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Tissue Engineering. Part C, Methods

Mohammad Houshmand, Masoud Soleimani, Amir Atashi, Giuseppe Saglio, Mohammad Abdollahi, Mahin Nikougoftar Zarif
Bone marrow niche is a major contributing factor in leukemia development and drug resistance in acute myeloid leukemia (AML) patients. Although mimicking leukemic bone marrow niche relies on two-dimensional (2D) culture conditions, it cannot recapitulate complex bone marrow structure that causes introduction of different three-dimensional (3D) scaffolds. Simultaneously, microfluidic platform by perfusing medium culture mimic interstitial fluid flow, along with 3D scaffold would help for mimicking bone marrow microenvironment...
January 13, 2017: Tissue Engineering. Part C, Methods
Anna Diez-Escudero, Montserrat Espanol, Edgar B Montufar, Gemma Di Pompo, Gabriela Ciapetti, Nicola Baldini, Maria-Pau Ginebra
This paper presents the application of dual Focused Ion Beam-Scanning Electron Microscopy (FIB-SEM) imaging for pre-clinical testing of calcium-phosphates with osteoclast precursor cells and how this high resolution imaging technique is able to reveal microstructural changes at a level of detail previously not possible. Calcium phosphate substrates, having similar compositions but different microstructures, were produced using low and high temperature processes(biomimetic calcium deficient hydroxyapatite and stoichiometric sintered hydroxyapatite respectively)...
January 12, 2017: Tissue Engineering. Part C, Methods
Wen-Yue Liu, Shi-Gang Lin, Ru-Yi Zhuo, Yuan-Yuan Xie, Wei Pan, Xian-Feng Lin, Fei-Xia Shen
Women younger than 40 years may face early menopause because of premature ovarian failure (POF). The cause of POF can be idiopathic or iatrogenic, especially the cancer-induced oophorectomy and chemo- or radiation therapy. The current treatments, including hormone replacement therapy (HRT) and cryopreservation techniques, have increased risk of ovarian cancer and may reintroduce malignant cells after autografting. Decellularization technique has been regarded as a novel regenerative medicine strategy for organ replacement, wherein the living cells of an organ are removed, leaving the extracellular matrix (ECM) for cellular seeding...
January 10, 2017: Tissue Engineering. Part C, Methods
Mark Colin Allenby, Ruth Misener, Nicki Panoskaltsis, Athanasios Mantalaris
Three-dimensional imaging techniques provide spatial insight to environmental and cellular interactions and are implemented in various fields, including tissue engineering, but have been restricted by limited quantification tools which misrepresent or underutilize the cellular phenomena captured. This study develops image post-processing algorithms pairing complex Euclidean metrics with Monte Carlo simulations in order to quantitatively assess cell and microenvironment spatial distributions while utilizing, for the first time, the entire 3D image captured...
January 9, 2017: Tissue Engineering. Part C, Methods
Aranzazu Villasante, Alessandro Marturano, Samuel T Robinson, Zen Liu, X Edward Guo, Gordana Vunjak-Novakovic
Ewing's sarcoma (ES) is a poorly differentiated pediatric tumor of aggressive behavior characterized by propensity to metastasize to bone. Interactions between the tumor and bone cells orchestrate a vicious cycle in which tumor cells induce osteoclast differentiation and activation to cause osteolytic lesions, broken bones, pain and hypercalcemia. The lack of controllable models that can recapitulate osteolysis in ES impedes the development of new therapies and limits our understanding of how tumor cells invade bone...
January 9, 2017: Tissue Engineering. Part C, Methods
Junnan Tang, Adam Vandergriff, Zegen Wang, Michael Hensley, Jhon Cores, Tyler Allen, Phuong-Uyen Dinh, Jinying Zhang, Thomas Caranasos, Ke Cheng
Layering a regenerative polymer scaffold on the surface of the heart, termed as a cardiac patch, has been proven to be effective in preserving cardiac function after myocardial infarction. However, the placement of such a patch on the heart usually needs open-chest surgery, which is traumatic therefore prevents the translation of this strategy into the clinic. We sought to device a novel method to apply a cardiac patch by spray painting in situ polymerizable biomaterials onto the heart with a minimally invasive procedure...
January 9, 2017: Tissue Engineering. Part C, Methods
Xunxun Lin, Wenbo Wang, Wen Jie Zhang, Zhi-Yong Zhang, Guangdong Zhou, Yilin Cao, Wei Liu
Synthetic polymers such as polyglycolic acid (PGA) fibers are the traditional tissue engineering scaffolds that are widely used for engineering a variety of soft tissues. However, the major disadvantage of this polymer material is its released acidic degradation products that trigger inflammatory response and fibrotic process, which affects the biocompatibility and the quality of the engineered tissues. In this study, the effect of hyaluronic acid (HA) coating on improving PGA biocompatibility was explored...
January 5, 2017: Tissue Engineering. Part C, Methods
Wenlong Liu, Xiuli Dan, Ting Wang, William W Lu, Haobo Pan
The development of an optimal animal model that could provide fast assessments of the interaction between bone and orthopedic implants is essential for both preclinical and theoretical researches in the design of novel biomaterials. Compared with other animal models, mice have superiority in accessing the well-developed transgenic modification techniques (e.g., cell tracing, knockoff, knockin, and so on), which serve as powerful tools in studying molecular mechanisms. In this study, we introduced the establishment of a mouse model, which was specifically tailored for the assessment of bone-implant interaction in a load-bearing bone marrow microenvironment and could potentially allow the molecular mechanism study of biomaterials by using transgenic technologies...
October 31, 2016: Tissue Engineering. Part C, Methods
Rahel D May, Adel Tekari, Daniela A Frauchiger, Anna Krismer, Lorin M Benneker, Benjamin Gantenbein
Low back pain (LBP) is an increasing global health problem associated with intervertebral disc (IVD) trauma and degeneration. Current treatment options include surgical interventions with partial unsatisfactory outcomes reported such as failure to relieve LBP, nonunions, nerve injuries, or adjacent segment disease. Cell-based therapy and tissue engineered IVD constructs supplemented with transfected disc cells that incorporate factors enhancing matrix synthesis represent an appealing approach to regenerate the IVD...
January 2017: Tissue Engineering. Part C, Methods
Chi Ma, Xiaohua Liu
Nanofibrous architecture presents unique biophysical cues to facilitate cellular responses and is considered an indispensable feature of a biomimetic three-dimensional (3D) scaffold and cell carrier. While electrospinning is a widely used method to prepare natural extracellular matrix-like nanofibers, it faces significant challenges to incorporate nanofibrous architecture into well-defined macroporous 3D scaffolds or injectable microspheres. Here we report a nonelectrospinning approach that is effective at generating nanofibers from a variety of synthetic and natural biodegradable polymers and integrating these nanofibers into (1) 3D scaffolds with constructive geometry and designed internal macropore structures; and (2) injectable microspheres...
January 2017: Tissue Engineering. Part C, Methods
Minu Karthika Ganesan, Richard Finsterwalder, Heide Leb, Ulrike Resch, Karin Neumüller, Rainer de Martin, Peter Petzelbauer
The response of blood vessels to physiological and pathological stimuli partly depends on the cross talk between endothelial cells (EC) lining the luminal side and smooth muscle cells (SMC) building the inner part of the vascular wall. Thus, the in vitro analysis of the pathophysiology of blood vessels requires coculture systems of EC and SMC. We have developed and validated a modified three-dimensional sandwich coculture (3D SW-CC) of EC and SMC using open μ-Slides with a thin glass bottom allowing direct imaging...
January 2017: Tissue Engineering. Part C, Methods
Deniz Öztürk-Kaloglu, David Hercher, Philipp Heher, Katja Posa-Markaryan, Simon Sperger, Alice Zimmermann, Susanne Wolbank, Heinz Redl, Ara Hacobian
Monitoring of cell differentiation is a crucial aspect of cell-based therapeutic strategies depending on tissue maturation. In this study, we have developed a noninvasive reporter system to trace murine skeletal muscle differentiation. Either a secreted bioluminescent reporter (Metridia luciferase) or a fluorescent reporter (green fluorescent protein [GFP]) was placed under the control of the truncated muscle creatine kinase (MCK) basal promoter enhanced by variable numbers of upstream MCK E-boxes. The engineered pE3MCK vector, coding a triple tandem of E-Boxes and the truncated MCK promoter, showed twentyfold higher levels of luciferase activation compared with a Cytomegalovirus (CMV) promoter...
January 2017: Tissue Engineering. Part C, Methods
Selcan Guler, Pezhman Hosseinian, Halil Murat Aydin
Decellularization of tissues and organs has high potential to obtain unique conformation and composition as native tissue structure but may result in weakened tissue mechanical strength. In this study, poly(glycerol-sebacate) (PGS) elastomers were combined with decellularized aorta fragments to investigate the changes in mechanical properties. PGS prepolymer was synthesized via microwave irradiation and then in situ crosslinked within the decellularized aorta extracellular matrix (ECM). Tensile strength (σ) values were found comparable as 0...
January 2017: Tissue Engineering. Part C, Methods
Li Min Tay, Christian Wiraja, David C Yeo, Yingnan Wu, Zheng Yang, Yon Jin Chuah, Eng Hin Lee, Yuejun Kang, Chenjie Xu
Chondrogenic differentiation of human mesenchymal stem cells (MSCs) in three-dimensional hydrogel holds promise as a method for repairing injured articular cartilage. Given MSC plasticity (its potential to mature into alternative lineages), nondestructive monitoring is critical for the optimization of chondrogenic differentiation conditions and the evaluation of the final product. However, conventional validation/assessments of the differentiation process (i.e., quantitative reverse transcription polymerase chain reaction [qRT-PCR] and histology) are end-point assays requiring disruption of the sample...
January 2017: Tissue Engineering. Part C, Methods
Hannah K Wilson, Madeline G Faubion, Michael K Hjortness, Sean P Palecek, Eric V Shusta
The blood-brain barrier (BBB) maintains brain homeostasis but also presents a major obstacle to brain drug delivery. Brain microvascular endothelial cells (BMECs) form the principal barrier and therefore represent the major cellular component of in vitro BBB models. Such models are often used for mechanistic studies of the BBB in health and disease and for drug screening. Recently, human induced pluripotent stem cells (iPSCs) have emerged as a new source for generating BMEC-like cells for use in in vitro human BBB studies...
December 2016: Tissue Engineering. Part C, Methods
Cornelia Schneider, Johannes Lehmann, Gerjo J V M van Osch, Florian Hildner, Andreas Teuschl, Xavier Monforte, David Miosga, Patrick Heimel, Eleni Priglinger, Heinz Redl, Suanne Wolbank, Sylvia Nürnberger
Natural extracellular matrix-derived biomaterials from decellularized allogenic tissues are of increasing interest for tissue engineering because their structure and composition provide a complexity that is not achievable with current manufacturing techniques. The prerequisite to bring allogenic tissue from bench to bedside as a functional biomaterial is the full removal of cells while preserving most of its native characteristics such as structure and composition. The exceptionally dense structure of articular cartilage, however, poses a special challenge for decellularization, scaffold preparation, and reseeding...
December 2016: Tissue Engineering. Part C, Methods
Luke R Jennings, Helen E Colley, Jane Ong, Foti Panagakos, James G Masters, Harsh M Trivedi, Craig Murdoch, Simon Whawell
Tissue-engineered oral mucosal equivalents (OME) are being increasingly used to measure toxicity, drug delivery, and to model oral diseases. Current OME mainly comprise normal oral keratinocytes (NOK) cultured on top of a normal oral fibroblasts-containing matrix. However, the commercial supply of NOK is limited, restricting widespread use of these mucosal models. In addition, NOK suffer from poor longevity and donor-to-donor variability. Therefore, we constructed, characterized, and tested the functionality of OME based on commercial TERT2-immortalized oral keratinocytes (FNB6) to produce a more readily available alternative to NOK-based OME...
December 2016: Tissue Engineering. Part C, Methods
Pedram Akbari, Stephen D Waldman, Evan J Propst, Sharon L Cushing, Joanna F Weber, Herman Yeger, Walid A Farhat
Traditional methods of cartilage tissue engineering rely on the use of scaffolds. Although successful chondrogenesis has been reported in scaffold-based constructs, the use of exogenous materials has limited their application due to eliciting host immunogenic responses and potentially resulting in construct failure. As a result, tissue engineering approaches, which aim to generate scaffold-free cartilaginous constructs, have become of particular interest. Here, we generated stable three-dimensional scaffold-free cartilaginous constructs by cultivating expanded pediatric nasal chondrocyte multilayers in a slow turning lateral vessel bioreactor system under chemically defined media...
December 2016: Tissue Engineering. Part C, Methods
Farrah A Monibi, Chantelle C Bozynski, Keiichi Kuroki, Aaron M Stoker, Ferris M Pfeiffer, Seth L Sherman, James L Cook
Decellularized scaffolds composed of extracellular matrix (ECM) hold promise for repair and regeneration of the meniscus, given the potential for ECM-based biomaterials to aid in stem cell recruitment, infiltration, and differentiation. The objectives of this study were to decellularize canine menisci to fabricate a micronized, ECM-derived scaffold and to determine the cytocompatibility and repair potential of the scaffold ex vivo. Menisci were decellularized with a combination of physical agitation and chemical treatments...
December 2016: Tissue Engineering. Part C, Methods
Todd A Goldstein, Casey J Epstein, John Schwartz, Alex Krush, Dan J Lagalante, Kevin P Mercadante, David Zeltsman, Lee P Smith, Daniel A Grande
Numerous studies have shown the capabilities of three-dimensional (3D) printing for use in the medical industry. At the time of this publication, basic home desktop 3D printer kits can cost as little as $300, whereas medical-specific 3D bioprinters can cost more than $300,000. The purpose of this study is to show how a commercially available desktop 3D printer could be modified to bioprint an engineered poly-l-lactic acid scaffold containing viable chondrocytes in a bioink. Our bioprinter was used to create a living 3D functional tissue-engineered cartilage scaffold...
December 2016: Tissue Engineering. Part C, Methods
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