Name: mechanical stimulation of chondrocyte-agarose hydrogels
Uploaded: 2012-10-27T19:00:00
Description: Watch this Scientific Journal Video about Mechanical Stimulation of Chondrocyte-agarose Hydrogels at JoVE.com

1. Isolering av Primary Artikulære chondrocytes Høste 10-15 full tykkelse brusk skiver fra artikulære flater av dyr leddene (f.eks metacarpal-phalangeal felles av skjelettet kyr hentet fra en lokal abbatoir). <ol><li> Place brusk skiver i en 100 mm petriskål og Inkuber i 20 ml 0,5% protease i Hams F-12 (w / v) i 2 timer ved 37 ° C. Skyll tre ganger i Ham F-12 dyrkningsmedier og inkuber med 20 ml 0,15% kollagenase A i Ham F-12 dyrkningsmedier natten ved 37 ° C. </li><li> Filt...

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<html> Den beskrevne fremgangsmåte for å anvende kontrollerte mekaniske stimuli til celle foret agarose hydrogeler tillater direkte undersøkelse på effekten av dynamiske trykkrefter på kondrocytt metabolisme. Bruken av den tilpassede-testing rigg i forbindelse med låseringene tilgjengelig sideveis begrensning for constructs å unngå potensielle problemer for utvalg tipping. Bruken av dødvekt-lasting platens sikret ved sett mannskaper sikrer direkte kontakt med konstruksjoner tross potensielle forskjeller i utvalg...

<table border="1"><tbody><tr><td> Navn av reagenset eller utstyr </td><td> Selskapet </td><td> Katalognummer </td><td> Kommentarer (valgfritt) </td></tr><tr><td> Hams F-12 </td><td> Thermo Fisher Scientific </td><td> SH3001002 </td><td></td></tr><tr><td> Kollagenase A </td><td> Sigma Aldrich Ltd </td><td> C0130 </td><td></td></tr><tr><td> Protease </td><td> Sigma Aldrich Ltd </td><td> P5147 </td><td></td></tr><tr><td> Føtalt bovint serum </td><td> Sigma Aldrich Ltd </td><td> F1051 </td><td></td></tr><tr><td> Askorbat </td><td> Sigma Aldrich Ltd </td><td> A4034 </td><td></td></tr><tr><td> Antibiotika / antimykotika </td><td> Sigma Aldrich Ltd </td><td> A5955 </td><td></td></tr><tr><td> HEPES </td><td> Bioshop Canada Ltd </td><td> HEP001 </td><td></td></tr><tr><td> Trypan blå </td><td> Sigma Aldrich Ltd </td><td> 93595 </td><td></td></tr><tr><td> Reichert Bright-Line Hemacytometer </td><td> Häusser Scientific </td><td> 1490 </td><td></td></tr><tr><td> Quant-iT PicoGreen </td><td> Invitrogen </td><td> P7589 </td><td></td></tr><tr><td> Papain fra papayalateks </td><td> Sigma Aldrich Ltd </td><td> P3125 </td><td></td></tr><tr><td> Ammoniumacetat </td><td> Sigma Aldrich Ltd </td><td> A1542 </td><td></td></tr><tr><td> Ethyldiaminetetraacetic Acid </td><td> Sigma Aldrich Ltd </td><td> E9884 </td><td></td></tr><tr><td> DL-Dithiothreitol </td><td> Sigma Aldrich Ltd </td><td> 43819 </td><td></td></tr><tr><td> Lavt smeltepunkt agarose, type VII </td><td> Sigma Aldrich Ltd </td><td> A9045 </td><td></td></tr> &lt;tr&gt; <td> Mesh-skjerm (200) Filter </td><td> Sigma Aldrich Ltd </td><td> S4145 </td><td></td></tr><tr><td> Mach-1 mikromekanisk Tester </td><td> Biomomentum Inc. </td><td> V500cs </td><td></td></tr><tr><td> Komprimering Laster Jig </td><td> Spesialbygde </td><td></td><td> Tilsvarende produkt kan leveres av Biomomentum Inc. </td></tr><tr><td> Falk 24 Vel Kultur Plate </td><td> Thermo Fisher Scientific </td><td> B353047 </td><td></td></tr><tr><td> β-væskescintillasjonsteller </td><td> Beckman Coulter </td><td> LS6500 </td><td></td></tr><tr><td> [3H] Proline </td><td> Perkin-Elmer </td><td> NET323005MC </td><td></td></tr><tr><td> [35S] Svovel </td><td> Perkin-Elmer </td><td> NEX041005MC </td><td></td></tr></tbody></table>

mechanical stimulation of chondrocyte-agarose hydrogels

Leddbrusk lider fra en begrenset kapasitet når reparasjon skadet ved mekaniske fornærmelse eller degradert av sykdom, slik som osteoartritt. For å bøte på denne mangelen, har flere medisinske intervensjoner blitt utviklet. En slik metode er å gjenoppstår det skadede området med vev-konstruert brusk, men den utviklet vev vanligvis mangler de biokjemiske egenskaper og holdbarhet av innfødt brusk, spørrende sin langsiktige overlevelsesevne. Dette begrenser anvendelsen av bruskvev engineering til reparasjon av små kontaktpunkter defekter, avhengig av den omgivende vev for å beskytte det implanterte materiale. For å forbedre egenskapene til den utviklede vev, er mekanisk stimulering en populær metode benyttes til å forbedre syntesen av brusk ekstracellulær matriks, så vel som de resulterende mekaniske egenskaper konstruert vev. Mekanisk stimulering gjelder krefter til vevet konstruerer analogt til de erfarne in vivo. Detteer basert på premisset om at den mekaniske miljø, delvis regulerer utvikling og vedlikehold av innfødte vev 1,2. Den hyppigst anvendt form av mekanisk stimulering i bruskvev engineering er dynamisk kompresjon ved fysiologisk stammer av ca 5-20% ved en frekvens på 1 Hz 1,3. Flere studier har undersøkt effekten av dynamisk kompresjon og har vist at det å ha en positiv effekt på chondrocyttransplantasjon metabolisme og biosyntese, til slutt påvirker funksjonelle egenskaper utviklet vev 4-8. I denne artikkelen illustrerer vi metoden til mekanisk stimulere chondrocyttransplantasjon-agarose hydrogel konstruerer under dynamisk kompresjon og analysere endringer i biosyntesen gjennom biokjemiske og radioisotop analyser. Denne metoden kan også bli lett modifisert for å vurdere eventuelle potensielt induserte endringer i cellulær respons som et resultat av mekaniske stimuli.

Watch this Scientific Journal Video about Mechanical Stimulation of Chondrocyte-agarose Hydrogels at JoVE.com

Mechanical Stimulation of Chondrocyte-agarose Hydrogels

Biosyntesen av brusk ekstracellulær matriks ved kondrocytter kan påvirkes ved anvendelse av mekaniske stimuli. Denne metoden beskriver teknikken for å bruke dynamiske kompresjons stammer til chondrocytes innkapslet i 3D konstruksjoner og evaluering av induserte endringer i chondrocyttransplantasjon metabolisme.

Mekanisk stimulering av chondrocyttransplantasjon-agarose Hydrogeler

Articular cartilage suffers from a limited repair capacity when damaged by mechanical insult or degraded by disease, such as osteoarthritis. To remedy ...

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