Name: a full skin defect model to evaluate vascularization of biomaterials in vivo
Uploaded: 2014-08-28T14:00:00
Description: Watch this Scientific Journal Video about A Full Skin Defect Model to Evaluate Vascularization of Biomaterials In Vivo at JoVE.com

Integra dermal regenerering skabelon er venligst leveret af Integra LifeSciences Corporation. Kilder til midlerne til støtte for arbejdet: Dette arbejde blev delvist finansieret af CIRM-BMBF Tidlig Translationel II Award og FONDAP Center for Genome regulering både JTE (Nr 15090007.).

1. Fremstilling af stilladser <ol><li> Generer prøver af stilladser ved hjælp af 12 mm biopsi slag. </li><li> At introducere bioaktive molekyler eller celler i stillads, dræne stilladser ved forsigtigt at klemme dem med en steril gaze. Derefter rehydrere stilladser ved tilsætning af 160 pi af en opløsning indeholdende de bioaktive molekyler eller celler af interesse. Dobbelt-tjek succes bioaktivering med celler gennem metaboliske assays, såsom MTT assays. </li><li> Hvis det er nødvendigt, fastsætte forbindelse...

<ol>
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G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vessel+segmentation+and+analysis+in+laboratory+skin+transplant+micro-angiograms.">Vessel segmentation and analysis in laboratory skin transplant micro-angiograms.</a> Proceedings of the Eighteenth IEEE Symposium on Computer-Based Medical Systems. , 21-26 (2005).</li><li>Danner, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+human+sweat+gland-derived+stem+cells+for+enhancing+vascularization+during+dermal+regeneration.">The use of human sweat gland-derived stem cells for enhancing vascularization during dermal regeneration.</a> Journal of Investigative Dermatology. 132 (6), 1707-1716 (2012).</li><li>Shaterian, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Real+Time+Analysis+of+the+Kinetics+of+Angiogenesis+and+Vascular+Permeability+in+an+Animal+Model+of+Wound+Healing.">Real Time Analysis of the Kinetics of Angiogenesis and Vascular Permeability in an Animal Model of Wound Healing.</a> Burns. 35 (6), 811-817 (2009).</li><li>McDonald, D. 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Der er behov for at etablere vellykkede tilgange til at forbedre blodperfusion i manipuleret væv konstruktioner, som kræver udvikling af nye pålidelige metoder til at studere vaskulariseringen processer inden for biomaterialer. Almindelige metoder til fremstilling stillads vaskularisering ex vivo synlig indbefatter anvendelse af mikroskopi, som giver en høj opløsning værktøj. I de fleste tilfælde, selv om denne metode er begrænset til analyse af små vævs områder, og har tendens til at være dyrt og ...

I de seneste årtier har tissue engineering åbnet en ny terapeutisk mulighed for at erstatte vævsdefekter med kroppens egne celler 1. For at støtte den fysiologiske proces vævsregenerering er stilladser udformet som en bionedbrydelig struktur, der giver et scenarie, hvor celler fra sårlejet kan vokse og genoprette defekten 2,3. Utilstrækkelig vaskularisering anses for at være den største hindring, som holder tilbage den kliniske gennembrud Bioartificial stilladser 4. Med indvækst af celler, bliver væsentlig efterspørgsel efter næringsstoffer og ilt stiger, og vaskularisering af materialet. Utilstrækkelig eller forsinket vaskularisering kan derfor føre til central nekrose af væv-manipuleret 5. Desuden blodkar forsyner immunkompetente celler og fjerne de metaboliske rester i regenererende område. Høje infektion satser og lav regenerering er kunnogle af konsekvenserne af utilstrækkelig blodperfusion observeret i tissue engineering, der har til formål at afværges ved at øge vaskularisering af stilladser 6,7. Adskillige strategier, der sigter mod at forbedre vaskularisering fokus på den centrale rolle, biomaterialet selv og mikrostruktur af stilladset. Der er intensiv forskning bestræbelser på at udvikle nye tilgange i skiftende helingsprocessen fra reparation til regenerering, og dermed (gen) generere en væv med de nærmeste fysiologiske egenskaber til en, der skal genoprettes 8,9. Biomaterialer, der blev undersøgt og evalueret med hensyn til deres regenerative potentiale inkluderet kollagen, fibrin, chitosan og alginat 10,11. Disse biomaterialer kan bruges og kombineres som en rygrad til at bygge nye stilladser ved hjælp af forskellige strategier såsom væv decellularization, self-assembly, rapid prototyping og electrospinning 12. For at ENHstemmelse kroppens egen regenerationsevne kan stilladser blive bioaktiveres. Inkorporeringen af rekombinant angiogene vækstfaktorer 13 eller gen vektorer, der koder for sådanne faktorer, 14 har vist sig at forbedre vaskularisering af stilladset. Anvendelsen af stamceller er i vid udstrækning blevet vist at være en lovende strategi til forbedring vaskularisering, hvor mesenchymale stromale celler og endotheliale progenitorceller har fået mest opmærksomhed 15,16. Andre tilgange forsøger at bygge konstruktioner, der indeholder præfabrikerede netværk fartøj før transplantation 17. Trods intensiv indsats i stillads design og deres bio-aktivering, har ingen strategi forbedret vaskularisering ved et klinisk signifikant niveau, og med undtagelse af dermale udskiftninger i massive forbrændinger, er oversættelsen af gensplejsede materialer i klinisk rutine kun finder sted tøvende 18 . En af grundene til, at vaskulariseringaf kunstige vævskonstruktioner er stadig et uløst problem, er vanskeligheden ved at evaluere resultaterne af nye teknologier i in vivo-metoder. Selvom in vitro-forsøg kan give vigtige indsigter vaskulariseringen potentiale stilladser, er egnede dyremodeller der kræves for at studere centrale parametre såsom biokompatibilitet af materialet, sikkerhed og effekt af behandlingen, og af særlig betydning, at vaskularisering af vævet konstruere. Derfor er pålidelige værktøjer visualisere og kvantificere blodkar netværk in vivo er afgørende. I denne undersøgelse præsenterer vi en enkel og pålidelig metode, der tillader visualisering og kvantificering af det vaskulære netværk inde eksplanterede stillads. Denne metode er baseret på væv gennemlysning og digital segmentering. Da denne metode er ikke-invasiv, det giver mulighed for yderligere molekylære og histologiske analyser af målmaterialet.

<table border="0" cellpadding="0" cellspacing="0" width="1091">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:319px;">Name of Material/ Equipment</td>
			<td style="width:223px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:431px;">Comments/Description</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:319px;">Ethilon P-3 13 mm 3/8 circle 5-0</td>
			<td style="width:223px;">Ethicon, Norderstedt, Germany</td>
			<td style="width:119px;">698G</td>
			<td>Ethilon polyamid-6 precision point-reverse cutting suture</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:319px;">Biopsy punches (10 mm)</td>
			<td style="width:223px;">Xiomedics, Acuderm inc., Fort Lauderdale, FL, USA</td>
			<td style="width:119px;">P1050</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:319px;">Biopsy punches (12 mm)</td>
			<td style="width:223px;">Xiomedics, Acuderm inc., Fort Lauderdale, FL, USA</td>
			<td style="width:119px;">P1250</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Digital camera&#160;</td>
			<td style="width:223px;">Ricoh, Hannover, Germany</td>
			<td style="width:119px;">Cx1</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Gazin Mullkompresse&#160;</td>
			<td style="width:223px;">Lohmann und Rauscher, Neuwied, Germany</td>
			<td style="width:119px;">13622</td>
			<td>Sterile gauze (10 cm x 10 cm)</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Double-layer collagen-based scaffold (8 &#39;x 10&#39;)</td>
			<td style="width:223px;">Integra Life Science Corporation, Plainsboro, NJ, USA</td>
			<td style="width:119px;">88101</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Isoflurane, liquid-gas for inhalative anesthesia&#160;</td>
			<td style="width:223px;">Baxter, Unterschleissheim, Germany</td>
			<td style="width:119px;">100196040</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Pentobarbital, 16 g / 100 ml</td>
			<td style="width:223px;">Fa. Merial, Hallbergmoos</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:319px;">Nuri Nu/Nu Nude mice, CrLNU-Foxn1nu</td>
			<td style="width:223px;">Charles River, Sulzfeld, Germany</td>
			<td style="width:119px;">Strain code 088</td>
			<td>Athymic nude mice, 6 to 8 weeks of age and with a body weight between 20 to 25 g&#160;</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:319px;">Buprenorphine (0.3 mg/ml)</td>
			<td style="width:223px;">Essex Pharma GmbH, Munich, Germany</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Titanized mesh (15 cm x 15 cm), extralight</td>
			<td style="width:223px;">PFM Medical AG, K&#246;ln, Germany</td>
			<td style="width:119px;">6000029</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:319px;">Tissucol Duo S Immuno 2 ml</td>
			<td style="width:223px;">Baxter Germany GmbH, Unterschlei&#223;heim, Germany</td>
			<td style="width:119px;">B1332020110614</td>
			<td>Fibrin-thrombin solution&#160;</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Transparent adhesove drape (30.5 cm x 26 cm)</td>
			<td style="width:223px;">KCI Medical Products, Wimborne Dorset, UK</td>
			<td style="width:119px;">M6275009/10</td>
			<td></td>
		</tr>
	</tbody>
</table>

a full skin defect model to evaluate vascularization of biomaterials in vivo

Utilstrækkelig vaskularisering anses for at være en af ​​de vigtigste faktorer, der begrænser den kliniske succes for væv-manipuleret konstruktioner. For at vurdere nye strategier, der sigter mod at forbedre vaskularisering, er pålidelige metoder kræves for at gøre det i vækst af nye blodkar i bio-kunstige stilladser synlige og kvantificere resultaterne. Gennem de seneste par år, har vores gruppe indført et komplet hud defekt model, der gør det muligt direkte visualisering af blodkar ved gennemlysning og giver mulighed for kvantificering via digital segmentering. I denne model, en kirurgisk skaber fuld overfladefejl i ryggen af ​​mus og erstatter dem med det afprøvede materiale. Molekyler eller celler af interesse kan også indgå i sådanne materialer til at studere deres potentielle effekt. Efter en observationstid på ens egen valg, materialer eksplanteret til evaluering. Bilaterale sår giver muligheden for at gøre de interne sammenligninger thved minimering af artefakter mellem individer samt mindske antallet af dyr, der er nødvendige for undersøgelsen. I sammenligning med andre fremgangsmåder, vores metode giver en enkel, pålidelig og omkostningseffektiv analyse. Vi har implementeret denne model som en rutine værktøj til at udføre høj opløsning screening ved test vaskularisering af forskellige biomaterialer og bio-aktivering tilgange.

Der kan skabes en pålidelig bilateral fuld hud defekt i mus (figur 1), hvor huden kan erstattes af et biomateriale under undersøgelse (figur 2). Her er ingen større komplikationer under eller efter den operative procedure, hverken makroskopiske tegn på infektion eller udenlandsk organ reaktion. I sjældne tilfælde, bliver et stillads tabt, når en mus fjerner det. Sårkontraktion blev aldrig observeret (figur 3). Tissue gennemlysning tilladt klar visualisering af v...

Watch this Scientific Journal Video about A Full Skin Defect Model to Evaluate Vascularization of Biomaterials In Vivo at JoVE.com

A Full Skin Defect Model to Evaluate Vascularization of Biomaterials In Vivo

Vaskularisering er nøglen til tilgange i vellykket tissue engineering. Derfor er pålidelige teknologier nødvendige for at vurdere udviklingen af ​​vaskulære netværk i væv-konstruktioner. Her præsenterer vi en enkel og omkostningseffektiv metode til at visualisere og kvantificere vaskularisering in vivo.

En fuld huddefekten model til evaluering Vaskularisering biomaterialer<em&gt; In Vivo</em

Insufficient vascularization is considered to be one of the main factors limiting the clinical success of tissue-engineered constructs. In order to ...

Research

JoVE Journal

Bioengineering

A Full Skin Defect Model to Evaluate Vascularization of Biomaterials In Vivo

The Three-Dimensional Human Skin Reconstruct Model: a Tool to Study Normal  Skin and Melanoma Progression

Most in vitro studies in experimental skin biology have been done in 2-dimensional (2D) monocultures, while accumulating evidence suggests that cells ...

Design of a Cyclic Pressure Bioreactor for the Ex Vivo Study of Aortic Heart Valves

Design of a Cyclic Pressure Bioreactor for the Ex Vivo Study of Aortic Heart Valves

The aortic valve, located between the left ventricle and the aorta, allows for unidirectional blood flow, preventing backflow into the ventricle. Aortic ...

Generation of a Three-dimensional Full Thickness Skin Equivalent and Automated Wounding

Generation af en tre-dimensionel fuld tykkelse Skin Equivalent og Automated Anskydning

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models ...

Sandwich-like Microenvironments to Harness Cell/Material Interactions

Sandwich-lignende mikromiljøer at udnytte Cell / Materiale Interactions

Cell culture has been traditionally carried out on bi-dimensional (2D) substrates where cells adhere using ventral receptors to the biomaterial surface. ...

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

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

Tissue Engineering af Intrinsic Vaskularisering i en<em&gt; In vivo</em&gt; 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 ...

A Method for Evaluating Timeliness and Accuracy of Volitional Motor Responses to Vibrotactile Stimuli

En metode til evaluering Aktualitet og nøjagtighed viljesbestemt Motor Svar til Vibrotactile Stimuli

Artificial sensory feedback (ASF) systems can be used to compensate for lost proprioception in individuals with lower-limb impairments.  Effective design ...

Quantification of Strain in a Porcine Model of Skin Expansion Using Multi-View Stereo and Isogeometric Kinematics

Kvantificering af Strain i et porcint Model af hud Ekspansion Brug Multi-View Stereo og Isogeometric Kinematik

Tissue expansion is a popular technique in plastic and reconstructive surgery that grows skin in vivo for correction of large defects such as burns and ...

Visualizing Angiogenesis by Multiphoton Microscopy In Vivo in Genetically Modified 3D-PLGA/nHAp Scaffold for Calvarial Critical Bone Defect Repair

Visualizing Angiogenesis by Multiphoton Microscopy In Vivo in Genetically Modified 3D-PLGA/nHAp Scaffold for Calvarial Critical Bone Defect Repair

Visualisere angiogenese af Multiphoton mikroskopi In Vivo i genetisk modificerede 3D-PLGA/nHAp stillads til Calvarial kritiske knogle defekt reparation

The reconstruction of critically sized bone defects remains a serious clinical problem because of poor angiogenesis within tissue-engineered scaffolds ...

A Zebrafish Embryo Model for In Vivo Visualization and Intravital Analysis of Biomaterial-associated Staphylococcus aureus Infection

A Zebrafish Embryo Model for In Vivo Visualization and Intravital Analysis of Biomaterial-associated Staphylococcus aureus Infection

Zebrafisk Embryo Model for In Vivo visualisering og Intravital analyse af biomateriale-associerede Staphylococcus aureus infektion

Biomaterial-associated infection (BAI) is a major cause of the failure of biomaterials/medical devices. Staphylococcus aureus is one of the major ...

A Reliable and Reproducible Critical-Sized Segmental Femoral Defect Model in Rats Stabilized with a Custom External Fixator

En pålidelig og reproducerbar kritiske mellemstore Segmental Femoral defekt Model i rotter stabiliseret med en brugerdefineret ekstern fiksator

Orthopedic research relies heavily on animal models to study mechanisms of bone healing in vivo as well as investigate the new treatment techniques. ...