Background Lymphedema is a chronic condition characterized by progressive edema with complicated treatment. Recently, new treatment strategies inducing lymphangiogenesis were proposed. The aim of our study was to examine the effect of VEGF-C and ADSC (adipose derived stem cells) on lymphatic regeneration and drainage reestablishment in vascularized lymph node transfer model using a pedicled VLN groin flap. Materials and methods Female Lewis rats with groin VLN flaps were utilized as a lymphedema model. Group A served as the control. Group B received VEGF-C. Group C received both VEGF-C and ADSCs. Group D received ADSCs only. Lymphatic drainage reestablishment was evaluated by ultrasound-photoacoustic imaging after ICG injection. Results The fastest regeneration of elevated flaps was observed in Groups B and C in all monitored periods. After the first month, ICG positivity was detected in 14.3% of animals in Group A, 71.43% of animals in Group B (OR=15; p= 0.048), and 83.33% in Group C (OR=30; p=0.027). Contrary, the difference between control group and Group D (16.67%; p=0.905) was statistically insignificant. Administration of VEGF-C, ADCs + VEGF-C, and ADSC led to full flap regeneration after six months. The control group had the lowest percentage of ICG positivity at all monitored time points. Conclusion We found that the fastest regeneration occurred with the combination of the VLN flap and VEGF-C. The addition of ADSC had an insignificant effect in our study. Furthermore, we proved the feasibility of photoacoustic imaging as an assessment tool of the lymphatic drainage recovery in a VLNT model.

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