Summary
Lymphedema er ekstremitet hævelse forårsaget af lymfe dysfunktion. Vi beskriver en kronisk murin hale model af Lymphedema og den nye brug af væv nanotransfection teknologi (TNT) for genetisk last levering til halen.
Abstract
Lymphedema er ekstremitet hævelse forårsaget af lymfe dysfunktion. Den berørte lemmer forstørrer på grund af ophobning af væske, fedt og fibrose. Der er ingen kur mod denne sygdom. En mus hale model, der bruger en brændvidde fuld tykkelse hud excision nær bunden af halen, hvilket resulterer i hale hævelse, er blevet brugt til at studere Lymphedema. Men, denne model kan resultere i vaskulære omfatter og deraf følgende hale nekrose og tidlig hale hævelse opløsning, begrænse dens kliniske omsættelighed. Den kroniske murin hale Lymphedema model inducerer vedvarende Lymphedema over 15 uger og en pålidelig perfusion til halen. Forbedringer af den traditionelle murin hale Lymphedema model omfatter 1) præcis fuld tykkelse excision og lymfeklip ved hjælp af et kirurgisk mikroskop, 2) bekræftelse af postoperative arteriel og venøs perfusion ved hjælp af høj opløsning laser speckle, og 3) funktionel vurdering ved hjælp af indocyanin grøn nær infrarød laser lymfefiliography. Vi bruger også væv nanotransfection teknologi (TNT) for nye ikke-virale, transkutane, brændpunkt levering af genetisk last til musen hale vaskulatur.
Introduction
Lymphedema er ekstremitet hævelse forårsaget af lymfe dysfunktion. Det berørte lem forstørrer på grund af ophobning af væske, fedt og fibrose1. Lymphedema påvirker 250 millioner mennesker på verdensplan2,3,4. Det anslås, at 20-40% af patienter, der gennemgår behandling for solide maligniteter, såsom brystkræft, melanom, gynækologiske / urologiske tumorer, eller sarkomer, udvikle Lymphedema2,4,5. Sygelighed fra Lymphedema omfatter tilbagevendende infektioner, smerter, og deformitet6. Der er ingen kur mod denne progressive, livslange sygdom. Nuværende behandlinger er variaby effektiv7 og omfatter kompression, komplet dekongetiv behandling af fysioterapeuter, excisional procedurer, og mikrokirurgiske operationer, herunder vaskulære lymfeknude overførsel og lymfeknude bypass7,8,9,10,11,12,13,14. Den ideelle behandling for Lymphedema er endnu ikke opdaget.
Undersøgelse af mekanisme og behandling af Lymphedema har været begrænset. Der er en gennemsnitlig forsinket debut på et år efter lymfeskade15,16 og de fleste personer, der oplever iatrogen fornærmelse med stråling og kirurgi ikke udvikler Lymphedema4,6,17. Selvom store dyremodeller, herunder hunde, får og gris er blevet beskrevet18,19,20, har musehalemodellen været den mest anvendte på grund af lethed, omkostninger og reproducibilty. Musemodeller til undersøgelse af Lymphedema omfatter en halemodel, difteri-toksinmedieret lymfeablation og aksillære eller popliteal lymfeknude dissektion21,22,23,24,25,26. De fleste hale modeller bruger en fokal, fuld tykkelse hud excision med lymfekanal klipning, der udføres nær bunden af halen22, hvilket resulterer i hale hævelse og histologiske træk svarende til human Lymphedema24,27,28,29. Standard murine halemodellen forsvinder dog typisk spontant på så få som 20 dage og ledsages af periodisk halenekrose30. Lymphedema mus hale model udvider en vedvarende Lymphedema ud over 15 uger, viser bekræftet arteriel og venøs patency, og giver funktionelle lymfe dysfunktion vurdering.
En murine hale model af Lymphedema giver mulighed for evaluering af nye terapeutiske til behandling af Lymphedema. Genbaserede strategier er blevet brugt i musemodellen medieret af virale vektorer31,32. Vi bruger også en ny væv nanotransfection teknologi (TNT) for genetisk last levering til lymfødem musehale. TNT letter direkte, transkutan genlevering ved hjælp af en chip med nanokanaler i et hurtigt fokuseret elektrisk felt33,34,35,36. Modellen omfatter brug af TNT2.0 at give mulighed for fokal gen levering af potentielle gen-baserede therapeutics til lymfeskade stedet for musen hale35.
Protocol
Representative Results
Discussion
Lymphedema er kategoriseret som en primær (medfødt) eller sekundær (iatrogen lymfe) skade38,39. Sekundær Lymphedema omfatter 99% aftilfældene 39. Sekundær lymphedema er oftest forårsaget af infektion (filariasis) eller post-onkologisk behandling med lymfektomi eller stråling4,39. En translationel dyremodel er udfordrende for sekundær Lymphedema, da 70% af dyr behandlet med…
Declarações
The authors have nothing to disclose.
Acknowledgements
Dette arbejde blev støttet af tilskud finansiering fra American Association of Plastic Surgeons Academic Scholarship og Department of Defense W81XWH2110135 til AHH. Æstetisk Kirurgi Uddannelse og Research Foundation tilskud til MS NIH U01DK119099, R01NS042617 og R01DK125835 til CKS.
Materials
| Surgical Microscope | Leica, Wetzlar, Germany | MSV266 | |
| Adherent Dressing (Tegaderm) | 3M, St. Paul, Minn. | 1626W | |
| Laser speckle (Pericam PSI System ) | Perimed AB, Stockholm, Sweden) | PSIZ | |
| Near-infrared laser (LUNA) | Stryker (Formerly Novadaq Technologies, Toronto, Canada) | LU3000 | |
| C57BL/6 mice | Jackson Laboratories | 000664 | |
| Micro-Adson Forceps – 1×2 Teeth | Fine Science Tools (USA) Inc. | 11019-12 | |
| V-Hook | Fine Science Tools (USA) Inc. | 18052-12 | |
| Scalpel SS NO15 | Fischer Scientific | 29556 | |
| Disposable Needle 30GX1 | Fischer Scientific | 305128 | |
| Operating Scissors | Fischer Scientific | 12-460-796 | |
| Surgi-Or Jeweler's Forceps, Sklar 4-1/2 in | Fischer Scientific | 50-118-4255 | |
| Spring Scissors – Straight/Sharp-Sharp/8mm Cutting Edge | Fine Science Tools (USA) Inc. | 15024-10 | |
| Cardiogreen | Sigma | I2633-25MG | |
| IsosulfanBlue (Lymphazurin) 50 mg/5ml | Mylan | 67457-220-05 |
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