Objective To investigate the clinical efficacy of minimally invasive surgery combined with hypodermic needle comb puncture in gynecomastia. Methods A retrospective analysis of 86 patients with gynecomastia undergoing the surgery in our hospital from May 2010 to April 2015 was carried out. They were randomly divided into observation group and control group (including control A group and control B group). The observation group included 25 patients with gynecomastia who underwent the minimally invasive surgery combined with hypodermic needle comb puncture, the control A group (30 patients) and the control B group (31 patients) were conducted the minimally invasive surgery only and conventional open excision respectively. The patients were followed up for 6 months. The surgical duration, intraoperative blood loss, the size of incision, skin damage, wound infection, hematoma, time of hospital stay, breast deformation and breast tissue regeneration were compared. Results There were significantly less intraoperative blood loss (t = 11.359 and 9.781, P = 0.031 and 0.000) and lower skin damage rate (χ2 = 4.467 and 13.742, P = 0.021 and 0.000) in the observation group than in the control A group and the control B group. The surgical duration, the size of incision, the number of incision, wound infection, hematoma, and the time of hospital stay in the observation group had no differences from those in the control A group (t/χ2 = 0.221, 0.641, 0.237, 0.391, 2.641 and 5.384; P = 0.371, 0.550, 0.411, 0.079, 0.530 and 0.058), but were significantly different from those in the control B group (t/χ2 = 9.182, 4.610, 3.391, 7.282, 5.542 and 9.683; P = 0.003, 0.035, 0.001, 0.000, 0.042 and 0.000). None of the patients experienced breast tissue regeneration with ultrasonography after six months. Conclusions The minimally invasive surgery combined with hypodermic needle comb puncture for treatment of gynecomastia has the advantages of less blood loss, better cosmetic effect, shorter hospital stay and faster recovery.