Name: engineered vascularized muscle flap
Uploaded: 2016-01-11T09:30:00
Description: Watch this Scientific Journal Video about Engineered Vascularized Muscle Flap at JoVE.com

This research was supported by the FP7 European Research Council Grant 281501, ENGVASC.

Alle tierexperimentellen Studien wurden vom Ausschuss der Ethik der Tierversuche des Technion genehmigt. Für dieses Verfahren wurden athymischen nackten Mäusen verwendet, um eine immunologische Abstoßung zu vermeiden. Bei Verwendung eines anderen Typs von Maus sollten die Mäuse rasiert vor dem chirurgischen Eingriff und Verabreichung von Cyclosporin (oder eines anderen anti-Abstoßung Ersatz) empfohlen werden. 1. Gerüst Vorbereitung und Handy Einbetten <ol><li> Vorbereitung Gerüste b...

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Die Fortschritte im Tissue Engineering haben mit einer wachsenden Nachfrage nach Ersatzgewebe für den Wiederaufbau von verschiedenen Gewebetypen erfüllt. Eine Vielzahl von synthetischen und biologischen 1,17,18 14-16 Materialien sowie Herstellungsverfahren wurden für ihre Fähigkeit, diesen Anforderungen gerecht beurteilt. Trotz der Fortschritte in der klinischen Versorgung und im Tissue Engineering, die Wiederherstellung der vollen Dicke Bauchwanddefekten bleibt jedoch eine Herausforderung. Ein...

Bauchwanddefekte entstehen oft nach schweren Traumata, Krebsbehandlung, Verbrennungen und Entfernung infizierter mesh. Diese Fehler beinhalten häufig erhebliche Gewebeverlust, komplizierte chirurgische Eingriffe und präsentiert eine große Herausforderung für die plastische Wiederherstellung Chirurgen 1-4. Tissue Engineering Forscher der Suche nach neuen Quellen für künstliche Gewebe haben unterschiedliche Materialien, Zellquellen und Wachstumsfaktoren untersucht. Erfolgreiche Restaurationen aus verschiedenen Geweben, wie zum Beispiel 5,6 Trachea, der Blase 7, 8 Kornea, Knochenhaut 9 und 10 durch Implantation von technischen Geweben wurden bereits berichtet. Jedoch die Herstellung einer dicken vaskularisierten technisch hergestellte Gewebe, insbesondere zur Rekonstruktion von großen Defekten, bleibt eine große Herausforderung im Tissue Engineering. Eine der wichtigsten Faktoren, die die Dicke eines lebensfähigen Gewebekonstrukt wird die Steuerung der Sauerstoffversorgung auf seine Nachteiletituent Zellen. Beim Rückgriff auf Diffusion konstruieren Dicke ist auf der von einer sehr dünnen Schicht. Der maximale Abstand zwischen Sauerstoff- und Nährstoff-Zufuhr Kapillaren in vivo etwa 200 um, die mit der Diffusionsgrenzsauerstoff 11,12 korreliert. Unzureichende Vaskularisierung in Gewebeischämie führen und zu eskalieren zu Gewebeabbau oder Nekrose 13. Darüber hinaus müssen die für die Geweberekonstruktion verwendet ideales Material biokompatibel und nicht-immunogen sein. Es muss auch in der Lage, die weitere Integration von Wirtszellen mit dem Biomaterial und die strukturelle Integrität zu gewährleisten. Verschiedene biologische 14-16 und synthetische 1,17,18 Matrizen wurden zuvor zum Wiederaufbau von Gewebe untersucht, aber ihre Verwendung bleibt wegen des Mangels an wirksamen Blutversorgung, Infektionen oder unzureichende Gewebefestigkeit begrenzt. In dieser Studie wurde ein biokompatibles, zell embedded Gerüst der Food and Drug Administration (FDA) -zugelassene Poly-L-Milchsäure (PLLA) / Polymilchsäure-co-Glykolsäure (PLGA) umfasste, wurde um die Arteria und Vena femoralis (AV) Schiffe mit einer nackten Maus implantiert und getrennt von dem umgebenden Gewebe, wodurch Vaskularisierung nur aus den AV Gefäße. Eine Woche nach der Implantation war das Transplantat lebensfähigen, dick und gut vaskularisiert. Diese dicke vaskularisiertes Gewebe mit den AV Gefäße, wurde dann als gestielten Lappen zu einer Bauch voller Dicke Defekt in der gleichen Maus übertragen. Eine Woche nach der Übertragung wurde die Klappe lebensfähigen, vaskularisiert und gut mit dem umgebenden Gewebe integriert, wobei eine ausreichende Festigkeit, um Bauchorgane zu unterstützen. Somit ist die technisch stark, vaskularisierten Gewebelappen, wobei eine autologe Stiel, stellt ein neues Verfahren für die Reparatur von voller Dicke Bauchwanddefekte.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>small fine straight scissors</td>
			<td>Fine Science Tools (FST)</td>
			<td>14090-09</td>
			<td></td>
		</tr>
		<tr>
			<td>spring scissors</td>
			<td>Fine Science Tools (FST)</td>
			<td>15003-08</td>
			<td></td>
		</tr>
		<tr>
			<td>straight forceps with fine tip</td>
			<td>Fine Science Tools (FST)</td>
			<td>11251-20</td>
			<td></td>
		</tr>
		<tr>
			<td>serrated forceps&#160;</td>
			<td>Fine Science Tools (FST)</td>
			<td>11050-10</td>
			<td></td>
		</tr>
		<tr>
			<td>needle holder</td>
			<td>Fine Science Tools (FST)</td>
			<td>12500-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Small vessel cauterizer&#160;</td>
			<td>Fine Science Tools (FST)</td>
			<td>18000-00</td>
			<td></td>
		</tr>
		<tr>
			<td>Duratears</td>
			<td>Alcon</td>
			<td align="right">5686</td>
			<td></td>
		</tr>
		<tr>
			<td>Sedaxylan</td>
			<td>Euravet</td>
			<td>DJ03</td>
			<td></td>
		</tr>
		<tr>
			<td>Clorketam 1000</td>
			<td>Vetoquinol</td>
			<td>4A0726B</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>vetmarket</td>
			<td>B15100</td>
			<td></td>
		</tr>
		<tr>
			<td>4-0 silk sutures</td>
			<td>Assut sutures</td>
			<td align="right">647</td>
			<td></td>
		</tr>
		<tr>
			<td>6-0 polypropylene sutures</td>
			<td>Assut sutures</td>
			<td>9351F</td>
			<td></td>
		</tr>
		<tr>
			<td>8-0 silk sutures</td>
			<td>Assut sutures</td>
			<td align="right">684568</td>
			<td></td>
		</tr>
		<tr>
			<td>Insulin syringe (6mm needle)</td>
			<td>BD</td>
			<td align="right">324911</td>
			<td></td>
		</tr>
		<tr>
			<td>Vevo 2100 high-resolution ultrasound system</td>
			<td>VisualSonics inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>MS250 non-linear transducer</td>
			<td>VisualSonics inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Micromarker non-targeted contrast agent</td>
			<td>VisualSonics inc.</td>
			<td>VS-11694</td>
			<td></td>
		</tr>
		<tr>
			<td>tail vein catheter</td>
			<td>VisualSonics inc.</td>
			<td>VS-11912</td>
			<td></td>
		</tr>
		<tr>
			<td>Vevo 2100 software</td>
			<td>VisualSonics inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>fluorescein isothiocyanate-conjugated dextran</td>
			<td>Sigma</td>
			<td>FD500S</td>
			<td></td>
		</tr>
		<tr>
			<td>Matlab</td>
			<td>Mathworks, MA, USA</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Kimwipes</td>
			<td>Kimtech</td>
			<td align="right">34120</td>
			<td></td>
		</tr>
		<tr>
			<td>antigen unmasking solution</td>
			<td>Vector laboratories</td>
			<td>H-3300</td>
			<td></td>
		</tr>
		<tr>
			<td>anti-CD31 antibody</td>
			<td>Abcam&#160;</td>
			<td>ab28364</td>
			<td></td>
		</tr>
		<tr>
			<td>biotinylated goat anti-rabbit (secondary) antibody</td>
			<td>Vector laboratories</td>
			<td>BA-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>streptavidin-peroxidase</td>
			<td>Jackson&#160;</td>
			<td>016-030-084</td>
			<td></td>
		</tr>
		<tr>
			<td>Mayer&#39;s hamatoxylin solution</td>
			<td>Sigma-Aldrich</td>
			<td>MHS-16</td>
			<td></td>
		</tr>
		<tr>
			<td>aminoethylcarbazole (AEC) substrate kit</td>
			<td>Life technologies, Invitrogen&#160;</td>
			<td>00-2007</td>
			<td></td>
		</tr>
		<tr>
			<td>Vectamount</td>
			<td>Vector laboratories</td>
			<td>H-5501</td>
			<td></td>
		</tr>
	</tbody>
</table>

engineered vascularized muscle flap

One of the main factors limiting the thickness of a tissue construct and its consequential viability and applicability in vivo, is the control of oxygen supply to the cell microenvironment, as passive diffusion is limited to a very thin layer. Although various materials have been described to restore the integrity of full-thickness defects of the abdominal wall, no material has yet proved to be optimal, due to low graft vascularization, tissue rejection, infection, or inadequate mechanical properties. This protocol describes a means of engineering a fully vascularized flap, with a thickness relevant for muscle tissue reconstruction. Cell-embedded poly L-lactic acid/poly lactic-co-glycolic acid constructs are implanted around the mouse femoral artery and vein and maintained in vivo for a period of one or two weeks. The vascularized graft is then transferred as a flap towards a full thickness defect made in the abdomen. This technique replaces the need for autologous tissue sacrifications and may enable the use of in vitro engineered vascularized flaps in many surgical applications.

Graft Vaskularisation und Perfusion in vivo Die Transplantate wurden ein oder zwei Wochen vor ihrer Übertragung als axiale Klappen implantiert. Bei einer und zwei Wochen nach der Implantation, der groben Beobachtung des Transplantatbereich enthüllt lebensfähig und vaskularisierten Gewebetransplantate. Diese Transplantate nachgewiesen gefäß zu sein, wie durch positive Immunfärbung CD31 (1A) bestimmt ist, und stark durchbluteten, wie von FITC-Dextran-Injektion i...

Watch this Scientific Journal Video about Engineered Vascularized Muscle Flap at JoVE.com

Engineered Vascularized Muscle Flap

To date, thick tissue defects are typically reconstructed by applying autologous tissue flaps or engineered tissues. In this protocol, we present a new method for engineering vascularized tissue flap bearing an autologous pedicle, to serve as a substitute to autologous flaps.

Engineered Vaskularisierte Muskellappen

One of the main factors limiting the thickness of a tissue construct and its consequential viability and applicability in vivo, is the control of oxygen ...

Research

JoVE Journal

Bioengineering

Engineered 3D Silk-collagen-based Model of Polarized Neural Tissue

Engineered 3D Silk-Kollagen-basierten Modell von polarisiertem Neural Tissue

Despite huge efforts to decipher the anatomy, composition and function of the brain, it remains the least understood organ of the human body. To gain a ...

Forschung

Biotechnik

Applications of In Vivo Functional Testing of the Rat Tibialis Anterior for Evaluating Tissue Engineered Skeletal Muscle Repair

Applications of In Vivo Functional Testing of the Rat Tibialis Anterior for Evaluating Tissue Engineered Skeletal Muscle Repair

Anwendungen<em&gt; In Vivo</em&gt; Funktionstest der Ratte Vorderer Schienbeingewebe Skeletal Muscle Repair Engineered für die Bewertung

Despite the regenerative capacity of skeletal muscle, permanent functional and/or cosmetic deficits (e.g., volumetric muscle loss (VML) resulting from ...

Microfluidic Bioprinting for Engineering Vascularized Tissues and Organoids

Mikrofluidische Bioprinting für Engineering durchblutet, Gewebe und Organellen

Engineering vascularized tissue constructs and organoids has been historically challenging. Here we describe a novel method based on microfluidic ...

Tissue-Engineered Graft for Circumferential Esophageal Reconstruction in Rats

The use of biocompatible materials for circumferential esophageal reconstruction is a technically challenging task in rats and requires an optimal implant ...

Fabrication of Zero Mode Waveguides for High Concentration Single Molecule Microscopy

Herstellung von Zero Mode Waveguides für die hochkonzentrierte Einzelmolekülmikroskopie

In single molecule fluorescence enzymology, background fluorescence from labeled substrates in solution often limits fluorophore concentration to pico- to ...

Generation of Self-assembled Vascularized Human Skin Equivalents

Generierung von selbst zusammengesetzten vaskularisierten äquivalenten menschlicher Haut

Human skin equivalents (HSEs) are tissue engineered constructs that model epidermal and dermal components of human skin. These models have been used to ...

Assessing Functional Metrics of Skeletal Muscle Health in Human Skeletal Muscle Microtissues

Bewertung funktioneller Metriken der Skelettmuskelgesundheit in Mikrobeschwerden der menschlichen Skelettmuskulatur

Three-dimensional (3D) in vitro models of skeletal muscle are a valuable advancement in biomedical research as they afford the opportunity to study ...

Fibroblast Derived Human Engineered Connective Tissue for Screening Applications

Fibroblasten-abgeleitetes Human Engineered Connective Tissue für Screening-Anwendungen

Fibroblasts are phenotypically highly dynamic cells, which quickly transdifferentiate into myofibroblasts in response to biochemical and biomechanical ...

Engineered Lung Tissues Prepared from Decellularized Lung Slices

Technisches Lungengewebe aus dezellularisierten Lungenscheiben

There is a need for improved 3-dimensional (3D) lung models that recapitulate the architectural and cellular complexity of the native lung alveolus ex ...

A Reliable Porcine Fascio-Cutaneous Flap Model for Vascularized Composite Allografts Bioengineering Studies

Ein zuverlässiges schweinisches Faszien-Hautlappenmodell für vaskularisierte Komposit-Allografts Bioengineering-Studien

Vascularized Composite Allografts (VCA) such as hand, face, or penile transplant represents the cutting-edge treatment for devastating skin defects, ...