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An Electrical Stimulation Culture System for Daily Maintenance-Free Muscle Tissue Production 用于日常无维护肌肉组织生产的电刺激培养系统
Q1 ENGINEERING, BIOMEDICAL Pub Date : 2021-04-08 DOI: 10.34133/2021/9820505
Y. Akiyama, Akemi Nakayama, S. Nakano, Ryuichiro Amiya, Jun Hirose
Low-labor production of tissue-engineered muscles (TEMs) is one of the key technologies to realize the practical use of muscle-actuated devices. This study developed and then demonstrated the daily maintenance-free culture system equipped with both electrical stimulation and medium replacement functions. To avoid ethical issues, immortal myoblast cells C2C12 were used. The system consisting of gel culture molds, a medium replacement unit, and an electrical stimulation unit could produce 12 TEMs at one time. The contractile forces of the TEMs were measured with a newly developed microforce measurement system. Even the TEMs cultured without electrical stimulation generated forces of almost 2 mN and were shortened by 10% in tetanic contractions. Regarding the contractile forces, electrical stimulation by a single pulse at 1 Hz was most effective, and the contractile forces in tetanus were over 2.5 mN. On the other hand, continuous pulses decreased the contractile forces of TEMs. HE-stained cross-sections showed that myoblast cells proliferated and fused into myotubes mainly in the peripheral regions, and fewer cells existed in the internal region. This must be due to insufficient supplies of oxygen and nutrients inside the TEMs. By increasing the supplies, one TEM might be able to generate a force up to around 10 mN. The tetanic forces of the TEMs produced by the system were strong enough to actuate microstructures like previously reported crawling robots. This daily maintenance-free culture system which could stably produce TEMs strong enough to be utilized for microrobots should contribute to the advancement of biohybrid devices.
组织工程肌肉的低劳动生产率是实现肌肉驱动装置实用化的关键技术之一。本研究开发并演示了具有电刺激和培养基更换功能的日常免维护培养系统。为了避免伦理问题,使用了永生成肌细胞C2C12。由凝胶培养模具、培养基更换单元和电刺激单元组成的系统可以一次产生12个TEM。TEM的收缩力是用新开发的微力测量系统测量的。即使在没有电刺激的情况下培养的TEMs也会产生几乎2 mN,在强直性收缩时缩短10%。关于收缩力,在1 Hz是最有效的,破伤风的收缩力超过2.5 mN。另一方面,连续脉冲降低了TEM的收缩力。HE染色截面显示,成肌细胞主要在外周区域增殖并融合成肌管,而在内部区域存在较少的细胞。这一定是由于TEM内氧气和营养物质供应不足。通过增加供应,一个TEM可能能够产生高达10左右的力 mN。由该系统产生的TEM的强直力足够强,可以像以前报道的爬行机器人一样驱动微观结构。这种日常免维护的培养系统可以稳定地产生足以用于微型机器人的TEM,这将有助于生物混合装置的发展。
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引用次数: 11
Bioprinted Vascularized Mature Adipose Tissue with Collagen Microfibers for Soft Tissue Regeneration 胶原微纤维生物打印血管化成熟脂肪组织用于软组织再生
Q1 ENGINEERING, BIOMEDICAL Pub Date : 2021-03-13 DOI: 10.34133/2021/1412542
F. Louis, Marie Piantino, Hao Liu, D. Kang, Y. Sowa, S. Kitano, M. Matsusaki
The development of soft tissue regeneration has recently gained importance due to safety concerns about artificial breast implants. Current autologous fat graft implantations can result in up to 90% of volume loss in long-term outcomes due to their limited revascularization. Adipose tissue has a highly vascularized structure which enables its proper homeostasis as well as its endocrine function. Mature adipocytes surrounded by a dense vascular network are the specific features required for efficient regeneration of the adipose tissue to perform host anastomosis after its implantation. Recently, bioprinting has been introduced as a promising solution to recreate in vitro this architecture in large-scale tissues. However, the in vitro induction of both the angiogenesis and adipogenesis differentiations from stem cells yields limited maturation states for these two pathways. To overcome these issues, we report a novel method for obtaining a fully vascularized adipose tissue reconstruction using supporting bath bioprinting. For the first time, directly isolated mature adipocytes encapsulated in a bioink containing physiological collagen microfibers (CMF) were bioprinted in a gellan gum supporting bath. These multilayered bioprinted tissues retained high viability even after 7 days of culture. Moreover, the functionality was also confirmed by the maintenance of fatty acid uptake from mature adipocytes. Therefore, this method of constructing fully functional adipose tissue regeneration holds promise for future clinical applications.
由于人工乳房植入物的安全性问题,软组织再生的发展最近变得越来越重要。目前自体脂肪移植由于其有限的血运重建,在长期结果中可导致高达90%的体积损失。脂肪组织具有高度血管化的结构,使其具有适当的体内平衡和内分泌功能。成熟的脂肪细胞被致密的血管网络包围是脂肪组织植入后有效再生并进行宿主吻合所需的特定特征。最近,生物打印作为一种很有前途的解决方案被引入体外,在大规模组织中重建这种结构。然而,干细胞在体外诱导血管生成和脂肪生成分化时,这两种途径的成熟状态有限。为了克服这些问题,我们报告了一种使用支持浴生物打印获得完全血管化脂肪组织重建的新方法。首次将直接分离的成熟脂肪细胞包裹在含有生理性胶原微纤维(CMF)的生物墨水中,在结冷胶支撑浴中进行生物打印。这些多层生物打印组织在培养7天后仍保持较高的活力。此外,这种功能也被成熟脂肪细胞对脂肪酸摄取的维持所证实。因此,这种构建全功能脂肪组织再生的方法有望在未来的临床应用中得到应用。
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引用次数: 20
Cell-Based Biohybrid Sensor Device for Chemical Source Direction Estimation 用于化学源方向估计的基于细胞的生物混合传感器装置
Q1 ENGINEERING, BIOMEDICAL Pub Date : 2021-01-23 DOI: 10.34133/2021/8907148
H. Oda, K. Kihara, Y. Morimoto, S. Takeuchi
This paper describes a method to estimate the direction from which the signal molecule reaches the sensor by using living cells. In this context, biohybrid sensors that utilize a sophisticated sensing system of cells can potentially offer high levels of chemical-detection sensitivity and selectivity. However, biohybrid-sensor-based chemical-source-direction estimation has not received research attention because the cellular response to chemicals has not been examined in the context of directional information. In our approach, we fabricated a device that can limit the interface between the cell-laden hydrogel and the chemical solution of interest to enhance the time difference over which the chemical solution reaches the cells. Chemical detection by cells that express specific receptors is reflected as the fluorescence of the calcium indicator within the cells. Our device has eight chambers that each house 3D cell-laden collagen hydrogels facing circularly outward. The device also works as a cover to prevent chemicals from permeating the hydrogel from above. In our study, by observing the time course of the fluorescence emission of each chamber, we were able to successfully estimate the chemical-source direction within an error range of 7–13°. Our results suggest that a combination of microstructure devices embedded with living cells can be used to exploit cell functionalities to yield chemical-source directional information.
本文描述了一种利用活细胞来估计信号分子到达传感器的方向的方法。在这种情况下,利用复杂的细胞传感系统的生物混合传感器可以潜在地提供高水平的化学检测灵敏度和选择性。然而,基于生物混合传感器的化学源方向估计尚未受到研究关注,因为细胞对化学物质的反应尚未在方向信息的背景下进行研究。在我们的方法中,我们制造了一种设备,可以限制负载细胞的水凝胶和感兴趣的化学溶液之间的界面,以增强化学溶液到达细胞的时间差。表达特定受体的细胞的化学检测被反映为细胞内钙指示剂的荧光。我们的设备有八个腔室,每个腔室容纳环形向外的3D细胞胶原水凝胶。该装置还可以作为盖子,防止化学物质从上面渗透到水凝胶中。在我们的研究中,通过观察每个腔室的荧光发射的时间过程,我们能够在7-13°的误差范围内成功地估计化学源方向。我们的研究结果表明,嵌入活细胞的微结构器件的组合可以用于利用细胞功能来产生化学源方向信息。
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引用次数: 11
Design of Electrohydrodynamic Devices with Consideration of Electrostatic Energy 考虑静电能的电流体动力装置设计
Q1 ENGINEERING, BIOMEDICAL Pub Date : 2021-01-09 DOI: 10.34133/2021/5158282
Tasuku Sato, S. Sakuma, M. Hijikuro, S. Maeda, M. Anyoji, Y. Yamanishi
The importance of actuators that can be integrated with flexible robot structures and mechanisms has increased in recent years with the advance of soft robotics. In particular, electrohydrodynamic (EHD) actuators, which have expandable integrability to adapt to the flexible motion of soft robots, have received much attention in the field of soft robotics. Studies have deepened the understanding of steady states of EHD phenomena but nonsteady states are not well understood. We herein observe the development process of fluid in a microchannel adopting a Schlieren technique with the aid of a high-speed camera. In addition, we analyze the behavior of fluid flow in a microchannel that is designed to have pairs of parallel plate electrodes adopting a computational fluid dynamics technique. Results indicate the importance of considering flow generated by electrostatic energy, which tends to be ignored in constructing and evaluating EHD devices, and by the body force generated by the ion-drag force. By considering these effects, we estimate the development process of EHD flow and confirm the importance of considering the generation of vortices and their interactions inside the microchannel during the development of EHD devices.
近年来,随着软机器人技术的发展,能够与柔性机器人结构和机构集成的致动器变得越来越重要。特别是电液动力(EHD)作动器,由于其具有可扩展的可积性以适应软机器人的柔性运动,在软机器人领域受到了广泛的关注。研究加深了对EHD稳态现象的认识,但对非稳态现象的认识还不充分。本文借助高速摄像机,采用纹影技术观察了微通道中流体的发展过程。此外,我们还采用计算流体动力学技术分析了设计成一对平行板电极的微通道中的流体流动行为。结果表明,考虑静电能量产生的流动和离子阻力产生的体力的重要性,这在构建和评估EHD装置时往往被忽视。通过考虑这些影响,我们估计了EHD流动的发展过程,并确认了在EHD装置的开发过程中考虑微通道内涡的产生及其相互作用的重要性。
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引用次数: 6
3D Bioprinted Spheroidal Droplets for Engineering the Heterocellular Coupling between Cardiomyocytes and Cardiac Fibroblasts. 三维生物打印球形液滴用于设计心肌细胞与心脏成纤维细胞之间的异细胞耦合。
Q1 ENGINEERING, BIOMEDICAL Pub Date : 2021-01-01 Epub Date: 2021-12-28 DOI: 10.34133/2021/9864212
Raven El Khoury, Naveen Nagiah, Joel A Mudloff, Vikram Thakur, Munmun Chattopadhyay, Binata Joddar

Since conventional human cardiac two-dimensional (2D) cell culture and multilayered three-dimensional (3D) models fail in recapitulating cellular complexity and possess inferior translational capacity, we designed and developed a high-throughput scalable 3D bioprinted cardiac spheroidal droplet-organoid model with cardiomyocytes and cardiac fibroblasts that can be used for drug screening or regenerative engineering applications. This study helped establish the parameters for bioprinting and cross-linking a gelatin-alginate-based bioink into 3D spheroidal droplets. A flattened disk-like structure developed in prior studies from our laboratory was used as a control. The microstructural and mechanical stability of the 3D spheroidal droplets was assessed and was found to be ideal for a cardiac scaffold. Adult human cardiac fibroblasts and AC16 cardiomyocytes were mixed in the bioink and bioprinted. Live-dead assay and flow cytometry analysis revealed robust biocompatibility of the 3D spheroidal droplets that supported the growth and proliferation of the cardiac cells in the long-term cultures. Moreover, the heterocellular gap junctional coupling between the cardiomyocytes and cardiac fibroblasts further validated the 3D cardiac spheroidal droplet model.

由于传统的人类心脏二维(2D)细胞培养和多层三维(3D)模型无法再现细胞的复杂性,而且转化能力较差,我们设计并开发了一种具有心肌细胞和心脏成纤维细胞的高通量可扩展三维生物打印心脏球状液滴-类器官模型,可用于药物筛选或再生工程应用。这项研究帮助确定了将明胶-海藻酸盐基生物墨水生物打印和交联成三维球状液滴的参数。我们实验室在之前的研究中开发的扁平圆盘状结构被用作对照。我们对三维球状液滴的微观结构和机械稳定性进行了评估,发现它是理想的心脏支架。将成人人类心脏成纤维细胞和 AC16 心肌细胞混合在生物墨水中并进行生物打印。活死实验和流式细胞仪分析表明,三维球形液滴具有良好的生物相容性,可支持心脏细胞在长期培养中的生长和增殖。此外,心肌细胞和心脏成纤维细胞之间的异细胞间隙连接进一步验证了三维心脏球形液滴模型。
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引用次数: 0
Surgical Robot for Intraluminal Access: An Ex Vivo Feasibility Study. 腔内手术机器人:离体可行性研究。
Q1 ENGINEERING, BIOMEDICAL Pub Date : 2020-01-01 DOI: 10.34133/2020/8378025
Ryu Nakadate, Tsutomu Iwasa, Shinya Onogi, Jumpei Arata, Susumu Oguri, Yasuharu Okamoto, Tomohiko Akahoshi, Masatoshi Eto, Makoto Hashizume

Early-stage gastrointestinal cancer is often treated by endoscopic submucosal dissection (ESD) using a flexible endoscope. Compared with conventional percutaneous surgery, ESD is much less invasive and provides a high quality of life for the patient because it does not require a skin incision, and the organ is preserved. However, the operator must be highly skilled because ESD requires using a flexible endoscope with energy devices, which have limited degrees of freedom. To facilitate easier manipulation of these flexible devices, we developed a surgical robot comprising a flexible endoscope and two articulating instruments. The robotic system is based on a conventional flexible endoscope, and an extrapolated motor unit moves the endoscope in all its degrees of freedom. The instruments are thin enough to allow insertion of two instruments into the endoscope channel, and each instrument has a bending section that allows for up-down, right-left, and forward-backward motion. In this study, we performed an ex vivo feasibility evaluation using the proposed robotic system for ESD in a porcine stomach. The procedure was successfully performed by five novice operators without complications. Our findings demonstrated the feasibility of the proposed robotic system and, furthermore, suggest that even operators with limited experience can use this system to perform ESD.

早期胃肠癌通常采用柔性内窥镜进行内镜粘膜下剥离(ESD)治疗。与传统的经皮手术相比,ESD的侵入性小得多,因为它不需要皮肤切口,而且器官得到了保存,为患者提供了高质量的生活。然而,操作人员必须具有很高的技能,因为ESD需要使用带有能量装置的灵活内窥镜,而内窥镜的自由度有限。为了方便这些柔性装置的操作,我们开发了一个手术机器人,包括一个柔性内窥镜和两个关节器械。机器人系统是基于一个传统的柔性内窥镜,外推电机单元移动内窥镜在其所有的自由度。仪器足够薄,可以将两个仪器插入内窥镜通道,每个仪器都有一个弯曲部分,允许上下,左右和前后运动。在这项研究中,我们使用提出的机器人系统在猪胃中进行ESD的离体可行性评估。该手术由5名新手成功完成,无并发症。我们的研究结果证明了所提出的机器人系统的可行性,此外,即使是经验有限的操作人员也可以使用该系统执行ESD。
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引用次数: 19
Cyborg and Bionic Systems: Signposting the Future. 半机械人与仿生系统:未来的路标。
Q1 ENGINEERING, BIOMEDICAL Pub Date : 2020-01-01 DOI: 10.34133/2020/1310389
Toshio Fukuda
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引用次数: 20
Robotic Surgery in Gastrointestinal Surgery. 胃肠外科中的机器人手术。
Q1 ENGINEERING, BIOMEDICAL Pub Date : 2020-01-01 DOI: 10.34133/2020/9724807
Kenoki Ohuchida

Robotic surgery is expanding in the minimally invasive treatment of gastrointestinal cancer. In the field of gastrointestinal cancer, robotic surgery is performed using a robot-assisted surgery system. In this system, the robot does not operate automatically but is controlled by the surgeon. The surgery assistant robot currently used in clinical practice worldwide is the leader-follower type, including the da Vinci® Surgical System (Intuitive Surgical). This review describes the current state of robotic surgery in the treatment of gastrointestinal cancer and discusses the future development of robotic systems in gastrointestinal surgery.

机器人手术正在扩大胃肠道癌症的微创治疗。在胃肠道癌症领域,机器人手术是使用机器人辅助手术系统进行的。在这个系统中,机器人不是自动操作,而是由外科医生控制。目前世界范围内临床应用的手术辅助机器人为leader-follower型,包括达芬奇®Surgical System (Intuitive Surgical)。本文综述了机器人手术在胃肠道肿瘤治疗中的现状,并讨论了机器人系统在胃肠道手术中的未来发展。
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引用次数: 6
期刊
Cyborg and bionic systems (Washington, D.C.)
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