Supplementary Material for: Therapeutic Potential of Human Adipose-Derived Stem Cell Exosomes in Stress Urinary Incontinence – An in Vitro and in Vivo Study

Background/Aims: To evaluate whether local injection of exosomes derived from human adipose-derived stem cells (hADSCs) facilitates recovery of stress urinary incontinence (SUI) in a rat model. Methods: For the in vitro study, a Cell Counting Kit-8 (CCK-8) array and proteomic analysis were performed. For the in vivo study, female rats were divided into four groups: sham, SUI, adipose-derived stem cell (ADSC), and exosomes (n = 12 each). The SUI model was generated by pudendal nerve transection and vaginal dilation. Vehicle, hADSCs, or exosomes were injected into the peripheral urethra. After 2, 4, and 8 weeks, the rats underwent cystometrography and leak point pressure (LPP) testing, and tissues were harvested for histochemical analyses. Results: The CCK-8 experiment demonstrated that ADSC-derived exosomes could enhance the growth of skeletal muscle and Schwann cell lines in a dose-dependent manner. Proteomic analysis revealed that ADSC-derived exosomes contained various proteins of different signaling pathways. Some of these proteins are associated with the PI3K-Akt, Jak-STAT, and Wnt pathways, which are related to skeletal muscle and nerve regeneration and proliferation. In vivo experiments illustrated that rats of the exosome group had higher bladder capacity and LPP, and had more striated muscle fibers and peripheral nerve fibers in the urethra than rats of the SUI group. Both urethral function and histology of rats in the exosome group were slightly better than those in the ADSC group. Conclusions: Local injection of hADSC-derived exosomes improved functional and histological recovery after SUI.

背景与目的：探讨人脂肪间充质干细胞（human adipose-derived stem cells, hADSCs）来源的外泌体局部注射是否可促进大鼠压力性尿失禁（stress urinary incontinence, SUI）模型的功能恢复。方法：体外实验采用细胞计数试剂盒-8（Cell Counting Kit-8, CCK-8）检测及蛋白质组学分析；体内实验将雌性大鼠分为4组：假手术组、SUI模型组、脂肪间充质干细胞（adipose-derived stem cell, ADSC）组及外泌体组，每组12只。通过阴部神经切断联合阴道扩张术构建SUI大鼠模型，分别向尿道周围注射溶剂对照、hADSCs或外泌体。分别于术后2、4、8周对大鼠进行膀胱测压及漏尿点压力（leak point pressure, LPP）检测，并取材进行组织化学分析。结果：CCK-8实验结果显示，ADSC来源的外泌体可剂量依赖性促进骨骼肌细胞与施万细胞系的增殖。蛋白质组学分析表明，ADSC来源的外泌体含有多种参与不同信号通路的蛋白质，其中部分蛋白与PI3K-Akt、Jak-STAT及Wnt通路相关，这些通路均与骨骼肌及神经再生、增殖密切相关。体内实验结果显示，外泌体组大鼠的膀胱容量与漏尿点压力均高于SUI模型组，尿道内骨骼肌纤维与外周神经纤维数量更多；外泌体组大鼠的尿道功能与组织学表现均略优于ADSC组。结论：局部注射hADSCs来源的外泌体可改善SUI模型大鼠的功能与组织学恢复。