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Remote Ischemic Preconditioning to Improve Fat Graft Viability: In Vitro Culture And Gene Expression Analysis of Mature Adipocytes
Patricia Fuentes
1, Danielle Lehman
*2, Stefanie Bonini
2, David M. Merrick
3, Brannon Claytor
21FIU Herbert Wertheim College of Medicine, Miami, FL; 2Claytor Noone Plastic Surgery, Bryn Mawr, PA; 3Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA
IntroductionFat grafting is a popular procedure to improve body contour, volume, and overall aesthetics. However, studies have shown that graft volume can be reduced by up to 70%. Remote ischemic preconditioning (RIPC) provides a promising framework to improve cell viability by utilizing physiologic adapative responses to tissue ischemia. Therefore, the purpose of this study was to evaluate in vitro mature adipocyte viability after RIPC.
Methods A total of three patients undergoing elective abdominal liposuction were enrolled. For each patient, baseline 10 mL lipoaspirate was obtained from one side of the abdomen (control). A RIPC protocol was then applied using an orthopedic tourniquet to the lower extremity, consisting of three cycles of 10-minute ischemia followed by reperfusion. Subsequently, 10 mL lipoaspirate was collected from the contralateral abdomen (experimental). Both control and experimental samples were processed at two time points: immediately after harvest (T0) and after 24 hours (T24). Lipoaspirate was centrifuged and washed to isolate mature adipocytes. At T0, three replicates of 400µL aliquots of adipocytes were processed in TRIzol for RNA extraction and quantitative PCR (qPCR). Three replicates of 200µL aliquots were cultured for 24 hours. At T24, cultured samples were processed in TRIzol for qPCR analysis. A total of 22 genes were evaluated, including markers of adipocyte identity, oxidative stress, and apoptosis.
Results At T24, the experimental group has a significantly higher expression of ADIPOQ compared to control (p<0.001). Vascular endothelial growth factor (VEGF) showed consistent recovery in all three experimental patient samples 24 hours post-culturing compared to control samples after 24 hours (p<0.001). IL1b expression was significantly lower in experimental T24 compared to control T24 (p<0.001).
Conclusion Remote ischemic preconditioning may improve adipocyte viability and neovascularization, while reducing inflammation.
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