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Mouse Hind Limb Transplantation – A New Composite Tissue Allotransplantation (cta) Model Utilizing Non-suture Supermicrosurgery
Robert Sucher, Cheng-hong Lin, Yu-Chung Shih, Rami Zanoun, Irmgard Kronberger, Benson Pulikkottil, Rishi Jindal, Matthias Weinstock, Vijay Gorantla, Stefan Schneeberger, Johann Pratschke, WP Andrew Le
University of Pittsburgh Medical Center,
Division of Plastic and Reconstructive Surgery
2010-03-30
Presenter: Robert Sucher
Affidavit:
Director Name:
Author Category: Resident/Fellow
Presentation Category: Basic Science Research
Abstract Category: Hand
Purpose: Murine hind limb transplantation is an extremely demanding procedure. Since the mouse, however, would be a good model for the study of various CTA-related problems, we designed two new surgical techniques for orthotopic (ORT) and heterotopic (HET) hind limb transplantation.
Methods: BalbC hind-limbs were transplanted to BalbC/C57BL6 recipients using a non-suture-cuff-technique and superfine microforceps for revascularisation. ORT: Donor femoral vessels were anastomosed to recipient femoral vessels, sciatic nerve was approximated end-to-end and osteosynthesis performed using an intramedullary rod. HET/cervical: Donor femoral vessels of a reduced size osteomyocutaneous CTA was anastomosed to recipient common carotid artery and external jugular vein without nerve approximation.
Results: Both (ORT/HET) procedures could be performed with a high success rate (80%). Donor operation lasted 100±12min and recipient operation 114±27min(ORT); 54±16min (HET). All syngeneic grafts survived long-term (>100 days). CTLA4-Ig, treatment was unable to prolong allograft survival (7.5±0.53d and 7±1.20d respectively) compared to untreated controls (6±0.85d). However, FK506 (5mg/kg, and 2mg/kg) prolonged graft survival (>50d). H&E stains of HET-grafts showed muscular atrophy and normal skin whereas ORT-grafts revealed unaltered muscle, skin and bone histology. Functional evaluation of ORT grafts by means of walking-track-analysis and video-gait-kinematics revealed marked differences in terms of ankle range of motion and angular velocity in transplanted animals as compared to controls (p<0.01).
Conclusion: The ORT hind-limb tranplantat model seems to be best suited to study functional outcome and nerve regeneration in CTA. The technically less demanding HET/cervical model may be used to investigate basic immunology and ischemia reperfusion injury in reconstructive transplantation.
