Mikro-Bilgisayarlı Tomografi ile Çıplak Fareler içine İnsan Yağ Enjeksiyonu Canlılığına Değerlendirilmesi
Summary
Fat grafting is an essential technique for reconstructing soft tissue deficits. However, it remains an unpredictable procedure characterized by variable graft survival. Our goal was to devise a mouse model that utilizes a novel imaging method to compare volume retention between differing techniques of fat graft preparation and delivery.
Abstract
Lipotransfer vücutta yumuşak doku eksikliklerinin tedavisinde cerrahın araçları arasında önemli bir araçtır. Yağ o, hazır kolayca elde ucuz ve doğal biyo-uyumlu olarak ideal bir yumuşak doku dolgu olduğunu. 1 Ancak, gelişen popülariteye rağmen, yağ grefti 10 yerde değişen yayınlanan tutma oranları ile, öngörülemeyen sonuçlara ve değişken greft sağkalım tarafından engellenmektedir -80%. 1-3
Yağ grefti ile ilgili soruşturma kolaylaştırmak için, biz bu nedenle enjekte yağ hacmi tutma gerçek zamanlı analiz için izin veren bir hayvan modeli geliştirdik. Kısaca, küçük bir kesik, bir CD-1 çıplak farenin derisi yapılır ve işlenmiş lipoaspirate 200-400 ul kafatası üzerine yerleştirilir. kafa derisi çünkü yardımları kafatasında tarafından sağlanan mükemmel arka plan kontrast, nedeniyle yerli deri altı yağ onun yokluğunda alıcı yeri olarak seçilen veAnaliz süreci. Mikro-bilgisayarlı tomografi (mikro-CT) başlangıçta greft ve bundan sonra her iki haftada bir tarama için kullanılır. BT görüntüleri yeniden inşa edilir ve bir görüntüleme yazılımı greft hacimlerini ölçmek için kullanılır.
Geleneksel teknikler ex vivo fiziksel ölçümleri ile greft ağırlık ve hacim sadece tek bir değerlendirme sağlamak için çalışma hayvanı euthanizing zorunlu olan yağ grefti hacmini değerlendirmek için. Biyokimyasal ve histolojik karşılaştırmaları aynı şekilde ötenazi için çalışma hayvanı gerektirmiştir. Bu tarif görüntüleme tekniği çalışması hayvan kurban zorunda kalmadan görselleştirme ve objektif ilk aşılama sonrası çoklu zaman noktalarında hacmi miktarının avantajı sunuyor. tekniği büyük greftler, risk deri ve yağ nekrozu olarak enjekte edilmesi mümkün greft boyutu ile sınırlıdır. Bu yöntem yağ grefti canlılığını ve hacim tutma değerlendiren tüm çalışmalar için yarar vardır. Özellikle providi çok uygundurYağ grefti bir görsel temsilini ng ve zamanla hacim değişiklikleri takip.
Introduction
Soft tissue defects arise from a variety of causes including trauma, tumor resection, aging, and congenital anomaly. They can be debilitating for patients, and represent one of the most common, yet challenging problems for reconstructive surgeons. Many methods exist for addressing soft tissue deficiencies, such as local and free flaps, collagen injections, and synthetic fillers.4-8 However, since its first documented use by Neuber in 18931, autologous fat transfer remains the gold standard for the repair of soft tissue deficits, as it is ready available, easy and safe to harvest, and naturally compatible.1,2
Despite these advantages, autologous fat grafts suffer from unpredictable and variable survival, with retention rates ranging anywhere from 10-80% over time.1-3,9 In order to account for this expected loss of volume and symmetry, surgeons must often overcorrect when filling soft tissue defects, or perform multiple follow-up procedures.
Poorly vascularized graft beds are partly to blame for this tissue resorption. Additionally, the lack of a benchmark analysis method to compare graft survival may also contribute to the inconsistency in reported results. A precise method for measuring graft volume would reduce measurement error when evaluating retention rates. This in turn would help researchers more accurately identify the causative factors that affect graft survival. Although many laboratory animal models have facilitated both quantitative and qualitative assessment of human fat graft survival, most are based on histological and biochemical means and require sacrificing the study animal to yield a single measurement.3,10-12 Little has been reported on the use of imaging techniques to enumerate fat graft volume retention in vivo.
A handful of clinical studies have shown more effective measurement techniques using imaging. Magnetic Resonance Imaging (MRI) was employed by Hörl et al. to measure fat graft survival13, and CT was utilized by Har-Shai et al. and Fontdevila et al. in their analyses of volume retention after grafting in patients who suffered from HIV.14,15 Employing three-dimensional (3D) imaging software, Meier et al. measured volume retention in humans after autologous fat grafting by comparing images from the preoperative and postoperative period.16
Yet, a standardized method employing imaging to measure fat graft survival is lacking in basic science research. A high resolution imaging approach for assessing the volumes of fat grafts would allow not only for accurate and reproducible volume measurements, but also for repeated measurements allowing visualization of the evolution of fat graft survival in a real time fashion.
Protocol
Representative Results
Discussion
Bu noktaya kadar, pek çok araştırmacı yağ greftlerinin uzun süreli sağkalım ölçmek olmayan görüntüleme yöntemleri dayanıyordu, ancak bu yöntemler çalışma hayvanın kurban gerektirir ve sadece tek bir ölçüm elde var. 3,10-12 Çalışmamız temsil bir fare modelinde yağ aşı yaşamda kalması amacı, gerçek zamanlı kantitatif izin geliştirilmiş bir analiz metodu.
Bu süreçte kritik bu sağlam bağışıklık sistemine sahip fare kullanıldığında ortay…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
Bu çalışma, ACS Franklin H. Martin Fakültesi Araştırma Bursu, Hagey tarafından desteklenen Meşe Vakfı, Pediatrik Rejeneratif Tıp Hagey Laboratuvarı ve MTLDCW Ulusal Sağlık, Hibeler NIHR21DE019274, NIHR01DE019434, NIHR01DE021683 Enstitüsü ve NIHU01HL099776 tarafından desteklenen Pediatrik Rejeneratif Tıp ve Stanford Üniversitesi Çocuk Sağlığı Araştırma Enstitüsü Fakültesi Akademik Ödülü için laboratuvar. Mikro-CT İn Vivo Görüntüleme Yenilik Stanford Merkezi'nde gerçekleştirildi.
Materials
| Reagents and Materials | Manufacturer |
| SAL lipoaspirate | N/A |
| Centrifuge | Beckman Coulter, Inc., Pasadena, CA |
| 50 ml conical tubes | BD Biosciences, San Jose, CA |
| CD-1 nude mice (Crl:CD1-Foxn1nu) | Charles River Laboratories, Inc., Wilmington, MA |
| Isoflurane | Henry Schein, Dublin, OH |
| 2.5% Betadine | Purdue Pharma, L.P., Stamford, CT |
| 70% Ethanol solution | Gold Shield, Hayward, CA |
| 1cc luer-lock syringe | BD Biosciences, San Jose, CA |
| 14 gauge cannula | Shippert Medical, Centennial, CO |
| Forceps | Fine Science Tools, Heidelberg, Germany |
| Tenotomy scissors | Fine Science Tools, Heidelberg, Germany |
| 6-0 nylon suture | Ethicon, Blue Ash, OH |
| Phosphate buffered saline | Gibco, Carlsbad, CA |
| micro-CT scanner | Siemens Healthcare, Pleasanton, CA |
| Phantom | TriFoil Imaging, Northridge, CA |
| Imaging analysis software | IRW, Siemens Healthcare, Pleasanton, CA |
| Scale | Mettler-Toledo International, Inc., Columbus, OH |
Riferimenti
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