CAJ | 학술논문

目的探讨以兔口腔黏膜细胞与同种异体膀胱黏膜下脱细胞基质(BAMG)复合物构建组织工程化尿道的可行性.方法新西兰雄性兔24只,距尿道外口2.0 cm剥离尿道黏膜(2.0 cm×0.8 cm)后,随机分实验组和对照组,每组12只.切取实验组兔口腔黏膜组织分离细胞,在有灭活的3T3细胞培养皿上进行培养扩增,将培养获得的第2代口腔黏膜细胞种植于BAMG(2.2 cm×1.0 cm)上,植入实验组兔尿道缺损区域;对照组单纯采用无细胞植入的BAMG修复尿道.分别于术后1、2、6个月观察动物排尿情况,行尿道造影,8 F尿管插管确定有无狭窄;随后处死实验兔,取修复段尿道黏膜组织行组织学检查.结果细胞培养获得的口腔黏膜细胞形态均一,生长良好;组织形态学、扫描电镜观察见口腔黏膜细胞与BAMG具有良好的相容性.实验组兔术后1、2、6个月伤口愈合良好、排尿通畅,无尿瘘发生,组织学和尿道造影检查显示带细胞修复的尿道形态完整、清晰宽敞,无狭窄发生;术后6个月植入的口腔黏膜细胞仍然存在,并明显扩增.对照组兔则出现排尿困难、尿道狭窄,光镜下发现黏膜及黏膜下存在严重的炎症反应.结论兔口腔黏膜细胞与同种异体BAMG复合后,可成功用于尿道缺损的修复,构建组织工程化尿道.
목적 탐토이토구강점막세포여동충이체방광점막하탈세포기질(BAMG)복합물구건조직공정화뇨도적가행성.방법 신서란웅성토24지,거뇨도외구2.0 cm박리뇨도점막(2.0 cm×0.8 cm)후,수궤분실험조화대조조,매조12지.절취실험조토구강점막조직분리세포,재유멸활적3T3세포배양명상진행배양확증,장배양획득적제2대구강점막세포충식우BAMG(2.2 cm×1.0 cm)상,식입실험조토뇨도결손구역;대조조단순채용무세포식입적BAMG수복뇨도.분별우술후1、2、6개월관찰동물배뇨정황,행뇨도조영,8 F뇨관삽관학정유무협착;수후처사실험토,취수복단뇨도점막조직행조직학검사.결과 세포배양획득적구강점막세포형태균일,생장량호;조직형태학、소묘전경관찰견구강점막세포여BAMG구유량호적상용성.실험조토술후1、2、6개월상구유합량호、배뇨통창,무뇨루발생,조직학화뇨도조영검사현시대세포수복적뇨도형태완정、청석관창,무협착발생;술후6개월식입적구강점막세포잉연존재,병명현확증.대조조토칙출현배뇨곤난、뇨도협착,광경하발현점막급점막하존재엄중적염증반응.결론 토구강점막세포여동충이체BAMG복합후,가성공용우뇨도결손적수복,구건조직공정화뇨도.
Objective To investigate the feasibility of replacing urinary epithelial cells with oral keratinocytes by being seeded on bladder acellular matrix graft(BAMG). Methods Twenty-four male rabbits were randomly divided into 2 groups:experimental group and control group.A length of 2.0 cm and width of 0.8 cm penile urethral mucosal defect was induced in the anterior urethra 2.0 cm awav from the urinary meatus in all the rabbits.Oral keratinocytes were isolated from a small buecal mucosa(1.0 cm×0.4 cm)of the 12 rabbits in experimental group and seeded onto a culture dish with a feeder layer of 3T3 mouse fibroblasts inhibited by mitomycin(i3T3).Passage 2 oraI keratinocytes cuItured with i3T3 were expanded and seeded onto sterilized BAMG(2.2 cm×1.0 cm)to repair the de-fects of the urethra.Urethroplasty was performed with BAMG with no cell seeding in the controlgroup.Catheter examination and retrograde urethI ography was done in 1,2 and 6 months after graft-ing.The urethral grafts were harvested and analyzed by histological staining. Results Oral kerati-nocytes seeded onto a feeder layer of i3T3 had better morphous and amplification capability.The corn-patibility of the compound graft was assessed by HE staining and scanning electron microscope.Twelve rabbits in the experimental group could void without difficulty.Catheter examination and ret-rograde urethrogram revealed that a wide urethral caliber was maintained with no sign of strictures and fistula formation.Histological examination showed the grafted urethra was covered with smooth mu- cosa with no strictures at 1,2 and 6 months.The oral keratinocytes of the graft still existed even 6 months after grafting.On contrast,gross and urethrogram demonstrated urethral stricture in the con-trol group.And inflammatory reactions could be found in the area of the stricture through light micro-scope. Conclusions Oral keratinocytes have good compatibility with BAMG and the compound graft could be used for urethral reconstruction in the animal model.