Improved Cuff Technique and Intraoperative Detection of Vascular Complications for Hind Limb Transplantation in Mice

Abstract

BackgroundVascularized composite tissue allotransplantation (VCA) from a cadaveric donor has now become a clinical reality and the treatment modality of choice for patients with devastating injuries, deformities, and complex tissue defects. However, many VCA patients experience severe toxicities due to the strong immunosuppression required secondary to high antigenicity of the grafts. To improve immunosuppressive protocols for VCA, feasible and reliable preclinical models are necessary. The purpose of this study was to introduce new techniques to an established preclinical VCA model to accelerate future investigations.MethodsC57BL/6 (H-2b) and BALB/c (H-2d) mice were used to perform VCA as recipients and donors, respectively. Surgery time, success rate, associated complications, and mortality were analyzed. Blood flow in grafts was interrogated with laser speckle image (LSI).ResultsA nonsuture cuff technique was used with the abdominal aorta for end-to-end anastomosis. The cuff technique demonstrated efficiency for donor surgery (52 ± 10 minutes for donor vs. 45 ± 8 minutes for recipient surgery). Successful revascularization was achieved in 27 (90%) of 30 transplants. The majority of surgical complications occurred within 48 hours including artery occlusion, venous occlusion, cerebral stroke, and minor bleeding without mortality. LSI was useful in detecting intraoperative vascular complications with display patterns predictive of complication type.ConclusionsThe described techniques may facilitate a more efficient heterotopic hind limb transplantation mouse model of VCA.

Manipulating adrenergic stress receptor signaling to enhance immunosuppression and prolong survival of vascularized composite tissue transplants

“…Our previous work has established a reliable and consistent platform using a pre‐clinical murine model of hind limb VCA 37 to investigate novel therapies to prolong transplant survival. The transplanted graft consisted of skin, fat, muscle, bone and blood vessels (Figure S2A ), a complex combination of tissues similar to those which are often used in VCA.…”

“…as described in our previous published work. 37 Briefly, a donor's abdominal aorta and femoral vein were used for revascularization with a recipient's common carotid artery and external jugular vein, respectively, using a non‐suture cuff technique. We used BALB/c background strain as donors and C57BL/6 background strain as recipients because we have revealed that a BALB/c strain had a higher anatomical mutation rate on the Circle of Willis than C57BL/6 strain.…”

“…We used BALB/c background strain as donors and C57BL/6 background strain as recipients because we have revealed that a BALB/c strain had a higher anatomical mutation rate on the Circle of Willis than C57BL/6 strain. 37 …”

Evolution of the rat hind limb transplant as an experimental model of vascularized composite allotransplantation: Approaches and advantages

“…66 Sucher reported 62% success rates in orthotopic models and 90% in heterotopic models. 40 Kim et al 41 modified the cuff technique and used the abdominal aorta for anastomosis because its diameter is 2-3 times larger than the femoral artery. These publications present an elegant solution for super-microvessel (0.2-0.4 mm) anastomosis and renewed interest in mouse VCA models.…”

“…More recently, studies utilizing cuff techniques for vascular anastomoses in rodent orthotopic models have reported significantly reduced ischemic time for the procedure, offering another possible avenue for researchers interested in full hind limb transplantation. 40,41…”