Ischemia and reperfusion injury in superficial inferior epigastric artery-based vascularized lymph node flaps

Autoři: David P. Perrault aff001;  Gene K. Lee aff001;  Antoun Bouz aff001;  Cynthia Sung aff001;  Roy Yu aff001;  Austin J. Pourmoussa aff001;  Sun Young Park aff001;  Gene H. Kim aff002;  Wan Jiao aff001;  Ketan M. Patel aff001;  Young-Kwon Hong aff001;  Alex K. Wong aff001
Působiště autorů: Division of Plastic and Reconstructive Surgery and Department of Surgery, Keck School of Medicine of USC, Los Angeles, California, United States of America aff001;  Departments of Pathology and Dermatology, Keck School of Medicine of USC, Los Angeles, California, United States of America aff002
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article


Vascularized lymph node transfer (VLNT) is a promising treatment modality for lymphedema; however, how lymphatic tissue responds to ischemia has not been well defined. This study investigates the cellular changes that occur in lymph nodes in response to ischemia and reperfusion. Lymph node containing superficial epigastric artery-based groin flaps were isolated in Prox-1 EGFP rats which permits real time identification of lymphatic tissue by green fluorescence during flap dissection. Flaps were subjected to ischemia for either 1, 2, 4, or 8 hours, by temporarily occluding the vascular pedicle. Flaps were harvested after 0 hours, 24 hours, or 5 days of reperfusion. Using EGFP signal guidance, lymph nodes were isolated from the flaps and tissue morphology, cell apoptosis, and inflammatory cytokines were quantified and analyzed via histology, immunostaining, and rtPCR. There was a significant increase in collagen deposition and tissue fibrosis in lymph nodes after 4 and 8 hours of ischemia compared to 1 and 2 hours, as assessed by picrosirius red staining. Cell apoptosis significantly increased after 4 hours of ischemia in all harvest times. In tissue subject to 4 hours of ischemia, longer reperfusion periods were associated with increased rates of CD3+ and CD45+ cell apoptosis. rtPCR analysis demonstrated significantly increased expression of CXCL1/GRO-α with 2 hours of ischemia and increased PECAM-1 and TNF-α expression with 1 hour of ischemia. Significant cell death and changes in tissue morphology do not occur until after 4 hours of ischemia; however, analysis of inflammatory biomarkers suggests that ischemia reperfusion injury can occur with as little as 2 hours of ischemia.

Klíčová slova:

Apoptosis – Collagens – Fibrosis – Gene expression – Ischemia – Lymph nodes – Reperfusion – Reperfusion injury


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