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Perfusion decellularization

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https://www.readbyqxmd.com/read/28735385/preparation-of-decellularized-liver-scaffolds-and-recellularized-liver-grafts
#1
Yibin Chen, Sharon Geerts, Maria Jaramillo, Basak E Uygun
Severe liver disease is the 12th leading cause of death in the USA, with organ transplantation often being the only viable option for treatment. However, due to the shortage of viable donor livers, it is estimated that over 1200 patients died in 2015 while waiting for liver transplantation. This highlights the need for alternative sources of viable organs. In this study, we describe a method that provides the groundwork for the development of functional liver grafts. The approach described here is for removal of cells from intact livers and subsequently repopulating them with functional liver cells...
July 23, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28726588/reconstituting-mouse-lungs-with-conditionally-reprogrammed-human-bronchial-epithelial-cells
#2
Ryan LaRanger, Jennifer R Peters-Hall, Melissa Coquelin, Busola R Alabi, Chris Chen, Woodring E Wright, Jerry Shay
We developed methods for conditionally reprogramming (CR) primary human bronchial epithelial cells (HBECs) to extend their functional life span and permit their differentiation into both upper and lower airway lung epithelium. We also developed a bioreactor to support vascular perfusion and rhythmic breathing of decellularized mouse lungs reconstituted with CR HBECs isolated from patients with and without cystic fibrosis (CF). While conditionally reprogrammed cells only differentiate into an upper airway epithelium after 35 days at the air liquid interface, in reconstituted lungs these cells differentiate into upper airway bronchial epithelium and lower airway alveolar structures after 12 days...
July 20, 2017: Tissue Engineering. Part A
https://www.readbyqxmd.com/read/28665233/decellularization-of-whole-human-liver-grafts-using-controlled-perfusion-for-transplantable-organ-bioscaffolds
#3
Monique Verstegen, Jorke Willemse, Sjoerd van den Hoek, Gert-Jan Kremers, Theo M Luider, Nick A van Huizen, Francois E J A Willemssen, Herold J Metselaar, Jan Nm IJzermans, Luc J W van der Laan, Jeroen de Jonge
Liver transplantation is the only effective treatment for end-stage liver disease, but absolute donor shortage remains a limiting factor. Recent advances in tissue engineering focus on generation of native extracellular matrix (ECM) by decellularized complete livers in animal models. Though proof of concept has been reported for human livers, the current study aims to perform whole liver decellularization in a clinically relevant series using controlled machine perfusion. Here we describe a mild non-destructive decellularization protocol, effective in 11 discarded human whole liver grafts to generate constructs that reliably maintain hepatic architecture and ECM components using machine perfusion, while completely removing cellular DNA and RNA...
June 30, 2017: Stem Cells and Development
https://www.readbyqxmd.com/read/28570606/effects-of-two-different-decellularization-routes-on-the-mechanical-properties-of-decellularized-lungs
#4
Jessica Julioti Urbano, Renata Kelly da Palma, Flávia Mafra de Lima, Paula Fratini, Leticia Lopes Guimaraes, Juan J Uriarte, Letícia Heineck Alvarenga, Maria Angelica Miglino, Rodolfo de Paula Vieira, Renato Araujo Prates, Daniel Navajas, Ramon Farrè, Luis Vicente Franco Oliveira
Considering the limited number of available lung donors, lung bioengineering using whole lung scaffolds has been proposed as an alternative approach to obtain lungs suitable for transplantation. However, some decellularization protocols can cause alterations on the structure, composition, or mechanical properties of the lung extracellular matrix. Therefore, the aim of this study was to compare the acellular lung mechanical properties when using two different routes through the trachea and pulmonary artery for the decellularization process...
2017: PloS One
https://www.readbyqxmd.com/read/28569086/estimation-of-the-physiological-mechanical-conditioning-in-vascular-tissue-engineering-by-a-predictive-fluid-structure-interaction-approach
#5
Claudia Tresoldi, Elena Bianchi, Alessandro Filippo Pellegata, Gabriele Dubini, Sara Mantero
The in vitro replication of physiological mechanical conditioning through bioreactors plays a crucial role in the development of functional Small-Caliber Tissue-Engineered Blood Vessels. An in silico scaffold-specific model under pulsatile perfusion provided by a bioreactor was implemented using a fluid-structure interaction (FSI) approach for viscoelastic tubular scaffolds (e.g. decellularized swine arteries, DSA). Results of working pressures, circumferential deformations, and wall shear stress on DSA fell within the desired physiological range and indicated the ability of this model to correctly predict the mechanical conditioning acting on the cells-scaffold system...
August 2017: Computer Methods in Biomechanics and Biomedical Engineering
https://www.readbyqxmd.com/read/28550502/decellularization-methods-for-scaffold-fabrication
#6
Sweta K Gupta, Narayan C Mishra
Decellularization is the process of removal of native cells from tissue, leaving behind a three-dimensional (3D) ultrastructure of extracellular matrix (ECM) proteins while preserving the bioactivity and mechanics of the tissue. It offers a unique top-down approach for fabricating ECM based natural scaffold for tissue engineering application. Herein, this chapter presents the fabrication of decellularized scaffold employing different methods: whole organ perfusion, immersion and agitation, pressure gradient, and supercritical fluid...
May 27, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28536786/future-prospects-for-human-tissue-engineered-urethra-transplantation-decellularization-and-recellularization-based-urethra-regeneration
#7
Abdol-Mohammad Kajbafzadeh, Reza Abbasioun, Shabnam Sabetkish, Nastaran Sabetkish, Parvin Rahmani, Kamyar Tavakkolitabassi, Hamid Arshadi
To evaluate the histological characteristics of decellularized human urethra after transplantation into the rat omentum and compare in vivo cell seeding with perfusion-based and cell sheet urethral regeneration. Eight adult human male urethras accompanied with the surrounding corpus spongiosum were obtained. The tissues were decellularized with detergent-based method. The efficacy of decellularization and extracellular matrix preservation was evaluated by several techniques. Decellularized scaffolds were transplanted into the omentum of 12 male rats and located into the scrotum...
May 23, 2017: Annals of Biomedical Engineering
https://www.readbyqxmd.com/read/28480220/culture-of-ipscs-derived-pancreatic-%C3%AE-like-cells-in-vitro-using-decellularized-pancreatic-scaffolds-a-preliminary-trial
#8
Jian Wan, Yan Huang, Pengcheng Zhou, Yibing Guo, Cen Wu, Shajun Zhu, Yao Wang, Lei Wang, Yuhua Lu, Zhiwei Wang
Diabetes mellitus is a disease which has affected 415 million patients in 2015. In an effort to replace the significant demands on transplantation and morbidity associated with transplantation, the production of β-like cells differentiated from induced pluripotent stem cells (iPSCs) was evaluated. This approach is associated with promising decellularized scaffolds with natural extracellular matrix (ECM) and ideal cubic environment that will promote cell growth in vivo. Our efforts focused on combining decellularized rat pancreatic scaffolds with mouse GFP(+)-iPSCs-derived pancreatic β-like cells, to evaluate whether decellularized scaffolds could facilitate the growth and function of β-like cells...
2017: BioMed Research International
https://www.readbyqxmd.com/read/28457326/breast-cancer-cells-form-primary-tumors-on-ex%C3%A2-vivo-four-dimensional-lung-model
#9
Kristi A Pence, Dhruva K Mishra, Michael Thrall, Bhuvanesh Dave, Min P Kim
BACKGROUND: Breast cancer mortality is most common in cancer in women, and there are no ex vivo models that can capture the primary growth of tumor with fidelity to the in vivo tumor growth. In this study, we grew human breast cancer cell lines in an acellular lung matrix of the ex vivo four-dimensional lung model to determine if they form primary tumor and the extent to which they mimic the histology and characteristics of the human tumors. MATERIALS AND METHODS: Rat lungs were harvested, decellularized, and placed in a bioreactor...
April 2017: Journal of Surgical Research
https://www.readbyqxmd.com/read/28453800/coronary-sinus-injury-resulting-from-endarterectomy-during-multivessel-bypass-surgery
#10
Mehmet H Akay, Manish K Patel, Igor D Gregoric
Coronary sinus injury is a very rare complication of cardiac surgery and is usually related to coronary sinus perfusion catheter placement for retrograde cardioplegia infusion. It can be either a catheter-related perforation or high-perfusion pressure-related injury to the coronary sinus. Primary repair of the coronary sinus or over-sewing are two possible options to resolve this complication. Decellularized extracellular matrix from porcine intestinal submucosa is widely used as an approved material for repairing cardiac structures...
May 1, 2017: Interactive Cardiovascular and Thoracic Surgery
https://www.readbyqxmd.com/read/28396609/in-vitro-biomimetic-platforms-featuring-a-perfusion-system-and-3d-spheroid-culture-promote-the-construction-of-tissue-engineered-corneal-endothelial-layers
#11
Shanyi Li, Yuting Han, Hao Lei, Yingxin Zeng, Zekai Cui, Qiaolang Zeng, Deliang Zhu, Ruiling Lian, Jun Zhang, Zhe Chen, Jiansu Chen
Corneal endothelial cells (CECs) are very important for the maintenance of corneal transparency. However, in vitro, CECs display limited proliferation and loss of phenotype via endothelial to mesenchymal transformation (EMT) and cellular senescence. In this study, we demonstrate that continuous supplementary nutrition using a perfusion culture bioreactor and three-dimensional (3D) spheroid culture can be used to improve CEC expansion in culture and to construct a tissue-engineered CEC layer. Compared with static culture, perfusion-derived CECs exhibited an increased proliferative ability as well as formed close cell-cell contact junctions and numerous surface microvilli...
April 10, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28361431/modification-of-rat-lung-decellularization-protocol-based-on-dynamic-conductometry-of-working-solution
#12
E V Kuevda, E A Gubareva, I S Gumenyuk, A S Sotnichenko, I V Gilevich, R Z Nakokhov, T V Rusinova, T G Yudina, A N Red'ko, S N Alekseenko
We modified the protocol of obtaining of biological scaffolds of rat lungs based on dynamic recording of specific resistivity of working detergent solution (conductometry) during perfusion decellularization. Termination of sodium deoxycholate exposure after attaining ionic equilibrium plateau did not impair the quality of decellularization and preserved structural matrix components, which was confirmed by morphological analysis and quantitative assay of residual DNA.
March 2017: Bulletin of Experimental Biology and Medicine
https://www.readbyqxmd.com/read/28323545/creation-of-a-bioengineered-skin-flap-scaffold-with-a-perfusable-vascular-pedicle
#13
Bernhard J Jank, Jeremy Goverman, Jacques P Guyette, Jon M Charest, Mark Randolph, Glenn R Gaudette, Joshua R Gershlak, Martin Purschke, Emilia Javorsky, Rosalynn M Nazarian, David A Leonard, Curtis L Cetrulo, William G Austen, Harald C Ott
Full-thickness skin loss is a challenging problem due to limited reconstructive options, demanding 75 million surgical procedures annually in the United States. Autologous skin grafting is the gold standard treatment, but results in donor-site morbidity and poor aesthetics. Numerous skin substitutes are available on the market to date, however, none truly functions as full-thickness skin due to lack of a vascular network. The creation of an autologous full-thickness skin analogue with a vascular pedicle would result in a paradigm shift in the management of wounds and in reconstruction of full-thickness skin defects...
June 19, 2017: Tissue Engineering. Part A
https://www.readbyqxmd.com/read/28323411/engineering-an-injectable-muscle-specific-microenvironment-for-improved-cell-delivery-using-a-nanofibrous-extracellular-matrix-hydrogel
#14
Nikhil Rao, Gillie Agmon, Matthew T Tierney, Jessica L Ungerleider, Rebecca L Braden, Alessandra Sacco, Karen L Christman
Injection of skeletal muscle progenitors has the potential to be a minimally invasive treatment for a number of diseases that negatively affect vasculature and skeletal muscle, including peripheral artery disease. However, success with this approach has been limited because of poor transplant cell survival. This is primarily attributed to cell death due to extensional flow through the needle, the harsh ischemic environment of the host tissue, a deleterious immune cell response, and a lack of biophysical cues supporting exogenous cell viability...
April 25, 2017: ACS Nano
https://www.readbyqxmd.com/read/28291964/application-of-a-parallelizable-perfusion-bioreactor-for-physiologic-3d-cell-culture
#15
Dominik Egger, Sarah Spitz, Monica Fischer, Stephan Handschuh, Martin Glösmann, Benedikt Friemert, Monika Egerbacher, Cornelia Kasper
It is crucial but challenging to keep physiologic conditions during the cultivation of 3D cell scaffold constructs for the optimization of 3D cell culture processes. Therefore, we demonstrate the benefits of a recently developed miniaturized perfusion bioreactor together with a specialized incubator system that allows for the cultivation of multiple samples while screening different conditions. Hence, a decellularized bone matrix was tested towards its suitability for 3D osteogenic differentiation under flow perfusion conditions...
2017: Cells, Tissues, Organs
https://www.readbyqxmd.com/read/28252516/decellularization-of-the-porcine-ear-generates-a-biocompatible-nonimmunogenic-extracellular-matrix-platform-for-face-subunit-bioengineering
#16
Jérôme Duisit, Giuseppe Orlando, Donovan Debluts, Louis Maistriaux, Daela Xhema, Yann-Alex J de Bisthoven, Cesare Galli, Andrea Peloso, Catherine Behets, Benoît Lengelé, Pierre Gianello
OBJECTIVE: The purpose of this study was to assess whether perfusion-decellularization technology could be applied to facial grafts. BACKGROUND: Facial allotransplantation remains an experimental procedure. Regenerative medicine techniques allow fabrication of transplantable organs from an individual's own cells, which are seeded into extracellular matrix (ECM) scaffolds from animal or human organs. Therefore, we hypothesized that ECM scaffolds also can be created from facial subunits...
March 1, 2017: Annals of Surgery
https://www.readbyqxmd.com/read/28241887/exploiting-decellularized-cochleae-as-scaffolds-for-inner-ear-tissue-engineering
#17
Adam J Mellott, Heather E Shinogle, Jennifer G Nelson-Brantley, Michael S Detamore, Hinrich Staecker
BACKGROUND: Use of decellularized tissues has become popular in tissue engineering applications as the natural extracellular matrix can provide necessary physical cues that help induce the restoration and development of functional tissues. In relation to cochlear tissue engineering, the question of whether decellularized cochlear tissue can act as a scaffold and support the incorporation of exogenous cells has not been addressed. Investigators have explored the composition of the cochlear extracellular matrix and developed multiple strategies for decellularizing a variety of different tissues; however, no one has investigated whether decellularized cochlear tissue can support implantation of exogenous cells...
February 28, 2017: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/28222326/crossing-kingdoms-using-decellularized-plants-as-perfusable-tissue-engineering-scaffolds
#18
Joshua R Gershlak, Sarah Hernandez, Gianluca Fontana, Luke R Perreault, Katrina J Hansen, Sara A Larson, Bernard Y K Binder, David M Dolivo, Tianhong Yang, Tanja Dominko, Marsha W Rolle, Pamela J Weathers, Fabricio Medina-Bolivar, Carole L Cramer, William L Murphy, Glenn R Gaudette
Despite significant advances in the fabrication of bioengineered scaffolds for tissue engineering, delivery of nutrients in complex engineered human tissues remains a challenge. By taking advantage of the similarities in the vascular structure of plant and animal tissues, we developed decellularized plant tissue as a prevascularized scaffold for tissue engineering applications. Perfusion-based decellularization was modified for different plant species, providing different geometries of scaffolding. After decellularization, plant scaffolds remained patent and able to transport microparticles...
May 2017: Biomaterials
https://www.readbyqxmd.com/read/28170430/efficient-decellularization-for-tissue-engineering-of-the-tendon-bone-interface-with-preservation-of-biomechanics
#19
Kai Xu, Lara A Kuntz, Peter Foehr, Katharina Kuempel, Alexandra Wagner, Jutta Tuebel, Constantin V Deimling, Rainer H Burgkart
Interfaces between tendon/ligament and bone ("entheses") are highly specialized tissues that allow for stress transfer between mechanically dissimilar materials. Entheses show very low regenerative capacity resulting in high incidences of failure after surgical repair. Tissue engineering is a promising approach to recover functionality of entheses. Here, we established a protocol to decellularize porcine entheses as scaffolds for enthesis tissue engineering. Chemical detergents as well as physical treatments were investigated with regard to their efficiency to decellularize 2 mm thick porcine Achilles tendon entheses...
2017: PloS One
https://www.readbyqxmd.com/read/28162155/-three-dimensional-circulation-perfusion-culture-of-hepatocytes-in-the-liver-decellularizedscaffold
#20
L Wang, P C Zhou, S S Zhu, Y Wang, X J Fan, M Y Zhu, Z W Wang, H X Qian
Objective: The intact rat liver decellularized scaffolds were preparedand repopulated hepatocytes by continuous perfusion technology.Toprovideexperimental support for the application of decellularized liver scaffolds in liver engineering. Method: Decellularized liver scaffolds were obtained by perfusing method. The composition and structure was examined by HE, Masson, Sirius red stain and immunofluorescence. The ultrastructure was examined by scanning electron microscope (SEM). DNA content was used to confirm the effect of decellularization...
January 24, 2017: Zhonghua Yi Xue za Zhi [Chinese medical journal]
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