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Tissue Engineering. Part C, Methods

Joseph M Mansour, Mostafa Motavalli, James E Dennis, Thomas J Kean, Arnold I Caplan, Jim A Berilla, Jean F Welter
Previous investigations have shown that tissue-engineered articular cartilage can be damaged under a combination of compression and sliding shear. In these cases, damage was identified in histological sections after a test was completed. This approach is limited, in that it does not identify when damage occurred. This especially limits the utility of an assay for evaluating damage when comparing modifications to a tissue-engineering protocol. In this investigation, the feasibility of using ultrasound to detect damage as it occurs was investigated...
July 12, 2018: Tissue Engineering. Part C, Methods
Yanni Jia, Wenjing Li, Haoyun Duan, Zongyi Li, Qingjun Zhou, Weiyun Shi
Corneal endothelium is a single layer of hexagonal cells that maintains the corneal transparency and thickness through its barrier and pump function. For the treatment of corneal endothelial dysfunction, the transplantation of tissue-engineered corneal endothelium and direct injection of cultured corneal endothelial cells were developed because of the severe shortage of donor cornea worldwide. However, the technique difficulty or safety risk still remained. Here we report a novel mini-sheet injection for the cultured corneal endothelial cell transplantation and compare with the effects of single cells injection in rabbit model...
July 11, 2018: Tissue Engineering. Part C, Methods
Guangpeng Liu, Caihe Liao, Xi Chen, Yipin Xu, Jian Tan, Fang Han, Xinhai Ye
Skeletal muscle stem cell (SMSC) transplantation has shown great therapeutical potential in repairing muscle loss and dysfunction, but the muscle acquisition is usually a traumatic procedure causing pain and morbidity to the donor. In the present study, we investigated the feasibility of isolating SMSCs from human orbicularis oculi muscle (OOM), which is routinely discarded during ophthalmic cosmetic surgeries. OOM fragments were harvested from 18 female healthy donors undergoing upper eyelid blepharoplasties...
July 11, 2018: Tissue Engineering. Part C, Methods
Chiemi Nakajima, Kenji Kamimoto, Katsuhiro Miyajima, Masahito Matsumoto, Yasushi Okazaki, Kazuo Kobayashi-Hattori, Makoto Shimizu, Takumi Yamane, Yuichi Oishi, Ken Iwatsuki
Proper identification of pancreatic ducts is a major challenge for researchers performing partial duct ligation (PDL), because pancreatic ducts, which are covered with acinar cells, are translucent and thin. Although damage to pancreatic ducts may activate quiescent ductal stem cells, which may allow further investigation into ductal stem cells for therapeutic use, there is a lack of effective techniques to visualize pancreatic ducts. Here, we report a new method for identifying pancreatic ducts. First, we aimed to visualize pancreatic ducts using black, waterproof fountain pen ink...
July 11, 2018: Tissue Engineering. Part C, Methods
Deirdre Ej Anderson, Grace Pohan, Jaishankar Raman, Filip Konecny, Evelyn Kf Yim, Monica T Hinds
While clinical vascular grafting uses an end-to-side surgical method, researchers primarily use end-to-end implant techniques in preclinical models. This may be due in part to the limitations of using small animal models in research. The work presented here provides support and evidence for the improvement of vascular graft implant techniques by demonstrating the successful implantation of experimental grafts into both large and small animal models. Specifically, models of aortoiliac baboon (Papio anubus) bypass and common carotid rabbit (Oryctolagus cuniculus) bypass were used to test vascular grafts for thrombosis and vascular healing after 1 month using an end-to-side anastomosis grafting procedure...
July 7, 2018: Tissue Engineering. Part C, Methods
Mitchel R Stacy, Cameron A Best, Mark W Maxfield, Maolin Qiu, Yuji Naito, Hirotsugu Kurobe, Nathan Mahler, Kevin A Rocco, Albert Sinusas, Toshiharu Shinoka, Smita Sampath, Christopher K Breuer
<b>Objectives:</b> Tissue-engineered vascular grafts (TEVGs) have demonstrated potential for treating congenital heart disease (CHD); however, quantitative imaging for tracking functional and structural remodeling of TEVGs has not been applied. Therefore, we evaluated the potential of magnetic resonance (MR) imaging for assessing TEVG wall shear stress (WSS) and wall thickness in a large animal model. <b>Methods:</b> Cell-seeded (n=3) or unseeded (n=3) TEVGs were implanted as inferior vena cava interposition grafts in juvenile lambs...
July 6, 2018: Tissue Engineering. Part C, Methods
Zhuoming Liu, Rudell Screven, Lynne Boxer, Michael J Myers, Lax R Devireddy
In this article, we report on the development of a defined serum-free medium capable of supporting the culture expansion of mesenchymal stromal/stem cells (MSCs) from canine adipose tissue (canine Ad-MSCs). The potential benefits of serum-free media can only be utilized if cells cultured in serum-free media maintain the same functional characteristics as cells cultured in serum-containing media. Therefore, we analyze the characteristics of canine Ad-MSCs cultured in this serum-free medium or in serum-containing medium through evaluation of growth kinetics, clonogenic capacity, senescence, and differentiation capacity...
June 20, 2018: Tissue Engineering. Part C, Methods
Marco Vigan├▓, Carlotta Perucca Orfei, Laura de Girolamo, John R Pearson, Enrico Ragni, Paola De Luca, Alessandra Colombini
The use of biochemical inducers of mesenchymal stem cell (MSC) differentiation into tenogenic lineage represents an investigated aspect of tendon disorder treatment. Bone morphogenetic protein 12 (BMP-12) is a widely studied factor, representing along with ascorbic acid (AA) and basic fibroblast growth factor (bFGF) one of the most promising stimulus in this context so far. Quantitative gene expression of specific tenogenic marker is commonly used to assess the efficacy of these supplements. Nevertheless, the reliability of these data is strongly associated with the choice of stable housekeeping genes...
May 16, 2018: Tissue Engineering. Part C, Methods
Maurits Geert Laurent Olthof, Marianna A Tryfonidou, Mahrokh Dadsetan, Wouter Dhert, Michael J Yaszemski, Diederik H R Kempen, Lichun Lu
Local sustained delivery of bioactive molecules from biomaterials is a promising strategy to enhance bone regeneration. To optimize delivery vehicles for bone formation, the design characteristics are tailored with consequential effect on BMP-2 release and bone regeneration. Complying with the 3R principles, the growth factor release is often investigated in vitro using several buffers to mimic the in vivo physiological environment. However, this remains an unmet need. Therefore, this study investigates the correlation between the in vitro and in vivo (IVIVC) BMP-2 release from complex delivery vehicles in several commonly used in vitro buffers: cell culture model, phosphate buffered saline, and a strong desorption buffer...
May 14, 2018: Tissue Engineering. Part C, Methods
Romina Buchs, Bruno Lehner, Philippe Meuwly, Bruno Schnyder
Candida albicans frequently causes recurrent intimal infectious disease (ID). This demands the treatment of multiple phases of the infection. The objective of this study was to uncover the host-pathogen interaction using two-dimensional (2D) epithelium cell-barrier and three-dimensional (3D) subepithelium tissue cells of human mucosa. The 2D cell cultures assessed C. albicans adhesion. Addition of the antifungal drug Fluconazol did not inhibit the adhesion, despite its pathogen growth inhibition (minimal inhibitory concentration value 0...
July 2018: Tissue Engineering. Part C, Methods
Adam Mendez, Alexandra N Rindone, Namrata Batra, Pegah Abbasnia, Janaka Senarathna, Stacy Gil, Darian Hadjiabadi, Warren L Grayson, Arvind P Pathak
Tissue-engineered scaffolds are a powerful means of healing craniofacial bone defects arising from trauma or disease. Murine models of critical-sized bone defects are especially useful in understanding the role of microenvironmental factors such as vascularization on bone regeneration. Here, we demonstrate the capability of a novel multimodality imaging platform capable of acquiring in vivo images of microvascular architecture, microvascular blood flow, and tracer/cell tracking via intrinsic optical signaling (IOS), laser speckle contrast (LSC), and fluorescence (FL) imaging, respectively, in a critical-sized calvarial defect model...
July 2018: Tissue Engineering. Part C, Methods
Eleonora Rossi, Eva Mracsko, Adam Papadimitropoulos, Nima Allafi, Dieter Reinhardt, Arne Mehrkens, Ivan Martin, Irene Knuesel, Arnaud Scherberich
Triggering receptor expressed on myeloid cells-2 (TREM-2), a transmembrane receptor expressed by macrophages, microglia, and osteoclasts (OCs), plays a protective role in late-onset Alzheimer Disease (AD). To validate TREM-2 as a therapeutic target in AD, its potential secondary parallel effect on bone homeostasis should be clarified. However, animal models and monolayer cultures of human cells were shown poorly predictive of TREM-2 function in human. Therefore, this study aimed to engineer a tridimensional in vitro model using human progenitors differentiated into osteoblasts and OCs, recapitulating physiological bone homeostasis...
July 2018: Tissue Engineering. Part C, Methods
Eline E van Haaften, Tamar B Wissing, Marcel C M Rutten, Jurgen A Bulsink, Kujtim Gashi, Mathieu A J van Kelle, Anthal I P M Smits, Carlijn V C Bouten, Nicholas A Kurniawan
The success of cardiovascular tissue engineering (TE) strategies largely depends on the mechanical environment in which cells develop a neotissue through growth and remodeling processes. This mechanical environment is defined by the local scaffold architecture to which cells adhere, that is, the microenvironment, and by external mechanical cues to which cells respond, that is, hemodynamic loading. The hemodynamic environment of early developing blood vessels consists of both shear stress (due to blood flow) and circumferential stretch (due to blood pressure)...
July 2018: Tissue Engineering. Part C, Methods
Thomas Anthony Dixon, Eliad Cohen, Dana M Cairns, Maria Rodriguez, Juanita Mathews, Rod R Jose, David L Kaplan
The physical connection between motoneurons and skeletal muscle targets is responsible for the creation of neuromuscular junctions (NMJs), which allow electrical signals to be translated to mechanical work. NMJ pathology contributes to the spectrum of neuromuscular, motoneuron, and dystrophic disease. Improving in vitro tools that allow for recapitulation of the physiology of the neuromuscular connection will enable researchers to better understand the development and maturation of NMJs, and will help to decipher mechanisms leading to NMJ degeneration...
June 2018: Tissue Engineering. Part C, Methods
Hanis Hasmad, Mohd Reusmaazran Yusof, Zainul Rashid Mohd Razi, Ruszymah Bt Hj Idrus, Shiplu Roy Chowdhury
Fabrication of composite scaffolds is one of the strategies proposed to enhance the functionality of tissue-engineered scaffolds for improved tissue regeneration. By combining multiple elements together, unique biomimetic scaffolds with desirable physical and mechanical properties can be tailored for tissue-specific applications. Despite having a highly porous structure, the utility of electrospun fibers (EF) as scaffold is usually hampered by their insufficient mechanical strength. In this study, we attempted to produce a mechanically competent scaffold with cell-guiding ability by fabricating aligned poly lactic-co-glycolic acid (PLGA) fibers on decellularized human amniotic membrane (HAM), known to possess favorable tensile and wound healing properties...
June 2018: Tissue Engineering. Part C, Methods
Vahid Mirshafiee, Brendan A C Harley, Mary L Kraft
Characterization of the heterogeneity within stem cell populations, which affects their differentiation potential, is necessary for the design of artificial cultures for stem cell expansion. In this study, we assessed whether self-organizing maps (SOMs) of single-cell time-of-flight secondary ion mass spectrometry (TOF-SIMS) data provide insight into the spectral, and thus the related functional heterogeneity between and within three hematopoietic cell populations. SOMs were created of TOF-SIMS data from individual hematopoietic stem and progenitor cells (HSPCs), lineage-committed common lymphoid progenitors (CLPs), and fully differentiated B cells that had been isolated from murine bone marrow via conventional flow cytometry...
June 2018: Tissue Engineering. Part C, Methods
Mitchell R Ladd, Laura Y Martin, Adam Werts, Cait Costello, Chhinder P Sodhi, William B Fulton, John C March, David J Hackam
Short bowel syndrome (SBS) is a major cause of morbidity and mortality in the pediatric population, for which treatment options are limited. To develop novel approaches for the treatment of SBS, we now focus on the development of a tissue-engineered intestine (also known as an "artificial intestine"), in which intestinal stem cells are cultured onto an absorbable bioscaffold, followed by implantation into the host. To enhance the translational potential of these preclinical studies, we have developed three clinically relevant models in neonatal piglets, which approximate the size of the human infant and were evaluated after implantation and establishment of intestinal continuity over the long term...
June 2018: Tissue Engineering. Part C, Methods
Callie An Knuth, Caoimhe H Kiernan, Virginia Palomares Cabeza, Johannes Lehmann, Janneke Witte-Bouma, Derk Ten Berge, Pieter A Brama, Eppo B Wolvius, Elske M Strabbing, Maarten J Koudstaal, Roberto Narcisi, Eric Farrell
Mesenchymal stem cells/marrow stromal cells (MSCs) are attractive for applications ranging from research and development to use in clinical therapeutics. However, the most commonly studied MSCs, adult bone marrow MSCs (A-MSCs), are limited by significant donor variation resulting in inconsistent expansion rates and multilineage differentiation capabilities. We have recently obtained permission to isolate pediatric MSCs (P-MSCs) from surplus iliac crest bone chips. Here, we developed a simple and easily replicable isolation protocol yielding P-MSCs, which adhere to MSC defining guidelines...
June 2018: Tissue Engineering. Part C, Methods
Maaike V J Braham, Tilman Ahlfeld, A Rahul Akkineni, Monique C Minnema, Wouter J A Dhert, F Cumhur ├ľner, Catherine Robin, Anja Lode, Michael Gelinsky, Jacqueline Alblas
The bone marrow microenvironment is the preferred location of multiple myeloma, supporting tumor growth and development. It is composed of a collection of interacting subniches, including the endosteal and perivascular niche. Current in vitro models mimic either of these subniches. By developing a model combining both niches, this study aims to further enhance the ability to culture primary myeloma cells in vitro. Also, the dependency of myeloma cells on each niche was studied. A 3D bone marrow model containing two subniches was created using 3D bioprinting technology...
May 2018: Tissue Engineering. Part C, Methods
Brian C Syverud, Jonathan P Gumucio, Brittany L Rodriguez, Olga M Wroblewski, Shelby E Florida, Christopher L Mendias, Lisa M Larkin
The growing deficit in suitable tissues for patients awaiting organ transplants demonstrates the clinical need for engineered tissues as alternative graft sources. Demonstrating safety and efficacy by tracking the migration and fate of implanted cells is a key consideration required for approval of promising engineered tissues. Cells from transgenic animals that express green fluorescent protein (GFP) are commonly used for this purpose. However, GFP can create difficulties in practice due to high levels of green autofluorescence in many musculoskeletal tissues...
May 2018: Tissue Engineering. Part C, Methods
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