Objective To construct three-dimensional (3D) pre-vascularized microstructures and explore the promoting effect of human fibroblasts (HFs) on the sprout and migration of human umbilical vein endothelial cells (HUVECs) in 3D co-culture system. Methods HUVECs and HFs were cultured and the 3rd to 5th generation cells were selected for subsequent experiments. In 2D co-culture system, HFs were stained with PKH26 and the cell density was fixed, which co-cultured with HUVECs in different ratios (1∶4, 1∶1, 4∶1) and inoculation methods (HUVECs inoculation at 48 hours after HFs, direct mixed inoculation). Then the formation of vascular like structures was observed under fluorescence microscope. In 3D co-culture system, HUVECs and HFs were labeled with green fluorescent protein and red fluorescent protein by lentivirus transfection, respectively. They were inoculated on porous micro-carriers followed by dynamically culturing in rotating bottles to prepare HF, HUVEC, HF-EC, or HF&EC microstructures. The cell growth in microstructures was testing by low permeability crystal violet staining. Subsequently, the microstructures were embedded in fibrin gel and the cell growth and adhesion in HF and HUVEC microstructures were observed by laser confocal microscopy. Laser confocal microscope were also used to observe the sprouts of 4 kinds of microstructures, as well as the cell composition, the number and length of sprouts from HF-EC and HF&EC microstructures. HFs conditioned medium was prepared to observe its effect on sprouts of HUVEC microstructures with DMEM as control group. Results In 2D co-culture system, HFs pre-culturing was helpful to the formation and stability of vascular like structures, and the best effect was when the ratio of two kinds of cells was 1∶1. In 3D co-culture system, it was found that the cells grew well on micro-carriers and had the ability of pre-vascularization. HUVEC microstructures did not sprout, but HF, HF-EC, and HF&EC microstructures could which indicated a good vascularization ability. The HF-EC microstructures were superior to HF&EC microstructures in terms of sprouts length and number (P<0.05). The tubes sprouting from co-cultured group were composed of HFs and HUVECs, and HF microstructures migration preceded HUVEC microstructures always, and their migration trajectories were the same. HUVEC microstructures could sprout when cultured in HFs conditioned media. Conclusion HF-HUVEC pre-vascularized microstructures can be prepared by pre-culturing HFs before HUVECs and with the cell ratio at 1∶1 in a rotating bottle. In 3D co-culture system, HFs can promote and guide the sprout of HUVECs.
Objective To investigate effectiveness of the modified edge resection technique for composite labia minora and clitoral hood reduction with preserved microstructures. Methods A retrospective analysis was conducted on 36 female patients, who were diagnosed with composite hypertrophy of the labia minora and clitoral hood and admitted between September 2022 and December 2024. The patients’ ages ranged from 18 to 45 years (mean, 27.4 years). The primary surgical motivations included poor appearance alone (8 cases), functional impairment alone (14 cases), both poor appearance and functional impairment (12 cases), and psychological factors (2 cases). All patients were treated with the modified edge resection technique for composite labiaminora and clitoral hood reduction with preserved microstructures. The surgical technique emphasized precise adjustments to the clitoral-labial junction and optimization of the anterior labial structure to ensure a natural postoperative appearance and functional integrity. Postoperative follow-up assessed improvements in appearance and function of clitoral hood and labia minora, complications, and overall patient satisfaction. Results One patient exhibited suboptimal wound healing, while the remaining patients experienced no complications such as postoperative bleeding, hematoma, wound dehiscence, suture cutting, or labial edema and enlargement. Thirty patients were followed up with a duration of 1-6 months (mean, 2.4 months). In the early postoperative period, 2 patients perceived asymmetry of the bilateral labia minora; 1 underwent labial revision surgery, while the other achieved near-symmetry without intervention. At last follow-up, 25 patients experienced varying degrees of relief from preoperative functional impairments, while the remaining patients showed no improvement; 27 patients reported varying degrees of improvement in appearance, 2 reported no change, and 1 reported a worse appearance compared to preoperatively. Sixteen patients were very satisfied with the surgical results, 8 were satisfied, 5 were moderate satisfied, and 1 was dissatisfied, with a satisfaction rate of 80% (24/30). Conclusion The modified edge resection technique for composite labia minora and clitoral hood reduction with preserved microstructures which preserves and optimizes fine anatomical structures through precise adjustments at the clitoral-labial junction, achieves high patient satisfaction with both aesthetic and functional outcomes while minimizing postoperative complications.