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Perfusion decellularization

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https://www.readbyqxmd.com/read/28170430/efficient-decellularization-for-tissue-engineering-of-the-tendon-bone-interface-with-preservation-of-biomechanics
#1
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
#2
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]
https://www.readbyqxmd.com/read/28150744/engineering-an-endocrine-neo-pancreas-by-repopulation-of-a-decellularized-rat-pancreas-with-islets-of-langerhans
#3
H Napierala, K-H Hillebrandt, N Haep, P Tang, M Tintemann, J Gassner, M Noesser, H Everwien, N Seiffert, M Kluge, E Teegen, D Polenz, S Lippert, D Geisel, A Reutzel Selke, N Raschzok, A Andreou, J Pratschke, I M Sauer, B Struecker
Decellularization of pancreata and repopulation of these non-immunogenic matrices with islets and endothelial cells could provide transplantable, endocrine Neo- Pancreata. In this study, rat pancreata were perfusion decellularized and repopulated with intact islets, comparing three perfusion routes (Artery, Portal Vein, Pancreatic Duct). Decellularization effectively removed all cellular components but conserved the pancreas specific extracellular matrix. Digital subtraction angiography of the matrices showed a conserved integrity of the decellularized vascular system but a contrast emersion into the parenchyma via the decellularized pancreatic duct...
February 2, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28137992/coronary-sinus-injury-resulting-from-endarterectomy-during-multivessel-bypass-surgery
#4
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...
January 30, 2017: Interactive Cardiovascular and Thoracic Surgery
https://www.readbyqxmd.com/read/28117788/tunable-hydrogels-from-pulmonary-extracellular-matrix-for-3d-cell-culture
#5
Patrick A Link, Robert A Pouliot, Nabil S Mikhaiel, Bethany M Young, Rebecca L Heise
Here we present a method for establishing multiple component cell culture hydrogels for in vitro lung cell culture. Beginning with healthy en bloc lung tissue from porcine, rat, or mouse, the tissue is perfused and submerged in subsequent chemical detergents to remove the cellular debris. Histological comparison of the tissue before and after processing confirms removal of over 95% of double stranded DNA and alpha galactosidase staining suggests the majority of cellular debris is removed. After decellularization, the tissue is lyophilized and then cryomilled into a powder...
January 17, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28029762/prevascularization-of-decellularized-porcine-myocardial-slice-for-cardiac-tissue-engineering
#6
Pawan Kc, Mickey Shah, Jun Liao, Ge Zhang
Prevacularization strategies have been implemented in tissue engineering to generate microvasculature networks within a scaffold prior to implantation. Prevascularizing scaffolds will shorten the time of functional vascular perfusion with host upon implantation. In this study, we explored key variables affecting the interaction between decellularized porcine myocardium slices (dPMSs) and reseeded stem cells toward the fabrication of prevascularized cardiac tissue. Our results demonstrated that dPMS supports attachment of human mesenchymal stem cells (hMSCs) and rat adipose derived stem cells (rASCs) with high viability...
January 10, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28029279/recellularization-via-the-bile-duct-supports-functional-allogenic-and-xenogenic-cell-growth-on-a-decellularized-rat-liver-scaffold
#7
Wessam Hassanein, Mehmet C Uluer, John Langford, Jhade D Woodall, Arielle Cimeno, Urmil Dhru, Avraham Werdesheim, Joshua Harrison, Carlos Rivera-Pratt, Stephen Klepfer, Ali Khalifeh, Bryan Buckingham, Philip S Brazio, Dawn Parsell, Charlie Klassen, Cinthia Drachenberg, Rolf N Barth, John C LaMattina
Recent years have seen a proliferation of methods leading to successful organ decellularization. In this experiment we examine the feasibility of a decellularized liver construct to support growth of functional multilineage cells. Bio-chamber systems were used to perfuse adult rat livers with 0.1% SDS for 24 hours yielding decellularized liver scaffolds. Initially, we recellularized liver scaffolds using a human tumor cell line (HepG2, introduced via the bile duct). Subsequent studies were performed using either human tumor cells co-cultured with human umbilical vein endothelial cells (HUVECs, introduced via the portal vein) or rat neonatal cell slurry (introduced via the bile duct)...
January 2, 2017: Organogenesis
https://www.readbyqxmd.com/read/28019665/the-impact-of-left-ventricular-stretching-in-model-cultivations-with-neonatal-cardiomyocytes-in-a-whole-heart-bioreactor
#8
Jörn Hülsmann, Hug Aubin, Alexander Wehrmann, Artur Lichtenberg, Payam Akhyari
Here, we investigate the impact of integrated three-dimensional (3D) left ventricular (LV) stretching on myocardial maturation in a whole-heart bioreactor setting. Therefore, decellularized rat hearts were selectively repopulated with rodent neonatal cardiomyocytes (5•10(6) cells per heart) and cultured over 5 d. Continuous medium perfusion was maintained through the coronary artery system in a customized whole-heart bioreactor system with or without integrated biomechanical stimulation of LV. 3D repopulation effectiveness and cellular vitality were evaluated by repetitive metabolic WST-1 assays and 3D confocal microscopy analysis through fluorescent staining, also assessing cellular organization...
December 26, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27957815/evolution-of-graft-morphology-and-function-after-recellularization-of-decellularized-rat-livers
#9
Antje Butter, Khalid Aliyev, Karl-Herbert Hillebrandt, Nathanael Raschzok, Martin Kluge, Nicolai Seiffert, Peter Tang, Hendrik Napierala, Muhammad Imtiaz Ashraf, Anja Reutzel-Selke, Andreas Andreou, Johann Pratschke, Igor Maximilian Sauer, Benjamin Struecker
Decellularization of livers is a well-established procedure. Data on different reseeding techniques or the functional evolution and re-organization processes of repopulated grafts remains limited. We established a proprietary, customized bioreactor to repopulate decellularized rat livers (n = 21) with primary rat hepatocytes (150 x 10(6) cells) via the hepatic artery and to subsequently evaluate graft morphology and function during seven days of ex vivo perfusion. Grafts were analyzed at 1 h, 6 h, 12 h, 24 h, 3d, 5d and 7d after recellularization (all n = 3) by immuno-histologic evaluation, hepatocyte-related enzyme (AST, ALT, LDH) and albumin measurement in the perfusate...
December 13, 2016: Journal of Tissue Engineering and Regenerative Medicine
https://www.readbyqxmd.com/read/27943597/re-endothelialization-of-rat-lung-scaffolds-through-passive-gravity-driven-seeding-of-segment-specific-pulmonary-endothelial-cells
#10
Michelle E Scarritt, Nicholas C Pashos, Jessica M Motherwell, Zachary R Eagle, Brian J Burkett, Ashley N Gregory, Ricardo Mostany, Daniel J Weiss, Diego F Alvarez, Bruce A Bunnell
Effective re-endothelialization is critical for the use of decellularized scaffolds for ex vivo lung engineering. Current approaches yield insufficiently re-endothelialized scaffolds that hemorrhage and become thrombogenic upon implantation. Herein, gravity-driven seeding coupled with bioreactor culture facilitated widespread distribution and engraftment of endothelial cells throughout rat lung scaffolds. Initially, human umbilical vein endothelial cells (HUVECs) were seeded into the pulmonary artery by either gravity-driven, variable flow perfusion seeding or pump-driven, pulsatile flow perfusion seeding...
December 12, 2016: Journal of Tissue Engineering and Regenerative Medicine
https://www.readbyqxmd.com/read/27942003/biochemical-and-cellular-determinants-of-renal-glomerular-elasticity
#11
Addie E Embry, Hamid Mohammadi, Xinying Niu, Liping Liu, Borren Moe, William A Miller-Little, Christopher Y Lu, Leslie A Bruggeman, Christopher A McCulloch, Paul A Janmey, R Tyler Miller
The elastic properties of renal glomeruli and their capillaries permit them to maintain structural integrity in the presence of variable hemodynamic forces. Measured by micro-indentation, glomeruli have an elastic modulus (E, Young's modulus) of 2.1 kPa, and estimates from glomerular perfusion studies suggest that the E of glomeruli is between 2 and 4 kPa. F-actin depolymerization by latrunculin, inhibition of acto-myosin contractility by blebbistatin, reduction in ATP synthesis, and reduction of the affinity of adhesion proteins by EDTA reduced the glomerular E to 1...
2016: PloS One
https://www.readbyqxmd.com/read/27924607/fabrication-and-characterization-of-decellularized-periodontal-ligament-cell-sheet-constructs
#12
Amro Farag, Cédryck Vaquette, Dietmar W Hutmacher, P Mark Bartold, Saso Ivanovski
Decellularized tissue-engineered constructs have the potential to promote regeneration by providing a biomimetic extracellular matrix that directs tissue-specific regeneration when implanted in situ. Recently, the use of cell sheets has shown promising results in promoting periodontal regeneration. Here, we describe the fabrication of decellularized periodontal cell sheets with intact extracellular matrix structural and biological properties. Melt electro spun polycaprolactone (PCL) scaffolds are used as a carrier for the inherently fragile cell sheets, to provide support during the processes of decellularization...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27911373/the-mouse-isolated-perfused-kidney-technique
#13
Jan Czogalla, Frank Schweda, Johannes Loffing
The mouse isolated perfused kidney (MIPK) is a technique for keeping a mouse kidney under ex vivo conditions perfused and functional for 1 hr. This is a prerequisite for studying the physiology of the isolated organ and for many innovative applications that may be possible in the future, including perfusion decellularization for kidney bioengineering or the administration of anti-rejection or genome-editing drugs in high doses to prime the kidney for transplantation. During the time of the perfusion, the kidney can be manipulated, renal function can be assessed, and various pharmaceuticals administered...
November 17, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27911030/acellular-endocardium-as-a-novel-biomaterial-for-the-intima-of-tissue-engineered-small-caliber-vascular-grafts
#14
Feng Wang, Xin Guan, TianYi Wu, JianOu Qiao, ZhaoQing Han, JinLong Wu, XiaoWei Yu, QingJun You
We aimed to investigate whether acellular endocardium can be used as a useful biomaterial for the intima of engineered small-caliber vascular grafts. Fresh endocardium was harvested from the swine left atrium and was decellularized by digestion with the decellularization solution of Triton X-100 and SDS containing DNase I and RNase A. Surface morphological characteristics and Young's modulus were evaluated. To analyze the effect of mechanical characteristics on cell adhesion, the decellularized endocardium was stiffened with 2...
December 2016: Artificial Organs
https://www.readbyqxmd.com/read/27894293/lung-bioengineering-physical-stimuli-and-stem-progenitor-cell-biology-interplay-towards-biofabricating-a-functional-organ
#15
REVIEW
Paula N Nonaka, Juan J Uriarte, Noelia Campillo, Vinicius R Oliveira, Daniel Navajas, Ramon Farré
A current approach to obtain bioengineered lungs as a future alternative for transplantation is based on seeding stem cells on decellularized lung scaffolds. A fundamental question to be solved in this approach is how to drive stem cell differentiation onto the different lung cell phenotypes. Whereas the use of soluble factors as agents to modulate the fate of stem cells was established from an early stage of the research with this type of cells, it took longer to recognize that the physical microenvironment locally sensed by stem cells (e...
November 28, 2016: Respiratory Research
https://www.readbyqxmd.com/read/27884776/inverted-orientation-improves-decellularization-of-whole-porcine-hearts
#16
Po-Feng Lee, Eric Chau, Rafael Cabello, Alvin T Yeh, Luiz C Sampaio, Andrea S Gobin, Doris A Taylor
: In structurally heterogeneous organs, such as heart, it is challenging to retain extracellular matrix integrity in the thinnest regions (eg, valves) during perfusion decellularization and completely remove cellular debris from thicker areas. The high inflow rates necessary to maintain physiologic pressure can distend or damage thin tissues, but lower pressures prolong the process and increase the likelihood of contamination. We examined two novel retrograde decellularization methods for porcine hearts: inverting the heart or venting the apex to decrease inflow rate...
November 21, 2016: Acta Biomaterialia
https://www.readbyqxmd.com/read/27871686/eliminating-the-organ-transplant-waiting-list-the-future-with-perfusion-decellularized-organs
#17
REVIEW
Dominique Seetapun, Jeffrey J Ross
No abstract text is available yet for this article.
November 18, 2016: Surgery
https://www.readbyqxmd.com/read/27862079/discarded-livers-find-a-new-life-engineered-liver-grafts-using-hepatocytes-recovered-from-marginal-livers
#18
Basak E Uygun, Maria-Louisa Izamis, Maria Jaramillo, Yibin Chen, Gavrielle Price, Sinan Ozer, Martin L Yarmush
Treatment for end-stage liver failure is restricted by the critical shortage of donor organs; about 4000 people die in the USA while waiting for a transplantable organ. This situation has been a major driving force behind the rise of tissue engineering to build artificial tissues/organs. Recent advancements in creating transplantable liver grafts using decellularized liver scaffolds bring the field closer to clinical translation. However, a source of readily available and highly functional adult hepatocytes in adequate numbers for regenerative liver therapies still remains unclear...
November 8, 2016: Artificial Organs
https://www.readbyqxmd.com/read/27826794/the-challenge-in-using-mesenchymal-stromal-cells-for-recellularization-of-decellularized-cartilage
#19
REVIEW
Zhao Huang, Owen Godkin, Gundula Schulze-Tanzil
Some decellularized musculoskeletal extracellular matrices (ECM)s derived from tissues such as bone, tendon and fibrocartilaginous meniscus have already been clinical use for tissue reconstruction. Repair of articular cartilage with its unique zonal ECM architecture and composition is still an unsolved problem, and the question is whether allogenic or xenogeneic decellularized cartilage ECM could serve as a biomimetic scaffold for this purpose.Hence, this survey outlines the present state of preparing decellularized cartilage ECM-derived scaffolds or composites for reconstruction of different cartilage types and of reseeding it particularly with mesenchymal stromal cells (MSCs)...
November 8, 2016: Stem Cell Reviews
https://www.readbyqxmd.com/read/27805597/study-of-viral-vectors-in-a-three-dimensional-liver-model-repopulated-with-the-human-hepatocellular-carcinoma-cell-line-hepg2
#20
Thomas Hiller, Viola Röhrs, Eva-Maria Dehne, Anke Wagner, Henry Fechner, Roland Lauster, Jens Kurreck
This protocol describes the generation of a three-dimensional (3D) ex vivo liver model and its application to the study and development of viral vector systems. The model is obtained by repopulating the extracellular matrix of a decellularized rat liver with a human hepatocyte cell line. The model permits studies in a vascularized 3D cell system, replacing potentially harmful experiments with living animals. Another advantage is the humanized nature of the model, which is closer to human physiology than animal models...
October 24, 2016: Journal of Visualized Experiments: JoVE
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