Figure 6 - uploaded by Boyang Zhang
Content may be subject to copyright.
Surgical anastomosis of the cardiac tissue. a,b, Surgical anastomosis of the AngioChip cardiac tissue on the rat femoral vessels in the configuration of artery-to-artery graft (a) and artery-to-vein graft (b). Blood perfusion was established immediately after anastomosis. Papers were placed under the implants during imaging for better visual contrast. c–j, Cross-section of the non-endothelialized rat cardiac tissue implants 1 week after surgery without (c–f; n = 3) or with direct anastomosis (g–j; n = 4) in the configuration of artery-to-vein graft. The sections were stained with Masson’s trichrome (c,d,g,h), for smooth muscle actin (e,i), and for troponin T (f,j; red). Scale bars, 200 μm (c,g), 100 μm (d,e,h,i) and 50 μm (f,j). k, Image of a cardiac tissue implant on rat hindlimb 1 week after surgery with direct anastomosis in the configuration of artery-to-vein graft. White dotted line outlines the AngioChip implant. l, Quantification of the area stained for smooth muscle actin (average ± s.d.). m–p, Histology cross-sections of an AngioChip cardiac tissue endothelialized with Lewis rat primary vein endothelial cells and implanted with direct surgical anastomosis after 1 week (n = 3). The sections were stained with Masson’s trichrome (m,n), and for CD31 (o,p). Scale bars, 200 μm (m), 100 μm (n,o) and 50 μm (p). Red dashed boxes in c,g,m,o indicate the imaging location of panels d,h,n,p, respectively.
Source publication
We report the fabrication of a scaffold (hereafter referred to as AngioChip) that supports the assembly of parenchymal cells on a mechanically tunable matrix surrounding a perfusable, branched, three-dimensional microchannel network coated with endothelial cells. The design of AngioChip decouples the material choices for the engineered vessel netwo...
Contexts in source publication
Context 1
... scaolds were connected to the femoral vessels on the hindlimbs of adult Lewis rats, in artery-to-artery ( Fig. 6a) and artery-to-vein (Fig. 6b) mode, to demonstrate two dierent configurations of direct surgical anastomosis. The inlet and outlet (inner dimensions of 100 µm ⇥ 200 µm and outer dimensions of 300 µm ⇥ 400 µm) were connected to femoral vessels ( Fig. 6) with surgical cus. Similar citric acid-based polymers 28 have been shown to be ...
Context 2
... scaolds were connected to the femoral vessels on the hindlimbs of adult Lewis rats, in artery-to-artery ( Fig. 6a) and artery-to-vein (Fig. 6b) mode, to demonstrate two dierent configurations of direct surgical anastomosis. The inlet and outlet (inner dimensions of 100 µm ⇥ 200 µm and outer dimensions of 300 µm ⇥ 400 µm) were connected to femoral vessels ( Fig. 6) with surgical cus. Similar citric acid-based polymers 28 have been shown to be antithrombotic in vascular grafts ...
Context 3
... connected to the femoral vessels on the hindlimbs of adult Lewis rats, in artery-to-artery ( Fig. 6a) and artery-to-vein (Fig. 6b) mode, to demonstrate two dierent configurations of direct surgical anastomosis. The inlet and outlet (inner dimensions of 100 µm ⇥ 200 µm and outer dimensions of 300 µm ⇥ 400 µm) were connected to femoral vessels ( Fig. 6) with surgical cus. Similar citric acid-based polymers 28 have been shown to be antithrombotic in vascular grafts and to support EC growth in vivo 30,31 . The animals were heparinized only during surgery. In both configurations, blood perfusion was established immediately, even in the absence of ECs (Fig. 6a,b), although ...
Context 4
... µm) were connected to femoral vessels ( Fig. 6) with surgical cus. Similar citric acid-based polymers 28 have been shown to be antithrombotic in vascular grafts and to support EC growth in vivo 30,31 . The animals were heparinized only during surgery. In both configurations, blood perfusion was established immediately, even in the absence of ECs (Fig. 6a,b), although artery-to-artery mode was more technically challenging owing to the higher pressure (Supplementary Movie 7). Blood pulsation was also observed (Supplementary Movie 7), more noticeably in the artery-to-artery configuration. Erythrocytes were observed only in the networks of tissues implanted with direct anastomosis (Fig. ...
Context 5
... of ECs (Fig. 6a,b), although artery-to-artery mode was more technically challenging owing to the higher pressure (Supplementary Movie 7). Blood pulsation was also observed (Supplementary Movie 7), more noticeably in the artery-to-artery configuration. Erythrocytes were observed only in the networks of tissues implanted with direct anastomosis (Fig. 6c,d,g,h,k and Supplementary Figs 21 and 22). One week after the implantation of the AngioChip cardiac tissues, native angiogenesis also took place around the implants (Supplementary Fig. 23). The presence of smooth muscle actin (SMA)-positive cells was merely 2% in the isolated neonatal rat heart cells 4 ; the significant SMA staining ...
Context 6
... (Fig. 6c,d,g,h,k and Supplementary Figs 21 and 22). One week after the implantation of the AngioChip cardiac tissues, native angiogenesis also took place around the implants (Supplementary Fig. 23). The presence of smooth muscle actin (SMA)-positive cells was merely 2% in the isolated neonatal rat heart cells 4 ; the significant SMA staining (Fig. 6e,i and Supplementary Fig. 24) suggested the penetration of mural cells or myofibroblasts into the implanted tissues consistent with the healing response 50 (Fig. 6l). Troponin T immunostaining demonstrated some elongated cardiomyocytes intertwining within the lattice of the AngioChips (Fig. 6f,j and Supplementary Fig. 25). AngioChip ...
Context 7
... the implants (Supplementary Fig. 23). The presence of smooth muscle actin (SMA)-positive cells was merely 2% in the isolated neonatal rat heart cells 4 ; the significant SMA staining (Fig. 6e,i and Supplementary Fig. 24) suggested the penetration of mural cells or myofibroblasts into the implanted tissues consistent with the healing response 50 (Fig. 6l). Troponin T immunostaining demonstrated some elongated cardiomyocytes intertwining within the lattice of the AngioChips (Fig. 6f,j and Supplementary Fig. 25). AngioChip cardiac tissues endothelialized with Lewis rat primary vein ECs were also implanted with direct anastomosis in the artery- to-vein configuration. One week later, ECs ...
Context 8
... Fig. 25). AngioChip cardiac tissues endothelialized with Lewis rat primary vein ECs were also implanted with direct anastomosis in the artery- to-vein configuration. One week later, ECs coating the lumen were observed in 50% of the microchannels from three dierent implants (2/5, 4/5, 1/5 of visible lumen for implant 1, 2, 3, respectively; Fig. 6m-p). Histologically, 85% of AngioChip lumen were blood clot free after 1 week in vivo ( Supplementary Figs 21 and 22). Specifically, in non-EC-coated AngioChips 3/3, 3/3, 4/5 and 3/5 of visible lumens were blood clot free in the four implants and in the EC-coated groups 3/5, 5/5 and 5/5 were blood clot free in the three implants. The ...
Similar publications
We developed a device that can quickly apply versatile electrical stimulation (ES) signals to cells suspended in microfluidic channels and measure extracellular field potential simultaneously. The device could trap cells onto the surface of measurement electrodes for ES and push them to the downstream channel after ES by increasing pressure for con...
Citations
... Advancements in tissue engineering technologies toward developing organs suitable for transplantation therapy can potentially facilitate the repair of defective organs [1]. Current methods for generating three-dimensional (3D) myocardial tissue include development of the tissue by mixing an extracellular matrix such as collagen with myocardial cells before gelation [2][3][4], seeding decellularized tissue with myocardial cells [5], and using a 3D bioprinter [6,7]. On the other hand, research on myocardial tissue engineering using cell sheets [8,9] and cell spheroids [10,11] without extracellular matrix has been conducted. ...
Despite the development of three-dimensional (3D) tissues that promises remarkable advances in myocardial therapies and pharmaceutical research, vascularization is required for the repair of damaged hearts using cardiac tissue engineering. In this study, we developed a method for rapid generation of a 3D cardiac tissue, with extremely high engraftment efficiency, by stacking cardiomyocyte sheets using fibrin as an adhesive. Cell sheets were created by peeling off confluent cultured cells from a culture dish grafted with a polymer that induced surface hydrophilicity in response to low temperatures. The high engraftment rate was attributed to the retention of the adhesive protein. The multistacked vascularized cell sheets prepared using fibrin, when transplanted into the subcutaneous tissue and at myocardial infarction site in rats, yielded a transplanted 3D myocardial tissue. Furthermore, multilayered cardiomyocyte sheets were transplanted twice at 1 week intervals to create a 3D myocardial tissue. Our data suggest that fibrin-based rapidly layered cell sheets can advance tissue-engineered transplantation therapy and should aid the development of next-generation tissue-engineered products in the fields of regenerative medicine and drug screening.
... Strategies to vascularize 3D engineered tissues can generally be categorized as top-down or bottom-up [11,15]. Top-down approaches involve creating vasculature using methods such as 3D printing of bio-inks or sacrificial materials [16][17][18], laser-mediated hydrogel ablation [19,20] or layer-by-layer assembly strategies [11,21,22]. Despite exquisite control over vascular network geometry and architecture, these methods cannot currently achieve the 5-20 μm diameter of capillaries. ...
Vasculogenic assembly of 3D capillary networks remains a promising approach to vascularizing tissue-engineered grafts, a significant outstanding challenge in tissue engineering and regenerative medicine. Current approaches for vasculogenic assembly rely on the inclusion of supporting mesenchymal cells alongside endothelial cells, co-encapsulated within vasculo-conducive materials such as low-density fibrin hydrogels. Here, we established a material-based approach to circumvent the need for supporting mesenchymal cells and report that the inclusion of synthetic matrix fibers in dense (>3 mg mL⁻¹) 3D fibrin hydrogels can enhance vasculogenic assembly in endothelial cell monocultures. Surprisingly, we found that the addition of non-cell-adhesive synthetic matrix fibers compared to cell-adhesive synthetic fibers best encouraged vasculogenic assembly, proliferation, lumenogenesis, a vasculogenic transcriptional program, and additionally promoted cell-matrix interactions and intercellular force transmission. Implanting fiber-reinforced prevascularized constructs to assess graft-host vascular integration, we demonstrate additive effects of enhanced vascular network assembly during in vitro pre-culture, fiber-mediated improvements in endothelial cell survival and vascular maintenance post-implantation, and enhanced host cell infiltration that collectively enabled graft vessel integration with host circulation. This work establishes synthetic matrix fibers as an inexpensive alternative to sourcing and expanding secondary supporting cell types for the prevascularization of tissue constructs.
... Additionally, high-frequency ultrasound adds to these systems' capabilities, enabling non-invasive quantitative studies of systolic kinetics in cardiac chips [58]. To address the inherent softness of biomaterials like hydrogels in constructing vascular-rich organ chips that mimic muscle contraction, bio-scaffolds can be embedded into vessels at the microscale [59]. ...
Cardiac organoids offer sophisticated 3D structures that emulate key aspects of human heart development and function. This review traces the evolution of cardiac organoid technology, from early stem cell differentiation protocols to advanced bioengineering approaches. We discuss the methodologies for creating cardiac organoids, including self-organization techniques, biomaterial-based scaffolds, 3D bioprinting, and organ-on-chip platforms, which have significantly enhanced the structural complexity and physiological relevance of in vitro cardiac models. We examine their applications in fundamental research and medical innovations, highlighting their potential to transform our understanding of cardiac biology and pathology. The integration of multiple cell types, vascularization strategies, and maturation protocols has led to more faithful representations of the adult human heart. However, challenges remain in achieving full functional maturity and scalability. We critically assess the current limitations and outline future directions for advancing cardiac organoid technology. By providing a comprehensive analysis of the field, this review aims to catalyze further innovation in cardiac tissue engineering and facilitate its translation to clinical applications.
... Perfusion was established immediately after surgery and it was reported that the vessels remained clot-free up till one week after surgery. In addition, native angiogenesis was also reported to be observed around the implant with endothelial cells subsequently coating the lumen in the implants [74] . Such surgeries might be more invasive than traditional treatment methods. ...
Vascular networks have an important role to play in transporting nutrients, oxygen, metabolic wastes and maintenance of homeostasis. Bioprinting is a promising technology as it is able to fabricate complex, specific multi-cellular constructs with precision. In addition, current technology allows precise depositions of individual cells, growth factors and biochemical signals to enhance vascular growth. Fabrication of vascularized constructs has remained as a main challenge till date but it is deemed as an important stepping stone to bring organ engineering to a higher level. However, with the ever advancing bioprinting technology and knowledge of biomaterials, it is expected that bioprinting can be a viable solution for this problem. This article presents an overview of the biofabrication of vascular and vascularized constructs, the different techniques used in vascular engineering such as extrusion-based, droplet-based and laser-based bioprinting techniques, and the future prospects of bioprinting of artificial blood vessels.
... Other fabrication techniques, including injectionmouldeing, 46 soft lithography, 56,57 and standard photolithography, 52 have also been employed to create pre-formed vascular channels on plastic substrates. Notably, the AngioChip platform, 50,71 which combines 3D stamping with LoC technology, has been utilised to produce biodegradable scaffolds with branched microchannel networks, enhancing vascularisation. This platform has been further developed into the InVADE system, which supports multiplexed perfusion and parallel screening of up to 20 3D tissues within a 96-well plate format. ...
Organoids are three‐dimensional (3D) cellular models designed to replicate human tissues and organs while preserving their physiological complexity and functionality. Among these, vascularised organoids represent a groundbreaking advancement in 3D tissue engineering, incorporating vascular networks into engineered tissues to more accurately mimic the in vivo tumour microenvironment. These models offer significantly improved physiological relevance compared to conventional two‐dimensional cultures or animal models, positioning them as invaluable tools in cancer research. Despite their potential, the rapid proliferation of techniques and materials for developing vascularised organoids presents challenges for researchers navigating this dynamic field. This systematic review provides a comprehensive examination of methodologies for fabricating vascularised organoids, with a focus on strategies that enhance vascularisation and support organoid growth. It critically evaluates the materials used, emphasising those that effectively mimic the extracellular matrix and facilitate vascular network formation. Key advancements in engineered organoids models are highlighted, emphasising their potential for studying interactions between vasculature and cancer cells, conducting drug screening, and understanding cytokine regulation. In summary, this review provides an in‐depth overview of the current landscape of vascularised organoid fabrication and functionality, addressing challenges and opportunities within the field. A detailed understanding of the scope and future trajectories is essential for advancing organoid development and expanding their applications in both basic cancer research and clinical practice.
Key points
Comparative analysis: Evaluation of organoids, animal models, and 2D models, highlighting their respective strengths and limitations in replicating physiological conditions and studying disease processes.
Vascularisation techniques: Comparative evaluation of vascularised organoid fabrication methods, emphasising their efficiency, scalability and ability to replicate physiological vascular networks.
Material selection: Thorough evaluation of materials for vascularised organoid culture system, focusing on those that effectively mimic the extracellular matrix and support vascular network formation.
Applications: Overview of organoid applications in basic cancer research and clinical settings, with an emphasis on their potential in drug discovery, disease modelling and exploring complex biological processes.
... Establishing standardized protocols will be crucial for ensuring interoperability and reproducibility. [18][19][20] • Data Integration and Management: The vast amount of data generated by robotic microfluidic experiments requires advanced computational tools for efficient analysis, storage, and retrieval. ...
... Such models have been used to study several aspects of vasculature physiology including angiogenesis and vasculogenesis 32,33 , effects of shear stress and fluid dynamics on vascular blood flow 34,35 . They have also been applied to investigate pathological conditions such vasculature associated with tumour 28,[36][37][38] and other pathophysiological conditions [39][40][41][42][43] , and to investigate outcomes of NP interactions with vascular ECs 17,[44][45][46][47][48][49][50] . However, these studies have been conducted using genomically stable endothelial cell models which do not mimic genomic instability in conditions of endothelial dysfunction, endothelial aging and angiogenic blood vessels. ...
... In-situ bioprinting presents significant advantages, including the capacity for real-time customization, and adaptability to patient-specific needs [8]. Additionally, this technique can operate in restricted or hard-to-reach locations, providing accurate and consistent performance in tissue engineering applications [4,9,10]. ...
Bioprinting has the potential to revolutionize tissue engineering and regenerative medicine, offering innovative solutions for complex medical challenges and addressing unmet clinical needs. However, traditional in vitro bioprinting techniques face significant limitations, including difficulties in fabricating and implanting scaffolds with irregular shapes, as well as limited accessibility for rapid clinical application. To overcome these challenges, in-situ bioprinting has emerged as a groundbreaking approach that enables the direct deposition of cells, biomaterials, and bioactive factors onto damaged organs or tissues, eliminating the need for pre-fabricated 3D constructs. This method promises a personalized, patient-specific approach to treatment, aligning well with the principles of precision medicine. The success of in-situ bioprinting largely depends on the advancement of bioinks, which are essential for maintaining cell viability and supporting tissue development. Recent innovations in hand-held bioprinting devices and robotic arms have further enhanced the flexibility of in-situ bioprinting, making it applicable to various tissue types, such as skin, hair, muscle, bone, cartilage, and composite tissues. This review examines in-situ bioprinting techniques, the development of smart, multifunctional bioinks, and their essential properties for promoting cell viability and tissue growth. It highlights the versatility and recent advancements in in-situ bioprinting methods and their applications in regenerating a wide range of tissues and organs. Furthermore, it addresses the key challenges that must be overcome for broader clinical adoption and propose strategies to advance these technologies toward mainstream medical practice.
... They may serve as a starting point for initial screenings and proof-of-concept studies. Models exhibiting architectures of medium complexity including organoids [70,71] and more advanced engineered tissues such as those generated from bioprinting techniques [72,73], vascularized 3D engineered tissues [53,74], and tissues used within heart-on-a-chip constructs [65,75,76] can better disclose multicellular crosstalk, cell-matrix interactions, vasculature interplay, and tissue development. Moreover, these models offer enhanced control over architectural matrix design, including the alignment of CMs through patterning structures [48]. ...
Heart failure is characterized by intricate myocardial remodeling that impairs the heart’s pumping and/or relaxation capacity, ultimately reducing cardiac output. It represents a major public health burden, given its high prevalence and associated morbidity and mortality rates, which continue to challenge healthcare systems worldwide. Despite advancements in medical science, there are no treatments that address the disease at its core. The development of three-dimensional engineered in vitro models that closely mimic the (patho)physiology and drug responses of the myocardium has the potential to revolutionize our insights and uncover new therapeutic avenues. Key aspects of these models include the precise replication of the extracellular matrix structure, cell composition, micro-architecture, mechanical and electrical properties, and relevant physiological and pathological stimuli, such as fluid flow, mechanical load, electrical signal propagation, and biochemical cues. Additionally, to fully capture heart failure and its diversity in vivo, it is crucial to consider factors such as age, gender, interactions with other organ systems and external influences—thereby recapitulating unique patient and disease phenotypes. This review details these model features and their significance in heart failure research, with the aim of enhancing future platforms that will deepen our understanding of the disease and facilitate the development of novel, effective therapies.
... Researchers at Boston University have developed a novel cardiac chip platform using two-photon polymerisation technology, which can generate 3D printed structures at the scale of biological tissues and with sub-micrometer resolution, making it well suited for simulating natural environments. 15 This 3D bioprinting constructs microfluidic chips in a layer-by-layer fashion, allowing for the creation of complex 3D structures. This integrated manufacturing method can complete the fabrication of microfluidic chips in one step, preventing assembly errors. ...
Cardiovascular diseases are a leading cause of death worldwide, and effective treatment for cardiac disease has been a research focal point. Although the development of new drugs and strategies has never ceased, the existing drug development process relies primarily on rodent models such as mice, which have significant shortcomings in predicting human responses. Therefore, human-based in vitro cardiac tissue models are considered to simulate physiological and functional characteristics more effectively, advancing disease treatment and drug development. The microfluidic device simulates the physiological functions and pathological states of the human heart by culture, thereby reducing the need for animal experimentation and enhancing the efficiency and accuracy of the research. The basic framework of cardiac chips typically includes multiple functional units, effectively simulating different parts of the heart and allowing the observation of cardiac cell growth and responses under various drug treatments and disease conditions. To date, cardiac chips have demonstrated significant application value in drug development, toxicology testing, and the construction of cardiac disease models; they not only accelerate drug screening but also provide a new research platform for understanding cardiac diseases. In the future, with advancements in functionality, integration, and personalised medicine, cardiac chips will further simulate multiorgan systems, becoming vital tools for disease modelling and precision medicine. Here, we emphasised the development history of cardiac organ chips, highlighted the material selection and construction strategy of cardiac organ chip electrodes and hydrogels, introduced the current application scenarios of cardiac organ chips, and discussed the development opportunities and prospects for their of biomedical applications.
