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Impact of microchannel width on axons for brain-on-chip applications† 微通道宽度对脑芯片应用中轴突的影响
IF 6.1 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-23 DOI: 10.1039/D4LC00440J
Katarina Vulić, Giulia Amos, Tobias Ruff, Revan Kasm, Stephan J. Ihle, Joël Küchler, János Vörös and Sean Weaver

Technologies for axon guidance for in vitro disease models and bottom up investigations are increasingly being used in neuroscience research. One of the most prevalent patterning methods is using polydimethylsiloxane (PDMS) microstructures due to compatibility with microscopy and electrophysiology which enables systematic tracking of axon development with precision and efficiency. Previous investigations of these guidance platforms have noted axons tend to follow edges and avoid sharp turns; however, the specific impact of spatial constraints remains only partially explored. We investigated the influence of microchannel width beyond a constriction point, as well as the number of available microchannels, on axon growth dynamics. Further, by manipulating the size of micron/submicron-sized PDMS tunnels we investigated the space restriction that prevents growth cone penetration showing that restrictions smaller than 350 nm were sufficient to exclude axons. This research offers insights into the interplay of spatial constraints, axon development, and neural behavior. The findings are important for designing in vitro platforms and in vivo neural interfaces for both fundamental neuroscience and translational applications in rapidly evolving neural implant technologies.

用于体外疾病模型和自下而上研究的轴突导向技术正越来越多地应用于神经科学研究。最普遍的图案化方法之一是使用聚二甲基硅氧烷(PDMS)微结构,因为它与显微镜和电生理学兼容,可以精确高效地系统跟踪轴突的发育。以前对这些引导平台的研究注意到,轴突倾向于沿着边缘发展并避免急转弯;然而,空间限制的具体影响仍只得到了部分探讨。我们研究了收缩点以外的微通道宽度以及可用微通道数量对轴突生长动态的影响。此外,通过操纵微米/亚微米级 PDMS 通道的尺寸,我们研究了阻止生长锥穿透的空间限制,结果显示小于 350 纳米的限制足以将轴突排除在外。这项研究为空间限制、轴突发育和神经行为的相互作用提供了见解。这些发现对于设计体外平台和体内神经接口非常重要,既可用于基础神经科学,也可用于快速发展的神经植入技术的转化应用。
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引用次数: 0
Self-actuated microfluidic chiplet for two-stage multiplex nucleic acid amplification assay 用于两级多重核酸扩增检测的自驱动微流控芯片组
IF 6.1 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-22 DOI: 10.1039/d4lc00752b
Felix Ansah, Marziyeh Hajialyani, Fatemeh Ahmadi, Yuming Gu, Ergun Alperay Tarım, Michael G Mauk, Gordon Awandare, Haim H Bau
Effective diagnosis of comorbidities and infectious diseases that present similar symptoms requires point-of-need assays capable of co-detecting and differentiating among multiple co-endemic pathogens to enable timely, precision medicine and effective control measures. We previously developed a two-stage isothermal amplification assay dubbed Penn-RAMP to address this need. Penn-RAMP’s first stage comprises a Recombinase Polymerase Amplification (RPA), which amplifies all targets of interest in a single reaction chamber for a short duration. The RPA amplicons are then aliquoted into multiple Loop-Mediated Isothermal Amplification (LAMP) reaction chambers, each customized with pre-dried primers to amplify a single target or a group of targets. To enable Penn-RAMP at the point of need, we describe here a self-actuated Penn-RAMP chiplet that accommodates the Penn-RAMP assay. Our chiplet employs temperature-controlled phase change valves and capillary valves to self-aliquot first-stage amplicons into multiple (five) second-stage reaction chambers and to seal these chambers. The functionality of our device is demonstrated by co-detecting plant pathogens. The analytical performance of our chiplet is comparable to that of the benchtop Penn-RAMP assay and surpasses that of standalone LAMP assays. Our self-actuated chiplet can be operated standalone with purified nucleic acids or as the downstream amplification module of a sample preparation cassette.
要有效诊断症状相似的合并症和传染病,需要能够共同检测和区分多种共同流行病原体的需求点检测方法,以便及时采取精准医疗和有效控制措施。为了满足这一需求,我们之前开发了一种两阶段等温扩增检测方法,称为宾夕法尼亚大学-RAMP。Penn-RAMP的第一阶段包括重组酶聚合酶扩增(RPA),在一个反应室中对所有感兴趣的目标进行短时间扩增。然后,RPA 扩增子被等分到多个环路介导等温扩增(LAMP)反应室中,每个反应室都有定制的预干燥引物,用于扩增单个或一组目标。为了在需要时实现宾夕法尼亚-RAMP,我们在此介绍一种可用于宾夕法尼亚-RAMP 分析的自驱动宾夕法尼亚-RAMP 芯片。我们的芯片采用温控相变阀和毛细管阀,将第一阶段的扩增子自流到多个(五个)第二阶段的反应室中,并密封这些反应室。我们的装置通过联合检测植物病原体证明了其功能。我们的芯片的分析性能与台式宾夕法尼亚-RAMP 分析法相当,并超过了独立的 LAMP 分析法。我们的自驱动芯片可与纯化核酸一起独立运行,也可作为样品制备盒的下游扩增模块。
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引用次数: 0
Zonal Patterning of Extracellular Matrix and Stromal Cell Populations Along a Perfusable Cellular Microchannel 细胞外基质和基质细胞群在可灌注细胞微通道上的带状分布
IF 6.1 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-21 DOI: 10.1039/d4lc00579a
Brea Chernokal, Bryan J Ferrick, Jason P Gleghorn
The spatial organization of biophysical and biochemical cues in the extracellular matrix (ECM) in concert with reciprocal cell-cell signaling is vital to tissue patterning during development. However, elucidating the role an individual microenvironmental factor plays using existing textit{in vivo} models is difficult due to their inherent complexity. In this work, we have developed a microphysiological system to spatially pattern the biochemical, biophysical, and stromal cell composition of the ECM along an epithelialized 3D microchannel. This technique is adaptable to multiple hydrogel compositions and scalable to the number of zones patterned. We confirmed that the methodology to create distinct zones resulted in a continuous, annealed hydrogel with regional interfaces that did not hinder the transport of soluble molecules. Further, the interface between hydrogel regions did not disrupt microchannel structure, epithelial lumen formation, or media perfusion through an acellular or cellularized microchannel. Finally, we demonstrated spatially patterned tubulogenic sprouting of a continuous epithelial tube into the surrounding hydrogel confined to local regions with stromal cell populations, illustrating spatial control of cell-cell interactions and signaling gradients. This easy-to-use system has wide utility for modeling three-dimensional epithelial and endothelial tissue interactions with heterogeneous hydrogel compositions and/or stromal cell populations to investigate their mechanistic roles during development, homeostasis, or disease.
细胞外基质(ECM)中的生物物理和生物化学线索的空间组织与细胞-细胞之间的相互信号传递对发育过程中的组织形态至关重要。然而,由于其固有的复杂性,利用现有的体内模型来阐明单个微环境因素所起的作用非常困难。在这项工作中,我们开发了一种微观生理学系统,可沿着上皮化三维微通道对 ECM 的生化、生物物理和基质细胞组成进行空间模式化。该技术可适应多种水凝胶成分,并可根据图案化区域的数量进行扩展。我们证实,创建不同区域的方法可产生连续的退火水凝胶,其区域界面不会阻碍可溶性分子的运输。此外,水凝胶区域之间的界面不会破坏微通道结构、上皮腔的形成或通过无细胞或细胞化微通道的介质灌注。最后,我们展示了连续上皮管向周围水凝胶的空间模式化管状萌发,这种萌发局限于基质细胞群的局部区域,说明了细胞-细胞相互作用和信号梯度的空间控制。这种易于使用的系统在模拟三维上皮和内皮组织与异质水凝胶成分和/或基质细胞群的相互作用方面具有广泛用途,可用于研究它们在发育、稳态或疾病过程中的机理作用。
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引用次数: 0
Integrated device for plasma separation and nucleic acid extraction from whole blood toward point-of-care detection of bloodborne pathogens† 从全血中分离血浆和提取核酸的集成装置,用于血源性病原体的床旁检测
IF 6.1 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-18 DOI: 10.1039/D4LC00571F
Abigail G. Ayers, Christia M. Victoriano and Samuel K. Sia

Sample preparation presents a major challenge in point-of-care (POC) diagnostic assays, including ones requiring whole blood as the starting specimen. This study presents an integrated sample preparation device – which we call PRECISE – that performs both plasma separation and nucleic acid extraction, enabling streamlined sample preparation from whole blood requiring only a commercially available blood collection tool and a syringe, and no other external equipment or electricity. Plasma separation is performed using a dual-membrane filter (which filters out blood components while limiting membrane clogging) integrated into the cartridge, and nucleic acid extraction is performed by users moving magnets (to mix the samples, and along a guided track). The plasma filtration demonstrated recovery on par with lab-based centrifugation, and the extraction module showed performance similar to benchtop-based magnetic bead extraction. A sample-to-result demonstration on 50 μL of whole blood spiked with virions of hepatitis C virus (HCV), operating the PRECISE cartridge in 16 minutes followed by benchtop PCR, showed a limit of detection (∼6770 IU mL−1) on the order of the minimal requirements of target product profile for POC HCV detection. Future work on the PRECISE cartridge, building on POC accessibility and fast sample preparation demonstrated in this work, may enable detection of bloodborne pathogens from whole-blood specimens collected at the POC.

样品制备是护理点 (POC) 诊断测定中的一大挑战,包括需要全血作为起始样本的测定。本研究介绍了一种集成的样品制备设备(我们称之为 PRECISE),它既能进行血浆分离,又能进行核酸提取,从而简化了全血样品的制备过程,只需一个市售的采血工具和一个注射器,无需其他外部设备或电力。血浆分离是利用集成在血盒中的双膜过滤器(既能过滤血液成分,又能限制膜堵塞)进行的,核酸提取则是由用户移动磁铁(沿导轨混合样本)进行的。血浆过滤的回收率与实验室离心法相当,提取模块的性能与台式磁珠提取相似。对 50 μL 丙型肝炎病毒(HCV)加标全血进行的样品到结果演示,在 16 分钟内操作 PRECISE 血盒,然后进行台式 PCR,结果显示检测限(∼6770 IU mL-1)达到了 POC HCV 检测对目标产物特征的最低要求。PRECISE 血盒的未来工作将以本研究中展示的 POC 可及性和快速样品制备为基础,实现从 POC 采集的全血标本中检测血源性病原体。
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引用次数: 0
Placental microphysiological systems: new advances on promising platforms that mimic the microenvironment of the human placenta 胎盘微生理系统:模拟人类胎盘微环境的前景广阔的平台的新进展
IF 6.1 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-17 DOI: 10.1039/d4lc00500g
Inês M. Gonçalves, Muhammad Afzal, Nithil Kennedy, Ana Moita, Rui Lima, Serge Ostrovidov, Takeshi Hori, Yuji Nashimoto, Hirokazu Kaji
One of the most complex human physiological processes to study is pregnancy. Standard animal models, as well as two-dimensional models, lack the complexity and biological relevance required to accurately study such a physiological process. Recent studies have focused on the development of three-dimensional models based on microfluidic systems, designated as placental microphysiological systems (PMPSs). PMPS devices provide a model of the placental barrier through culturing relevant cell types in specific arrangements and media to mimic the in vivo environment of the maternal–fetal circulation. Here, recent developments of PMPS models for embryo uterine implantation, preeclampsia evaluation, and toxicological screening are presented. Studies that use bioprinting techniques are also discussed. Lastly, recent developments in endometrium microphysiological systems are reviewed. All these presented models showed their superiority compared to standard models in recapitulating the biological environment seen in vivo. However, several limitations regarding the types of cells and materials used for these systems were also widely reported. Despite the need for further improvements, PMPS models contribute to a better understanding of the biological mechanisms surrounding pregnancy and the respective pathologies.
怀孕是人类最复杂的生理过程之一。标准的动物模型和二维模型缺乏准确研究这种生理过程所需的复杂性和生物学相关性。最近的研究重点是开发基于微流控系统的三维模型,即胎盘微生理系统(PMPS)。PMPS 装置通过在特定排列和培养基中培养相关细胞类型来模拟母胎循环的体内环境,从而提供胎盘屏障模型。本文介绍了用于胚胎子宫植入、子痫前期评估和毒理学筛选的 PMPS 模型的最新进展。此外,还讨论了使用生物打印技术的研究。最后,还回顾了子宫内膜微生理系统的最新发展。与标准模型相比,所有介绍的这些模型在重现体内生物环境方面都显示出其优越性。然而,这些系统所使用的细胞类型和材料方面的一些局限性也被广泛报道。尽管还需要进一步改进,但 PMPS 模型有助于更好地了解与妊娠有关的生物机制和相关病理。
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引用次数: 0
A nanofluidic exchanger for harvesting saline gradients energy 用于收集盐水梯度能量的纳米流体交换器
IF 6.1 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-15 DOI: 10.1039/d4lc00544a
Saranath Sripriya, Cyril Picard, Vincent Larrey, Frank Fournel, Elisabeth Charlaix
The energy of saline gradients is a very promising source of non-intermittent renewable energy, the exploitation of which is hampered by the lack of viable technology. The most investigated harvesting methods rely on selective transport of ions or water molecules through semi-permeable or ion-selective membranes, which demonstrate limited power densities of the order of a few W/m2. While in the last decade single nanofluidic objects such as nanopores of nanotubes have opened up very promising prospects with power density capabilities in the kW or even MW/m2, scale-up efforts face serious issues, as concentration polarization phenomena result in a massive loss of performance. We propose here a concept of nanofluidic exchanger for power generation from saline gradients, focused on designing a nanoscale flow able to harvest the power at the output of the nanopores. We study analytically and numerically a simple exchanger made of a selective nanoslit fed by a nanofluidic assembly. One specific feature of such an exchanger relies on the non-linear ion fluxes through the nanoslit analytically expressed from the integration of the Poison-Nernt Planck equation. Such an elemental brick could be massively parallelized in stackable electricity-generating layers using standard technologies of the semi-conductors industry. We demonstrate here a scheme for rationalizing the choice of the exchanger parameters, taking into account the transport properties at all scales. The full numerical resolution of three-dimensional device shows that net power densities of 300 W/m2 and more can be achieved.
盐梯度能量是一种非常有前景的非间歇性可再生能源,但由于缺乏可行的技术,其开发利用受到了阻碍。研究最多的收集方法依赖于离子或水分子通过半透膜或离子选择性膜的选择性传输,其功率密度有限,仅为几瓦/平方米。过去十年中,单个纳米流体(如纳米管的纳米孔)的功率密度已达到千瓦甚至兆瓦/平方米,前景十分广阔,但由于浓度极化现象会导致性能大幅下降,因此扩大规模的工作面临严重问题。我们在此提出了一种利用盐水梯度发电的纳米流体交换器概念,重点是设计一种能够在纳米孔输出端收集电能的纳米级流动。我们通过分析和数值计算研究了一种简单的交换器,该交换器由纳米流体组件提供的选择性纳米液滴组成。这种交换器的一个具体特征是,通过对 Poison-Nernt Planck 方程的积分,分析得出通过纳米缝隙的非线性离子通量。利用半导体工业的标准技术,这种元素砖可以在可堆叠的发电层中大规模并行化。我们在此展示了一种合理选择交换器参数的方案,其中考虑到了所有尺度上的传输特性。三维设备的全数值分辨率表明,净功率密度可以达到 300 W/m2 或更高。
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引用次数: 0
Real-time impedance-activated dielectrophoretic actuation for reconfigurable manipulation of single flowing particles† 实时阻抗激活介电致动,实现单个流动粒子的可重构操纵
IF 6.1 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-14 DOI: 10.1039/D4LC00622D
Alexis Lefevre, Cristian Brandi, Adele De Ninno, Filippo Ruggiero, Enrico Verona, Michaël Gauthier, Paolo Bisegna, Aude Bolopion and Federica Caselli

This work presents an innovative all-electrical platform for selective single-particle manipulation. The platform combines microfluidic impedance cytometry for label-free particle characterization and dielectrophoresis for contactless multi-way particle separation. The microfluidic chip has a straightforward coplanar electrode layout and no particle pre-focusing mechanism is required. An original online algorithm analyzes the impedance signals of each incoming particle and regulates in real time the dielectrophoretic voltages according to a desired control logic. As a proof-of-concept, three operation modes are demonstrated on a mixture of 8, 10, and 12 μm diameter beads: (i) particle position swapping across the channel axis, irrespective of particle size, (ii) size-based particle separation, irrespective of particle position, and (iii) sorting of a selected sequence of particles. As a perspective, the versatility of impedance cytometry and dielectrophoresis, and the possibility of configuring alternative control logics, hold promise for advanced particle and cell manipulation.

这项研究提出了一种用于选择性单粒子操作的创新型全电平台。该平台结合了用于无标记颗粒表征的微流体阻抗细胞仪和用于非接触式多向颗粒分离的介电泳技术。微流控芯片采用直接的共面电极布局,无需粒子预聚焦机制。一种独创的在线算法可分析每个进入粒子的阻抗信号,并根据所需的控制逻辑实时调节介电泳电压。作为概念验证,在直径分别为 8、10 和 12 微米的微珠混合物上演示了三种操作模式:(i) 颗粒位置跨通道轴交换,与颗粒大小无关;(ii) 基于大小的颗粒分离,与颗粒位置无关;(iii) 选定颗粒序列的分类。从这个角度来看,阻抗细胞仪和介质电泳的多功能性以及配置其他控制逻辑的可能性为先进的粒子和细胞操作带来了希望。
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引用次数: 0
Compact lens-free imager using thin-film transistor for long-term quantitative monitoring of stem cell culture and cardiomyocyte production 使用薄膜晶体管的紧凑型无透镜成像仪,用于长期定量监测干细胞培养和心肌细胞生产
IF 6.1 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-14 DOI: 10.1039/d4lc00528g
Taishi Kakizuka, Tohru Natsume, Takeharu Nagai
With advancements in human induced pluripotent stem cell (hiPSC) technology, there is an increasing demand for quality control techniques to manage the long-term process of target cell production effectively. While monitoring systems designed for use within incubators are promising for assessing culture quality, existing systems still face challenges in terms of compactness, throughput, and available metrics. To address these limitations, we have developed a compact and high-throughput lens-free imaging device named INSPCTOR. The device is as small as a standard culture plate, which allows for the installation of multiple units within an incubator. INSPCTOR utilises a large thin-film transistor image sensor, enabling simultaneous observation of six independent culture environments, each approximately 1 cm2. With this device, we successfully monitored the confluency of hiPSC cultures and identified the onset timing of epithelial-to-mesenchymal transition during mesodermal induction. Additionally, we quantified the beating frequency and conduction of hiPSC-derived cardiomyocytes by using high-speed imaging modes. This enabled us to identify the onset of spontaneous beating during differentiation and assess chronotropic responses in drug evaluations. Moreover, by tracking beating frequency over 10 days of cardiomyocyte maturation, we identified week-scale and daily-scale fluctuations, the latter of which correlated with cellular metabolic activity. The metrics derived from this device would enhance the reproducibility and quality of target cell production.
随着人类诱导多能干细胞(hiPSC)技术的发展,对质量控制技术的需求日益增加,以有效管理目标细胞生产的长期过程。虽然专为培养箱设计的监测系统在评估培养质量方面大有可为,但现有系统在紧凑性、吞吐量和可用指标方面仍面临挑战。为了解决这些局限性,我们开发了一种名为 INSPCTOR 的紧凑型高通量无透镜成像设备。该设备与标准培养板一样小,可在培养箱中安装多个装置。INSPCTOR 采用大型薄膜晶体管图像传感器,可同时观测六个独立的培养环境,每个环境约 1 平方厘米。利用该装置,我们成功监测了 hiPSC 培养物的汇合情况,并确定了中胚层诱导过程中上皮向间质转化的开始时间。此外,我们还利用高速成像模式量化了 hiPSC 衍生心肌细胞的跳动频率和传导能力。这使我们能够确定分化过程中自发跳动的起始时间,并在药物评估中评估促时变反应。此外,通过跟踪 10 天的心肌细胞成熟过程中的跳动频率,我们确定了周范围和日范围的波动,后者与细胞代谢活动相关。该装置得出的指标将提高靶细胞生产的可重复性和质量。
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引用次数: 0
Acoustic enrichment of sperm for in vitro fertilization† 体外受精精子的声学富集
IF 6.1 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-11 DOI: 10.1039/D4LC00604F
Chunqiu Zhang, Ning Rong, Ziyi Lin, Peng-Qi Li, Jingyao Shi, Wei Zhou, Lili Niu, Fei Li, Rongxin Tang, Lei Li and Long Meng

Assisted reproductive technology (ART) has emerged as a crucial method in modern medicine for tackling infertility. However, the success of fertilization depends on the quality and quantity of sperm, often necessitating invasive surgical intervention, which presents challenges for non-invasive in vitro fertilization. Acoustic microfluidics technology has found widespread application across various biological contexts. In this paper, we propose to introduce a novel approach using asymmetric acoustic streaming generated by a single interdigital transducer (IDT) to enhance sperm concentration and improve fertilization in vitro, particularly in cases of moderate oligozoospermia. The concentration of particles increased approximately 6-fold in the central region after acoustic enrichment. Moreover, sperm motility was significantly improved without additional DNA fragmentation, and all the oocytes remained viable after 5 min of acoustic enrichment. Notably, acoustic enrichment accelerated fertilization and embryo development, leading to a higher fertilization rate and faster cleavage speed. Specifically, within 36 hours, the multiple-cell embryo ratio was significantly increased compared to the control group. This finding further validates the feasibility and non-invasiveness of acoustic enrichment for sperm fertilization in vitro. This work provides a promising tool for in vitro fertilization, holding significant implications for assisted reproduction.

辅助生殖技术(ART)已成为现代医学解决不孕不育问题的重要方法。然而,受精成功与否取决于精子的质量和数量,通常需要进行侵入性手术干预,这给无创体外受精带来了挑战。声学微流控技术已在各种生物领域得到广泛应用。在本文中,我们提出了一种新方法,利用单个趾间换能器(IDT)产生的不对称声流来提高精子浓度,改善体外受精,尤其是在中度少精症的情况下。声波富集后,中央区域的颗粒浓度增加了约 6 倍。此外,精子的运动能力也得到了明显改善,而且不会造成额外的 DNA 断裂,所有卵母细胞在声学富集 5 分钟后仍能存活。值得注意的是,声学富集加速了受精和胚胎发育,使受精率更高,裂殖速度更快。具体来说,与对照组相比,36 小时内多细胞胚胎比率明显增加。这一发现进一步验证了声学富集技术在体外精子受精方面的可行性和非侵入性。这项工作为体外受精提供了一种前景广阔的工具,对辅助生殖具有重要意义。
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引用次数: 0
Rapid low-cost assembly of modular microvessel-on-a-chip with benchtop xurography† 利用台式 Xurography 技术低成本快速组装模块化微血管芯片
IF 6.1 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-07 DOI: 10.1039/D4LC00565A
Shashwat S. Agarwal, Marcos Cortes-Medina, Jacob C. Holter, Alex Avendano, Joseph W. Tinapple, Joseph M. Barlage, Miles M. Menyhert, Lotanna M. Onua and Jonathan W. Song

Blood and lymphatic vessels in the body are central to molecular and cellular transport, tissue repair, and pathophysiology. Several approaches have been employed for engineering microfabricated blood and lymphatic vessels in vitro, yet traditionally these approaches require specialized equipment, facilities, and research training beyond the capabilities of many biomedical laboratories. Here we present xurography as an inexpensive, accessible, and versatile rapid prototyping technique for engineering cylindrical and lumenized microvessels. Using a benchtop xurographer, or a cutting plotter, we fabricated modular multi-layer poly(dimethylsiloxane) (PDMS)-based microphysiological systems (MPS) that house endothelial-lined microvessels approximately 260 μm in diameter embedded within a user-defined 3-D extracellular matrix (ECM). We validated the vascularized MPS (or vessel-on-a-chip) by quantifying changes in blood vessel permeability due to the pro-angiogenic chemokine CXCL12. Moreover, we demonstrated the reconfigurable versatility of this approach by engineering a total of four distinct vessel-ECM arrangements, which were obtained by only minor adjustments to a few steps of the fabrication process. Several of these arrangements, such as ones that incorporate close-ended vessel structures and spatially distinct ECM compartments along the same microvessel, have not been widely achieved with other microfabrication strategies. Therefore, we anticipate that our low-cost and easy-to-implement fabrication approach will facilitate broader adoption of MPS with customizable vascular architectures and ECM components while reducing the turnaround time required for iterative designs.

体内的血液和淋巴管是分子和细胞运输、组织修复和病理生理学的核心。体外微制造血管和淋巴管工程已经采用了多种方法,但这些方法无一例外地需要专业设备、设施和研究培训,超出了大多数生物医学实验室的能力范围。在这里,我们介绍了一种成本低廉、易于使用且用途广泛的快速原型制造技术--Xurography,用于制造圆柱形和腔化微血管。我们使用台式 Xurography 或切割绘图仪制造了基于模块化多层聚二甲基硅氧烷(PDMS)的微生理系统(MPS),该系统容纳直径约 260 微米的内皮衬里微血管,并嵌入用户定义的三维细胞外基质(ECM)中。我们通过量化促血管生成趋化因子 CXCL12 引起的血管通透性变化,验证了血管化 MPS(或芯片上的血管)。此外,我们通过对制造过程中的一个或两个步骤进行微调,设计出三种不同的血管-ECM 排列方式,证明了这种方法的可重构多功能性。其中有几种排列方式是其他微制造策略无法轻易实现的,例如在同一微血管中加入近端血管结构和空间上不同的 ECM 区室。因此,我们预计我们的低成本、易实施的制造方法将有助于更广泛地获得具有可调血管结构和 ECM 成分的 MPS,同时缩短迭代设计所需的周转时间。
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引用次数: 0
期刊
Lab on a Chip
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