Name: osmotic drug delivery to ischemic hindlimbs and perfusion of vasculature with microfil for micro-computed tomography imaging
Uploaded: 2013-06-29T13:00:00
Description: Watch this Scientific Journal Video about Osmotic Drug Delivery to Ischemic Hindlimbs and Perfusion of Vasculature with Microfil for Micro-Computed Tomography Imaging at JoVE.com

Forfatterne vil gerne takke Keith Michel af MD Anderson Mindre Husdyrs Imaging Facility for sin tekniske bistand med mikro-CT-scanning, Edward TH Yeh, MD, til kirurgisk assistance, og Rebecca Bartow, ph.d., til redaktionel assistance. Dette arbejde blev støttet delvist af American Heart Association.

1.. Priming den osmotiske pumpe <ol><li> Brug sterile teknikker (f.eks handsker, laminar flow hætte), når udarbejdelsen af pumperne. Fjern pumpen og strømningsmoderatoren fra emballagen. Vej den tomme pumpe og optage vægten (mg). </li><li> Fyld pumpen langsomt og undgå at skabe luftbobler, ved hjælp af en lille 1cc sprøjte og stump spids 27G påfyldningsrør. Når opløsningen når toppen af ​​pumpen, stop påfyldning. </li><li> Tør overskydende opløsning og vejer fyldt pumpe. For de fleste vandi...

<ol>
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Her præsenterer vi en metode til osmotisk stof / stof levering i en musemodel af bagben iskæmi. Herudover beskriver vi en støbeteknik, hvor vi har brugt Microfil at frembringe en 3D-rekonstruktion til analyse af det vaskulære netværk. Niveauet eller sværhedsgraden af ​​iskæmi varierer efter hvor arteriel ligering / excision er lavet. Vi skabte en dobbelt ligation i fælles femoralarterie proksimalt til oprindelsen af ​​den dybtliggende femoris og epigastrica arterier, denne ti...

Perifere kredsløbsforstyrrelser (PAD) er en aterosklerotisk sygdom, der forårsager utilstrækkelig blodforsyning i benene 4.. Det påvirker 8-12.000.000 amerikanere, og aktuelle medicinske behandlinger kun tilbyde begrænset lindring 5,6. Nye terapeutiske midler, der forbedrer blodcirkulationen i benene vil ikke kun begrænse sygdomsprogression, men også øge livskvaliteten. Forekomsten af ​​PAD er højere hos personer over 50 år, så lokale farmakologiske terapi er en mere ønskelig behandlingsmetode, fordi den reducerede nyre-og leverfunktion ofte ses hos ældre patienter kan mindske drug metabolisme og øge bivirkninger ved systemisk administration. Således har vi skabt en musemodel af PAD at undersøge, om lokalt administrerede agenter afhjælpe hindlimb iskæmi ved at fremme angiogenese og mikrovaskulære remodeling. Specifikt har vi brugt en kateter osmotisk pumpe til kontinuerligt levere det terapeutiske middel tiliskæmisk lårmuskel af mus. Ved hjælp af vores delivery system, vi var i stand til at opretholde optimale koncentrationer af lægemidlet i det lokale miljø, denne tilgang giver mulighed for passende stof bioaktivitet undgår mulige systemiske bivirkninger, og overvinder den ulempe begrænsede lokale stof adgangen forbundet med systemisk administration. Derudover at vurdere, om lokalt administrerede agenter fremme revaskularisering, brugte vi avancerede støbning og high-definition imaging teknikker, der muliggør kvantificering af ændringer i microvasculature. Kollektivt, at kombinationen af metoder, der anvendes i denne video artikel er nyttigt i prækliniske studier til at hjælpe på forståelsen farmakologisk induceret revaskularisering i PAD patienter 7-9.

<table border="1">
		<tbody>
		 <tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Reagent/Material</td>
		 </tr>
		 <tr>
			<td>Surgical tools</td>
			<td>Fine Science Tools</td>
			<td></td>
			<td>Type: Tool</td>
		 </tr>
		 <tr>
			<td>Puritan sterile cotton swabs</td>
			<td>Fisher Scientific</td>
			<td>22-029-499</td>
			<td>Type: Tool</td>
		 </tr>
		 <tr>
			<td>Betadine (povidone-iodine)</td>
			<td>Fisher Scientific</td>
			<td>19-065534</td>
			<td>Type: Reagent</td>
		 </tr>
		 <tr>
			<td>70% Alcohol pads</td>
			<td>Fisher Scientific</td>
			<td>NC9926371</td>
			<td>Type: Reagent</td>
		 </tr>
		 <tr>
			<td>Phosphate buffered saline</td>
			<td>Lonza</td>
			<td>17-516F</td>
			<td>Type: Reagent</td>
		 </tr>
		 <tr>
			<td>6-0 prolene suture</td>
			<td>Cardinal Health</td>
			<td>8709</td>
			<td>Type: Tool</td>
		 </tr>
		 <tr>
			<td>8-0 prolene suture</td>
			<td>Cardinal Health</td>
			<td>2775</td>
			<td>Type: Tool</td>
		 </tr>
		 <tr>
			<td>Depilatory cream</td>
			<td>Nair</td>
			<td></td>
			<td>Type: Tool</td>
		 </tr>
		 <tr>
			<td>Osmotic pump</td>
			<td>ALZET</td>
			<td>1002</td>
			<td>Type: Tool, 14 day release</td>
		 </tr>
		 <tr>
			<td>Vinyl catheter</td>
			<td>ALZET</td>
			<td>7760</td>
			<td>Type: Tool</td>
		 </tr>
		 <tr>
			<td>Heparinized saline (0.9%)</td>
			<td>Baxter</td>
			<td>2B0944</td>
			<td>Type: Reagent</td>
		 </tr>
		 <tr>
			<td>Neutral buffered formalin</td>
			<td>Richard-Allan Scientific</td>
			<td>5705</td>
			<td>Type: Reagent</td>
		 </tr>
		 <tr>
			<td>Microfil (silicone rubber contrast agent)</td>
			<td>Flowtech</td>
			<td>MV-112</td>
			<td>Type Reagent, Microfil White</td>
		 </tr>
		 <tr>
			<td>Cal-Ex II (formic acid solution)</td>
			<td>Fisher Scientific</td>
			<td>CS511-1D</td>
			<td>Type: Reagent</td>
		 </tr>
		 <tr>
			<td>Buprenex</td>
			<td>CIII</td>
			<td>7571</td>
			<td>Type: Analgesic</td>
		 </tr>
		 <tr>
			<td>Bupivicaine</td>
			<td>Hospira, Inc.</td>
			<td>381</td>
			<td>Type: Analgesic</td>
		 </tr>
		 <tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Equipment</td>
		 </tr>
		 <tr>
			<td>Dissecting microscope</td>
			<td>Carl Zeiss Microimaging</td>
			<td>Zeiss Stemi 2000-C</td>
			<td>Type:Equipment</td>
		 </tr>
		 <tr>
			<td>Laser Doppler perfusion imager</td>
			<td>Perimed Inc.</td>
			<td>Periscan PIM3 </td>
			<td>Type:Equipment</td>
		 </tr>
		 <tr>
			<td>Micro-CT imaging system</td>
			<td>GE Healthcare</td>
			<td>Explore Locus SP </td>
			<td>Type:Equipment</td>
		 </tr>
		</tbody>
 </table>

osmotic drug delivery to ischemic hindlimbs and perfusion of vasculature with microfil for micro-computed tomography imaging

Prækliniske forskning i dyremodeller af perifer arteriel sygdom spiller en afgørende rolle i at teste effektiviteten af ​​terapeutiske midler, der skal stimulere mikrocirkulationen. Valget af levering metode til disse midler er vigtigt, fordi indgivelsesvejen dybt påvirker bioaktiviteten og effekten af disse agenter 1,2. I denne artikel viser vi, hvordan man lokalt indgive et stof i iskæmiske baglemmer ved hjælp af en kateteriseres osmotisk pumpe. Denne pumpe kan levere et fast volumen af ​​vandig opløsning kontinuerligt i et fastsat tidsrum. Vi har også præsentere vores musemodel af ensidig bagben iskæmiinduceret af ligering af den fælles femoralarterie proksimalt til oprindelsen af ​​Profunda femoris og epigastrica arterier i venstre bagben. Endelig beskriver vi in vivo kanyle og ligering af den infrarenale abdominale aorta og perfusion af bagbenet vaskulatur med Microfil, en silicone røntgenfaste casting agent. Microfil kan perfundere og udfylde hele vaskulære seng (arteriel og venøs), og fordi vi har ligeret den store vaskulære kanal til udgang, kan midlet tilbageholdes i karrene til fremtidig ex vivo billeddannelse med brug af lille prøve mikro-CT 3 .

Animationen viser den osmotiske pumpe indsættelse og bagben iskæmi kirurgi beskrevet i protokollen. Figur 1 viser laser Doppler billeder af perfusion af bagbenet, bekræfter iskæmi. Efter vaskulær støbning med Microfil, viser en 3D mikro-CT billede af det vaskulære netværk, Microfil kan fylde skibene effektivt (figur 2A), men at diskontinuiteter kan forekomme (fig. 2B) på grund af forskellige proceduremæssige faktorer (fx luftbobler, manglende tryk, vi...

Watch this Scientific Journal Video about Osmotic Drug Delivery to Ischemic Hindlimbs and Perfusion of Vasculature with Microfil for Micro-Computed Tomography Imaging at JoVE.com

Osmotic Drug Delivery to Ischemic Hindlimbs and Perfusion of Vasculature with Microfil for Micro-Computed Tomography Imaging

Vi viser her<em&gt; In vivo</em&gt; Indsættelse af en osmotisk pumpe til konstant lokale stof levering og etablering af bagbenet iskæmi i en musemodel. Desuden er bagbenet kar perfunderet med Microfil, en silicone stråleuigennemtrængelig agent, for at forberede mikro-computertomografi (mikro-CT).

Osmotisk Drug Delivery til Iskæmisk bagbensknogler og perfusion af kar med Microfil for Micro-computertomografi Imaging

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to ...

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Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ...

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The aim of this protocol presents an optimized procedure for the purification and cultivation of pBECs and to establish in vitro blood-brain barrier (BBB) ...

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Structural remodeling is a common consequence of chronic pathological stresses imposed on the heart. Understanding the architectural and compositional ...