Use Of Autologous Oral Micro-mucosa Graft To Repair The Wound And Its Clinical Application In Vaginal Reconstruction

Part 1 Experimental study of autologous oral micro-mucosa graft on wound repairObjective:To investigate the survival and epithelization of the autologous oral micro-mucosa graft to the wound,and identify its mucosal process and healing pacing.We analyzed the morphology and distribution of the micro-mucosa on the wound and explored the method of accelerating wound epithetization.Materials and methods:1.We contive an animal model of full-thickness skin defect wound(3×3cm²) and use autologous micro-mucosa graft to repair the defect with area expansion ratio of 1;9. The wounds were randomly divided into 5-minute groups(groupâ… ),8-minute groups (groupâ…¡),10-minute groups(groupâ…¢),15-minute groups(groupâ…£) and blank control groups(groupâ…¤) depending on the different periods of mincing time.We applied MTT measurement in each group of mucosa cell viability.The morphology and distribution of the micro-mucosa were observed on the wound.Macroscopy and histological samples by HE and immunohistochemical staining were examined on the 7^th,14^th and 21^st day after grafting.SEM and TEM examination were taken on the 21^st day.2.The mincing time was set to 8 minutes when we observe the impact on the healing process.The Same size wounds were randomly divided into blank control group (group A),micro-mucosa graft group(group B),BMSCs + micro-mucosa transplantation group(group C),BMSCs transplantation group(group D),micro-mucosa combined with micro-skin transplantation group(group E).Macroscopy and histological samples by HE and immunohistochemical staining were examined on the 7th,14th and 21st day after grafting.Results:1.Mucosal particles of all the experimental groups had different index in terms of quantity,area,perimeter,equivalent diameter and shape factor.All the total circumference of micro-mucosa in 4 groups were statistically analyzed and had significant difference(P＜0.05).2.On the 7^th day postoperation,the micro-mucosa survived and grew outward in groupâ…¡,â…¢andâ…£,however,the survived micro-mucoa in groupâ… seemed isolated.The wound healing rates of groupâ…¡was over 70%while groupâ…¤was granulation tissue. On the 14^th day postoperation,the neogenesis mucosa became to integrate into films in groupâ…¡and the wound healing rate was more than 85%and higher than the other three groups(P＜0.05).The neogenesis mucosa was not seen to integrate in groupâ… ,â…¢andâ…£. On the 21^st day postoperation,the wounds of all experimental groups have covered by mucosa.The neogenesis mucosa showed smooth and thick texture in groupâ… ,â…¡andâ…¢while the neogenesis mucosa in groupâ…£seemed thinner than them.The wound healing rate of groupâ… andâ…¢was lower than groupâ…¡(P＜0.05) but higher than groupâ…£(P＜0.05).3.Epithelial cell layers and polarity of all the groups increased gradually over time postoperatively.Groupâ…¡had the most epithelial cell layers and best polarity. Extended-spectrum keratin-positive staining of each group strengthened gradually with time.Groupâ…¡had the strongest positive expression and complete consecutive.4.As for the comparison of mucosal cell viability,groupâ… achieved the highest average OD value but had no significance with groupâ…¡(P＞0.05).Comparison between other groups got P＜0.05.5.The wound healing rates of group E(micro-mucosa and micro-skin composite graft) were significantly higher than all the other groups postoperation(P＜0.05).the junction between skin and mucous membrane can be clearly distinguished by HE staining.The mucous membrane had more epithelial cell layers,deep rete pegs and parakeratosis.The skin had obvious keratinized layer.Conclusions:1.Micro-mucosa graft to the wound can survive and grow to be a complete piece of flake.The epithelium becomes complete and achieves good quality on the 21st day postoperation.2.The morphology and distribution of the micro-mucosa on the wound have effects on the healing process after transplantation.The healing process goes fast and the neogenesis epithelium gets good quality when the average diameter,size and the density of the micro-mucosa is about 0.7mm,0.46mm² and 25/cm² respectively.3.The combination of micro-mucosa and micro-skin graft can accelerate the speed of wound epithelialization,however,both of them remain the histological characteristics of their own after transplantation. Part 2 Clinical research on vaginoplasty using autologous micro-mucosaObjective:The purpose of our study was to describe the procedure and outcome of creating neovagina lined with autologous micro-mucosa.To evaluate the neovagina by the terms of morphology,histology,microenvironment as well as the function and identify the significance of this method.Materials and methods:Between June 2006 and September 2008,12 patients underwent vaginaoplasty using autologous micro-mucosa graft from the oral cavity,whose age ranged from 19 to 30 years(mean 24.6 years).11 of them were congenital absence of vagina,and the remaining one complained about vaginal stenosis after vaginal reconstruction 7 years ago. The oral mucosa graft was taken from the bilateral cheek and inner lips and minced into 0.7 mm in size and was transplanted to the cavity which is dissected between the bladder and the rectum.The follow-up includes measuring the depth and width of the neovagina, colposcopy,colpography,histological detection of the vaginal wall as well as secretion analysis.The function of the neovagina was assessed by female sexual function questionnaire in the patients having sexual life 6 months after operation and compared with normal female sexual function.Results:1.The operation was performed successfully in all twelve cases.The operative time was about 112 minutes(90～140 min) and operative blood loss was 80-100 mL.The average length of stay was about 14 days(10～19 days).The time of epithelialization was about 14 days.All cases were followed up 4-26 months(average 14 months).The donor sites healed uneventfully with no change in mouth opening.2.The perineal area was not disturbed,and a neovagina formed with a depth of 8cm (6-10 cm) and a width of two fingers.The lining is pink-colored and smooth and is confirmed as nonkeratizing squamous stratified mucosa.Vaginal plica was also visible from the orifice.The cream-colored viscous fluid was found in the neovaginal cavity and its PH value was about 6.2(5.5～7.0).3.Of the twelve patients,eight were married.Seven of them answered the questionnaire and satisfied with their sexual life without pain and bleeding.There were significantly differences between the vaginoplasty group and normal control group on the total score and orgasm score(P＜0.05),while no differences were found on desire, arousal,lubrication,relationship satisfaction and comfort score(P＞0.05).Conclusions: 1.Vaginal reconstruction with autologous micro-mucosa graft is an easy,minimally invasive and useful method.The donor sites and perineal area were not disturbed.2.The neovagina lined with soft,smooth and pliable mucosa has adequate depth and width.3.Histologically,the neovaginal lining is confirmed as nonkeratinized squamous epithelium which is similar to the normal vagina.4.The neovagina achieves good function without any lubricants during sexual life.