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Long-term effects of muscle-derived stem cell therapy on the regeneration of the urethra of female rats

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Abstract

Introduction and hypothesis

The aim was to analyze the long-term effects of muscle-derived stem cells (MDSCs) therapy in traumatized urethras of female rats regarding messenger ribonucleic acid (mRNA) expression of collagens 1 and 3, Ngf and Ki67; and the mRNA and protein expression of Myh11 and Myh2.

Methods

Muscle-derived stem cells were injected into the tail vein of rats 3 days after trauma by vaginal distention. Urethras were analyzed from 30 animals divided into three groups: control without injury or treatment, trauma (30 days post-injury), and MDSC (30 days post-injury who received MDSC therapy). Real-time quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry were performed. The Kruskal–Wallis and ANOVA tests were used with p < 0.05 indicating significance.

Results

We detected increased Myh11 and Myh2 mRNA expression in the trauma group compared with the control group (p = 0.03 and p = 0.04 respectively). Ki67 and Col1a1 genes were overexpressed in the MDSC group compared with both the trauma (p = 0.02 and p = 0.008 respectively) and the control group (p = 0.01 and p = 0.03 respectively). Col3a1 gene was upregulated in the MDSC compared with the control group (p = 0.03). Ngf mRNA level was lower in the MDSC group than in the trauma group (p = 0.002). Myh11, Myh2, and Desmin proteins were overexpressed in the MDSC compared with the trauma group (1.5-fold, p = 0.01; 1.5-fold, p = 0.04; 1.3-fold, p = 0.01 respectively).

Conclusions

Muscle-derived stem cell therapy may have had long-term structural and molecular effects on the injured urethra of female rats, particularly on markers of cell proliferation, neural growth factor, extracellular matrix, and muscle content. This study suggests that MDSC therapy acted mainly to produce urethral sphincter regeneration marked by increased immunohistochemical expression of the proteins desmin, smooth muscle Myh11, and skeletal muscle Myh2.

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Funding

Conselho Nacional de Desenvolvimento Científico e Tecnológico- CnPq 17/2008.

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Authors and Affiliations

  1. Department of Gynecology, Universidade Federal de São Paulo, São Paulo, Brazil

    Andreisa P. M. Bilhar, Maria A. T. Bortolini, Alexandre B. Sé, Suellen M. Feitosa & Rodrigo A. Castro

  2. Department of Gynecology, Maternidade Escola Assis Chateaubriand, Universidade Federal do Ceará, Fortaleza, Brazil

    Andreisa P. M. Bilhar

  3. Department of Neurology, Universidade de São Paulo, São Paulo, Brazil

    Edmar Zanoteli

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  1. Andreisa P. M. Bilhar
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  2. Maria A. T. Bortolini
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Correspondence to Maria A. T. Bortolini.

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Conflicts of interest

Dr. Bortolini receives funding from the IUGA to cover expenses associated with attending the IUGA Annual Meeting as her role as an Associate Editor of International Urogynecology Journal. The other authors had no conflicts to declare.

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Bilhar, A.P.M., Bortolini, M.A.T., Sé, A.B. et al. Long-term effects of muscle-derived stem cell therapy on the regeneration of the urethra of female rats. Int Urogynecol J 33, 965–975 (2022). https://doi.org/10.1007/s00192-021-04708-y

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