Autologous in vitro cultured vaginal tissue for vaginoplasty in women with Mayer-Rokitansky-Küster-Hauser syndrome: anatomic and functional results.

STUDY OBJECTIVE: To present the procedure and the results of a technique in which in vitro autologous cell cultures were used for the canal lining in patients with Mayer-Rokitansky-Küster-Hauser syndrome (MRKHS) subjected to vaginoplasty with a modified Abbè-McIndoe technique. MRKHS is a rare anomaly characterized by vaginal agenesis with variable müllerian duct abnormalities. The Abbè-McIndoe procedure is 1 of the most frequent surgical treatments adopted in these women. In the last decades, several modifications have been introduced by different authors, mostly changing the lining material, but no consensus has been reached on what material should be used for the neovagina canal wall lining. DESIGN: A pilot study (Canadian Task Force classification II-1). SETTING: Policlinico Umberto I, "Sapienza" University of Rome. PATIENTS: A consecutive series of 23 women with MRKHS underwent neovaginoplasty with autologous vaginal tissue as the graft material between 2006 and 2013. INTERVENTIONS: Each patient with MRKHS was subjected to a full-thickness mucosal biopsy from the vaginal vestibule. After enzymatic dissociation, cells were inoculated onto collagen IV-coated plates and cultured for 2 to 3 weeks. The patients were subjected to vaginoplasty with a modified Abbè-McIndoe technique with autologous in vitro cultured vaginal tissue. Patients underwent clinical follow-up visits at 1, 3, 6, and 12 months after surgery and every year thereafter. Anatomic, functional, and sexual results were assessed. MEASUREMENTS AND MAIN RESULTS: In all cases, the vagina appeared normal in length and depth. A vaginal cytology and a vaginal biopsy obtained at the 3-month follow-up visit revealed physiological vaginal tissue. All 23 patients completed the Female Sexual Function Index questionnaire at 12 months after surgery. The results showed a total score of 27.2. These results indicate a satisfactory quality of sexual life. CONCLUSION: The modified Abbè-McIndoe technique with autologous vaginal tissue appears to be safe and feasible. This technique allows normal and satisfying sexual intercourse. Larger series with longer follow-ups will be necessary to confirm if this technique represents the ideal procedure for vaginal agenesis.

Characterization of human vaginal mucosa cells for autologous in vitro cultured vaginal tissue transplantation in patients with MRKH syndrome.

Mayer-Rokitansky-Küster-Hauser (MRKH) is a rare syndrome characterized by congenital aplasia of the uterus and vagina. The most common procedure used for surgical reconstruction of the neovagina is the McIndoe vaginoplasty, which consists in creation of a vaginal canal covered with a full-thickness skin graft. Here we characterized the autologous in vitro cultured vaginal tissue proposed as alternative material in our developed modified McIndoe vaginoplasty in order to underlie its importance in autologous total vaginal replacement. To this aim human vaginal mucosa cells (HVMs) were isolated from vaginal mucosa of patients affected by MRKH syndrome and characterized with respect to growth kinetics, morphology, PAS staining, and expression of specific epithelial markers by immunofluorescence, Western blot, and qRT-PCR analyses. The presence of specific epithelial markers along with the morphology and the presence of mucified cells demonstrated the epithelial nature of HMVs, important for an efficient epithelialization of the neovagina walls and for creating a functional vaginal cavity. Moreover, these cells presented characteristics of effective proliferation as demonstrated by growth kinetics assay. Therefore, the autologous in vitro cultured vaginal tissue might represent a highly promising and valid material for McIndoe vaginoplasty.

Gene expression profile of patients with Mayer-Rokitansky-Küster-Hauser syndrome: new insights into the potential role of developmental pathways.

Mayer-Rokitansky-Küster-Hauser syndrome (MRKHS) is a rare disease characterized by congenital aplasia of uterus and vagina. Although many studies have investigated several candidate genes, up to now none of them seem to be responsible for the aetiology of the syndrome. In our study, we identified differences in gene expression profile of in vitro cultured vaginal tissue of MRHKS patients using whole-genome microarray analysis. A group of eight out of sixteen MRKHS patients that underwent reconstruction of neovagina with an autologous in vitro cultured vaginal tissue were subjected to microarray analysis and compared with five healthy controls. Results obtained by array were confirmed by qRT-PCR and further extended to other eight MRKHS patients. Gene profiling of MRKHS patients delineated 275 differentially expressed genes, of which 133 downregulated and 142 upregulated. We selected six deregulated genes (MUC1, HOXC8, HOXB2, HOXB5, JAG1 and DLL1) on the basis of their fold change, their differential expression in most patients and their relevant role in embryological development. All patients showed upregulation of MUC1, while HOXB2 and HOXB5 were downregulated, as well as Notch ligands JAG1 and DLL1 in the majority of them. Interestingly, HOXC8 was significantly upregulated in 47% of patients, with a differential expression only in MRKHS type I patients. Taken together, our results highlighted the dysregulation of developmental genes, thus suggesting a potential alteration of networks involved in the formation of the female reproductive tract and providing a useful clue for understanding the pathophysiology of MRKHS.

Vaginal reconstruction with the Abbè-McIndoe technique: from dermal grafts to autologous in vitro cultured vaginal tissue transplant.

Vaginal agenesis represents the most common anomaly of the lower female genital tract. Surgical treatments have gradually evolved from aggressive procedures to minimally invasive techniques. The Abbè-McIndoe procedure is one of the most frequent surgical procedures used. The original Abbè-McIndoe procedure consisted of the surgical creation of a vagina in between the bladder and the rectum and the successive lining with a dermal graft. In the last decades different authors have introduced several modifications, mostly changing the lining material. Amniotic membranes, inert materials, and oral mucosa have all been used to improve the short- and long-term results. Recently, we have reported the use of autologous in vitro grown vaginal tissue as lining material with highly promising results. In this review, we discuss the improvements achieved using this minimally invasive procedure and discuss the advantages and disadvantages of the different materials.

Early prolapse of a neovagina created with self-dilatation and treated with sacrospinous ligament suspension in a patient with Mayer-Rokitansky-Küster-Hauser syndrome: a case report.

OBJECTIVE: To report a case of a patient who had an early prolapse of the neovagina, while using Frank's dilators method, that was successfully treated with sacrospinous ligament suspension. DESIGN: Case report. SETTING: Tertiary-care university hospital. PATIENT(S): A 22-year-old woman with Mayer-Rokitansky-Küster-Hauser syndrome. INTERVENTION(S): Frank's vaginal dilators method. MAIN OUTCOME MEASURE(S): Vaginal elongation by self-dilatation. Early prolapse of the neovagina. Surgical repair with vaginal sacrospinous ligament suspension. RESULT(S): A few months after the beginning of the treatment the patient had a prolapse of the neovagina. On pelvic examination, the total vaginal length was 5 cm with eversion of the vaginal apex 3 cm beyond the hymen with strain. Vaginal length was not satisfactory for the patient yet; therefore, the vaginal dilation program was continued to reach the prefixed vaginal length. Two months later the total vaginal length was 8 cm with eversion of the vaginal apex 6 cm beyond the hymen with strain. The patient underwent a sacrospinous ligament suspension. Twenty months after surgery, the patient had no symptoms and was highly satisfied with the result. CONCLUSION(S): Prolapse of a neovagina may occur early after conservative treatment. If the initial prefixed goal of vaginal length has not been reached when prolapse develops, the vaginal dilatation program may be continued. In our case vaginal sacrospinous ligament fixation was successful, with good anatomic and functional results.