中国医师杂志
中國醫師雜誌
중국의사잡지
JOURNAL OF CHINESE PHYSICIAN
2012年
8期
1009-1012
,共4页
申复进%洛若愚%张蔚%梁华%蒋艳萍
申複進%洛若愚%張蔚%樑華%蔣豔萍
신복진%락약우%장위%량화%장염평
膀胱%细胞外基质%阴道/细胞学%肌细胞,平滑肌%阴道/畸形%兔
膀胱%細胞外基質%陰道/細胞學%肌細胞,平滑肌%陰道/畸形%兔
방광%세포외기질%음도/세포학%기세포,평활기%음도/기형%토
Urinary bladder%Extracellular matrix%Vagina/cytology%Myocytes,smooth muscle%Vagina/abnormalities%Rabbits
目的 探讨膀胱无细胞基质(BACM)复合阴道平滑肌细胞修复兔阴道缺损的可行性,寻求理想的阴道修复材料.方法 将24只成年新西兰雌兔按随机数字表法分3组,每组8只,A组用于制备BACM和阴道无细胞基质(VACM),分别在B、C两组新西兰兔获取1 cm2阴道前壁组织,分离培养免阴道平滑肌细胞,免疫组化染色鉴定.以1 × 107个细胞/cm2种植于BACM和VACM上,体外培养5d后形成VACM-平滑肌细胞复合物[对照组(C组)]和BACM-平滑肌细胞复合物[实验组(C组)],用于修复兔中段阴道缺损,于移植后3、6、12周观察阴道修复情况.结果 制备的BACM和VACM均为白色透明状,HE染色和扫描电镜观察为规则排列的网状胶原纤维,无细胞碎片；体外培养的阴道平滑肌细胞为梭形,呈典型的“峰和谷”样,免疫组化染色示α-肌动蛋白阳性,种植到BACM和VACM上5h后,均可见部分细胞黏附于无细胞基质上,随时间推移,细胞逐渐增多并伸展成梭形,实验组和对照组移植后3周均可见基质表面已形成完整的上皮层,部分平滑肌细胞长入,移植后6周形成多层复合上皮,平滑肌束明显增多,移植后12周其组织结构与正常阴道组织结构相似,阴道造影证实修复部位阴道宽敞,未见明显狭窄和瘢痕形成,两组组织修复再生情况一致.结论 BACM和VACM复合阴道平滑肌细胞修复阴道缺损的组织再生规律基本相同,但BACM来源广泛,具有良好的组织相容性,是一种较好的修复阴道缺损的支架材料.
目的 探討膀胱無細胞基質(BACM)複閤陰道平滑肌細胞脩複兔陰道缺損的可行性,尋求理想的陰道脩複材料.方法 將24隻成年新西蘭雌兔按隨機數字錶法分3組,每組8隻,A組用于製備BACM和陰道無細胞基質(VACM),分彆在B、C兩組新西蘭兔穫取1 cm2陰道前壁組織,分離培養免陰道平滑肌細胞,免疫組化染色鑒定.以1 × 107箇細胞/cm2種植于BACM和VACM上,體外培養5d後形成VACM-平滑肌細胞複閤物[對照組(C組)]和BACM-平滑肌細胞複閤物[實驗組(C組)],用于脩複兔中段陰道缺損,于移植後3、6、12週觀察陰道脩複情況.結果 製備的BACM和VACM均為白色透明狀,HE染色和掃描電鏡觀察為規則排列的網狀膠原纖維,無細胞碎片；體外培養的陰道平滑肌細胞為梭形,呈典型的“峰和穀”樣,免疫組化染色示α-肌動蛋白暘性,種植到BACM和VACM上5h後,均可見部分細胞黏附于無細胞基質上,隨時間推移,細胞逐漸增多併伸展成梭形,實驗組和對照組移植後3週均可見基質錶麵已形成完整的上皮層,部分平滑肌細胞長入,移植後6週形成多層複閤上皮,平滑肌束明顯增多,移植後12週其組織結構與正常陰道組織結構相似,陰道造影證實脩複部位陰道寬敞,未見明顯狹窄和瘢痕形成,兩組組織脩複再生情況一緻.結論 BACM和VACM複閤陰道平滑肌細胞脩複陰道缺損的組織再生規律基本相同,但BACM來源廣汎,具有良好的組織相容性,是一種較好的脩複陰道缺損的支架材料.
목적 탐토방광무세포기질(BACM)복합음도평활기세포수복토음도결손적가행성,심구이상적음도수복재료.방법 장24지성년신서란자토안수궤수자표법분3조,매조8지,A조용우제비BACM화음도무세포기질(VACM),분별재B、C량조신서란토획취1 cm2음도전벽조직,분리배양면음도평활기세포,면역조화염색감정.이1 × 107개세포/cm2충식우BACM화VACM상,체외배양5d후형성VACM-평활기세포복합물[대조조(C조)]화BACM-평활기세포복합물[실험조(C조)],용우수복토중단음도결손,우이식후3、6、12주관찰음도수복정황.결과 제비적BACM화VACM균위백색투명상,HE염색화소묘전경관찰위규칙배렬적망상효원섬유,무세포쇄편；체외배양적음도평활기세포위사형,정전형적“봉화곡”양,면역조화염색시α-기동단백양성,충식도BACM화VACM상5h후,균가견부분세포점부우무세포기질상,수시간추이,세포축점증다병신전성사형,실험조화대조조이식후3주균가견기질표면이형성완정적상피층,부분평활기세포장입,이식후6주형성다층복합상피,평활기속명현증다,이식후12주기조직결구여정상음도조직결구상사,음도조영증실수복부위음도관창,미견명현협착화반흔형성,량조조직수복재생정황일치.결론 BACM화VACM복합음도평활기세포수복음도결손적조직재생규률기본상동,단BACM래원엄범,구유량호적조직상용성,시일충교호적수복음도결손적지가재료.
Objective To evaluate the feasibility of bladder extracellular matrix (BACM) seeding with autologous cultured vaginal smooth muscle cells (VSMC) repaired rabbit vaginal defect.Methods This study included 24 female rabbits.BACM and vaginal acellular matrix (VACM) were obtained from 8 rabbits by decellularization process.The other 16 rabbits were randomly divided into experimental and control groups.Vaginal tissue biopsies ( ～ 1 cm2) were harvested from female New Zealand rabbits.VSMC were cultured and stained with a-smooth muscle actin antibodies. Cultured VSMC cells were seeded on BACM ( experimental group) or VACM ( control group) at a cell density of 1 × 107 cells/cm2.The cell-seeded matrixes were cultured for 5days.In experimental group of 8 rabbits,a 2 cm segment of vagina was resected and replaced with BACM seeding with VSMC.Then the regenerative segment was studied with histological technique by hematoxylin-eosin staining after 3,6 and 12 weeks postoperative aud vaginography was performed at 12 weeks postoperative.The 8 rabbits in control group underwent the exact same procedure as above but the vaginal defect was repaired with VACM seeding with VSMC,Results The prepared BACM and VACM were transparent,HE staining and scanning electron microscopy showed the two acellular matrix was both consisted of abundant network of fibers with the regular arrangement,without cellular debris.Primary culture of VSMC was successfully established and passaged,and was uniformly spindle-shaped in the confluent state and showed a characteristic hill and valley'formation.Immunohistochemical staining showed VSMC in culture stained positively with a-smooth muscle actin antibodies.VSMC began to adhere to the BACM 5 hour after implanting and the number of the adhered cells increased with time.Cells gradually expanded and showed the typical morphology of smooth muscle cells.Three weeks after implantation in vivo,the luminal surface of matrix was completely covered by vaginal epithelial tissue,a layer of smooth muscle cells was formed in the outer surface of the matrix.Multilayered vaginal epithelial and improved development of organized muscle bundles was observed after 6 weeks.The regenerative tissue was equivalent to the normal vaginal tissue at 12 weeks postoperatively.Vaginography demonstrated the maintenance of full patency and a wide vaginal caliber without fibrosis and graft rejection.There was no significant difference in all evaluated items between experimental and control groups.Conclusions BACM has the same regenerative process as VACM in the replacement of vaginal defect.Moreover,BACM has wider source and appears to be an suitable material for vaginal replacement.
