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Perfusion decellularization

Rajesh Guruswamy Damodaran, Patrick Vermette
Diabetes Mellitus involves the loss of function and/or absolute numbers of insulin-producing β-cells in pancreatic islets. Islet transplantation is currently being investigated as a potential cure, and advances in tissue engineering methods can be used to improve pancreatic islets survival and functionality. Transplanted islets experience anoikis, hypoxia and inflammation-mediated immune response, leading to early damage and subsequent failure of the graft. Recent development in tissue engineering enables the use of decellularized organs as scaffolds for cell therapies...
March 2, 2018: Journal of Tissue Engineering and Regenerative Medicine
Jörn Hülsmann, Hug Aubin, Fabian Oberle, Nicolas Schuetterle, Shahbaz T Bandesha, Makoto Iijima, Artur Lichtenberg, Payam Akhyari
Whole-organ engineering - based on the functional repopulation of acellular whole-organ scaffolds derived from perfusion-based in toto decellularization of the specific organ system - is one of the most promising fields in tissue engineering. However, to date, we still have hardly any insights into the process of perfusion-based scaffold generation itself, with human-scale scaffolds usually obtained by adoption of small animal decellularization models, although those organs are of decreased biomass and potentially different biological characteristics...
February 21, 2018: Biomedical Materials
Dominique Manikowski, Birgit Andrée, Esther Samper, Clémence Saint-Marc, Ruth Olmer, Peter Vogt, Sarah Strauß, Axel Haverich, Andres Hilfiker
In natural tissues, the nutrition of cells and removal of waste products is facilitated by a dense capillary network which is generated during development. This perfusion system is also indispensable for tissue formation in vitro. Nutrition depending solely on diffusion is not sufficient to generate tissues of clinically relevant dimensions, which is a core aim in tissue engineering research. In this study, the establishment of a vascular network was investigated in a self-assembling approach employing endothelial and mural cells...
February 13, 2018: Vascular Pharmacology
Melva Suarez Muñoz, Davide Confalonieri, Heike Walles, Elisabeth M W M van Dongen, Gudrun Dandekar
Tissue engineering is a promising field, focused on developing solutions for the increasing demand on tissues and organs regarding transplantation purposes. The process to generate such tissues is complex, and includes an appropriate combination of specific cell types, scaffolds, and physical or biochemical stimuli to guide cell growth and differentiation. Microcarriers represent an appealing tool to expand cells in a three-dimensional (3D) microenvironment, since they provide higher surface-to volume ratios and mimic more closely the in vivo situation compared to traditional two-dimensional methods...
February 7, 2018: Journal of Visualized Experiments: JoVE
Amro Farag, Saeed M Hashimi, Cedryck Vaquette, Fabio Z Volpato, Dietmar W Hutmacher, Saso Ivanovski
OBJECTIVES: Decellularization aims to harness the regenerative properties of native extracellular matrix. The objective of this study was to evaluate different methods of decellularization of periodontal ligament cell sheets whilst maintaining their structural and biological integrity. DESIGN: Human periodontal ligament cell sheets were placed onto melt electrospun polycaprolactone (PCL) membranes that reinforced the cell sheets during the various decellularization protocols...
January 31, 2018: Archives of Oral Biology
Sung Min Park, Seran Yang, Se-Min Rye, Seong Wook Choi
BACKGROUND: Decellularized animal organs have been used as scaffolds for tissue engineering. To make a properly functioning scaffolds, the extracellular matrix (ECM) components must be preserved after decellularization. Because pulsatile flow is known to be beneficial for tissue perfusion, pulsatile perfusion of a detergent might decrease the exposure time of the tissues to the detergent used for decellularization. Using Energy Equivalent Pressure (EEP) as a pulsatility parameter, the effect of pulsatile flow in decellularization process is studied...
February 1, 2018: Biomedical Engineering Online
Matthew J Robertson, Benjamin Soibam, Jacqueline G O'Leary, Luiz C Sampaio, Doris A Taylor
Liver-like organoids that recapitulate the complex functions of the whole liver by combining cells, scaffolds, and mechanical or chemical cues are becoming important models for studying liver biology and drug metabolism. The advantages of growing cells in three-dimensional constructs include enhanced cell-cell and cell-extracellular matrix interactions and preserved cellular phenotype including, prevention of de-differentiation. In the current study, biomimetic liver constructs were made via perfusion decellularization of rat liver, with the goal of maintaining the native composition and structure of the extracellular matrix...
2018: PloS One
Yibing Guo, Cen Wu, Liancheng Xu, Yang Xu, Li Xiaohong, Zhu Hui, Lu Jingjing, Yuhua Lu, Zhiwei Wang
Vascularization remains a large obstacle for creating a functional pancreas-tissue equivalent for transplantation. In this study, a pre-vascularized pancreatic decellularized scaffold was prepared through endothelializing with endothelial progenitor cells (EPCs) in a bioreactor, and the ability to regenerate new blood vessels was detected in vivo. Initially, pancreases of Sprague-Dawley (SD) rats were perfused with 1% Triton X-100 and 0.1% ammonium hydroxide to remove the cellular components while the intact vascular network was preserved...
January 24, 2018: Journal of Artificial Organs: the Official Journal of the Japanese Society for Artificial Organs
Christopher A Neal, Jennifer G Nelson-Brantley, Michael S Detamore, Hinrich Staecker, Adam J Mellott
In mammals, mechanosensory hair cells that facilitate hearing lack the ability to regenerate, which has limited treatments for hearing loss. Current regenerative medicine strategies have focused on transplanting stem cells or genetic manipulation of surrounding support cells in the inner ear to encourage replacement of damaged stem cells to correct hearing loss. Yet, the extracellular matrix (ECM) may play a vital role in inducing and maintaining function of hair cells, and has not been well investigated. Using the cochlear ECM as a scaffold to grow adult stem cells may provide unique insights into how the composition and architecture of the extracellular environment aids cells in sustaining hearing function...
January 1, 2018: Journal of Visualized Experiments: JoVE
Mattia Francesco Maria Gerli, Jacques Paul Guyette, Daniele Evangelista-Leite, Brian Burns Ghoshhajra, Harald Christian Ott
Muscle and fasciocutaneous flaps taken from autologous donor sites are currently the most utilized approach for trauma repair, accounting annually for 4.5 million procedures in the US alone. However, the donor tissue size is limited and the complications related to these surgical techniques lead to morbidities, often involving the donor sites. Alternatively, recent reports indicated that extracellular matrix (ECM) scaffolds boost the regenerative potential of the injured site, as shown in a small cohort of volumetric muscle loss patients...
2018: PloS One
H Kojima, K Yasuchika, K Fukumitsu, T Ishii, S Ogiso, Y Miyauchi, R Yamaoka, T Kawai, H Katayama, E Y Yoshitoshi-Uebayashi, S Kita, K Yasuda, N Sasaki, J Komori, S Uemoto
Tissue decellularization produces a three-dimensional scaffold that can be potentially used to fabricate functional liver grafts following recellularization. An inappropriate cell distribution and clotting during blood perfusion hinder the practical use of recellularized livers. Here, we aimed to establish a seeding method for the optimal distribution of parenchymal and endothelial cells, and to evaluate the effect of liver sinusoidal endothelial cells (LSECs) in the decellularized liver. Primary rat hepatocytes and LSECs were seeded into decellularized whole-liver scaffolds via the biliary duct and portal vein, respectively...
January 16, 2018: American Journal of Transplantation
Matthias Becker, Janita A Maring, Barbara Oberwallner, Benjamin Kappler, Oliver Klein, Volkmar Falk, Christof Stamm
Acellular extracellular matrix preparations are useful for studying cell-matrix interactions and facilitate regenerative cell therapy applications. Several commercial extracellular matrix products are available as hydrogels or membranes, but these do not possess tissue-specific biological activity. Because perfusion decellularization is usually not possible with human heart tissue, we developed a 3-step immersion decellularization process. Human myocardial slices procured during surgery are first treated with detergent-free hyperosmolar lysis buffer, followed by incubation with the ionic detergent, sodium dodecyl sulfate, and the process is completed by exploiting the intrinsic DNase activity of fetal bovine serum...
December 4, 2017: Journal of Visualized Experiments: JoVE
Zurab Kakabadze, Anne Kakabadze, David Chakhunashvili, Lika Karalashvili, Ekaterine Berishvili, Yogeshwar Sharma, Sanjeev Gupta
Tissue engineering with scaffolds to form transplantable organs is of wide interest. Decellularized tissues have been tested for this purpose, although supplies of healthy donor tissues, vascular recellularization for perfusion and tissue homeostasis in engineered organs pose challenges. We hypothesized decellularized human placenta will be suitable for tissue engineering. The universal availability and unique structures of placenta for accommodating tissue, including presence of embedded vessels, were major attractions...
December 6, 2017: Hepatology: Official Journal of the American Association for the Study of Liver Diseases
L C P C Leonel, C M F C Miranda, T M Coelho, G A S Ferreira, R R Caãada, M A Miglino, S E Lobo
Biological biomaterials for tissue engineering purposes can be produced through tissue and/or organ decellularization. The remaining extracellular matrix (ECM) must be acellular and preserve its proteins and physical features. Placentas are organs of great interest because they are discarded after birth and present large amounts of ECM. Protocols for decellularization are tissue-specific and have not been established for canine placentas yet. This study aimed at analyzing a favorable method for decellularization of maternal and fetal portions of canine placentas...
November 17, 2017: Brazilian Journal of Medical and Biological Research, Revista Brasileira de Pesquisas Médicas e Biológicas
Rodrigo da Silva Nunes Barreto, Patricia Romagnolli, Andrea Maria Mess, Maria Angelica Miglino
Technically produced scaffolds are common to establish transplantable tissues for Regenerative Medicine, but also biological ones that are closer to the natural condition become of interest. Placentas are promising, because they represented available, complete organs with rich extracellular matrix (ECM) and well developed vasculature that easily could build anastomoses to a host´s organ. Only placentas from larger animal models such as the bovine meet the dimensions large enough for most organs, but are not adequately described yet...
November 21, 2017: Journal of Tissue Engineering and Regenerative Medicine
Andrea Peloso, Antonio Citro, Valeria Corradetti, Szandra Brambilla, Graziano Oldani, Francesco Calabrese, Tommaso Dominioni, Marcello Maestri, Lorenzo Cobianchi
Whole-organ decellularization is recently gaining interest in the transplantation field as strategy to obtain acellular scaffold only composed by extracellular matrix. These structures, that still remain organ-specific in terms of biological cues and tridimensional morphology could be then recellularized with patient's autologous cells. The final result should be a nwe transplantable autologous organ that should by-pass, at the same time, the problem of organ shortage and secondly the consequences related to the immunosuppression need...
November 21, 2017: Methods in Molecular Biology
Sarah Rajabi, Sara Pahlavan, Mohammad Kazemi Ashtiani, Hassan Ansari, Saeed Abbasalizadeh, Forough Azam Sayahpour, Fahimeh Varzideh, Sawa Kostin, Nasser Aghdami, Thomas Braun, Hossein Baharvand
Bioengineering of whole hearts using human embryonic stem cells (hESCs)-derived cardiovascular progenitor cells (CPCs) and natural matrices is a promising approach to overcome organ donor shortage threatening millions of patients awaiting for heart transplantation. Here, we developed a novel strategy for generation of heart constructs by repopulating engineered decellularized rat hearts using hESCs-derived CPCs. Careful expansion of CPCs in a scalable stirred-suspension bioreactor combined with step-wise seeding (60 million cells in 3 steps of 20 million per 1...
November 1, 2017: Biomaterials
Erik Elebring, Vijay K Kuna, Niclas Kvarnström, Suchitra Sumitran-Holgersson
Despite progress in the field of decellularization and recellularization, the outcome for pancreas has not been adequate. This might be due to the challenging dual nature of pancreas with both endocrine and exocrine tissues. We aimed to develop a novel and efficient cold-perfusion method for decellularization of porcine pancreas and recellularize acellular scaffolds with human fetal pancreatic stem cells. Decellularization of whole porcine pancreas at 4°C with sodium deoxycholate, Triton X-100 and DNase efficiently removed cellular material, while preserving the extracellular matrix structure...
January 2017: Journal of Tissue Engineering
Ramon Farré, Jordi Otero, Isaac Almendros, Daniel Navajas
Lung biofabrication is a new tissue engineering and regenerative development aimed at providing organs for potential use in transplantation. Lung biofabrication is based on seeding cells into an acellular organ scaffold and on culturing them in an especial purpose bioreactor. The acellular lung scaffold is obtained by decellularizing a non-transplantable donor lung by means of conventional procedures based on application of physical, enzymatic and detergent agents. To avoid immune recipient's rejection of the transplanted bioengineered lung, autologous bone marrow/adipose tissue-derived mesenchymal stem cells, lung progenitor cells or induced pluripotent stem cells are used for biofabricating the bioengineered lung...
November 1, 2017: Archivos de Bronconeumología
Matteo Ghiringhelli, Alessandro Zenobi, Stefano Brizzola, Fulvio Gandolfi, Valentino Bontempo, Sandro Rossi, Tiziana A L Brevini, Fabio Acocella
The development of new approaches for organ transplantation has become crucial in the last years. In particular, organ engineering, involving the preparation of acellular matrices that provide a natural habitat for reseeding with an appropriate population of cells, is an attractive although technically demanding approach. We here describe a method that allows for the derivation of functional in vitro hepatic organoids and that does not require a previous selection of all the parenchymal hepatocytes and non-parenchymal cells, namely, Kupffer cells, liver endothelial cells, and hepatic stellate cells...
November 4, 2017: Methods in Molecular Biology
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