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Tissue Engineering. Part C, Methods

Katherine Anne Cummins, Alexandra Leigh Crampton, David K Wood
Changes to the cellular microenvironment are an integral characteristic of numerous pathologies including cancer, fibrosis, and autoimmune disease. Current <i>in vitro</i> methodologies available to study 3D tissue remodeling are ill-suited for high-throughput studies as they are not scalable for large-scale experiments. Combining droplet microfluidics and patterned low-adhesion culture surfaces, we have engineered a workflow to incorporate cell-ECM interactions in a versatile and high-throughput platform that is compatible with existing high-throughput liquid handling systems, enables long-term experiments (>1 month), and is well-suited for traditional and novel biological measurements...
November 15, 2018: Tissue Engineering. Part C, Methods
Luis Antonio Diaz-Gomez, Brandon T Smith, Panayiotis D Kontoyiannis, Sean M Bittner, Anthony J Melchiorri, Antonios G Mikos
In this work, we present a printing method to fabricate scaffolds consisting of multimaterial segmented fibers. Particularly, we developed a reproducible printing process to create single fibers with multiple discrete compositions and control over the distribution of particulate ceramics-namely hydroxyapatite (HA) and β-tricalcium phosphate (TCP)-within poly(ε-caprolactone)-based composite scaffolds. Tensile testing revealed the mechanical integrity of individual segmented fibers was preserved compared to non-segmented fibers, and microcomputed tomography and thermal analysis confirmed the homogeneous distribution of ceramics incorporated in the fiber compositions...
November 13, 2018: Tissue Engineering. Part C, Methods
Tiago Lazzaretti Fernandes, Heitor A Kimura, Carla Cristina Gomes Pinheiro, Kazunori Shimomura, Norimasa Nakamura, José R M Ferreira, Andreas H Gomoll, Arnaldo José Hernandez, Daniela Franco Bueno
BACKGROUND: cartilage restoration is a desperately needed bridge for patients with symptomatic cartilage lesions. Chondral lesion is a pathology with high prevalence, reaching as much as 63% of general population and 36% among athletes. Despite Autologous Chondrocyte Implantation (ACI) versatility, it still fails to fully reproduce hyaline articular cartilage characteristics. Mesenchymal stem cells (MSCs) may be isolated from various known tissues, including discarded fragments at arthroscopy such as synovial membrane...
November 9, 2018: Tissue Engineering. Part C, Methods
Takeshi Hori, Osamu Kurosawa
In vitro 3D cultures of hepatocytes are increasingly being used to assess human hepatic metabolism and toxicity in drug development. Here, we developed an in vitro 3D cell culture method with a microstructured mesh sheet and applied it to culturing human hepatoma HepG2 cells. The micromesh sheet is constituted of fine mesh strands and apertures that are each much larger than a single cell in size. Proliferating on a micromesh sheet, HepG2 cells spread out in a planar manner and then formed a multilayered cell sheet, so that cell-cell adhesion was dominant over cell-substrate adhesion as being different from 2D cultures...
November 9, 2018: Tissue Engineering. Part C, Methods
Simona Walker, Jessy Schönfelder, Sems-Malte Tugtekin, Christiane Wetzel, Michael C Hacker, Michaela Barbara Schulz-Siegmund
Pericardial scaffolds have a wide spectrum of clinical applications ranging from patches for vascular reconstruction and abdominal wall defects to bioprosthetic heart valves. The current gold standard of tissue preparation involves disinfection and crosslinking using glutaraldehyde. However, glutaraldehyde-associated toxicity as well as rapid calcification and premature graft failure represent the major modes of failure <sup>1</sup>. Therefore, a variety of alternative strategies for tissue conservation have been pursued...
November 9, 2018: Tissue Engineering. Part C, Methods
Elahe Hadavi, Jeroen Leijten, Jenny Brinkmann, Pascal Jonkheijm, Marcel Karperien, Aart van Apeldoorn
This research deals with finding a proper bioengineering strategy for the creation of improved β-cell replacement therapy in type 1 diabetes. It specifically deals with the microenvironment of β-cells and its relationship to their endocrine function.
November 7, 2018: Tissue Engineering. Part C, Methods
Roberto Gaetani, Soraya Aouad, Lea L Demaddalena, Heinz Straessle, Monika Dzieciatkowska, Matthew Wortham, Hugh Rt Bender, Kim-Vy Nguyen-Ngoc, Geert W Schmid-Schoenbein, Steven C George, Christopher C W Hughes, Maike Sander, Kirk C Hansen, Karen L Christman
Different approaches have investigated the effects of the different extracellular matrices (ECMs) and 3D culture on islets function, showing encouraging results. Ideally, the proper scaffold should mimic the biochemical composition of the native tissue as it drives numerous signaling pathways involved in tissue homeostasis and functionality. Tissue-derived decellularized biomaterials can preserve the ECM composition of the native tissue making it an ideal scaffold for 3D tissue engineering applications. However, the decellularization process may affect the retention of specific components, and the choice of the proper detergent is fundamental in preserving the native ECM composition...
November 6, 2018: Tissue Engineering. Part C, Methods
Yuki Ota, Naosuke Kamei, Takahiro Tamura, Nobuo Adachi, Mitsuo Ochi
Magnetic cell delivery system, termed as magnetic targeting, has been developed for minimal invasive cell transplantation. Cells are magnetically labeled with nano-sized iron particles. However, the temporal change in iron particles in vivo has not been clarified. In addition, the influence of magnetic resonance imaging (MRI) after magnetic targeting therapy has never been investigated. In the present study, magnetically labeled bone marrow mesenchymal stem cells (MSCs) were injected into rabbit knee joint with an articular cartilage defect, with or without existence of the magnetic field (m-MSC group, MSC group)...
November 6, 2018: Tissue Engineering. Part C, Methods
Michella Hendrika Hagmeijer, Lucienne A Vonk, Jan-Willem Kouwenhoven, Roel Custers, Ronald Bleys, Aaron J Krych, Daniel Saris
PURPOSE: To test the technical aspects and feasibility of seeding a combination of meniscus cells isolated from a rapid digestion protocol and mesenchymal stromal cells (MSCs) (20: 80 ratio) into a meniscus scaffold for the development of a one-stage arthroscopic procedure for meniscus regeneration. METHODS: A cadaveric study was performed using nine fresh frozen human cadaveric knee joints. Two different arthroscopic cell-seeding methods were applied to the Collagen Meniscus Implant (CMI®) as carrier scaffold: either 1) seeding before arthroscopic surgical implantation of the scaffold, or 2) after implantation of the scaffold...
November 6, 2018: Tissue Engineering. Part C, Methods
Kyung Whan Yoo, Ning Li, Vishruti Makani, Ravi N Singh, Anthony Atala, Baisong Lu
This article describes a method for producing microRNA (miRNA)-enriched extracellular vesicles in large quantities. It enables in vivo delivery of specific miRNA for therapeutic applications.
November 2, 2018: Tissue Engineering. Part C, Methods
Niloufar Khosravi, Vanessa C Mendes, Ghata Nirmal, Safa Majeed, Ralph S DaCosta, John E Davies
These new experimental methods allow us to image, and quantify, angiogenesis and perivascular cell dynamics in the endosseous healing compartment. As such, the method is capable of providing a new perspective on, and unique information regarding, healing that occurs around orthopedic and dental implants.
November 2, 2018: Tissue Engineering. Part C, Methods
Daniel E Gorman, Tong Wu, Sarah E Gilpin, Harald C Ott
The careful study of cell-based lung repair and regeneration ex vivo may one day provide us with the necessary tools to create a patient-derived alternative to cadaveric donor lungs for transplantation. Many parameters must be monitored and optimized in order to advance this aim. The use of rat lungs as a small-scale model for lung regeneration is an efficient way to develop the key improvements required for optimal tissue repair and regeneration. Here we report the use of a novel high-throughput, automated, multi-channel lung bioreactor system which allows for culture and analysis of rodent scale isolated lungs...
October 26, 2018: Tissue Engineering. Part C, Methods
Shahram Ghanaati, Sarah Al-Maawi, Carlos Herrera-Vizcaino, Gutemberg Gomes Alves, Mônica Diuana Calasans-Maia, Robert Sader, C James Kirkpatrick, Joseph Choukroun, Halvard Bönig, Carlos Fernando de Almeida Barros Mourão
Blood concentrates are obtained by centrifugation of the patient's own peripheral blood. These concentrates revealed increasing importance in the regenerative medicine and tissue engineering. Many limitations are faced in preclinical research concerning the volume of blood needed to produce platelet-rich fibrin (PRF) matrices (10 ml). This study evaluated for the first time whether it is possible to obtain three different centrifugation protocols to obtain injectable PRF matrices (i-PRF) using a small blood volume of 3 ml...
October 25, 2018: Tissue Engineering. Part C, Methods
Xiao Zhang, Chenjun Zhai, Hao Fei, Yang Liu, Zhen Wang, Chunyang Luo, Jiyong Zhang, Yanzi Ding, Tao Xu, Weimin Fan
Bioactive scaffolds are used in cartilage regeneration for providing an environment conducive to chondrogenic differentiation and proliferation of stem cells in vitro and in vivo. This study aimed to develop a composite scaffold comprising silk fibroin (SF) and cartilage extracellular matrix (ECM), which could not only preserve the biological activity of the original ECM, which promotes chondrogenic differentiation of bone-marrow-derived mesenchymal stem cells (BMSCs), but also could provide great mechanical strength...
October 23, 2018: Tissue Engineering. Part C, Methods
Anna Lange-Consiglio, Barbara Lazzari, Flavia Pizzi, Alessandra Stella, Alessia Girani, Arianna Quintè, Fausto Cremonesi, Emanuele Capra
Conditioned medium (CM) and microvesicles (MVs) are produced using different protocols: CM is collected following 12-96 hours of cell culture without renewal of tissue culture medium, whilst MVs are collected after overnight cell culture. For future comparative studies in regenerative medicine looking at efficacy of CM and MVs, it is important to understand how the quality of cell secretions is affected by culture. The aim of this study was to evaluate whether the duration of culture influences the micro-RNAs (miRNAs) cargo of equine amniotic-derived cells (AMCs) and their MVs...
September 20, 2018: Tissue Engineering. Part C, Methods
Katsuhisa Matsuura, Kyoji Ito, Nobuaki Shiraki, Shoen Kume, Nobuhisa Hagiwara, Tatsuya Shimizu
Pluripotent stem cells including induced pluripotent stem (iPS) cells are promising cell sources for regenerative medicine to replace injured tissues and tissue engineering technologies enable engraftment of functional iPS cell-derived cells in vivo for prolong periods. However, the risk of tumor formation is a concern for the use of iPS cells. Bioengineered tissues provide a suitable environment for cell survival, which require vigorous efforts to eliminate remaining iPS cells and prevent tumor formation. We recently reported three iPS cell elimination strategies including methionine-free medium, TRPV1 activation through 42 °C cultivation, and dinaciclib, a cyclin-dependent kinase 1/9 inhibitor...
September 20, 2018: Tissue Engineering. Part C, Methods
Xianqi Li, Ni Li, Kai Chen, Sakae Nagasawa, Michiko Yoshizawa, Hideaki Kagami
Following the discovery of the primary culture of neural stem cells, the spheroid culture has been recognized as one of the selective culture methods for somatic stem cells. Since then, various methods were reported to generate spheroids, which can enrich the potent stem cell population. However, the fundamental factors affecting spheroid formation remain unclear. In this study, we focused on the surface property of the culture dishes, in particular, hydrophobicity. Primary mouse skin culture cells were prepared with conventional two-dimensional culture, and then, the cells were transferred to culture dishes with varying hydrophobicity, which was confirmed with the water contact angles...
September 20, 2018: Tissue Engineering. Part C, Methods
Kali L Manning, Andrew H Thomson, Jeffrey R Morgan
The field of tissue engineering is developing new additive manufacturing technologies to fabricate 3D living constructs for use as in vitro platforms for the testing of drugs and chemicals, or to restore lost function in vivo. In this article, we describe the funnel-guide (FG), a new additive manufacturing strategy for the noncontact manipulation and positioning of multicellular microtissues and we show that the FG can be used to build macrotissues layer by layer. We used agarose micromolds to self-assemble cells into toroid-shaped and honeycomb-shaped microtissues, and observed that when falling in cell culture medium, the microtissues spontaneously righted themselves to a horizontal orientation...
September 20, 2018: Tissue Engineering. Part C, Methods
Charles Blackshear, Mimi Rosealie Borrelli, Ethan Zi Shen, Ryan Chase Ransom, Natalie Narie Chung, Stephanie Vistnes, Dre Irizarry, Rahim Nazerali, Arash Momeni, Michael T Longaker, Derrick C Wan
Significant advances in our understanding of human obesity, endocrinology, and metabolism have been made possible by murine comparative models, in which anatomically analogous fat depots are utilized; however, current research has questioned how truly analogous these depots are. In this study, we assess the validity of the analogy from the perspective of cellular architecture. Whole tissue mounting, confocal microscopy, and image reconstruction software were employed to characterize the three-dimensional structure of the inguinal fat pad in mice, gluteofemoral fat in humans, and subcutaneous adipose tissue of the human abdominal wall...
September 14, 2018: Tissue Engineering. Part C, Methods
Jakob Schmid, Sascha Schwarz, Robert Meier-Staude, Stefanie Sudhop, Hauke Clausen-Schaumann, Matthias Schieker, Robert Huber
Bioreactor systems facilitate three-dimensional (3D) cell culture by coping with limitations of static cultivation techniques. To allow for the investigation of proper cultivation conditions and the reproducible generation of tissue-engineered grafts, a bioreactor system, which comprises the control of crucial cultivation parameters in independent-operating parallel bioreactors, is beneficial. Furthermore, the use of a bioreactor as an automated cell seeding tool enables even cell distributions on stable scaffolds...
October 2018: Tissue Engineering. Part C, Methods
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