Department of Urology, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
Eur Urol. 2011 Jan;59(1):155-63. doi: 10.1016/j.eururo.2010.10.038. Epub 2010 Oct 26.
Stem cell therapy is a promising therapeutic strategy for stress urinary incontinence (SUI). However, its current efficacy is insufficient.
We designed a stem cell transplantation system that contains autologous adipose-derived stem cells (ADSC) and controlled-release nerve growth factor (NGF). We evaluated whether this system could enhance the therapeutic efficacy of ADSCs by periurethral coinjection in SUI rats.
DESIGN, SETTING, AND PARTICIPANTS: We first tested for the presence of NGF receptors in rat ADSCs and observed the effect of NGF on ADSCs in vitro and in vivo. NGF was encapsulated within poly(lactic-co-glycolic acid-PLGA) microspheres (PLGA/NGF) to control its release. SUI was created in rats, and ADSCs were harvested, cultured from fat tissue, and retained for later transplantation. SUI rats then received different forms of periurethral injection therapy. Their urodynamic index was monitored. Eight weeks after injection, the SUI rats were sacrificed and their urethra removed for histologic evaluation.
Forty SUI rats were allocated to five groups for receiving periurethral injection with phosphate-buffered saline (PBS), ADSC, ADSC+PLGA, ADSC+NGF, or ADSC+PLGA/NGF. Bladder capacities, abdominal leak point pressure (ALPP), and retrograde urethral perfusion pressure (RUPP) were reassessed at 2, 6, and 8 wk after injection.
The rat SUI model was generated by bilateral pudendal nerve transection (PNT). Real-time polymerase chain reaction (RT-PCR) and western blotting detected the NGF receptor Ark-A. The regeneration of muscles and peripheral nerves was evaluated by Masson's trichrome and immunohistochemical staining.
Results revealed the presence of the NGF receptor Trk-A on rat ADSCs. Short-term observations showed that NGF could improve ADSCs' viability in vitro and in vivo. ADSCs delivered intramuscularly into the urethra in combination with PLGA/NGF resulted in significant improvements in ALPP and RUPP as well as the amount of muscle and ganglia. There was a significant difference between the ADSC+PLGA/NGF group and other groups.
Periurethral coinjection of autologous ADSCs with controlled-release NGF may be a potential strategy for SUI treatment.
干细胞疗法是治疗压力性尿失禁（SUI）的一种很有前途的治疗策略。然而，其目前的疗效还不够。
我们设计了一种包含自体脂肪来源干细胞（ADSC）和控释神经生长因子（NGF）的干细胞移植系统。我们评估了通过尿道周围 coinjection 这种系统是否可以增强 SUI 大鼠中 ADSC 的治疗效果。
设计、设置和参与者：我们首先检测了大鼠 ADSC 中 NGF 受体的存在，并观察了 NGF 对体外和体内 ADSC 的影响。将 NGF 包裹在聚乳酸-共-羟基乙酸（PLGA）微球（PLGA/NGF）中以控制其释放。SUI 是在大鼠中创建的，然后从脂肪组织中提取、培养 ADSC 并保留用于以后的移植。然后，SUI 大鼠接受不同形式的尿道周围注射治疗。监测他们的尿动力学指数。注射后 8 周，处死 SUI 大鼠并取出尿道进行组织学评估。
40 只 SUI 大鼠被分配到五个组，接受尿道周围注射磷酸盐缓冲盐水（PBS）、ADSC、ADSC+PLGA、ADSC+NGF 或 ADSC+PLGA/NGF。在注射后 2、6 和 8 周重新评估膀胱容量、腹压漏点压（ALPP）和逆行尿道灌注压（RUPP）。
通过双侧阴部神经切断术（PNT）生成大鼠 SUI 模型。实时聚合酶链反应（RT-PCR）和蛋白质印迹检测 NGF 受体 Ark-A。通过 Masson 三色和免疫组织化学染色评估肌肉和周围神经的再生。
结果表明大鼠 ADSC 上存在 NGF 受体 Trk-A。短期观察表明，NGF 可以提高 ADSC 的体外和体内活力。将 ADSC 肌肉内注射到尿道中，结合 PLGA/NGF，可显著提高 ALPP 和 RUPP 以及肌肉和神经节的数量。ADSC+PLGA/NGF 组与其他组之间存在显著差异。
尿道周围共注射自体 ADSC 和控释 NGF 可能是治疗 SUI 的一种有潜力的策略。
