Stimulating Vaginal Repair in Rats Through Skeletal Muscle–Derived Stem Cells Seeded on Small Intestinal Submucosal Scaffolds

Abstract

OBJECTIVES Grafts are used for vaginal repair after prolapse, but their use to carry stem cells to regenerate vaginal tissue has not been reported. In this study, we investigated whether 1) muscle-derived stem cells (MDSC) grown on small intestinal submucosa (SIS) generate smooth-muscle cells (SMC) in vitro and upon implantation in a rat model of vaginal defects; 2) express markers applicable to the in-vivo detection of vaginal endogenous stem cells; and 3) stimulate the repair of the vagina. METHODS Mouse MDSC grown on monolayer, SIS, or polymeric mesh, were tested for cell differentiation by immunocytochemistry, Western blot and real-time polymerase chain reaction (PCR). Stem cell markers were screened by DNA microarrays followed by real-time PCR, immunocytochemistry, and Western blot. Rats that underwent hysterectomy and partial vaginectomy were left as such or implanted in the vagina with 4’,6-Diamidino-2-Phenylindole (DAPI)–labeled MDSC on SIS, or SIS without MDSC, immunosuppressed, and killed at 2–8 weeks. Immunofluorescence, hematoxylin-eosin, and Masson trichrome were applied to tissue sections. RESULTS Muscle-derived stem cell cultures on monolayer and on scaffolds differentiate into SMC, as shown by α-smooth muscle actin (ASMA), calponin, and smoothelin markers. Muscle-derived stem cells express embryonic stem cell markers Oct-4 and nanog. Dual DAPI/ASMA fluorescence indicated MDSC conversion to SMC. Muscle-derived stem cells/SIS stimulated vaginal tissue repair, including keratin-5 positive epithelium formation and prevented fibrosis at 4 and 8 weeks. Oct-4+ putative endogenous stem cells were identified. CONCLUSION Muscle-derived stem cells/SIS implants stimulate vaginal tissue repair in the rat, thus autologous MDSC on scaffolds may be a promising approach for the treatment of vaginal repair.

Temporal changes in the biomechanical properties of endometrial mesenchymal stem cell seeded scaffolds in a rat model

“…This study highlights detailed differences in specific tensile properties between cellseeded and non-cell-seeded scaffolds, with enhanced collagen organization for scaffolds implanted with eMSCs. In agreement with our findings, others have found increased total collagen content for cell-seeded scaffolds following implantation [12,13], with enhanced stimulation of tissue growth [19]. Contrary to our findings, others have found cell-seeded meshes to possess higher tensile strength [12] and increased linear region stiffness [12,32] compared to unseeded synthetic meshes.…”

“…For reconstruction of the vagina due to trauma or a congenital defect using a tissue engineering approach, epithelial cells [16,17] and smooth muscle cells [13,16] have been explored. Similarly, repairing damaged vaginal tissue due to POP, vaginal fibroblasts [18], musclederived stem cells [19], and mesenchymal stem cells (MSCs) [14] have been used. MSCs have a particular advantage over differentiated cells, largely due to their ability to control inflammation and encourage repair caused by injury, through complex paracrine immunomodulatory pathways involving both the innate and adaptive immune responses [20].…”

In vivo determination of muscle-derived stem cells in rat corpus cavernosum

“…The present study detected the expression of Sca-1 and Oct4 in cavernous tissue and cells and the results were similar to the findings from non-corpus cavernosum-derived MDSCs reported by Nieponice et al (2008) and Ho et al (2009). The techniques presented in the present study can be used with previously established methods to identify corpus cavernosum stem cells and improve in their identification rate.…”

Muscle-derived stem cells: isolation, characterization, differentiation, and application in cell and gene therapy

“…This study revealed that the injection of MDSCs into the anal sphincter improved functional properties of fecal incontinence. Furthermore, Ho et al (2009) demonstrated that autologous MDSCs grown on small intestinal submucosa generated SMCs and, upon implantation into a rat model of vaginal defects, stimulated vaginal tissue repair. Thus, autologous MDSCs on scaffolds represent a promising approach for the treatment of vaginal repair.…”