Name: determination of the transport rate of xenobiotics and nanomaterials across the placenta using the ex vivo human placental perfusion model
Uploaded: 2013-06-18T13:00:00
Description: Watch this Scientific Journal Video about Determination of the Transport Rate of Xenobiotics and Nanomaterials Across the Placenta using the ex vivo Human Placental Perfusion Model at JoVE.com

Dette arbejde er støttet af den schweiziske nationalbank Foundation, (NRP 64 programmet ikke give 4064 til 131.232).

1.. Forberedelse perfusionssystemet <ol><li> Opsæt perfusion system bestående af et vandbad, en perfusionskammer, to kolonner for iltning, to peristaltiske pumper, to boble fælder, to flow varmeapparater og en tryksensor (Figur 1). Forbinde disse komponenter med rørsektioner sammensat af silikone og polyvinylklorid materialer ifølge skemaet i figur 2. Endelig er der to kredsløb, der repræsenterer den føtale og maternelle kredsløb, hhv. </li><li> Tænd vandbad, strømmen varme...

<ol>
	<li>Ala-Kokko, T. I., Myllynen, P., Vahakangas, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ex+vivo+perfusion+of+the+human+placental+cotyledon:+implications+for+anesthetic+pharmacology.">Ex vivo perfusion of the human placental cotyledon: implications for anesthetic pharmacology.</a> Int. J. Obstet. Anesth. 9, 26-38 (2000).</li><li>Panigel, M., Pascaud, M., Brun, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radioangiographic+study+of+circulation+in+the+villi+and+intervillous+space+of+isolated+human+placental+cotyledon+kept+viable+by+perfusion.">Radioangiographic study of circulation in the villi and intervillous space of isolated human placental cotyledon kept viable by perfusion.</a> J. Physiol. (Paris). 59, 277 (1967).</li><li>Schneider, H., Panigel, M., Dancis, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transfer+across+the+perfused+human+placenta+of+antipyrine,+sodium+and+leucine.">Transfer across the perfused human placenta of antipyrine, sodium and leucine.</a> Am. J. Obstet. Gynecol. 114, 822-828 (1972).</li><li>Enders, A. C., Blankenship, T. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+placental+structure.">Comparative placental structure.</a> Adv. Drug Deliv. Rev. 38, 3-15 (1999).</li><li>Takata, K., Hirano, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanism+of+glucose+transport+across+the+human+and+rat+placental+barrier:+a+review.">Mechanism of glucose transport across the human and rat placental barrier: a review.</a> Microsc. Res. Tech. 38, 145-152 (1997).</li><li>Saunders, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transplacental+transport+of+nanomaterials.">Transplacental transport of nanomaterials.</a> Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol. 1, 671-684 (2009).</li><li>Buerki-Thurnherr, T., von Mandach, U., Wick, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Knocking+at+the+door+of+the+unborn+child:+engineered+nanoparticles+at+the+human+placental+barrier.">Knocking at the door of the unborn child: engineered nanoparticles at the human placental barrier.</a> Swiss Med. Wkly. 142, w13559 (2012).</li><li>Gendron, M. P., Martin, B., Oraichi, D., Berard, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Health+care+providers'+requests+to+Teratogen+Information+Services+on+medication+use+during+pregnancy+and+lactation.">Health care providers' requests to Teratogen Information Services on medication use during pregnancy and lactation.</a> Eur. J. Clin. Pharmacol. 65, 523-531 (2009).</li><li>Burns, L., Mattick, R. P., Lim, K., Wallace, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methadone+in+pregnancy:+treatment+retention+and+neonatal+outcomes.">Methadone in pregnancy: treatment retention and neonatal outcomes.</a> Addiction. 102, 264-270 (2007).</li><li>von Mandach, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug+use+in+pregnancy.">Drug use in pregnancy.</a> Ther. Umsch. 62, 29-35 (2005).</li><li>Malek, A., Obrist, C., Wenzinger, S., von Mandach, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+cocaine+and+heroin+on+the+placental+transfer+of+methadone.">The impact of cocaine and heroin on the placental transfer of methadone.</a> Reprod. Biol. Endocrinol. 7, 61 (2009).</li><li>Hutson, J. R., Garcia-Bournissen, F., Davis, A., Koren, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+human+placental+perfusion+model:+a+systematic+review+and+development+of+a+model+to+predict+in+vivo+transfer+of+therapeutic+drugs.">The human placental perfusion model: a systematic review and development of a model to predict in vivo transfer of therapeutic drugs.</a> Clin. Pharmacol. Ther. 90, 67-76 (2011).</li><li>International Organization for Standardization (ISO). <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technical+Specification+(ISO/TS)+27687.">Technical Specification (ISO/TS) 27687.</a> Nanotechnologies â€“ Terminology and definitions for nano-objects â€“ Nanoparticles, nanofibre and nanoplate. , (2008).</li><li>Pietroiusti, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Health+implications+of+engineered+nanomaterials.">Health implications of engineered nanomaterials.</a> Nanoscale. 4, 1231-1247 (2012).</li><li>Latzin, P., Roosli, M., Huss, A., Kuehni, C. E., Frey, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Air+pollution+during+pregnancy+and+lung+function+in+newborns:+a+birth+cohort+study.">Air pollution during pregnancy and lung function in newborns: a birth cohort study.</a> Eur. Respir. J. 33, 594-603 (2009).</li><li>Lacasana, M., Esplugues, A., Ballester, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exposure+to+ambient+air+pollution+and+prenatal+and+early+childhood+health+effects.">Exposure to ambient air pollution and prenatal and early childhood health effects.</a> Eur. J. Epidemiol. 20, 183-199 (2005).</li><li>Menezes, V., Malek, A., Keelan, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanoparticulate+drug+delivery+in+pregnancy:+placental+passage+and+fetal+exposure.">Nanoparticulate drug delivery in pregnancy: placental passage and fetal exposure.</a> Curr. Pharm. Biotechnol. 12, 731-742 (2011).</li><li>Muhlemann, K., Menegus, M. A., Miller, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytomegalovirus+in+the+perfused+human+term+placenta+in+vitro.">Cytomegalovirus in the perfused human term placenta in vitro.</a> Placenta. 16, 367-373 (1995).</li><li>Wick, P., 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=Barrier+capacity+of+human+placenta+for+nanosized+materials.">Barrier capacity of human placenta for nanosized materials.</a> Environ. Health Perspect. 118, 432-436 (2010).</li><li>Dancis, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Why+perfuse+the+human+placenta.">Why perfuse the human placenta.</a> Contrib Gynecol. Obstet. 13, 1-4 (1985).</li><li>May, K., 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=Perfusion+of+human+placenta+with+hemoglobin+introduces+preeclampsia-like+injuries+that+are+prevented+by+alpha1-microglobulin.">Perfusion of human placenta with hemoglobin introduces preeclampsia-like injuries that are prevented by alpha1-microglobulin.</a> Placenta. 32, 323-332 (2011).</li><li>Guller, 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=Protein+composition+of+microparticles+shed+from+human+placenta+during+placental+perfusion:+Potential+role+in+angiogenesis+and+fibrinolysis+in+preeclampsia.">Protein composition of microparticles shed from human placenta during placental perfusion: Potential role in angiogenesis and fibrinolysis in preeclampsia.</a> Placenta. 32, 63-69 (2011).</li><li>Challier, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Criteria+for+evaluating+perfusion+experiments+and+presentation+of+results.">Criteria for evaluating perfusion experiments and presentation of results.</a> Contrib. Gynecol. Obstet. 13, 32-39 (1985).</li><li>Kraemer, J., Klein, J., Lubetsky, A., Koren, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perfusion+studies+of+glyburide+transfer+across+the+human+placenta:+implications+for+fetal+safety.">Perfusion studies of glyburide transfer across the human placenta: implications for fetal safety.</a> Am. J. Obstet. Gynecol. 195, 270-274 (2006).</li><li>leal, J. K., 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=Modification+of+fetal+plasma+amino+acid+composition+by+placental+amino+acid+exchangers+in+vitro.">Modification of fetal plasma amino acid composition by placental amino acid exchangers in vitro.</a> J. Physiol. 582, 871-882 (2007).</li><li>athiesen, L., 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=Quality+assessment+of+a+placental+perfusion+protocol.">Quality assessment of a placental perfusion protocol.</a> Reprod. Toxicol. 30, 138-146 (2010).</li><li>Myllynen, P., 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=Preliminary+interlaboratory+comparison+of+the+ex+vivo+dual+human+placental+perfusion+system.">Preliminary interlaboratory comparison of the ex vivo dual human placental perfusion system.</a> Reprod Toxicol. 30, 94-102 (2010).</li><li>Malek, A., Sager, R., Schneider, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maternal-fetal+transport+of+immunoglobulin+G+and+its+subclasses+during+the+third+trimester+of+human+pregnancy.">Maternal-fetal transport of immunoglobulin G and its subclasses during the third trimester of human pregnancy.</a> Am. J. Reprod. Immunol. 32, 8-14 (1994).</li><li>Prouillac, C., Lecoeur, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+the+placenta+in+fetal+exposure+to+xenobiotics:+importance+of+membrane+transporters+and+human+models+for+transfer+studies.">The role of the placenta in fetal exposure to xenobiotics: importance of membrane transporters and human models for transfer studies.</a> Drug Metab. Dispos. 38, 1623-1635 (2010).</li><li>Poulsen, M. S., Rytting, E., Mose, T., Knudsen, L. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+placental+transport:+correlation+of+in+vitro+BeWo+cell+permeability+and+ex+vivo+human+placental+perfusion.">Modeling placental transport: correlation of in vitro BeWo cell permeability and ex vivo human placental perfusion.</a> Toxicol. In Vitro. 23, 1380-1386 (2009).</li><li>Mathiesen, L., Rytting, E., Mose, T., Knudsen, L. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transport+of+benzo[alpha]pyrene+in+the+dually+perfused+human+placenta+perfusion+model:+effect+of+albumin+in+the+perfusion+medium.+Basic+Clin.">Transport of benzo[alpha]pyrene in the dually perfused human placenta perfusion model: effect of albumin in the perfusion medium. Basic Clin.</a> Pharmacol. Toxicol. 105, 181-187 (2009).</li></ol>

Under den dobbelte recirkulerende perfusion viste her, er der flere andre eksperimentelle konfigurationer muligt, afhængigt af det spørgsmål, der skal besvares. Særligt åbne placenta perfusioner er almindeligt anvendt til at vurdere stoffet clearance ved steady state koncentration 3.. Den recirkulerende perfusion set-up kan også anvendes til at bekræfte aktiv transport af endogene eller eksogene stoffer. For denne fremgangsmåde den samme koncentration af miljøfremmede skal tilføjes til den maternell...

Moderkagen er et komplekst organ, der er ansvarlig for udveksling af oxygen, kuldioxid, næringsstoffer og affaldsprodukter og på samme tid i stand til at holde de to blodkredsløb hos moderen og den voksende fosteret adskilt fra hinanden. Derudover forhindrer afvisning af barnet den maternelle immunsystemet og udskiller hormoner til at opretholde graviditet. Den cellulære barriere er dannet af cytotrophoblastceller som fusionerer og danner en sand syncytium uden laterale cellemembraner 4,5. Hele placenta er opdelt i flere kimblade, som indeholder en føtal villøs træet og repræsenterer en funktionel enhed af moderkagen. Studiet af placentabarrieren funktion blev intensiveret med opdagelsen af ​​de thalidomid inducerede misdannelser i 1960&#39;erne. Af indlysende grunde translokation studier med gravide kvinder kan ikke udføres. Derfor har forskellige alternative modeller blevet udviklet 6,7 </sup&gt;. Den mest lovende og nok mest klinisk relevant model er den ex vivo humane placentaperfusion udviklet af Panigel og medarbejdere 2,3. Mange kvinder udsættes for forskellige miljøfremmede stoffer såsom narkotika eller miljøforurening under deres graviditet 8.. For nogle lægemidler, som allerede blev administreret regelmæssigt under graviditeten, kan in vivo-undersøgelser udføres ved sammenligning af den maternale blodkoncentration med det i navlestrengsblod. Men generelt er der kun begrænsede oplysninger om farmakokinetik og-dynamik i fosteret og teratogenicitet af disse stoffer. For eksempel opiater som heroin let krydser placentabarrieren og kan føre til intrauterin vækst begrænsning for tidlig fødsel eller spontan abort 9,10. Så i tilfælde af manglende afholdenhed under graviditeten en substitutionsterapi med metadon anbefales. Exvivo human placentaperfusion model viste, at overførslen af metadon i den føtale cirkulation er ubetydelig 11, som korrelerer godt med den beregnede navlestrengsblod-til-maternale blodkoncentration forholdet efter levering 12.. Nanoteknologi er et voksende område især i medicin. Så under naturligt forekommende fine (&lt;2,5 um i diameter) og ultrafine partikler (&lt;0,1 um i diameter) i dampe af skovbrande, vulkanudbrud og ørken støv, udsættelse for manipuleret nanomaterialer (mindst én dimension &lt;0,1 um 13 ) er stigende. Dette rejste spørgsmål om toksikologiske potentiale af industrielt fremstillede nanomaterialer. Selvom ingen menneskelig risiko kunne bevises endnu, er der væsentlige eksperimentelle studier tyder på, at nanopartikler kan forårsage uønskede biologiske reaktioner, der fører til toksikologiske resultater 14. For nylig nogle undersøgelser, at prænatal udsættelse forluftforureningen er knyttet til en højere respiratorisk behov og betændelse i luftvejene hos nyfødte og børn 15,16. Derudover kan små nanopartikler kan anvendes som bærere af lægemidler til specifikt behandle fosteret eller moderen. Derfor bliver det tydeligt, at omfattende undersøgelser af forskellige miljøfremmede stoffer eller nanomaterialer og deres evne til at krydse placentabarrieren er påkrævet. En egentlig overblik over de igangværende undersøgelser om placenta gennemtrængelighed til industrielt fremstillede nanomaterialer er sammenfattet i Menezes et al. 2011 17 og Buerki-Thurnherr et al. 2012 7.. Den ex vivo dual recirkulerende human placentaperfusion model giver en kontrolleret og pålideligt system til at studere den placentale transport af forskellige endogene og eksogene forbindelser 3,11,12,18,19 og en lang række andre funktioner i moderkagen som mekanismer, der er ansvarlige for udvikling af patologiske tilstande som præeklampsi &lt;sup&gt; 20-22. I denne protokol fokuserer vi primært på opsætning, håndtering og metode, der tillader studiet af ophobning, effekter og translokation satser for en bred vifte af miljøfremmede stoffer eller nanopartikler.

<table border="1">
	<tbody>
 <tr>
 <td>Name of the Reagent</td>
 <td>Company</td>
 <td>Catalogue Number</td>
 <td>Comments (optional)</td>
 </tr>
 <tr>
 <td>NCTC-135 medium</td>
 <td>ICN Biomedicals, Inc.</td>
 <td>10-911-22C</td>
 <td>could be replaced by Medium 199 from Sigma (M3769)</td>
 </tr>
 <tr>
 <td>Sodium chloride (NaCl)</td>
 <td>Sigma-Aldrich, Fluka</td>
 <td>71381</td>
 <td></td>
 </tr>
 <tr>
 <td>Potassium chloride (KCl)</td>
 <td>Hospital pharmacy</td>
 <td></td>
 <td>also possible: Sigma (P9541)</td>
 </tr>
 <tr>
 <td>Monosodium phosphate (NaH2PO4 &middot; H2O)</td>
 <td>Merck</td>
 <td>106346</td>
 <td></td>
 </tr>
 <tr>
 <td>Magnesium sulfate (MgSO4 &middot; H2O)</td>
 <td>Sigma-Aldrich, Fluka</td>
 <td>63139</td>
 <td></td>
 </tr>
 <tr>
 <td>Calcium chloride (CaCl, anhydrous)</td>
 <td>Merck</td>
 <td>102388</td>
 <td></td>
 </tr>
 <tr>
 <td>D(+) Glucose (anhydrous)</td>
 <td>Sigma-Aldrich, Fluka</td>
 <td>49138</td>
 <td></td>
 </tr>
 <tr>
 <td>Sodium bicarbonate (NaHCO3)</td>
 <td>Merck</td>
 <td>106329</td>
 <td></td>
 </tr>
 <tr>
 <td>Dextran from Leuconostoc spp.</td>
 <td>Sigma-Aldrich</td>
 <td>31389</td>
 <td></td>
 </tr>
 <tr>
 <td>Bovine serum albumin (BSA)</td>
 <td>Applichem</td>
 <td>A1391</td>
 <td></td>
 </tr>
 <tr>
 <td>Amoxicilline (Clamoxyl)</td>
 <td>GlaxoSmithKline AG</td>
 <td>2021101A</td>
 <td></td>
 </tr>
 <tr>
 <td>Sodium heparin</td>
 <td>B. Braun Medical AG</td>
 <td>3511014</td>
 <td></td>
 </tr>
 <tr>
 <td>Sodium hydoxide (NaOH) pellets</td>
 <td>Merck</td>
 <td>106498</td>
 <td>CAUTION: corrosive</td>
 </tr>
 <tr>
 <td>Ortho-phosphoric acid 85% (H3PO4)</td>
 <td>Merck</td>
 <td>100573</td>
 <td>CAUTION: corrosive</td>
 </tr>
 <tr>
 <td>Maternal gas mixture: 95% synthetic air, 5% CO2</td>
 <td>PanGas AG</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Fetal gas mixture: 95% N2, 5% CO2</td>
 <td>PanGas AG</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Antipyrine (N-methyl-14C)</td>
 <td>American Radiolabeled Chemicals, Inc.</td>
 <td>ARC 0108-50 &mu;Ci</td>
 <td>CAUTION: radioactive material (specific activity: 55mCi/mmol)</td>
 </tr>
 <tr>
 <td>Scintillation cocktail (IrgaSafe Plus)</td>
 <td>Zinsser Analytic GmbH</td>
 <td>1003100</td>
 <td></td>
 </tr>
 <tr>
 <td>Polystyrene particles 80 nm</td>
 <td>Polyscience, Inc.</td>
 <td>17150</td>
 <td></td>
 </tr>
 <tr>
 <td>Polystyrene particles 500 nm</td>
 <td>Polyscience, Inc.</td>
 <td>17152</td>
 <td></td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>EQUIPMENT</td>
 </tr>
 <tr>
 <td>Water bath</td>
 <td>VWR</td>
 <td>462-7001</td>
 <td></td>
 </tr>
 <tr>
 <td>Thermostat</td>
 <td>IKA-Werke GmbH &amp; Co. KG</td>
 <td>3164000</td>
 <td></td>
 </tr>
 <tr>
 <td>Peristaltic pumps</td>
 <td>Ismatec</td>
 <td>ISM 833</td>
 <td></td>
 </tr>
 <tr>
 <td>Bubble traps (glass)</td>
 <td>UNI-GLAS Laborbedarf</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Flow heater</td>
 <td>UNI-GLAS Laborbedarf</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Pressure sensor + Software for analyses</td>
 <td>MSR Electronics GmbH</td>
 <td>145B5</td>
 <td></td>
 </tr>
 <tr>
 <td>Notebook</td>
 <td>Hewlett Packard</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Miniature gas exchange oxygenator</td>
 <td>Living Systems Instrumentation</td>
 <td>LSI-OXR</td>
 <td></td>
 </tr>
 <tr>
 <td>Tygon Tube (ID: 1.6 mm; OD: 4.8 mm)</td>
 <td>Ismatec</td>
 <td>MF0028</td>
 <td></td>
 </tr>
 <tr>
 <td>Tubes for pumps (PharMed BPT; ID: 1.52 mm)</td>
 <td>Ismatec</td>
 <td>SC0744</td>
 <td></td>
 </tr>
 <tr>
 <td>Blunt cannulae (&Oslash; 0.8 mm)</td>
 <td>Polymed Medical Center</td>
 <td>03.592.81</td>
 <td></td>
 </tr>
 <tr>
 <td>Blunt cannulae (&Oslash; 1.2 mm)</td>
 <td>Polymed Medical Center</td>
 <td>03.592.90</td>
 <td></td>
 </tr>
 <tr>
 <td>Blunt cannulae (&Oslash; 1.5 mm)</td>
 <td>Polymed Medical Center</td>
 <td>03.592.94</td>
 <td></td>
 </tr>
 <tr>
 <td>Blunt cannulae (&Oslash; 1.8 mm)</td>
 <td>Polymed Medical Center</td>
 <td>03.952.82</td>
 <td></td>
 </tr>
 <tr>
 <td>Parafilm</td>
 <td>VWR</td>
 <td>291-1212</td>
 <td></td>
 </tr>
 <tr>
 <td>Perfusion chamber with tissue holder (plexiglass)</td>
 <td>Internal technical department</td>
 <td></td>
 <td>Similar equipment is available from Hemotek Limited, UK</td>
 </tr>
 <tr>
 <td>Surgical suture material (PremiCron)</td>
 <td>B. Braun Medical AG</td>
 <td>C0026005</td>
 <td></td>
 </tr>
 <tr>
 <td>Winged Needle Infusion Set (21G Butterfly)</td>
 <td>Hospira, Inc.</td>
 <td>ASN 2102</td>
 <td></td>
 </tr>
 <tr>
 <td>Multidirectional stopcock (Discofix C-3)</td>
 <td>B. Braun Medical AG</td>
 <td>16494C</td>
 <td></td>
 </tr>
 <tr>
 <td>Surgical scissors</td>
 <td>B. Braun Medical AG</td>
 <td>BC304R</td>
 <td></td>
 </tr>
 <tr>
 <td>Dissecting scissors</td>
 <td>B. Braun Medical AG</td>
 <td>BC162R</td>
 <td></td>
 </tr>
 <tr>
 <td>Needle holder</td>
 <td>B. Braun Medical AG</td>
 <td>BM200R</td>
 <td></td>
 </tr>
 <tr>
 <td>Dissecting forceps</td>
 <td>B. Braun Medical AG </td>
 <td>BD215R</td>
 <td></td>
 </tr>
 <tr>
 <td>Automated blood gas system </td>
 <td>Radiometer Medical ApS</td>
 <td>ABL800 FLEX</td>
 <td></td>
 </tr>
 <tr>
 <td>Multi-mode microplate reader</td>
 <td>BioTek</td>
 <td>Synergy HT</td>
 <td></td>
 </tr>
 <tr>
 <td>Liquid scintillation analyzer</td>
 <td>GMI, Inc.</td>
 <td>Packard Tri-Carb 2200</td>
 <td></td>
 </tr>
 <tr>
 <td>Scintillation tubes 5.5 ml</td>
 <td>Zinsser Analytic GmbH</td>
 <td>3020001</td>
 <td></td>
 </tr>
 <tr>
 <td>Tissue Homogenizer</td>
 <td>OMNI, Inc.</td>
 <td>TH-220</td>
 <td></td>
 </tr>
 <tr>
 <td>pH meter + electrode</td>
 <td>VWR</td>
 <td>662-2779 </td>
 <td></td>
 </tr>
	 </tbody>
 </table>

determination of the transport rate of xenobiotics and nanomaterials across the placenta using the ex vivo human placental perfusion model

Årtier siden den humane placenta blev anset for at være en uigennemtrængelig barriere mellem mor og ufødte barn. Men opdagelsen af ​​thalidomid-inducerede fosterskader og mange senere studier bagefter viste det modsatte. I dag flere skadelige fremmedstoffer som nikotin, blev heroin, metadon eller narkotika samt miljøforurening beskrevet at overvinde denne barriere. Med den stigende brug af nanoteknologi, er moderkagen forventes at komme i kontakt med nye nanopartikler enten ved et uheld ved eksponering eller forsætligt i tilfælde af potentielle nanomedical applikationer. Data fra dyreforsøg ikke kan ekstrapoleres til mennesker, fordi moderkagen er den mest artsspecifikke pattedyr organ 1.. Derfor ex vivo dual recirkulerende human placentaperfusion, udviklet af Panigel et al. I 1967 2 og løbende modificeret af Schneider et al. I 1972 3, tjener kan som en fremragende model to studere overførsel af xenobiotika eller partikler. Her fokuserer vi på ex vivo dual recirkulerende human placentaperfusion protokol og dens videre udvikling at erhverve reproducerbare resultater. Den efterbyrd blev opnået efter informeret samtykke af mødrene fra ukomplicerede sigt graviditeter gennemgår kejsersnit. De fosteret og moderen fartøjer med en intakt kimbladene blev kanyleret og perfunderet i mindst fem timer. Som model partikel fluorescensmærkede polystyrenpartikler med størrelser på 80 og 500 nm i diameter sattes til moderens kredsløb. De 80 nm partikler var i stand til at passere placentabarrieren og giver et perfekt eksempel for et stof, der overføres over placenta til fostret, mens de 500 nm partikler blev bibeholdt i placenta væv eller moderens kredsløb. Den ex vivo humane placentaperfusion model er en af de få modeller, der giver pålidelige oplysninger omtransport adfærd miljøfremmede stoffer på et vigtigt væv barriere, som leverer forudsigende og kliniske relevante data.

Figur 4A viser perfusion profiler af små polystyrenpartikler (80 nm), som blev transporteret over placenta sammenlignet med større polystyrenpartikler (500 nm), som ikke blev overført til fosterets rum. Hvert datapunkt repræsenterer middelværdien partikelkoncentrationen til det givne tidspunkt på mindst 3 uafhængige forsøg. For polystyren nanopartikler placentaoverførsel er størrelse-afhængig 19. Efter 3 timer moderkage perfusion allerede 20-30% af de oprindeligt tilsatte 80 nm pol...

Watch this Scientific Journal Video about Determination of the Transport Rate of Xenobiotics and Nanomaterials Across the Placenta using the ex vivo Human Placental Perfusion Model at JoVE.com

Determination of the Transport Rate of Xenobiotics and Nanomaterials Across the Placenta using the ex vivo Human Placental Perfusion Model

Den<em&gt; Ex vivo</em&gt; Dobbelt recirkulerende human placental perfusion model kan bruges til at undersøge overførslen af ​​xenobiotika og nanopartikler over den humane placenta. I denne video protokol beskriver vi udstyr og teknikker, der kræves for en vellykket gennemførelse af en moderkage perfusion.

Bestemmelse af transport Rate af Xenobiotics og nanomaterialer over placenta ved hjælp af<em&gt; Ex vivo</em&gt; Menneskelige placentaperfusion Model

Decades ago the human placenta was thought to be an impenetrable barrier between mother and unborn child. However, the discovery of thalidomide-induced ...

Research

JoVE Journal

Bioengineering

Determination of the Transport Rate of Xenobiotics and Nanomaterials Across the Placenta using the ex vivo Human Placental Perfusion Model

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The foreign body reaction occurs when a synthetic surface is introduced to the body. It is characterized by adsorption of blood proteins and the ...

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En ny Ex Vivo Model for evaluering af endoskopisk submukøse injektion materielle ydeevne

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