Korean J Urol.  2008 May;49(5):432-438. 10.4111/kju.2008.49.5.432.

Effects of Mesenchymal Stem Cells on Stress Incontinence in a Rat Model

• Son KC
• Kim SO
• Joo SY
• Ahn YK
• Lee JH
• Kwon DD

• KMID: 1204524
• DOI: http://doi.org/10.4111/kju.2008.49.5.432

Abstract

PURPOSE: Several study trials have used stem cells to treat stress incontinence in an animal model. In this study, we compared injecting either periurethral mesenchymal stem cells(MSC) or normal saline(C) to increase the leak point pressure(LPP) and closing pressure(CP) in a rat model of stress urinary incontinence.
MATERIALS AND METHODS
Sprague Dawley rats(250g each, 12 weeks old) were divided into the MSC group(n=5) and group C(n=5). They were anesthetized and the pudendal nerve was transected bilaterally via a ventral incision in order to denervate the external urethral sphincter. The MSCs were obtained from both femurs of Sprague Dawley rats(150g each, 6 weeks, n=10). After 1 week, the MSCs were stained by 4'-6-diamidino- 2-phenylindole(DAPI), which was injected into both sites of the proximal external urethra(n=1.5x10(6)). At 3 weeks after injection, cystometry was performed and this was followed by cord transection at the T9-10 level with the rat under anesthesia. Visually identified LPP and CP measurements were evaluated with using a vertical tilt/intravesical pressure clamp. The urethral tissues of the rats were harvested for histology.
RESULTS
Both the LPP and CP measurements were significantly higher in the MSC group when compared with that of the C group(p<0.05). The mean LPP of the MSC group and group C was 42.3+/-2.1cmH2O and 25.8+/-1.7cmH2O, respectively. The mean CP of the MSC group and group C was 31.7+/-2.5cmH2O and 21.3+/-1.1cmH2O, respectively. The existence of DAPI-stained MSCs in the injected periurethral tissue was verified by histology after the completion of the study.
CONCLUSIONS
Injection of MSCs into the periurethal tissue after transection of the bilateral pudendal nerve in rats led to an increase in the LPP and CP. This finding suggests that MSCs can be used as one of the potentially effective cell therapies for stress urinary incontinence.

Keyword

Urinary incontinence; Mesenchymal stem cells; Urodynamics

MeSH Terms

Rats
Animals

Figure

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  Fig. 1 Schematic figure of mesenchymal stem cell isolation from rat femur. Mesenchymal stem cells (MSC) were obtained by flushing the bone marrow cavity of the femurs and tibiae of rats with a 22-gauge syringe filled with phosphate-buffered saline (PBS).

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  Fig. 2 Western blot analysis of mesenchymal stem cells (MSC) protein. Western blot of protein from MSC show the results of CD31, CD34, CD45 negative and CD90 positive.

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  Fig. 3 Vertical tilt/intravesical pressure clamp model of stress urinary incontinence. An intravesical catheter for leak point pressure (LPP) measurement was inserted into the bladder after spinal cord transection at the T9-T10 level.

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  Fig. 4 Graph of urodynamic results in both group. After 3 weeks of mesenchymal stem cells (MSC) injection or saline injection, cystometry was similar in both groups. Leak point pressure (LPP) and closing pressure (CP) in MSC group were higher than that of C group. (A, B; cystometry; C, D; LPP and CP).

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  Fig. 5 Hematoxylin/eosin staining of the injected periurethral tissue of female rat. The black circles indicate the injection area of saline (A) and mesenchymal stem cells (MSC) (C). There are increased muscle masses in the proximal urethral sphincter of MSC group (C and D) compare with that of C group (A and B). In addition, the muscle fibers of C group were relatively atrophied in contrast with that of MSC group (magnification; A, C; ×40; B, D; ×400).

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  Fig. 6 H&E stain and fluorescence microscopic photograph of mesenchymal stem cells (MSC) injected periurethral tissue of female rat. After 3 weeks from injection of 4',6-diamidino-2-phenylindole (DAPI) stained stem cell, under fluorescence microscopy, DAPI was excited with ultraviolet light and was detected by blue emission (A: H&E, B: Fluorescent stain, ×400).

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