Plastic and Reconstructive Surgery • March 2014 Supplement

105 A Non-Suture Cuff Technique for LymphVein Anastomosis in a Rat Model Dedi Tong, MD1; Shan Zhu, MD1; Yong Huang, PhD2; Lehao Wu, MD1; Zuhaib Ibrahim, MD1; Qi Mao, MD1; Jin Li, MD1; Damon S Cooney, MD1; W P Andrew Lee, MD1; Gerald Brandacher, MD1 1 Johns Hopkins University School of Medicine, Baltimore, MD, 2Department of Electrical and Computer Engineering, Johns Hopkins University, Baltimore, MD, Baltimore, MD Purpose: Lymph-vein anastomosis has been an effective way to treat lymphedema. However, the extremely thin and fragile wall of both the lymphatic vessels and vein impose great technical microsurgical challenges. In this pilot project we explored a novel cuff-based non-suture technique for lymph-vein anastomosis in a rat model, aiming to improve the success rate and reduce operation time. Method: A total of 15 Lewis rats (age: 6–8 weeks, body weight: 252.8±3.78g) underwent lymph-vein anastomosis. The left cervical branch of the lymphatic duct that commence into the thoracic duct and the facial vein were freed and dissected. Lymph-vein anastomosis was performed in an endto-end fashion using a polyimide tube (0.368361mm as inner diameter, 0.41148mm as outer diameter), which was prepared into a wedge-like cuff. After the cuff was mounted on the lymph duct, the duct was then inverted and held in place with a circular tie. Next the facial vein was pulled over the lymph stent and anastomosis was completed with a second nylon tie over the cuff. All anastomoses were evaluated by histology on post-op day 7. Results: Total operating time averaged about 2.3±0.28 hours. Lymph-vein anastomosis could be completed within 16.8±4.3 minutes. All 15 procedures in this series were successful. Histology on post-operative day 7 showed patent anastomosis in all cases with both vein and lymph endothelia smooth and intact, no evidence of thrombus formation. Conclusion: This novel non-suture cuff based technique is a safe, efficient and reliable method for lymph-vein anastomosis, which might offer a promising approach that could be utilized in clinical lymphedema surgery.

106 Development of a Chimeric Free Flap Allograft Using the LGR6+ Epithelial Stem Cell Denver M Lough, MD, PhD1; Damon Cooney, MD, PhD2; Shaun Mendenhall, MD3; Joel Reichensperger, BS3; Lisa Cox, BS3; Nicole Cosenza, MS3; Nathan Wetter, MD3; Carrie Harrison, BS3; Michael Neumeister, MD3 1 SIU School of Medicine/Johns Hopkins, Springfield, IL, 2Johns Hopkins, Baltimore, MD, 3SIU School of Medicine, Springfield, IL Purpose: Reconstructive Transplantation (RT) and Reconstructive Microsurgery have seen enormous growth over the last decade with regards to both surgical technique and post operative monitoring and therapy. Understanding the nature of the chronic cellular rejection cascade seen in RT, in that it primarily initiates with the epithelium, we seek to apply our knowledge of the LGR epithelial stem cell (ESC) system, an epithelial stem cell capable of developing the skin, hair shaft and sweat gland, in developing the first chimeric free flap allograft. From this LGR recipient seeded chimeric allograft free flap, we intend to determine if the slow repopulation of the recipient epithelium from its own autologous stem cell niche, while in the presence of donor tissues, can lead to more successful transplantation as well as enhanced tolerance and freedom from rejection. Methods: Using populations of age matched Sprague Dawley and Lewis rat species within a DIEP based abdominal wall transplant model, we transplanted a donor partially de-epithelialized (reticular dermis) composite tissue allograft containing abdominal wall to a recipient abdominal wall defect. Following inset of the flap, a recipient-derived LGR GFP+ ESC seeded acellular matrix was grafted onto the flap’s exposed dermis and bolstered in place. Recipients received either LGR6+ ESC graft, standard skin grafts and either rapamycin-based mono-therapy or normal saline as control. Specimens were examined for histological signs of rejection using published protocols as well as additional assays utilizing: immunofluorescent up-regulation of apoptosis, tunnel assay augmentation and recipient T-cell infiltrate. Additionally highthroughput gene arrays examining mTOR, cytokine, adhesion molecules, WNT, NFKB and NOTCH pathways were utilized to determine alterations in key markers ie: IL-12 IFN-γ, IL-4, and IL-10, IL-1, IL-4, and TNF-α etc. Results: Recipients receiving the LGR6+ ESC seeded graft when compared to skin graft controls had significantly lower levels of inflammatory markers and augmented levels of freedom from rejection. Additionally, LGR6+ ESC chimeric flaps were able to develop nascent hair follicles, which the skin controls were not capable of producing.

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Abstract 105: a non-suture cuff technique for lymph-vein anastomosis in a rat model.

Abstract 105: a non-suture cuff technique for lymph-vein anastomosis in a rat model. - PDF Download Free
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