Cannon Tracy W, Lee Ji Youl, Somogyi George, Pruchnic Ryan, Smith Christopher P, Huard Johnny, Chancellor Michael B
Department of Urology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
Urology. 2003 Nov;62(5):958-63. doi: 10.1016/s0090-4295(03)00679-4.
To study the physiologic outcome of allogenic transplant of muscle-derived progenitor cells (MDPCs) in the denervated female rat urethra.
MDPCs were isolated from muscle biopsies of normal 6-week-old Sprague-Dawley rats and purified using the preplate technique. Sciatic nerve-transected rats were used as a model of stress urinary incontinence. The experimental group was divided into three subgroups: control, denervated plus 20 microL saline injection, and denervated plus allogenic MDPCs (1 to 1.5 x 10(6) cells) injection. Two weeks after injection, urethral muscle strips were prepared and underwent electrical field stimulation. The pharmacologic effects of d-tubocurare, phentolamine, and tetrodotoxin on the urethral strips were assessed by contractions induced by electrical field stimulation. The urethral tissues also underwent immunohistochemical staining for fast myosin heavy chain and CD4-activated lymphocytes.
Urethral denervation resulted in a significant decrease of the maximal fast-twitch muscle contraction amplitude to only 8.77% of the normal urethra and partial impairment of smooth muscle contractility. Injection of MDPCs into the denervated sphincter significantly improved the fast-twitch muscle contraction amplitude to 87.02% of normal animals. Immunohistochemistry revealed a large amount of new skeletal muscle fiber formation at the injection site of the urethra with minimal inflammation. CD4 staining showed minimal lymphocyte infiltration around the MDPC injection sites.
Urethral denervation resulted in near-total abolishment of the skeletal muscle and partial impairment of smooth muscle contractility. Allogenic MDPCs survived 2 weeks in sciatic nerve-transected urethra with minimal inflammation. This is the first report of the restoration of deficient urethral sphincter function through muscle-derived progenitor cell tissue engineering. MDPC-mediated cellular urethral myoplasty warrants additional investigation as a new method to treat stress urinary incontinence.
研究肌肉来源的祖细胞(MDPCs)异体移植至去神经支配的雌性大鼠尿道后的生理结果。
从6周龄正常Sprague-Dawley大鼠的肌肉活检组织中分离出MDPCs,并采用预铺板技术进行纯化。将坐骨神经切断的大鼠用作压力性尿失禁模型。实验组分为三个亚组:对照组、去神经支配加注射20微升生理盐水组、去神经支配加异体MDPCs(1至1.5×10⁶个细胞)注射组。注射两周后,制备尿道肌肉条并进行电场刺激。通过电场刺激诱导的收缩来评估筒箭毒碱、酚妥拉明和河豚毒素对尿道条的药理作用。尿道组织还进行了快肌球蛋白重链和CD4激活淋巴细胞的免疫组织化学染色。
尿道去神经支配导致最大快肌收缩幅度显著降低,仅为正常尿道的8.77%,且平滑肌收缩力部分受损。将MDPCs注射到去神经支配的括约肌中可显著提高快肌收缩幅度,达到正常动物的87.02%。免疫组织化学显示在尿道注射部位有大量新的骨骼肌纤维形成,炎症轻微。CD4染色显示MDPC注射部位周围淋巴细胞浸润极少。
尿道去神经支配导致骨骼肌几乎完全丧失,平滑肌收缩力部分受损。异体MDPCs在坐骨神经切断的尿道中存活2周,炎症轻微。这是通过肌肉来源的祖细胞组织工程恢复尿道括约肌功能缺陷的首次报道。MDPC介导的细胞尿道肌成形术作为一种治疗压力性尿失禁的新方法值得进一步研究。