>Trial registration: </strong>NCT03348215. Registered 20 November 2017.</p>","PeriodicalId":72425,"journal":{"name":"BMC biomedical engineering","volume":"1 ","pages":"20"},"PeriodicalIF":0.0,"publicationDate":"2019-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s42490-019-0020-1","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38454818","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 7
Telemetry-controlled simultaneous stimulation-and-recording device (SRD) to study interhemispheric cortical circuits in rat primary somatosensory (SI) cortex. 遥测控制同步刺激记录装置(SRD)研究大鼠初级体感皮层(SI)半球间皮层回路。
Pub Date : 2019-08-08 eCollection Date: 2019-01-01 DOI: 10.1186/s42490-019-0019-7
John T Ramshur, Bashir I Morshed, Amy L de Jongh Curry, Robert S Waters

Background: A growing need exists for neuroscience platforms that can perform simultaneous chronic recording and stimulation of neural tissue in animal models in a telemetry-controlled fashion with signal processing for analysis of the chronic recording data and external triggering capability. We describe the system design, testing, evaluation, and implementation of a wireless simultaneous stimulation-and-recording device (SRD) for modulating cortical circuits in physiologically identified sites in primary somatosensory (SI) cortex in awake-behaving and freely-moving rats. The SRD was developed using low-cost electronic components and open-source software. The function of the SRD was assessed by bench and in-vivo testing.

Results: The SRD recorded spontaneous spiking and bursting neuronal activity, evoked responses to programmed intracortical microstimulation (ICMS) delivered internally by the SRD, and evoked responses to external peripheral forelimb stimulation.

Conclusions: The SRD is capable of wireless stimulation and recording on a predetermined schedule or can be wirelessly synchronized with external input as would be required in behavioral testing prior to, during, and following ICMS.

背景:对神经科学平台的需求日益增长,这些平台可以在遥测控制的方式下对动物模型中的神经组织进行同步慢性记录和刺激,并通过信号处理来分析慢性记录数据和外部触发能力。我们描述了一种无线同步刺激和记录设备(SRD)的系统设计、测试、评估和实现,用于调节清醒行为和自由运动大鼠初级体感(SI)皮层生理识别部位的皮层回路。SRD是使用低成本电子元件和开源软件开发的。通过台架试验和体内试验评估SRD的功能。结果:SRD记录了自发的尖峰和破裂神经元活动,诱发了SRD内部传递的程序性皮质内微刺激(ICMS)的反应,以及外部外周前肢刺激的反应。结论:SRD能够按照预定的时间表进行无线刺激和记录,也可以在ICMS之前、期间和之后的行为测试中与外部输入进行无线同步。
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引用次数: 3
Perfusion culture maintained with an air-liquid interface to stimulate epithelial cell organization in renal organoids in vitro. 以气液界面维持灌注培养,刺激体外肾类器官上皮细胞组织。
Pub Date : 2019-07-23 eCollection Date: 2019-01-01 DOI: 10.1186/s42490-019-0017-9
Sachiko Sekiya, Tetsutaro Kikuchi, Tatsuya Shimizu

Background: Organoids derived from induced pluripotent stem (iPS) or embryonic stem (ES) cells have been evaluated as in vitro models of development and disease. However, maintaining these cells under long-term static culture conditions is difficult because of nutrition shortages and waste accumulation. To overcome these issues, perfusion culture systems are required for organoid technology. A system with a stable microenvironment, nutrient availability, and waste removal will accelerate organoid generation. The aim of this study was to develop a novel perfusion system for renal organoids by maintaining the air-liquid interface with a device fabricated using a 3D printer.

Results: Our results revealed slow flow at the organoid cultivation area based on microbead movement on the membrane, which depended on the perfusion rate under the membrane. Moreover, the perfused culture medium below the organoids via a porous membrane diffused throughout the organoids, maintaining the air-liquid interface. The diffusion rates within organoids were increased according to the flow rate of the culture medium under the membrane. The perfused culture medium also stimulated cytoskeletal and basement membrane re-organization associated with promotion tubular formation under 2.5 μL/min flow culture. In contrast, tubules in organoids were diminished at a flow rate of 10 μL/min.

Conclusions: Our liquid-air interface perfusion system accelerated organization of the renal organoids. These results suggest that suitable perfusion conditions can accelerate organization of epithelial cells and tissues in renal organoids in vitro.

背景:来自诱导多能干细胞(iPS)或胚胎干细胞(ES)的类器官已被评估为发育和疾病的体外模型。然而,由于营养短缺和废物积累,在长期静态培养条件下维持这些细胞是困难的。为了克服这些问题,类器官技术需要灌注培养系统。一个具有稳定的微环境、营养可利用性和废物清除的系统将加速类器官的产生。本研究的目的是通过使用3D打印机制造的设备来维持气液界面,从而开发一种新的肾类器官灌注系统。结果:基于膜上微珠运动的类器官培养区血流缓慢,这与膜下灌注速率有关。此外,在类器官下方灌注的培养基通过多孔膜扩散到整个类器官,保持了气液界面。随着膜下培养基流速的增加,类器官内的扩散速率也随之增加。在2.5 μL/min的流速下,灌注的培养基还能促进细胞骨架和基底膜的重组,促进小管的形成。在10 μL/min流速下,类器官小管减少。结论:我们的液气界面灌注系统加速了肾类器官的组织。提示适宜的灌注条件能促进体外肾类器官上皮细胞和组织的形成。
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引用次数: 7
Skin substitutes are more potent than dermal or epidermal substitutes in stimulating endothelial cell sprouting. 皮肤替代品在刺激内皮细胞发芽方面比真皮或表皮替代品更有效。
Pub Date : 2019-07-17 eCollection Date: 2019-01-01 DOI: 10.1186/s42490-019-0018-8
Hanneke N Monsuur, Ester M Weijers, Susan Gibbs, Lenie J van den Broek

Background: Therapy resistant ulcers are wounds that remain open for a long time period and often arise from chronic venous disease, prolonged pressure or diabetes. For healing of chronic wounds, revitalization of the inert wound bed, which is achieved by angiogenic sprouting of new blood vessels is of great importance. An alternative treatment option to conventional therapies is the use of skin substitutes: dermal (DS), epidermal (ES) or bi-layered skin substitutes (SS). The aim of this study was to determine the mode of action of an autologous SS, ES and DS with regards to endothelial cell proliferation, migration and angiogenic sprouting into a fibrin hydrogel.

Results: SS consists of a fully differentiated epidermis expanding over the acellular donor dermis (AD) which has become repopulated with fibroblasts. DS is the same construct as SS but without the epidermis and ES is the same construct as SS but without the fibroblasts. As a control, AD was used throughout. It was found that the bi-layered SS was the most potent substitute in inducing migration and sprouting of endothelial cells. The cross talk between dermis and epidermis resulted in the strongest induction of sprouting via VEGF and uPAR. ES stimulated sprouting more than DS again via VEGF and uPAR. The slight induction of sprouting mediated by DS was not mediated by VEGF, but was in part stimulated through uPAR.

Conclusion: This in vitro study supports our clinical observations that a bi-layered SS is a strong stimulator of angiogenesis and therefore has the potential to revitalize an inert wound bed.

背景:治疗抵抗性溃疡是指长时间保持开放的伤口,通常由慢性静脉疾病、长期压力或糖尿病引起。对于慢性伤口的愈合,通过新生血管的新生来激活惰性伤口床是非常重要的。常规疗法的替代治疗选择是使用皮肤替代品:真皮(DS),表皮(ES)或双层皮肤替代品(SS)。本研究的目的是确定自体SS、ES和DS对内皮细胞增殖、迁移和血管生成萌生成纤维蛋白水凝胶的作用模式。结果:SS由一个完全分化的表皮组成,表皮扩张在脱细胞供体真皮(AD)上,该真皮已被成纤维细胞重新填充。DS与SS结构相同,但没有表皮;ES与SS结构相同,但没有成纤维细胞。作为对照,自始至终使用AD。结果表明,双层SS是诱导内皮细胞迁移和发芽的最有效替代物。真皮和表皮间的相互作用导致VEGF和uPAR对发芽的诱导作用最强。通过VEGF和uPAR, ES比DS更能刺激发芽。DS介导的微诱导发芽不是由VEGF介导的,而是部分通过uPAR刺激的。结论:这项体外研究支持了我们的临床观察,即双层SS是一种强大的血管生成刺激剂,因此有可能使惰性伤口床恢复活力。
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引用次数: 2
Overcoming the limitations of locally administered oncolytic virotherapy. 克服局部给药溶瘤病毒治疗的局限性。
Pub Date : 2019-07-01 eCollection Date: 2019-01-01 DOI: 10.1186/s42490-019-0016-x
JinWoo Hong, Chae-Ok Yun

Adenovirus (Ad) has been most extensively evaluated gene transfer vector in clinical trials due to facile production in high viral titer, highly efficient transduction, and proven safety record. Similarly, an oncolytic Ad, which replicates selectively in cancer cells through genetic modifications, is actively being evaluated in various phases of clinical trials as a promising next generation therapeutic against cancer. Most of these trials with oncolytic Ads to date have employed intratumoral injection as the standard administration route. Although these locally administered oncolytic Ads have shown promising outcomes, the therapeutic efficacy is not yet optimal due to poor intratumoral virion retention, nonspecific shedding of virion to normal organs, variable infection efficacy due to heterogeneity of tumor cells, adverse antiviral immune response, and short biological activity of oncolytic viruses in situ. These inherent problems associated with locally administered Ad also holds true for other oncolytic viral vectors. Thus, this review will aim to discuss various nanomaterial-based delivery strategies to improve the intratumoral administration efficacy of oncolytic Ad as well as other types of oncolytic viruses.

腺病毒(Ad)由于其易于生产、高病毒滴度、高效转导和已证实的安全记录,在临床试验中被广泛评价为基因转移载体。同样,一种溶瘤性Ad,通过基因修饰在癌细胞中选择性复制,正在临床试验的各个阶段被积极评估为有希望的下一代癌症治疗方法。迄今为止,大多数溶瘤性广告的试验都采用肿瘤内注射作为标准给药途径。尽管这些局部给药的溶瘤病毒已显示出良好的效果,但由于肿瘤内病毒粒子潴留差、病毒粒子向正常器官的非特异性脱落、肿瘤细胞异质性导致的感染效果不稳定、抗病毒免疫反应不良以及溶瘤病毒在原位的生物活性短,治疗效果尚不理想。这些与局部给药Ad相关的固有问题也适用于其他溶瘤病毒载体。因此,本综述旨在讨论各种基于纳米材料的递送策略,以提高溶瘤性Ad和其他类型溶瘤病毒的肿瘤内给药效果。
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引用次数: 23
A high content, phenotypic 'scar-in-a-jar' assay for rapid quantification of collagen fibrillogenesis using disease-derived pulmonary fibroblasts. 高含量、表型 "瓶中疤痕 "检测法,利用疾病衍生肺成纤维细胞快速量化胶原纤维生成。
Pub Date : 2019-06-28 eCollection Date: 2019-01-01 DOI: 10.1186/s42490-019-0014-z
Robert B Good, Jessica D Eley, Elaine Gower, Genevieve Butt, Andrew D Blanchard, Andrew J Fisher, Carmel B Nanthakumar

Background: Excessive extracellular matrix (ECM) deposition is a hallmark feature in fibrosis and tissue remodelling diseases. Typically, mesenchymal cells will produce collagens under standard 2D cell culture conditions, however these do not assemble into fibrils. Existing assays for measuring ECM production are often low throughput and not disease relevant. Here we describe a robust, high content, pseudo-3D phenotypic assay to quantify mature fibrillar collagen deposition which is both physiologically relevant and amenable to high throughput compound screening. Using pulmonary fibroblasts derived from patients with idiopathic pulmonary fibrosis (IPF), we developed the 'scar-in-a-jar' assay into a medium-throughput phenotypic assay to robustly quantify collagen type I deposition and other extracellular matrix (ECM) proteins over 72 h.

Results: This assay utilises macromolecular crowding to induce an excluded volume effect and enhance enzyme activity, which in combination with TGF-β1 stimulation significantly accelerates ECM production. Collagen type I is upregulated approximately 5-fold with a negligible effect on cell number. We demonstrate the robustness of the assay achieving a Z prime of approximately 0.5, and % coefficient of variance (CV) of < 5 for the assay controls SB-525334 (ALK5 inhibitor) and CZ415 (mTOR inhibitor). This assay has been used to confirm the potency of a number of potential anti-fibrotic agents. Active compounds from the 'scar-in-a-jar' assay can be further validated for other markers of ECM deposition and fibroblast activation such as collagen type IV and α-smooth muscle actin exhibiting a 4-fold and 3-fold assay window respectively.

Conclusion: In conclusion, we have developed 'scar -in-a-jar is' into a robust disease-relevant medium-throughput in vitro assay to accurately quantify ECM deposition. This assay may enable iterative compound profiling for IPF and other fibroproliferative and tissue remodelling diseases.

背景:过多的细胞外基质(ECM)沉积是纤维化和组织重塑疾病的标志性特征。通常情况下,间充质细胞会在标准二维细胞培养条件下产生胶原蛋白,但这些胶原蛋白不会组装成纤维。现有的测量 ECM 产量的检测方法通常通量较低,且与疾病无关。在这里,我们描述了一种稳健、高含量、伪三维表型检测方法,用于量化成熟纤维状胶原沉积,这种方法既与生理相关,又适合高通量化合物筛选。利用特发性肺纤维化(IPF)患者的肺成纤维细胞,我们将 "罐中疤痕 "试验发展成了一种中等通量表型试验,可在 72 小时内对 I 型胶原沉积和其他细胞外基质(ECM)蛋白进行稳健的定量分析:结果:该检测方法利用大分子拥挤诱导排斥体积效应并增强酶活性,结合 TGF-β1 刺激,显著加快了 ECM 的生成。I 型胶原上调约 5 倍,对细胞数量的影响微乎其微。我们证明了该检测方法的稳健性,其 Z 值约为 0.5,方差系数 (CV) 为 Conclusion:总之,我们将 "罐中疤痕 "开发成了一种稳健的疾病相关介质通量体外检测方法,可准确量化 ECM 沉积。这种检测方法可用于 IPF 及其他纤维增生性疾病和组织重塑疾病的迭代化合物分析。
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BMC biomedical engineering
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