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Perfusion decellularization

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https://www.readbyqxmd.com/read/28922323/face-graft-scaffold-production-in-a-rat-model
#1
Jérôme Duisit, Hadrien Amiel, Giuseppe Orlando, Adeline Dedriche, Catherine Behets, Pierre Gianello, Benoît Lengelé
BACKGROUND: As a route towards face bioengineering, our group previously reported the production of a complete scaffold production by perfusion-decellularization of a porcine ear subunit graft and partial recellularization. In order to extend the scaffold to the whole face and to down-scale it, we applied our findings to a rodent hemi-facial graft model. METHODS: After euthanasia, seven full-thickness rat hemi-face grafts were harvested with the CCA and the EJV as pedicle, and cannulated...
September 14, 2017: Plastic and Reconstructive Surgery
https://www.readbyqxmd.com/read/28895502/characterization-of-the-epicardial-adipose-tissue-in-decellularized-human-scaled-whole-hearts-implications-for-the-whole-heart-tissue-engineering
#2
Payam Akhyari, Fabian Oberle, Joern Huelsmann, Hans Heid, Stefan Lehr, Andreas Barbian, Sentaro Nakanishi, Hug Aubin, Alexander Jenke, Artur Lichtenberg
Whole-organ engineering is an innovative field of regenerative medicine with growing translational perspectives. Recent reports suggest the feasibility of decellularization and repopulation of entire human size hearts. However, little is known about the susceptibility of epicardial adipose tissue (EAT) to decellularization. Here, human size hearts of ovine donors were subjected to perfusion-based decellularization using detergent solutions. Upon basic histological evaluation and total DNA measurement myocardial regions prove largely decellularized while EAT demonstrated cellular remnants, further confirmed by transmission electron microscopy...
September 12, 2017: Tissue Engineering. Part A
https://www.readbyqxmd.com/read/28886732/optimized-protocol-for-whole-organ-decellularization
#3
A Schmitt, R Csiki, A Tron, B Saldamli, J Tübel, K Florian, S Siebenlist, E Balmayor, R Burgkart
BACKGROUND: The idea of tissue decellularization to gain matrices for tissue engineering is promising. The aim of the present study is to establish a safe and reproducible protocol for solid tissue decellularization that prevents the architecture of the matrix with the inherent vascular network. METHODS: The study was performed in rat kidneys which were decellularized by a SDS-based perfusion protocol. Perfusion time and SDS concentration were systematically changed to obtain the shortest and most gentle protocol that leads to complete decellularization...
September 8, 2017: European Journal of Medical Research
https://www.readbyqxmd.com/read/28863041/decellularized-rat-lung-scaffolds-using-sodium-lauryl-ether-sulfate-for-tissue-engineering
#4
Jinhui Ma, Zhihai Ju, Jie Yu, Yeru Qiao, Chenwei Hou, Chen Wang, Feilong Hei
Perfusion decellularization with detergents is effective to maintain the architecture and proteins of extracellular matrix (ECM) for use in the field of lung tissue engineering (LTE). However, it is unclear which detergent is ideal to produce an acellular lung scaffold. In this study, we obtained two decellularized rat lung scaffolds using a novel detergent sodium lauryl ether sulfate (SLES) and a conventional detergent sodium dodecyl sulfate (SDS). Both decellularized lung scaffolds were assessed by histology, immunohistochemistry, scanning electron microscopy, DNA quantification, sulfated glycosaminoglycans (GAGs) quantification and western blot...
August 31, 2017: ASAIO Journal: a Peer-reviewed Journal of the American Society for Artificial Internal Organs
https://www.readbyqxmd.com/read/28828244/freeze-thaw-decellularization-of-the-trabecular-meshwork-in-an-ex-vivo-eye-perfusion-model
#5
Yalong Dang, Susannah Waxman, Chao Wang, Adrianna Jensen, Ralitsa T Loewen, Richard A Bilonick, Nils A Loewen
OBJECTIVE: The trabecular meshwork (TM) is the primary substrate of outflow resistance in glaucomatous eyes. Repopulating diseased TM with fresh, functional TM cells might be a viable therapeutic approach. Decellularized TM scaffolds have previously been produced by ablating cells with suicide gene therapy or saponin, which risks incomplete cell removal or dissolution of the extracellular matrix, respectively. We hypothesized that improved trabecular meshwork cell ablation would result from freeze-thaw cycles compared to chemical treatment...
2017: PeerJ
https://www.readbyqxmd.com/read/28776178/protocols-for-rat-uterus-isolation-and-decellularization-applications-for-uterus-tissue-engineering-and-3d-cell-culturing
#6
Arvind Manikantan Padma, Tom Tristan Tiemann, Ahmed Baker Alshaikh, Randa Akouri, Min Jong Song, Mats Hellström
Sophisticated culturing conditions are required to grow cells in a three-dimensional (3D) environment. Cells then require a type of scaffold rich in proteins, growth factors, and signaling molecules that simulates their natural environment. Tissues from all species of animals have an organ-specific extracellular matrix (ECM) structure that plays a key role in cell proliferation and migration. Hence, the scaffold composition plays a significant role for any successful 3D cell culturing system. We developed a whole rat uterus ECM scaffold by the perfusion of detergents and ionic solutions through the vascular system of an isolated normal rat uterus in a process termed "decellularization...
August 4, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28758007/re-epithelialization-of-whole-porcine-kidneys-with-renal-epithelial-cells
#7
Nafiseh Poornejad, Evan Buckmiller, Lara Schaumann, Haonan Wang, Jonathan Wisco, Beverly Roeder, Paul Reynolds, Alonzo Cook
Decellularized porcine kidneys were recellularized with renal epithelial cells by three methods: perfusion through the vasculature under high pressure, perfusion through the ureter under high pressure, or perfusion through the ureter under moderate vacuum. Histology, scanning electron microscopy, confocal microscopy, and magnetic resonance imaging were used to assess vasculature preservation and the distribution of cells throughout the kidneys. Cells were detected in the magnetic resonance imaging by labeling them with iron oxide...
January 2017: Journal of Tissue Engineering
https://www.readbyqxmd.com/read/28742686/bioengineering-a-human-face-graft-the-matrix-of-identity
#8
Jérôme Duisit, Louis Maistriaux, Adriano Taddeo, Giuseppe Orlando, Virginie Joris, Emmanuel Coche, Catherine Behets, Jan Lerut, Chantal Dessy, Giulio Cossu, Esther Vögelin, Robert Rieben, Pierre Gianello, Benoît Lengelé
OBJECTIVE: During the last decade, face allotransplantation has been shown to be a revolutionary reconstructive procedure for severe disfigurements. However, offer to patients remains limited due to lifelong immunosuppression. To move forward in the field, a new pathway in tissue engineering is proposed. BACKGROUND: Our previously reported technique of matrix production of a porcine auricular subunit graft has been translated to a human face model. METHODS: 5 partial and 1 total face grafts were procured from human fresh cadavers...
July 24, 2017: Annals of Surgery
https://www.readbyqxmd.com/read/28735385/preparation-of-decellularized-liver-scaffolds-and-recellularized-liver-grafts
#9
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
#10
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
#11
Monique M A 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 N M 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. Although proof of concept has been reported for human livers, this study aims to perform whole liver decellularization in a clinically relevant series using controlled machine perfusion. In this study, we describe a mild nondestructive 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...
September 15, 2017: Stem Cells and Development
https://www.readbyqxmd.com/read/28570606/effects-of-two-different-decellularization-routes-on-the-mechanical-properties-of-decellularized-lungs
#12
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
#13
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
#14
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
#15
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
#16
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
#17
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
#18
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
#19
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
#20
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
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