ObjectiveTo evaluate the effectiveness of acellular dermal matrix (ADM) with autologous buccal micro mucosa and micro skin graft in vaginoplasty. MethodsA retrospective analysis was made on the clinical data of 67 patients with vaginal agenesis treated between July 2006 and June 2013. ADM and mixed particles were used in 20 cases (ADM group) and mixed particles graft in 47 cases (control group) in vaginoplasty. There was no significant difference in age between 2 groups (t=0.233, P=0.816). The depth, diameter, and volume of neovagina, epithelization time, stent needing time, and female sexual function index (FSFI) score were compared between 2 groups. ResultsThere was no significant difference in operation time and amount of bleeding between 2 groups (t=-1.922, P=0.059; t=0.398, P=0.692). The patients were followed up 11-38 months (mean, 16.08 months). Fifteen cases in ADM group and 29 cases in control group had sexual life after operation. Bleeding after operation occurred in 6 cases (2 in ADM group and 4 in control group). No stenosis was observed. Difference in epithelization time was not statistically significant (t=-1.938, P=0.057). However, the stent needing time of ADM group was significantly shorter than that of control group (t=7.020, P=0.000). The neovagina was ideal in wetness degree, smoothness, flexibility, and hairlessness during follow-up. The depth, diameter, and volume of vagina had no significant difference between 2 groups (P>0.05) at last follow-up, which were close to normal vagina. The other patients had normal sexual function except 1 patient whose FSFI score was less than 23; no statistically significant difference was found in FSFI score between 2 groups (P>0.05). ConclusionOn the basis of mixed particles grafting, the ADM could improve trestle structure for resisting contracture. The effectiveness is better than merely mixed particles graft. The procedure has satisfactory anatomical and functional results.
Objective To investigate a method of repairing hypospadias by combining buccal mucosal graft with scrotal flap and its therapeutic effect. Methods From March 2002 to December 2007, 42 patients with hypospadias underwent primary urethral reconstruction using buccal mucosal graft and scrotal flap. The patients ranged in age from 18 months to 18 years. There were 21 cases of penoscrotal type, 12 cases of scrotal type and 9 cases of perineal type. Among them,8 cases were at initial operation, and 34 cases suffered from the failure of hypospadias repair 6-19 months (average 10 months) after initial operation. During operation, the defect of urethra was 3-7 cm (average 4.2 cm) when the penis was straightened; the buccal mucosa (3.0 cm × 1.2 cm-7.0 cm × 1.5 cm) was transplanted to the tunica albuginea in the ventral aspect of the penis, and was paired with the scrotal flap (3.0 cm × 1.5 cm-7.0 cm × 1.5 cm) to repair urethra. Results The incision of 38 cases healed by first intention, and no compl ication occurred. At 7 days after operation, 4 cases had urinary fistula at either coronary sulcus or anastomotic stoma, one of which spontaneously closed 2 months after operation and the rest 3 recovered by repairing urinary fistula 6 months after operation. All patients were followed for 3-48 months (average 18 months). Urination was smooth, the reconstructed urethral opening was at the tip of glans peins without retraction and with apperance similar to the normal urethral opening. The appearance of penis and scrotum was satisfying, and the penis was straightened completely. Conclusion Combined buccal mucosal graft and scrotal flap, with considerable tissue for uretha tract reconstruction and low incidence rate of urethral stricture, is one of the effective methods to repair hypospadias.
Objective To investigate the growth of the tissue engineered mucosa after its heterotransplantation. Methods The epithelial cells and fibroblasts were isolated from a postoperative tissue of the 3month patient with labialcleft. The epithelial cells and fibroblasts were separately seeded on the polylactic／glycolic acid copolymer membrane, and then they were exposed to the air-liquid interface. Seven volunteer patients, whose traumatic beds were repaired with the tissue engineered oral mucosa. The biopsy tissue from one of the seven patients was observed under light icroscope 18 and 30 days after transplantation, respectively. Results The tissue engineered oral mucosa having 5-6 layers anticytoeratin staining positively cells in the epithelial layer and 3-7 layers anti-Cytoeratin staining negatively cells in the subepithelial layer grew well after the he terotransplantation. No differ ence could be found between the transplanting and normal areas. At 18 days, the epithelial layer and lamina propria grew well and the fibroblasts were found; at 30 days, collagen was obviously observed. The structures in both the transplanting and the normal areas were similar. Conclusion The tissue engineered oral mucosa can grow well after the heterotransplantation.
【摘要】目的探讨颌下腺移位术对预防急性放射性口腔黏膜反应及口干燥症的临床效果。方法2007年7月2009年6月间选择40例患者进行前瞻性临床对照研究。治疗组20例,在放疗前将颌下腺移位至颊下区。对照组20例不行颌下腺移位术。观察放疗中两组急性口腔黏膜反应,测定放疗前后唾液分泌量的变化,放疗后3个月进行口干燥程度问卷调查。结果治疗组急性口腔黏膜反应明显轻于对照组(Plt;0.05)。治疗组放疗后3个月移位术侧颌下腺摄取、排泌功能均明显较对照好,两组比较有统计学意义(Plt;0.05)。结论颌下腺移位术预防鼻咽癌放疗后口干燥症的临床近期疗效较好,可改善鼻咽癌患者放疗后的生活质量。
Objective To observe the therapeutic effect of thermosensitive hydrogel containing curcumin-vitamin E (VE) complex (hereinafter referred to as “curcumin-VE hydrogel”) on radiation-induced oral mucositis in mice. Methods Curcumin-VE hydrogel was prepared using the synthesized curcumin-VE complex as the carrier and poloxam as the substrate. The structure of curcumin-VE complex was characterized by Fourier transform infrared spectrometer, the microstructure of curcumin-VE hydrogel was determined by scanning electron microscope, and the gelation temperature was determined by rheometer, gel swelling and degradation were tested and gel adhesion was determined using a universal testing machine. Thirty healthy male BALB/C mice with specific pathogen free grade were randomly divided into three groups, with ten mice in each group. The radiation group and radiation+hydrogel group were modeled by a single high dose of radiation (25 Gy), while the control group had anesthesia but no radiation. The control group and radiation group were given daily feed and water 7 days after radiation. In addition to daily feed and water, the radiation+hydrogel group was given curcumin-VE hydrogel twice a day. The mice were sacreficed on the 8th day after radiation. The weight changes of each group were recorded after radiation. The ulceration area of tongue was measured by toluidine blue. The tongue of mouse were pathologically observed. The activities of superoxide dismutase, catalase (CAT), and glutathione peroxidase and the level of malondialdehyde in tongue tissue were determined. The levels of tumor necrosis factor α (TNF-α), interleukin (IL)-1β and IL-6 in tongue tissue were determined by enzyme linked immunosorbent assay. The distribution and positive expression of phosphorylated histone H2AX (γ-H2AX) and nuclear factor-erythroid 2-related factor 2 were determined by immunohistochemistry. Results Curcumin-VE hydrogel had a porous network structure and the gelation temperature was 30℃, the swelling rate was close to 300%, the gel degradation rate was up to 95% after 48 h, and the adhesion strength was 12.748 kPa. Compared with the radiation group, the weight of mice in the radiation+hydrogel group increased (P<0.05), the ulcer area decreased (P<0.05); the activity of CAT increased (P<0.05); the levels of TNF-α, IL-1β and IL-6 decreased (P<0.05); the expression of γ-H2AX was down-regulated (P<0.05). Conclusion Curcumin-VE hydrogel can delay or weaken the process of radiation-induced oral mucositis by reducing the DNA damage caused by radiation, inhibiting the production of reactive oxygen species, and effectively reducing the level of inflammation in tongue tissue.
Objective To investigate the possibility of culturing human oral keratinocyte using autologous serum in order to provide theoretical and technical foundation for clinical application of tissue engineering oral mucosa epithelium.Methods The human oral keratinocytes were cultured by the medium containing different concentrations of autologous serum(10%,20%,30%)and fetalbovine serum (10%), respectively. The growth conditions for the cell and the mucosa epithelium in the groups were observed, the cell growth curves were drawn, and the population doubling time (PDT) was counted. Results The results showed that the human oral keratinocyte could proliferate well in the medium containing autologous serum or fetal bovine serum. The differences in the 24hour clone rate and PDT were not significant. Both the area and the thickness of the cultured oral epithelium increased with the increase of the autologous serum concentration, and the difference between autologous serum and fetal bovine serum was significant, especially with the medium containing 20% autologous serum( P＜0.05) . The human nature of the cultured epithelium was demonstrated by the immunofluorescent mouse anti-HLA antigen. Conclusion The autologous serum can replace the fetal bovine serum to culture the oral keratinocyte well, and the cultured oral mucosa epithelium can be better differentiated in the autologous serum than in the fetal bovine serum.
Objective To investigate the feasibil ity of replacing urinary epithel ial cells with oral mucosa cell to reconstruct tissue engineered urethra by being seeded on bladder acellular matrix graft (BAMG). Methods Eighteen male New Zealand rabbits, aged 10 weeks, weighing 0.3-0.5 kg, were used in this study. Oral mucosa cell of 12 rabbits were isolated and seeded onto a culture dish with a feeder layer of 3T3 and a culture dish without 3T3, respectively. The morphologic change and growth condition of oral mucosa cells were observed by inverted phase contrast microscope after 2 days of seeding. The quantity of oral mucosa cells was counted using cell counting meter; the cell growth curve was drawn and the immunofluorescence staining with broad-spectrum keratin antibody was carried out. The bladders taken from the rest 6 rabbits were decelluled to make BAMG and the tissue of 1 cm × 1 cm was randomly selected to observe the effect of acellularization. The second passage oral mucosa cells cultured with 3T3 were appl ied to steril ized BAMG to obtain a issueengineered mucosa. The tissue-engineered mucosa was assessed using HE staining and scanning electron microscope after being cultured for 1 week. Results Oral mucosa cells seeded onto a feeder layer of 3T3 could be passaged for 7 or 8 generations with homogeneous forms and full function. Oral mucosa cells cultured without 3T3 could only be subcultured for 2 generations before aging and had multiple shapes and different sizes. Oral mucosa cells cultured by the two methods both started logarithmic growth on the 8th day and reached the peak value on the 14th day, which was indicated by the cell growth curve. However, more cells could be obtained through oral mucosa cells cultured with 3T3 than those cultured without 3T3. Oral mucosa cells manifestated green colour fluorescence cultured with or without 3T3. After the cells were removed, the BAMG presented as a porous membrane. The HE staining showed that the effect of acellularization was good and there were no cells at BAMG. The second passage oral mucosa cells cultured with 3T3 were expanded and seeded onto steril ized BAMG to obtain a tissue-engineered mucosa. Good compatibil ity of the compound graft was assessed using HE staining and scanning electron microscope. HE staining and scanning electron microscope showed that oral mucosa cells had good biocompatibil ity with BAMG after the tissue engineered mucosa was cultured for 1 week. Conclusion Oral mucosa cells of rabbit can be cultured in vitro and attain magnitude quantities. Oral mucosa cell also have good biocompatibil ity with BAMG and the compound graft could be a new material for urethral reconstruction.