Ringraziamo BBSRC per il finanziamento di un dottorato di ricerca per Bye Mr. Frazer.

<ol>
	<li>Canton, I., McKean, R., Charnley, M., Blackwood, K., Fiorica, C., Ryan, A., MacNeil, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+an+Ibuprofen-releasing+biodegradable+PLA/PGA+electrospun+scaffold+for+tissue+regeneration.">Development of an Ibuprofen-releasing biodegradable PLA/PGA electrospun scaffold for tissue regeneration.</a> Biotechnology and bioengineering. 105, 396-408 (2010).</li><li>Blackwood, K., McKean, R., Canton, I., Freeman, C., Franklin, K., Cole, A., Brook, I., Farthing, P., Rimmer, S., Haycock, J., Ryan, A., MacNeil, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+biodegradable+electrospun+scaffolds+for+dermal+replacement.">Development of biodegradable electrospun scaffolds for dermal replacement.</a> Biomaterials. 29, 3091-3104 (2008).</li><li>Yang, F., Maurugan, R., Wang, S., Ramakrishna, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrospinning+of+nano/micro+scale+poly(L-lactic+acid)+aligned+fibers+and+their+potential+in+neural+tissue+engineering.">Electrospinning of nano/micro scale poly(L-lactic acid) aligned fibers and their potential in neural tissue engineering.</a> Biomaterials. 26, 2603-2610 (2005).</li><li>Sittichokechaiwut, A., Edwards, J. H., Scutt, A. M., Reilly, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Short+bouts+of+mechanical+loading+are+as+effective+as+dexamethasone+at+inducing+matrix+production+by+human+bone+marrow+mesenchymal+stem+cell.">Short bouts of mechanical loading are as effective as dexamethasone at inducing matrix production by human bone marrow mesenchymal stem cell.</a> Eur. Cell Mater. 20, 45-57 (2010).</li><li>Sill, T. J., von Recum, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrospinning:+applications+in+drug+delivery+and+tissue+engineering.">Electrospinning: applications in drug delivery and tissue engineering.</a> Biomaterials. 29 (13), 1989-2006 (2008).</li><li>Deitzel, J., Kleinmeyer, J., Harris, D., Beck Tan, N. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+processing+variables+on+the+morphology+of+electrospun+nanofibers+and+textiles.">The effect of processing variables on the morphology of electrospun nanofibers and textiles.</a> Polymer. 42, 261-272 (2001).</li><li>Fridrikh, S., Yu, J., Brenner, M., Rutledge, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlling+the+fiber+diameter+during+electrospinning.">Controlling the fiber diameter during electrospinning.</a> Physical review letters. 90, 1-4 (2003).</li><li>Fong, H., Chun, I., Reneker, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beaded+nanofibers+formed+during+electrospinning.">Beaded nanofibers formed during electrospinning.</a> Polymer. 40 (16), 4585-4592 (1999).</li><li>Selim, M., Bullock, A. J., Blackwood, K. A., Chapple, C. R., MacNeil, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developing+biodegradable+scaffolds+for+tissue+engineering+of+the+urethra.">Developing biodegradable scaffolds for tissue engineering of the urethra.</a> BJU Int. 107 (2), 296-302 (2010).</li><li>Tong, H. -. W., Wang, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+investigation+into+the+influence+of+electrospinning+parameters+on+the+diameter+and+alignment+of+poly(hydroxybutyrate-co-hydroxyvalerate)+fibers.">An investigation into the influence of electrospinning parameters on the diameter and alignment of poly(hydroxybutyrate-co-hydroxyvalerate) fibers.</a> Journal of Applied Polymer Science. 120 (3), 1694-1706 (2011).</li><li>Tong, H. -. W., Wang, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrospinning+of+poly(hydroxybutyrate-co-hydroxyvalerate)+fibrous+tissue+engineering+scaffolds+in+two+different+electric+fields.">Electrospinning of poly(hydroxybutyrate-co-hydroxyvalerate) fibrous tissue engineering scaffolds in two different electric fields.</a> Polymer Engineering &amp; Science. 51 (7), 1325-1338 (2011).</li><li>Retzepi, M., Donos, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guided+Bone+Regeneration:+biological+principle+and+therapeutic+applications.">Guided Bone Regeneration: biological principle and therapeutic applications.</a> Clinical oral implants research. 21, 567-576 (2010).</li><li>Moreau, J., Caccamese, J., Coletti, D., Sauk, J., Fisher, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+engineering+solutions+for+cleft+palates.">Tissue engineering solutions for cleft palates.</a> Journal of oral maxillofacial. 65, 2503-2511 (2007).</li><li>Yang, F., Both, S., Yang, X., Walboomers, X., Jansen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+an+electrospun+nano-apatite/PCL+composite+membrane+for+GTR/GBR+application.">Development of an electrospun nano-apatite/PCL composite membrane for GTR/GBR application.</a> Acta biomaterialia. 5, 3295-3304 (2009).</li><li>Yoshimoto, H., Shin, Y., Terai, H., Vacanti, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+biodegradable+nanofiber+scaffold+by+electrospinning+and+its+potential+for+bone+tissue+engineering.">A biodegradable nanofiber scaffold by electrospinning and its potential for bone tissue engineering.</a> Biomaterials. 24, 2077-2082 (2003).</li><li>Telemeco, T., Ayres, C., Bowlin, G., Wnek, G., Boland, E., Cohen, N., Baumgarten, C., Mathews, J., Simpson, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+cellular+infiltration+into+tissue+engineering+scaffolds+composed+of+submicron+diameter+fibrils+produced+by+electrospinning.">Regulation of cellular infiltration into tissue engineering scaffolds composed of submicron diameter fibrils produced by electrospinning.</a> Acta biomaterialia. 1, 377-385 (2005).</li></ol>

Non ci sono conflitti di interesse dichiarati.

Electrospinning è una tecnica molto popolare per la produzione di scaffolds per l&#39;ingegneria tissutale. 14, 15, 16 Mentre è relativamente semplice da realizzare ponteggi elettrofilati di base per uso sperimentale la tecnica è complessa e sfaccettata con molte variabili. 6 Ci sono molti studi che descrivono come la electrospinning parametri determinano il patibolo prodotto. In questo studio l&#39;attenzione è rivolta alla post-produzione una notevole sfida per fare di architetture adeguate i...

<table border="1"><tbody><tr><td> Nome del reattivo </td><td> Azienda </td><td> Numero di catalogo </td><td> Comments </td></tr><tr><td> Poly lattico-co-glicolico </td><td> Sigma Aldrich </td><td></td><td></td></tr><tr><td> Poly acido lattico </td><td> Sigma Aldrich </td><td> 81273 </td><td> Viscosità intrinseca ~ 2.0dl / g </td></tr><tr><td> Poly ε-caprolattone </td><td> Sigma Aldrich </td><td></td><td></td></tr><tr><td> Poly idrossibutirrato-co-idrossivalerato 00:01 </td><td> Goodfellow </td><td> 578-446-59 </td><td> PHB88/PHV12 </td></tr><tr><td> Diclorometano </td><td> Sigma Aldrich o Fisher </td><td> 270997 o D/1850/17 </td><td> &gt; 99,8% contiene 50-150ppm amilene stabilizzatore </td></tr><tr><td> 50 palloncini multicolori </td><td> Wilkinson Hardware Stores Ltd. </td><td> 0105790 </td><td></td></tr><tr><td> Capra anti-IgG di coniglio (FC): FITC </td><td> AbDserotec </td><td> STAR121F </td><td></td></tr><tr><td> Coniglio anti-umano alfa elastina </td><td> AbDserotec </td><td> 4060-1060 </td><td></td></tr><tr><td> Tappo a vite GL45 PP 2 porte, pk / 2 </td><td> SLS </td><td> 1129750 </td><td></td></tr><tr><td> 4 &#39;,6-Diamidino-2-fenilindolo dicloridrato </td><td> Sigma Aldrich </td><td> 32670 </td><td></td></tr><tr><td> CellTracker verde CMFDA </td><td> Invitrogen </td><td> C7025 </td><td></td></tr><tr><td> CellTracker rosso CMTX </td><td> Invitrogen </td><td> C34552 </td><td></td></tr></tbody></table>

postproduction processing of electrospun fibres for tissue engineering

Electrospinning è un metodo comunemente usato e versatile per la produzione di ponteggi (spesso biodegradabile) per l&#39;ingegneria tissutale 3D. 1, 2, 3 Molti tessuti in vivo sottoposti a distensione biassiale in varia misura come la pelle, della vescica, pavimento pelvico e anche il palato duro come i bambini crescere. Nel produrre scaffold per questi scopi è necessario sviluppare scaffold di adeguate proprietà biomeccaniche (sia ottenuta senza o con cellule) e che sono sterile per uso clinico. Il focus di questo articolo non è come stabilire i parametri di electrospinning di base (in quanto vi è un&#39;ampia letteratura sul electrospinning), ma su come modificare spun messaggio ponteggi produzione per renderli adatti per scopi di ingegneria dei tessuti - qui spessore, proprietà meccaniche e la sterilizzazione (necessaria per uso clinico) sono considerati e si descrivono anche come le cellule possono essere coltivate sui ponteggi e sottoposto a deformazione biassiale per condizionare loro per applicazioni specifiche. Elettrofilatura tende a produrre fogli sottili, come il collettore elettrofilatura diventa rivestiti con fibre isolanti diventa un cattivo conduttore tale che le fibre non deposito su di esso. Quindi si descrive approcci per produrre strutture più spessi da calore o vapore ricottura aumentando la forza di ponteggi, ma non necessariamente l&#39;elasticità. Filatura sequenziali di scaffolds di polimeri diversi per raggiungere ponteggi complessi è anche descritto. Metodologie di sterilizzazione può influire negativamente sulla resistenza e l&#39;elasticità di ponteggi. Mettiamo a confronto tre metodi per il loro effetto sulle proprietà biomeccaniche su ponteggi elettrofilati di poli lattico-co-glicolico (PLGA). Imaging di cellule su ponteggi e la valutazione della produzione della matrice extracellulare (ECM) proteine ​​da parte delle cellule è descritto su ponteggi. Coltura di cellule su ponteggi in vitro in grado di migliorare la forza scaffold e elasticità, ma il tessuto di ingegneria alla letteraturare dimostra che le cellule spesso non riescono a produrre ECM appropriato quando coltivato in condizioni statiche. Ci sono alcuni sistemi commerciali disponibili che permettono di cellule di coltura su ponteggi in regime di condizionamento dinamici -. Un esempio è la ElectroForce Bose 3100 che può essere usato per esercitare un programma di condizionamento su cellule in impalcature tenute con grip meccaniche entro un supporto camera riempita 4 Un approccio ad un bioreattore coltura cellulare budget per la distorsione controllata in 2 dimensioni è descritto. Abbiamo dimostrato che le cellule possono essere indotte a produrre elastina in queste condizioni. Infine valutazione delle proprietà biomeccaniche di ponteggi trasformati in coltura con o senza cellule è descritto.

Watch this Scientific Journal Video about Postproduction Processing of Electrospun Fibres for Tissue Engineering at JoVE.com

Postproduction Processing of Electrospun Fibres for Tissue Engineering

Ponteggi elettrofilate possono essere trattati post-produzione per applicazioni di ingegneria tissutale. Qui si descrivono i metodi per la filatura ponteggi complessi (da spinning consecutivi), per rendere più spessi (da ponteggi multi-layering che utilizza il calore o vapore ricottura), per ottenere la sterilità (produzione asettica o di post produzione la sterilizzazione) e per il raggiungimento di adeguate proprietà biomeccaniche.

Elaborazione Postproduzione di fibre elettrofilate per l&#39;ingegneria tissutale

Electrospinning is a commonly used and versatile method to produce  scaffolds (often biodegradable) for 3D tissue engineering.1, 2, 3 Many tissues in vivo ...

postproduction-processing-of-electrospun-fibres-for-tissue-engineering

Research

JoVE Journal

Bioengineering

18.0K Views.  University of Sheffield. Ponteggi elettrofilate possono essere trattati post-produzione per applicazioni di ingegneria tissutale. Qui si descrivono i metodi per la filatura ponteggi complessi (da spinning consecutivi), per rendere più spessi (da ponteggi multi-layering che utilizza il calore o vapore ricottura), per ottenere la sterilità (produzione asettica o di post produzione la sterilizzazione) e per il raggiungimento di adeguate proprietà biomeccaniche.

Video: Postproduction Processing of Electrospun Fibres for Tissue Engineering

Fabrication of Biologically Derived Injectable Materials for Myocardial Tissue Engineering

This protocol provides methods for the preparation of an injectable extracellular matrix (ECM) gel for myocardial tissue engineering applications. Briefly, decellularized tissue is lyophilized, milled, enzymatically digested, and then brought to physiological pH. The lyophilization removes all water content from the tissue, resulting in dry ECM that can be ground into a fine powder with a small mill. After milling, the ECM powder is digested with pepsin to form an injectable matrix. After adjustment to pH 7.4, the liquid matrix material can be injected into the myocardium. Results of previous characterization assays have shown that matrix gels produced from decellularized pericardial and myocardial tissue retain native ECM components, including diverse proteins, peptides and glycosaminoglycans. Given the use of this material for tissue engineering, in vivo characterization is especially useful; here, a method for performing an intramural injection into the left ventricular (LV) free wall is presented as a means of analyzing the host response to the matrix gel in a small animal model. Access to the chest cavity is gained through the diaphragm and the injection is made slightly above the apex in the LV free wall. The biologically derived scaffold can be visualized by biotin-labeling before injection and then staining tissue sections with a horse radish peroxidase-conjugated neutravidin and visualizing via diaminobenzidine (DAB) staining. Analysis of the injection region can also be done with histological and immunohistochemical staining. In this way, the previously examined pericardial and myocardial matrix gels were shown to form fibrous, porous networks and promote vessel formation within the injection region.

Methods for preparing an injectable matrix gel from decellularized tissue and injecting it into rat myocardium in vivo are described.

Fabbricazione di derivazione biologica Materiali iniettabili per l&#39;ingegneria tissutale miocardica

This protocol provides methods for the preparation of an injectable extracellular matrix (ECM) gel for myocardial tissue engineering applications.  ...

Ricerca

Bioingegneria

Elastomeric PGS Scaffolds in Arterial Tissue Engineering

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and increasing obesity1. Currently, bypass surgery using autologous vessels, allografts, and synthetic grafts are known as a commonly used for arterial substitutes2. However, these grafts have limited applications when an inner diameter of arteries is less than 6 mm due to low availability, thrombotic complications, compliance mismatch, and late intimal hyperplasia3,4. To overcome these limitations, tissue engineering has been successfully applied as a promising alternative to develop small-diameter arterial constructs that are nonthrombogenic, robust, and compliant. Several previous studies have developed small-diameter arterial constructs with tri-lamellar structure, excellent mechanical properties and burst pressure comparable to native arteries5,6. While high tensile strength and burst pressure by increasing collagen production from a rigid material or cell sheet scaffold, these constructs still had low elastin production and compliance, which is a major problem to cause graft failure after implantation. Considering these issues, we hypothesized that an elastometric biomaterial combined with mechanical conditioning would provide elasticity and conduct mechanical signals more efficiently to vascular cells, which increase extracellular matrix production and support cellular orientation.
The objective of this report is to introduce a fabrication technique of porous tubular scaffolds and a dynamic mechanical conditioning for applying them to arterial tissue engineering. We used a biodegradable elastomer, poly (glycerol sebacate) (PGS)7 for fabricating porous tubular scaffolds from the salt fusion method. Adult primary baboon smooth muscle cells (SMCs) were seeded on the lumen of scaffolds, which cultured in our designed pulsatile flow bioreactor for 3 weeks. PGS scaffolds had consistent thickness and randomly distributed macro- and micro-pores. Mechanical conditioning from pulsatile flow bioreactor supported SMC orientation and enhanced ECM production in scaffolds. These results suggest that elastomeric scaffolds and mechanical conditioning of bioreactor culture may be a promising method for arterial tissue engineering.

Elastomeric PGS scaffolds with vascular smooth muscle cells cultured in a pulsatile flow bioreactor may lead to promising small-diameter arterial constructs with native ECM production in a relatively short culture period.

PGS elastomerico Ponteggi in Ingegneria tessuto arterioso

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and ...

Capillary Force Lithography for Cardiac Tissue Engineering

Cardiovascular disease remains the leading cause of death worldwide1. Cardiac tissue engineering holds much promise to deliver groundbreaking medical discoveries with the aims of developing functional tissues for cardiac regeneration as well as in vitro screening assays. However, the ability to create high-fidelity models of heart tissue has proven difficult. The heart&#8217;s extracellular matrix (ECM) is a complex structure consisting of both biochemical and biomechanical signals ranging from the micro- to the nanometer scale2. Local mechanical loading conditions and cell-ECM interactions have recently been recognized as vital components in cardiac tissue engineering3-5.
A large portion of the cardiac ECM is composed of aligned collagen fibers with nano-scale diameters that significantly influences tissue architecture and electromechanical coupling2. Unfortunately, few methods have been able to mimic the organization of ECM fibers down to the nanometer scale. Recent advancements in nanofabrication techniques, however, have enabled the design and fabrication of scalable scaffolds that mimic the in vivo structural and substrate stiffness cues of the ECM in the heart6-9.
Here we present the development of two reproducible, cost-effective, and scalable nanopatterning processes for the functional alignment of cardiac cells using the biocompatible polymer poly(lactide-co-glycolide) (PLGA)8 and a polyurethane (PU) based polymer. These anisotropically nanofabricated substrata (ANFS) mimic the underlying ECM of well-organized, aligned tissues and can be used to investigate the role of nanotopography on cell morphology and function10-14.
Using a nanopatterned (NP) silicon master as a template, a polyurethane acrylate (PUA) mold is fabricated. This PUA mold is then used to pattern the PU or PLGA hydrogel via UV-assisted or solvent-mediated capillary force lithography (CFL), respectively15,16. Briefly, PU or PLGA pre-polymer is drop dispensed onto a glass coverslip and the PUA mold is placed on top. For UV-assisted CFL, the PU is then exposed to UV radiation (&#955; = 250-400 nm) for curing. For solvent-mediated CFL, the PLGA is embossed using heat (120 &#176;C) and pressure (100 kPa). After curing, the PUA mold is peeled off, leaving behind an ANFS for cell culture. Primary cells, such as neonatal rat ventricular myocytes, as well as human pluripotent stem cell-derived cardiomyocytes, can be maintained on the ANFS2.

In this protocol, we demonstrate the fabrication of biomimetic cardiac cell culture substrata made from two distinct polymeric materials using capillary force lithography. The described methods provide a scalable, cost-effective technique to engineer the structure and function of macroscopic cardiac tissues for in vitro and in vivo applications.

Capillare Forza litografia per Cardiac Tissue Engineering

Cardiovascular disease remains the leading cause of death worldwide1. Cardiac tissue engineering holds much promise to deliver groundbreaking medical ...

Design of a Biaxial Mechanical Loading Bioreactor for Tissue Engineering

We designed a loading device that is capable of applying uniaxial or biaxial mechanical strain to a tissue engineered biocomposites fabricated for transplantation. While the device primarily functions as a bioreactor that mimics the native mechanical strains, it is also outfitted with a load cell for providing force feedback or mechanical testing of the constructs. The device subjects engineered cartilage constructs to biaxial mechanical loading with great precision of loading dose (amplitude and frequency) and is compact enough to fit inside a standard tissue culture incubator. It loads samples directly in a tissue culture plate, and multiple plate sizes are compatible with the system. The device has been designed using components manufactured for precision-guided laser applications. Bi-axial loading is accomplished by two orthogonal stages. The stages have a 50 mm travel range and are driven independently by stepper motor actuators, controlled by a closed-loop stepper motor driver that features micro-stepping capabilities, enabling step sizes of less than 50 nm. A polysulfone loading platen is coupled to the bi-axial moving platform. Movements of the stages are controlled by Thor-labs Advanced Positioning Technology (APT) software. The stepper motor driver is used with the software to adjust load parameters of frequency and amplitude of both shear and compression independently and simultaneously. Positional feedback is provided by linear optical encoders that have a bidirectional repeatability of 0.1 &mu;m and a resolution of 20 nm, translating to a positional accuracy of less than 3 &mu;m over the full 50 mm of travel. These encoders provide the necessary position feedback to the drive electronics to ensure true nanopositioning capabilities. In order to provide the force feedback to detect contact and evaluate loading responses, a precision miniature load cell is positioned between the loading platen and the moving platform. The load cell has high accuracies of 0.15% to 0.25% full scale.

We designed a novel mechanical loading bioreactor that can apply uniaxial or biaxial mechanical strain to a cartilage biocomposite prior to transplantation into an articular cartilage defect.

Progettazione di un biassiale Carico meccanico bioreattore per l&#39;ingegneria tissutale

We  designed a loading device that is capable of applying uniaxial or biaxial  mechanical strain to a tissue engineered biocomposites fabricated for  ...

Tissue Engineering of a Human 3D in vitro Tumor Test System

Cancer is one of the leading causes of death worldwide. Current therapeutic strategies are predominantly developed in 2D culture systems, which inadequately reflect physiological conditions in vivo. Biological 3D matrices provide cells an environment in which cells can self-organize, allowing the study of tissue organization and cell differentiation. Such scaffolds can be seeded with a mixture of different cell types to study direct 3D cell-cell-interactions. To mimic the 3D complexity of cancer tumors, our group has developed a 3D in vitro tumor test system.	
	Our 3D tissue test system models the in vivo situation of malignant peripheral nerve sheath tumors (MPNSTs), which we established with our decellularized porcine jejunal segment derived biological vascularized scaffold (BioVaSc). In our model, we reseeded a modified BioVaSc matrix with primary fibroblasts, microvascular endothelial cells (mvECs) and the S462 tumor cell line. For static culture, the vascular structure of the BioVaSc is removed and the remaining scaffold is cut open on one side (Small Intestinal Submucosa SIS-Muc). The resulting matrix is then fixed between two metal rings (cell crowns).	
	Another option is to culture the cell-seeded SIS-Muc in a flow bioreactor system that exposes the cells to shear stress. Here, the bioreactor is connected to a peristaltic pump in a self-constructed incubator. A computer regulates the arterial oxygen and nutrient supply via parameters such as blood pressure, temperature, and flow rate. This setup allows for a dynamic culture with either pressure-regulated pulsatile or constant flow.	
	In this study, we could successfully establish both a static and dynamic 3D culture system for MPNSTs. The ability to model cancer tumors in a more natural 3D environment will enable the discovery, testing, and validation of future pharmaceuticals in a human-like model.

Tissue Engineering of a Human 3D in vitro Tumor Test System

Methods to create human 3D tumor tissues as test systems are described. These technologies are based on a decellularized Biological Vascularized Scaffold (BioVaSc), primary human cells and a tumor cell line, which can be cultured under static as well as under dynamic conditions in a flow bioreactor.

Ingegneria dei tessuti di un Umano 3D<em&gt; In vitro</em&gt; Sistema Tumore test

Cancer is one of the leading causes of death worldwide.  Current therapeutic strategies are predominantly developed in 2D culture  systems, which ...

Electrospun Fibrous Scaffolds of Poly(glycerol-dodecanedioate) for Engineering Neural Tissues From Mouse Embryonic Stem Cells

For tissue engineering applications, the preparation of biodegradable and biocompatible scaffolds is the most desirable but challenging task. &#160;Among the various fabrication methods, electrospinning is the most attractive one due to its simplicity and versatility. Additionally, electrospun nanofibers mimic the size of natural extracellular matrix ensuring additional support for cell survival and growth. This study showed the viability of the fabrication of long fibers spanning a larger deposit area for a novel biodegradable and biocompatible polymer named&#160;poly(glycerol-dodecanoate) (PGD)1 by using a newly designed collector for electrospinning. PGD exhibits unique elastic properties with similar mechanical properties to nerve tissues, thus it is suitable for neural tissue engineering applications. The synthesis and fabrication set-up for making fibrous scaffolding materials was simple, highly reproducible, and inexpensive. In biocompatibility testing, cells derived from mouse embryonic stem cells could adhere to and grow on the electrospun PGD fibers. In summary, this protocol provided a versatile fabrication method for making PGD electrospun fibers to support the growth of mouse embryonic stem cell derived neural lineage cells.

Electrospun Fibrous Scaffolds of Polyglycerol-dodecanedioate for Engineering Neural Tissues From Mouse Embryonic Stem Cells

Synthesis and fabrication of electrospun long fibers spanning a larger deposit area via a newly designed collector from a novel biodegradable polymer named poly(glycerol-dodecanoate) (PGD) was reported. The fibers were able to support the growth of cells derived from mouse pluripotent stem cells.

Elettrofilate Fibrosa Ponteggi di Poli (glicerolo-dodecanedioate) per l&#39;ingegneria neurale tessuti da cellule staminali embrionali di topo

For tissue engineering applications, the preparation of biodegradable and biocompatible scaffolds is the most desirable but challenging task. &#160;Among ...

Engineering 3D Cellularized Collagen Gels for Vascular Tissue Regeneration

Synthetic materials are known to initiate clinical complications such as inflammation, stenosis, and infections when implanted as vascular substitutes. Collagen has been extensively used for a wide range of biomedical applications and is considered a valid alternative to synthetic materials due to its inherent biocompatibility (i.e., low antigenicity, inflammation, and cytotoxic responses). However, the limited mechanical properties and the related low hand-ability of collagen gels have hampered their use as scaffold materials for vascular tissue engineering. Therefore, the rationale behind this work was first to engineer cellularized collagen gels into a tubular-shaped geometry and second to enhance smooth muscle cells driven reorganization of collagen matrix to obtain tissues stiff enough to be handled.
The strategy described here is based on the direct assembling of collagen and smooth muscle cells (construct) in a 3D cylindrical geometry with the use of a molding technique. This process requires a maturation period, during which the constructs are cultured in a bioreactor under static conditions (without applied external dynamic mechanical constraints) for 1 or 2 weeks. The &#8220;static bioreactor&#8221; provides a monitored and controlled sterile environment (pH, temperature, gas exchange, nutrient supply and waste removal) to the constructs. During culture period, thickness measurements were performed to evaluate the cells-driven remodeling of the collagen matrix, and glucose consumption and lactate production rates were measured to monitor the cells metabolic activity. Finally, mechanical and viscoelastic properties were assessed for the resulting tubular constructs. To this end, specific protocols and a focused know-how (manipulation, gripping, working in hydrated environment, and so on) were developed to characterize the engineered tissues.

In this work, we present a technique for the rapid fabrication of living vascular tissues by direct culturing of collagen, smooth muscle cells and endothelial cells. In addition, a new protocol for the mechanical characterization of engineered vascular tissues is described.

Ingegneria 3D Cellularized collagene Gel per Vascolare Rigenerazione Tissutale

Synthetic materials are known to initiate clinical complications such as inflammation, stenosis, and infections when implanted as vascular substitutes. ...

Preparation of Thermoresponsive Nanostructured Surfaces for Tissue Engineering

Thermoresponsive poly(N-isopropylacrylamide) (PIPAAm)-immobilized surfaces for controlling cell adhesion and detachment were fabricated by the Langmuir-Schaefer method. Amphiphilic block copolymers composed of polystyrene and PIPAAm (St-IPAAms) were synthesized by reversible addition-fragmentation chain transfer (RAFT) radical polymerization. A chloroform solution of St-IPAAm molecules was gently dropped into a Langmuir-trough apparatus, and both barriers of the apparatus were moved horizontally to compress the film to regulate its density. Then, the St-IPAAm Langmuir film was horizontally transferred onto a hydrophobically modified glass substrate by a surface-fixed device. Atomic force microscopy images clearly revealed nanoscale sea-island structures on the surface. The strength, rate, and quality of cell adhesion and detachment on the prepared surface were modulated by changes in temperature across the lower critical solution temperature range of PIPAAm molecules. In addition, a two-dimensional cell structure (cell sheet) was successfully recovered on the optimized surfaces. These unique PIPAAm surfaces may be useful for controlling the strength of cell adhesion and detachment.

Nanoscaled sea-island surfaces composed of thermoresponsive block copolymers were fabricated by the Langmuir-Schaefer method for controlling spontaneous cell adhesion and detachment. Both the preparation of the surface and the adhesion and detachment of cells on the surface were visualized.

Preparazione di Thermoresponsive Nanostrutturati Superfici Tissue Engineering

Thermoresponsive poly(N-isopropylacrylamide) (PIPAAm)-immobilized surfaces for controlling cell adhesion and detachment were fabricated by the ...

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and trade one defect for another. Tissue engineering holds the promise to address this increasing demand. However, most tissue engineering strategies fail to generate stable and functional tissue substitutes because of poor vascularization. This paper focuses on an in vivo tissue engineering chamber model of intrinsic vascularization where a perfused artery and a vein either as an arteriovenous loop or a flow-through pedicle configuration is directed inside a protected hollow chamber. In this chamber-based system angiogenic sprouting occurs from the arteriovenous vessels and this system attracts ischemic and inflammatory driven endogenous cell migration which gradually fills the chamber space with fibro-vascular tissue. Exogenous cell/matrix implantation at the time of chamber construction enhances cell survival and determines specificity of the engineered tissues which develop. Our studies have shown that this chamber model can successfully generate different tissues such as fat, cardiac muscle, liver and others. However, modifications and refinements are required to ensure target tissue formation is consistent and reproducible. This article describes a standardized protocol for the fabrication of two different vascularized tissue engineering chamber models in vivo.

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

This is a guideline for constructing in vivo vascularized tissue using a microsurgical arteriovenous loop or a flow-through pedicle configuration inside a tissue engineering chamber. The vascularized tissues generated can be employed for organ regeneration and replacement of tissue defects, as well as for drug testing and disease modeling.

Ingegneria dei tessuti da parte intrinseca Vascolarizzazione in un<em&gt; In Vivo</em&gt; Camera ingegneria dei tessuti

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and ...

Isolation of Murine Adipose Tissue-derived Microvascular Fragments as Vascularization Units for Tissue Engineering

A functional microvascular network is of pivotal importance for the survival and integration of engineered tissue constructs. For this purpose, several angiogenic and prevascularization strategies have been established. However, most cell-based approaches include time-consuming in vitro steps for the formation of a microvascular network. Hence, they are not suitable for intraoperative one-step procedures. Adipose tissue-derived microvascular fragments (ad-MVF) represent promising vascularization units. They can be easily isolated from fat tissue and exhibit a functional microvessel morphology. Moreover, they rapidly reassemble into new microvascular networks after in vivo implantation. In addition, ad-MVF have been shown to induce lymphangiogenesis. Finally, they are a rich source of mesenchymal stem cells, which may further contribute to their high vascularization potential. In previous studies we have demonstrated the remarkable vascularization capacity of ad-MVF in engineered bone and skin substitutes. In the present study, we report on a standardized protocol for the enzymatic isolation of ad-MVF from murine fat tissue.

We present a protocol to isolate adipose tissue-derived microvascular fragments that represent promising vascularization units. They can be rapidly isolated, do not require in vitro processing and, thus, may be used for one-step prevascularization in different fields of tissue engineering.

L&#39;isolamento di Murine adiposo derivate dal tessuto microvascolare Frammenti come vascolarizzazione Unità per Tissue Engineering

A functional microvascular network is of pivotal importance for the survival and integration of engineered tissue constructs. For this purpose, several ...