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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
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 paper, we describe the funnel-guide, a new additive manufacturing strategy for the non-contact manipulation and positioning of multi-cellular microtissues and we show that the funnel-guide can be used to build macrotissues layer-by-layer. We used agarose micro-molds 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...
August 14, 2018: Tissue Engineering. Part C, Methods
David Ede, Nikki Davidoff, Alejandro Blitch, Niloofar Farhang, Robby D Bowles
Musculoskeletal tissues contain critical gradients in extracellular matrix (ECM) composition and cell types that allow for proper mechanical function of tissues and integration between adjacent tissues. However, properly controlling these patterns in engineered tissues is difficult and tissue engineering (TE) is presently in need of methods to generate integration zones for tissue anchoring, transition zones between tissues, and controlled ECM gradients for proper mechanical function. In this study, we present a novel method of using a microfluidic flow cell array (MFCA) to precisely control cell deposition onto TE constructs to produce tunable cell patterns on engineered constructs...
September 2018: Tissue Engineering. Part C, Methods
Qiliang Zhou, Xulu Ye, Qingsong Ran, Akihiko Kitahara, Yoshifumi Matsumoto, Masato Moriyama, Yoichi Ajioka, Yasuo Saijo
The outcomes of tracheal transplantation for the treatment of airway stenosis are unsatisfactory. We investigated the feasibility of regeneration of the trachea using a rat decellularized tracheal scaffold and mouse-induced pluripotent stem (iPS) cells for in vivo transplantation. The rat trachea was first decellularized using a detergent/enzymatic treatment method. We successfully established a centrifugation method that can transplant cells onto the luminal surface of the decellularized rat tracheal scaffold circumferentially...
September 2018: Tissue Engineering. Part C, Methods
Audrey Baze, Céline Parmentier, Delilah F G Hendriks, Tracey Hurrell, Bruno Heyd, Philippe Bachellier, Catherine Schuster, Magnus Ingelman-Sundberg, Lysiane Richert
Recent advances in the development of various culture platforms are promising for achieving more physiologically relevant in vitro hepatic models using primary human hepatocytes (PHHs). Previous studies have shown the value of PHHs three-dimensional (3D) spheroid models, cultured in low cell number (1330-2000 cells/3D spheroid), to study long-term liver function as well as pharmacological drug effects and toxicity. In this study, we report that only plateable PHHs aggregate and form compact 3D spheroids with a success rate of 79%, and 96% reproducibility...
September 2018: Tissue Engineering. Part C, Methods
William H Trousdale, Christopher G Salib, Nicolas Reina, Eric A Lewallen, Anthony Viste, Daniel J Berry, Mark E Morrey, Joaquin Sanchez-Sotelo, Andre J van Wijnen, Matthew P Abdel
INTRODUCTION: The inflammatory cascade and production of prostaglandins may play a role in the pathogenesis of arthrofibrosis, a debilitating condition after joint replacement and other orthopedic procedures. Cyclooxygenase 2 (COX-2) inhibitors may mitigate the inflammatory response and formation of arthrofibrosis, but oral delivery is associated with risk of systemic side effects in many patients. The nonsteroidal anti-inflammatory drug, celecoxib, may have therapeutic benefits for arthrofibrosis, but current methods for its local delivery (e...
September 2018: Tissue Engineering. Part C, Methods
Eva Dohle, Smriti Singh, Akihiro Nishigushi, Thorsten Fischer, Matthias Wessling, Martin Möller, Robert Sader, Jennifer Kasper, Shahram Ghanaati, C James Kirkpatrick
The development of an in vitro model resembling the alveolar-capillary barrier might be a highly beneficial tool to study lung physiology as well as the immune response of the lung to infection or after exposure to nanoparticles. This study is based on an in vitro alveolar barrier developed on a basement membrane mimic, composed of ultrathin nanofiber meshes generated via electrospinning using bioresorbable poly(ɛ-caprolactone). As cellular components, NCI H441, resembling the alveolar epithelial cells, and ISO-HAS-1, an endothelial cell line, were used to perform bipolar coculture experiments for a total cultivation period of 14 days...
September 2018: Tissue Engineering. Part C, Methods
Lin Fan, Chao Lin, Peng Zhao, Xuejun Wen, Guodong Li
The purpose of this study is to corroborate the feasibility of an injectable dextran hydrogel, in situ formed through a catalyst-free bioorthogonal reaction, for cartilage regeneration with adult chondrocytes or adipose-derived stem cells (ADSCs). To this end, an injectable dextran (dextran-AN) hydrogel encapsulating the rabbit adult chondrocytes or rabbit ADSCs (rADSCs) or human ADSCs (hADSCs) was successfully prepared by simply incubating the cells with dibenzocyclooctyne-dextran conjugate and azide-dextran conjugate under physiological conditions through a copper-free bioorthogonal reaction without using additional reagents...
September 2018: 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 (US) to detect damage as it occurs was investigated...
August 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. In this study, we report a novel mini-sheet injection for the cultured corneal endothelial cell transplantation and compare with the effects of single-cell injection in rabbit model...
August 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 this study, we investigated the feasibility of isolating SMSCs from human orbicularis oculi muscle (OOM), which is routinely removed and discarded during ophthalmic cosmetic surgeries. OOM fragments were harvested from 18 female healthy donors undergoing upper eyelid plasties...
August 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. In this study, we report a new method for identifying pancreatic ducts. First, we aimed to visualize pancreatic ducts using black, waterproof fountain pen ink...
August 2018: Tissue Engineering. Part C, Methods
Deirdre E J Anderson, Grace Pohan, Jaishankar Raman, Filip Konecny, Evelyn K F 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 anubis) 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...
August 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 J Sinusas, Toshiharu Shinoka, Smita Sampath, Christopher K Breuer
OBJECTIVES: 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. METHODS: Cell-seeded (n = 3) or unseeded (n = 3) TEVGs were implanted as inferior vena cava interposition grafts in juvenile lambs...
August 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
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...
July 2018: Tissue Engineering. Part C, Methods
Maurits G L Olthof, Marianna A Tryfonidou, Mahrokh Dadsetan, Wouter J A 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 bone morphogenetic protein-2 (BMP-2) release and bone regeneration. Complying with the 3R principles (Replacement, Reduction, and Refinement), 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...
July 2018: Tissue Engineering. Part C, Methods
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