Name: generation of alginate microspheres for biomedical applications
Uploaded: 2012-08-12T14:00:00
Description: Watch this Scientific Journal Video about Generation of Alginate Microspheres for Biomedical Applications at JoVE.com

Denne undersøgelse blev støttet af US Department of Veterans Affairs (Washington DC), tilskud 0852048 og 0731201, og 0854430 fra National Science Foundation (Arlington, VA), og tilskud RO1 DK080897 fra National Institutes of Health (Bethesda, MD) . Mr. Khanna modtaget støtte fra en generøs donation af Mr. Edward Ross og Dr. Monica Moya fra Bill &amp; Melinda Gates Foundation (Seattle, WA).

Protokollen her beskriver en tre-trins procedure til generering af flerlags alginat mikroperler (figur 1). Først bliver alginat mikroperler dannet (figur 2A). Denne procedure er beskrevet i afsnit 1 nedenfor. Celler eller proteiner kan tilsættes til mikroperlerne i dette trin, for at virke som et afgivelsessystem. Det næste trin involverer dannelsen af ​​en permselektiv lag på mikroperlerne og er beskrevet i afsnit 2. Det sidste trin indebærer dannelsen af ​​en ekstra algin...

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	<li>Darrabie, M. D., Kendall, W. F., Opara, E. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characteristics+of+Poly-L-+Ornithine-coated+alginate+microcapsules.">Characteristics of Poly-L- Ornithine-coated alginate microcapsules.</a> Biomaterials. 26, 6846-6852 (2005).</li><li>Yamagiwa, K., Kozawa, T., Ohkawa, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+alginate+composition+and+gelling+conditions+on+diffusional+and+mechanical+properties+of+calcium-alginate+gel+beads.">Effects of alginate composition and gelling conditions on diffusional and mechanical properties of calcium-alginate gel beads.</a> J. Chem. Eng. Jpn. 28, 462-467 (1995).</li><li>Amsden, B., Turner, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diffusion+characteristics+of+calcium+alginate+gels.">Diffusion characteristics of calcium alginate gels.</a> Biotechnol. Bioeng. 65, 605-610 (1999).</li><li>Uriel, S., Brey, E. M., Greisler, H. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sustained+low+levels+of+fibroblast+growth+factor-1+promote+persistent+microvascular+network+formation.">Sustained low levels of fibroblast growth factor-1 promote persistent microvascular network formation.</a> Am. J. Surg. 192, 604-609 (2006).</li><li>Moya, M. L., Lucas, S., Francis-Sedlak, M., Liu, X., Garfinkel, M. R., Huang, J. J., Cheng, M. H., Opara, E. C., Brey, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sustained+delivery+of+FGF-1+increases+vascular+density+in+comparison+to+bolus+administration.">Sustained delivery of FGF-1 increases vascular density in comparison to bolus administration.</a> Microvasc. Res. 78, 142-147 (2009).</li><li>Moya, M. L., Cheng, M. H., Huang, J. J., Francis-Sedlak, M. E., Kao, S. W., Opara, E. C., Brey, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+FGF-1+loaded+alginate+microbeads+on+neovascularization+and+adipogenesis+in+a+vascular+pedicle+model+of+adipose+tissue+engineering.">The effect of FGF-1 loaded alginate microbeads on neovascularization and adipogenesis in a vascular pedicle model of adipose tissue engineering.</a> Biomaterials. 31, 2816-2826 (2010).</li><li>Khanna, O., Moya, M. L., Greisler, H. P., Opara, E. C., Brey, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multilayered+Microcapsules+for+the+Sustained-Release+of+Angiogenic+Proteins+From+Encapsulated+Cells.">Multilayered Microcapsules for the Sustained-Release of Angiogenic Proteins From Encapsulated Cells.</a> Am. J. Surg. 200, 655-658 (2010).</li><li>Khanna, O., Moya, M. L., Opara, E. C., Brey, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+of+multilayered+alginate+microcapsules+for+the+sustained-release+of+fibroblast+growth+factor-1.">Synthesis of multilayered alginate microcapsules for the sustained-release of fibroblast growth factor-1.</a> J. Biomed. Mater. Res. A. 95, 632-640 (2010).</li></ol>

Alginat er en naturlig, surt polysaccharid udvundet af alger og består af enheder med 1,4 &#39;-β-D-mannuronsyre (M) og α-L-guluronsyre (G) 2,3. Simple gelering finder sted, når divalente kationer, såsom Ca2 +, Sr2 + eller Ba2 + interagerer med G-monomererne, der danner ioniske broer mellem hosliggende alginat kæder. Mikroperler af alginat har været anvendt til at levere en række proteiner, herunder fibroblast vækstfaktor-1 (FGF-1), nervevækstfaktor, leukæmi inhibere faktor, v...

<table border="1"><tbody><tr><td> Navn på reagenset </td><td> Firma </td><td> Katalog nummer </td><td> Kommentarer (valgfri) </td></tr><tr><td> Pronova ultrarent LVG alginat </td><td> NOVA-matrix </td><td> 4200006 </td><td> En række af alginat formuleringer er til rådighed. Valget af alginat påvirker de endelige egenskaber af mikroperler, herunder størrelse, mekaniske egenskaber og transport. Den anvendte sammensætning skal optimeres for en given anvendelse. </td></tr><tr><td> Pronova ultrarent LVM alginat </td><td> NOVA-matrix </td><td> 4200206 </td><td> En række af alginat formuleringer er til rådighed. Valget af alginat påvirker de endelige egenskaber af mikroperler, herunder størrelse, mekaniske egenskaber og transport. Den anvendte sammensætning skal optimeres for en given anvendelse. </td></tr><tr><td> Poly-L-ornithin-hydrochlorid </td><td> Sigma-Aldrich </td><td> P2533 </td><td></td></tr></tbody></table>

generation of alginate microspheres for biomedical applications

Alginat-baserede materialer har fået betydelig opmærksomhed til biomedicinske anvendelser på grund af deres hydrofile natur, biokompatibilitet og fysiske struktur. Anvendelser omfatter celleindkapsling, drug delivery, stamcelle kultur, og vævs-engineering scaffolds. Faktisk er kliniske forsøg øjeblikket udføres der øer er indkapslet i PLO overtrukne alginat mikroperler som en behandling af type I diabetes. Imidlertid stort antal øer kræves for effektivitet på grund af dårlige overlevelse efter transplantation. Evnen til lokalt at stimulere mikrovaskulære netværksdannelse omkring de indkapslede celler kan øge deres levedygtighed gennem forbedret transport af ilt, glucose og andre vitale næringsstoffer. Fibroblast vækstfaktor-1 (FGF-1) er en naturligt forekommende vækstfaktor, der er i stand til at stimulere blodkardannelse og forbedre iltindholdet i iskæmiske væv. Virkningen af ​​FGF-1 forøges, når den afgives med en Sustained måde snarere end en enkelt stor-bolusindgivelse. Den lokale langvarig frigivelse af vækstfaktorer fra ø indkapslingssystemer kan stimulere væksten af ​​blodkar direkte mod de transplanterede celler, som potentielt forbedrer funktionelle podede resultater. I denne artikel skitsere vi procedurer for udarbejdelse af alginat mikrosfærer til brug i biomedicinske anvendelser. Herudover beskriver vi en fremgangsmåde vi udviklet til generering af flerlags alginat mikroperler. Celler kan indkapsles i det indre alginat kerne og angiogene proteiner i den ydre alginat lag. Frigivelse af proteiner fra dette ydre lag vil stimulere dannelsen af ​​lokale mikrovaskulære net direkte mod de transplanterede øer.

Watch this Scientific Journal Video about Generation of Alginate Microspheres for Biomedical Applications at JoVE.com

Generation of Alginate Microspheres for Biomedical Applications

I de følgende afsnit, skitsere vi procedurer for udarbejdelse af alginat mikrosfærer til brug i biomedicinske anvendelser. Vi specifikt illustrerer en teknik til at skabe flerlaget alginat mikrosfærer til det dobbelte formål at celle-og protein indkapsling som en potentiel behandling af type 1 diabetes.

Generering af alginat mikrosfærer til biomedicinske anvendelser

Alginate-based materials have received considerable attention for biomedical applications because of their hydrophilic nature, biocompatibility, and ...

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