Name: heterotopic mucosal engrafting procedure for direct drug delivery to the brain in mice
Uploaded: 2014-07-16T14:00:00
Description: Watch this Scientific Journal Video about Heterotopic Mucosal Engrafting Procedure for Direct Drug Delivery to the Brain in Mice at JoVE.com

Denne undersøgelse blev finansieret af Mcihael J. Fox Foundation for Parkinsons Research 2011 reaktionsoperationer Innovations Awards Program. De finansieringskilderne havde ingen rolle i studie design, dataindsamling og-analyse, beslutning om at offentliggøre, eller udarbejdelse af manuskriptet.

Før operation sikre, at alle procedurer, der skal gøres, er godkendt af IACUC og eventuelle yderligere etiske eller juridiske myndigheder og bruge human behandling af dyr praksis. Dette omfatter brug af sterile kirurgi betingelser, bedøve musen ved at bruge IACUC godkendt metode, smøring mus øjne med dyrlægen salve under operationen, og give postsurgical pleje. Må ikke fortsætte med kirurgi, hvis der er nogen spørgsmål, om aspekter af proceduren er godkendt. Alle procedurer udføres heri, blev godkendt af Bost...

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Den sværeste skridt i proceduren, der er beskrevet heri, er den vellykket overdragelse af en tilstrækkelig størrelse slimhinde på hjernen overflade. Dette trin er væsentligt nemmere, hvis det høstede næseskillevæggen er stort nok og rengøres godt. Hvis den ventrale del af skillevæggen er afkortet, hvis en ny graft opnås. Boringen vinkel skal være vinkelret på musen hovedet for at sikre, at slimhinden ikke beskadiges af boret. Hvis en bredere end anbefalet boring sti taget det vil være meget sværere at fje...

Behandling af neurologiske og psykiatriske sygdomme er alvorligt hæmmet af tilstedeværelsen af blod-hjerne-barrieren (BBB), hvilket hindrer over 95% af alle potentielle farmaceutiske midler at nå centralnervesystemet 1-3. For eksempel glial neurotrofisk faktor (GDNF) har vist sig at være effektivt i behandling af Parkinsons sygdom, når det injiceres direkte i hjernen, men er ineffektiv, når den leveres systemisk, fordi det ikke kan trænge BBB 4-6. Talrige nærmede er blevet udviklet til at omgå dette problem. Forbedring i systemisk levering af neurotheraputics er blevet påvist ved hjælp af lægemiddelkonjugater indeholdende antistoffer er selektive for transportproteiner placeret på hjernen kapillærendotelet; men denne metode har ikke vist sig at være gældende for en bred vifte af lægemidler 7,8. Derudover har osmotisk åbning af BBB blevet anvendt klinikallierede, men denne fremgangsmåde lider af systemisk lægemiddel dosering i modsætning til en mere direkte levering til hjernen regionen af interesse 9. Væsentlig indsats har været lagt i at optimere transnasal levering i håb om direkte rettet mod hjernen 10-12. Selv om nogle succes er opnået, har afgørende resultater kun er opnået for lægemidler, der besidder endogene receptorer, såsom insulin 13,14. Desuden mekanismen for transnasal levering har været kontroversiel med tyder indirekte indrejse i hjernen via olfaktoriske neuron optagelse eller gennem blodbanen 11. Direct transkraniel levering ved hjælp af implanterbare katetre er opnået, men denne fremgangsmåde er meget invasiv og forbundet med mange komplikationer 15,16. Til dato er der ingen generel, minimalt invasiv metode til at levere forbindelser med høj molekylvægt i hjernen. Præsenteret heri er et murint kirurgisk procedureder skaber en semipermeabel grænseflade med hjernen. Dette opnås ved at transplantere en slimhindemembran eksplantatet 17 over en kirurgisk kraniotomi defekt i en mus. Ved hjælp af denne fremgangsmåde er det blevet vist, at opløselige forbindelser op til 500 kDa kan leveres ind i det centrale nervesystem (direkte ind i hjerneparenkymet samt i cerebrospinalvæske) i både tid og molekylvægt afhængig måde 18. Denne metode til at omgå BBB er en model for kraniet basen defekt reparationer hos mennesker, der bruger vaskulariserede slimhinde podninger til at reparere huller i kraniet efter transnasal endoskopisk kirurgi 19,20.

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			<td height="21" style="height:21px;width:247px;">Name of Material/ Equipment</td>
			<td style="width:220px;">Company</td>
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			<td style="width:263px;">Comments/Description</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:247px;">mice</td>
			<td style="width:220px;">Taconic</td>
			<td>C57BL/6</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Isoflurane</td>
			<td style="width:220px;">Piramal Healthcare</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Student Fine Scissors</td>
			<td style="width:220px;">Fine Science Tools</td>
			<td>91461-11</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:247px;">pneumatic drill</td>
			<td style="width:220px;">MTI Dental</td>
			<td style="width:119px;">333-CB</td>
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			<td height="21" style="height:21px;width:247px;">drill bit</td>
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			<td height="21" style="height:21px;width:247px;">forcepts</td>
			<td style="width:220px;">Fine Science Tools</td>
			<td>91106-12</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:247px;">0.9% Sodium Chloride Injection USP</td>
			<td style="width:220px;">Abbott Laboratories</td>
			<td>4925</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Polystyrene Detri Dish</td>
			<td style="width:220px;">Fisher</td>
			<td>08-757-12</td>
			<td>for temporarily storing graft</td>
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			<td height="21" style="height:21px;">Bead Sterilizer</td>
			<td style="width:220px;">Fine Science Tools</td>
			<td>18000-45</td>
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			<td height="21" style="height:21px;">Oxygen/Isoflurane System</td>
			<td style="width:220px;">SurgiVet</td>
			<td>V720100</td>
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			<td height="21" style="height:21px;">Temperature Control System</td>
			<td style="width:220px;">Physitemp</td>
			<td style="width:119px;">TCAT-2LV</td>
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			<td height="21" style="height:21px;">Small Animal Stereotaxic Instrument</td>
			<td style="width:220px;">KOPF</td>
			<td style="width:119px;">Model 940</td>
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			<td height="21" style="height:21px;">Eye Ointment</td>
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			<td height="21" style="height:21px;width:247px;">Cotton-Tipped Applicators</td>
			<td style="width:220px;">Fisher</td>
			<td>23-400-106</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">7.5% Provadone Iodine</td>
			<td style="width:220px;"></td>
			<td style="width:119px;"></td>
			<td>Betadine surgical scrub</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">70% Ethanol</td>
			<td style="width:220px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Surgical Blade Stainless</td>
			<td style="width:220px;">Feather</td>
			<td style="width:119px;">2976#10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Scalpel Handle - #3</td>
			<td style="width:220px;">Fine Science Tools</td>
			<td>10003-12</td>
			<td></td>
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			<td height="21" style="height:21px;">3% Hydogen Peroxide</td>
			<td style="width:220px;"></td>
			<td style="width:119px;"></td>
			<td>for cleaning the skull</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Vetbond Tissue Adhesive</td>
			<td style="width:220px;">3M</td>
			<td>1469SB</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">needles</td>
			<td>Becton, Dickinson and Company</td>
			<td>305176</td>
			<td>needle tip cut off and used as well</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:247px;">syringes</td>
			<td>Becton, Dickinson and Company</td>
			<td>309597</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:247px;">nitrile gloves</td>
			<td style="width:220px;">Denville Scientific Inc</td>
			<td>G4162</td>
			<td>for well closure and protection of graft</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">5-0 Nylon Suture Thread</td>
			<td style="width:220px;"></td>
			<td style="width:119px;"></td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Student Halsey Needle Holder</td>
			<td style="width:220px;">Fine Science Tools</td>
			<td>91201-13</td>
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			<td height="21" style="height:21px;width:247px;">cyanoacrylate adhesive</td>
			<td style="width:220px;"></td>
			<td style="width:119px;"></td>
			<td>commecially available super glue</td>
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		<tr height="21">
			<td height="21" style="height:21px;">Dental Cement Kit, 1 lb, Pink Opaque</td>
			<td style="width:220px;">Stoelting</td>
			<td>51458</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">isobutane (2-methylbutane)</td>
			<td style="width:220px;">Aldrich</td>
			<td>M32631</td>
			<td>for dry ice bath</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">dry ice</td>
			<td style="width:220px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
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heterotopic mucosal engrafting procedure for direct drug delivery to the brain in mice

Levering af terapeutiske midler ind i hjernen hæmmes af tilstedeværelsen af ​​blod-hjerne-barrieren (BBB), som begrænser passagen af ​​polære og forbindelser med høj molekylvægt fra blodbanen og ind i hjernevæv. Nogle direkte succes levering i mennesker er opnået via implantation af transkranielle katetre; men denne metode er meget invasiv og forbundet med mange komplikationer. En mindre invasiv alternativ ville være at dosere hjernen gennem et kirurgisk implanteret, semipermeabel membran, såsom den nasale mucosa, der anvendes til at reparere cranii manglerne efter endoskopisk transnasal tumor fjernelse kirurgi i mennesker. Drug overførsel selvom denne membran vil reelt omgå BBB og diffundere direkte ind i hjernen og cerebrospinalvæsken. Inspireret af denne fremgangsmåde blev en kirurgisk fremgangsmåde udviklede musene, der bruger en donor septal slimhinde indpodet i et ekstrakranialt kirurgisk BBB defekt. Denne model har vist sig effektivt attillade passage af forbindelser med høj molekylvægt i hjernen. Da mange lægemiddelkandidater er i stand til at krydse BBB, denne model er værdifuldt for at udføre præklinisk afprøvning af nye behandlingsformer for neurologiske og psykiatriske sygdomme.

Erhvervelse af en stor nok næseskillevæggen explant er afgørende for de efterfølgende trin. Dette kan opnås ved at bore ved placeringen på donormus kraniet er vist i figur 1a. Skæring ad denne vej vil producere en explant af tilstrækkelig størrelse, som vist i figur 1b. Hvis sådybden ikke er dyb nok, vil transplantatet være trunkerede og det vil være svært at opnå en tilstrækkelig stor membran til at dække hjernens overflade. Boring mere sideværts end den foreslåede st...

Watch this Scientific Journal Video about Heterotopic Mucosal Engrafting Procedure for Direct Drug Delivery to the Brain in Mice at JoVE.com

Heterotopic Mucosal Engrafting Procedure for Direct Drug Delivery to the Brain in Mice

Heterotop Mucosal transplantere Procedure for direkte Drug Delivery til Brain i mus

En musemodel af human endoskopisk cranii genopbygning er blevet udviklet, der skaber en semipermeabel grænseflade mellem hjernen og næsen ved hjælp af nasal slimhinde-podninger. Denne metode gør det muligt for forskere at undersøge levering til centralnervesystemet af højmolekylære terapeutika, der ellers er udelukket af blod-hjerne-barrieren, når de indgives systemisk.

Delivery of therapeutics into the brain is impeded by the presence of the blood-brain barrier (BBB) which restricts the passage of polar and high ...

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Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium

Intraluminale Drug Delivery til Mouse arteriovenøse Fistula Endothel

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The ...

Optimized LC-MS/MS Method for the High-throughput Analysis of Clinical Samples of Ivacaftor, Its Major Metabolites, and Lumacaftor in Biological Fluids of Cystic Fibrosis Patients

Optimeret LC-MS/MS metoden til høj overførselshastighed analyse af kliniske prøver af Ivacaftor, dets vigtigste metabolitter, og Lumacaftor i biologiske væsker af cystisk fibrose patienter

Defects in the cystic fibrosis trans-membrane conductance regulator (CFTR) are the cause of cystic fibrosis (CF), a disease with life-threatening ...

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

Forbedret metode til etablering af et In Vitro blod - hjerne barrieren Model baseret på svin hjernen Endothelial Cells

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) ...

Trans-Tympanic Drug Delivery for the Treatment of Ototoxicity

Trans-tympaniske Drug Delivery til behandling af ototoksicitet

The systemic administration of protective agents to treat drug-induced ototoxicity is limited by the possibility that these protective agents could ...

In Vivo Photolabeling of Cells in the Colon to Assess Migratory Potential of Hematopoietic Cells in Neonatal Mice

In Vivo Photolabeling of Cells in the Colon to Assess Migratory Potential of Hematopoietic Cells in Neonatal Mice

In Vivo Photolabeling af celler i tyktarmen til at vurdere vandrende potentiale af hæmatopoietisk celler i Neonatal mus

Enteric bacterial communities are established early in life and influence immune cell development and function. The neonatal microbiota is susceptible to ...

Novel Methods for Intranasal Administration Under Inhalation Anesthesia to Evaluate Nose-to-Brain Drug Delivery

Nye metoder for Intranasal Administration Under Inhalation anæstesi til at evaluere næse-til-hjerne Drug Delivery

Intranasal administration has been reported to be a potential pathway for nose-to-brain delivery of therapeutic agents that circumvents the blood-brain ...