CAJ | 학술논문

背景:有实验表明聚羟基乙酸支架材料已成功在裸鼠及大型哺乳动物体内构建形成了组织工程化软骨、骨、肌腱等组织,将血管内皮细胞与平滑肌细胞接种于聚羟基乙酸能否在裸鼠皮下形成血管样结构?目的:采用新生婴儿脐静脉血管内皮细胞与平滑肌细胞接种于片状聚羟基乙酸形成的复合物移植于裸鼠皮下,验证其形成血管样结构的可行性.设计:对比观察.单位:原上海第二医科大学组织工程重点实验室.材料:实验于2002-01/06在上海第二医科大学组织工程重点实验室完成.新生婴儿脐带来源于本院妇产科健康新生儿,经产妇知情同意,实验经过医院伦理委员会的批准.选用26只3～4周龄清洁级裸鼠,雌雄不拘,实验过程中对动物的处置符合动物伦理学标准.聚羟基乙酸购自Albany International Research Co..方法:将体外培养、扩增的新生婴儿脐静脉血管内皮细胞与平滑肌细胞接种于片状聚羟基乙酸材料上,形成细胞-材料复合物,将该复合物包绕于硅胶管使成管状结构后移植于20只裸鼠皮下为实验组,将未接种细胞的单纯聚羟基乙酸置入其余6只裸鼠皮下.主要观察指标:分别于细胞.生物材料复合物置入后第2,6周后取出两组移植物进行大体观察,并进行苏木精-伊红染色及免疫组织化学染色,检测Ⅷ因子及α-平滑肌肌动蛋白的表达.结果:裸鼠26只均进入结果分析.①大体观察结果:移植物置入2周后,两组移植物均有管状结构形成;置入6周后,对照组管形结构消失,围绕硅胶管有极薄的纤维组织包裹形成,抽去硅胶管后,纤维组织失去支撑,无法维持管型结构.实验组抽去硅胶管后,新生组织仍维持管型结构.②组织学观察及免疫组化检测结果:移植物置入2周后,两组均可在镜下观察到大量未完全降解的聚羟基乙酸,对照组内少有细胞成分,实验组内可见多量细胞存在;置入6周后,两组聚羟基乙酸成分基本消失,对照组只有菲薄的纤维结缔组织形成,实验组显示有新生血管组织形成,其最内层有Ⅷ因子阳性细胞覆盖,管壁内有大量细胞外基质及散在的α-平滑肌肌动蛋白阳性细胞存在.结论:血管内皮细胞、平滑肌细胞.聚羟基乙酸复合物置入裸鼠皮下可形成具有与正常血管组织学结构相似的血管. 揹景:有實驗錶明聚羥基乙痠支架材料已成功在裸鼠及大型哺乳動物體內構建形成瞭組織工程化軟骨、骨、肌腱等組織,將血管內皮細胞與平滑肌細胞接種于聚羥基乙痠能否在裸鼠皮下形成血管樣結構?目的:採用新生嬰兒臍靜脈血管內皮細胞與平滑肌細胞接種于片狀聚羥基乙痠形成的複閤物移植于裸鼠皮下,驗證其形成血管樣結構的可行性.設計:對比觀察.單位:原上海第二醫科大學組織工程重點實驗室.材料:實驗于2002-01/06在上海第二醫科大學組織工程重點實驗室完成.新生嬰兒臍帶來源于本院婦產科健康新生兒,經產婦知情同意,實驗經過醫院倫理委員會的批準.選用26隻3～4週齡清潔級裸鼠,雌雄不拘,實驗過程中對動物的處置符閤動物倫理學標準.聚羥基乙痠購自Albany International Research Co..方法:將體外培養、擴增的新生嬰兒臍靜脈血管內皮細胞與平滑肌細胞接種于片狀聚羥基乙痠材料上,形成細胞-材料複閤物,將該複閤物包繞于硅膠管使成管狀結構後移植于20隻裸鼠皮下為實驗組,將未接種細胞的單純聚羥基乙痠置入其餘6隻裸鼠皮下.主要觀察指標:分彆于細胞.生物材料複閤物置入後第2,6週後取齣兩組移植物進行大體觀察,併進行囌木精-伊紅染色及免疫組織化學染色,檢測Ⅷ因子及α-平滑肌肌動蛋白的錶達.結果:裸鼠26隻均進入結果分析.①大體觀察結果:移植物置入2週後,兩組移植物均有管狀結構形成;置入6週後,對照組管形結構消失,圍繞硅膠管有極薄的纖維組織包裹形成,抽去硅膠管後,纖維組織失去支撐,無法維持管型結構.實驗組抽去硅膠管後,新生組織仍維持管型結構.②組織學觀察及免疫組化檢測結果:移植物置入2週後,兩組均可在鏡下觀察到大量未完全降解的聚羥基乙痠,對照組內少有細胞成分,實驗組內可見多量細胞存在;置入6週後,兩組聚羥基乙痠成分基本消失,對照組隻有菲薄的纖維結締組織形成,實驗組顯示有新生血管組織形成,其最內層有Ⅷ因子暘性細胞覆蓋,管壁內有大量細胞外基質及散在的α-平滑肌肌動蛋白暘性細胞存在.結論:血管內皮細胞、平滑肌細胞.聚羥基乙痠複閤物置入裸鼠皮下可形成具有與正常血管組織學結構相似的血管.
배경:유실험표명취간기을산지가재료이성공재라서급대형포유동물체내구건형성료조직공정화연골、골、기건등조직,장혈관내피세포여평활기세포접충우취간기을산능부재라서피하형성혈관양결구?목적:채용신생영인제정맥혈관내피세포여평활기세포접충우편상취간기을산형성적복합물이식우라서피하,험증기형성혈관양결구적가행성.설계:대비관찰.단위:원상해제이의과대학조직공정중점실험실.재료:실험우2002-01/06재상해제이의과대학조직공정중점실험실완성.신생영인제대래원우본원부산과건강신생인,경산부지정동의,실험경과의원윤리위원회적비준.선용26지3～4주령청길급라서,자웅불구,실험과정중대동물적처치부합동물윤리학표준.취간기을산구자Albany International Research Co..방법:장체외배양、확증적신생영인제정맥혈관내피세포여평활기세포접충우편상취간기을산재료상,형성세포-재료복합물,장해복합물포요우규효관사성관상결구후이식우20지라서피하위실험조,장미접충세포적단순취간기을산치입기여6지라서피하.주요관찰지표:분별우세포.생물재료복합물치입후제2,6주후취출량조이식물진행대체관찰,병진행소목정-이홍염색급면역조직화학염색,검측Ⅷ인자급α-평활기기동단백적표체.결과:라서26지균진입결과분석.①대체관찰결과:이식물치입2주후,량조이식물균유관상결구형성;치입6주후,대조조관형결구소실,위요규효관유겁박적섬유조직포과형성,추거규효관후,섬유조직실거지탱,무법유지관형결구.실험조추거규효관후,신생조직잉유지관형결구.②조직학관찰급면역조화검측결과:이식물치입2주후,량조균가재경하관찰도대량미완전강해적취간기을산,대조조내소유세포성분,실험조내가견다량세포존재;치입6주후,량조취간기을산성분기본소실,대조조지유비박적섬유결체조직형성,실험조현시유신생혈관조직형성,기최내층유Ⅷ인자양성세포복개,관벽내유대량세포외기질급산재적α-평활기기동단백양성세포존재.결론:혈관내피세포、평활기세포.취간기을산복합물치입라서피하가형성구유여정상혈관조직학결구상사적혈관.
BACKGROUND: A lot of researches have proved that polyglicolide acid (PGA), as a stent material, has been successfully used to construct engineered tissues, such as cartilage, bone and tendons, in nude mice or even big mammal. Whether the incubation of vascular endothelial cells and smooth muscle cells in the polyglicolide acid may subcutaneously construct vessel-like structure in nude mice needs a further study.OBJECTIVE: To verify the feasibility of forming vessel-like structure in the nude mice by subcutaneous implantation of polyglicolide acid cocultured with vascular endothelial cells and smooth muscle cells derived from newborn umbilical vein.DESIGN: Contrast study.SETTING: Tissue Engineering Key Laboratory, Medical College, Shanghai Jiao Tong University.MATERIALS: This study was performed at Tissue Engineering Key Laboratory, Medical College, Shanghai Jiao Tong University from January to June 2002. Belly band was derived from newborn babies in our department of obstetrics & gynecology. The parturien provided the informed consent, and this study was approved by the local research ethics committee. Twenty-six nude mice (3-4 weeks old, clean grade, irrespective of gender) were selected in this study. The animal experiment received confirmed consent from the local ethic committee. Polyglicolide acid was provided by Albany International Research Co.METHODS: Vascular endothelial cells and smooth muscle cells derived from newborn umbilical vein were incubated on a piece of polyglicolide acid to produce cell-material compound. In addition, the compound covered around the silicone tube to form a tube-like structural substance. Subsequently, the tube-like structural substance was subcutaneously implanted in 20 nude mice, which were regarded as an experimental group. And then, polyglicolide acid alone was subcutaneously transplanted in the rest 6 nude mice, which was regarded as a control group.MAIN OUTCOME MEASURES: Gross observations of cell-material compound by 2 and 6 weeks after transplantation;HE staining and immunohistochemical staining.detection; expression of factor Ⅷ and α-smooth muscle actin.RESULTS: Twenty-six nude mice were included in the final analysis. ① Gross observation: At 2 weeks after implantation,both the experimental and control groups formed tubular structures, however, at 6 weeks after implantation, the tubular structure still remained in experimental group but not in the controls. ② Histological observation and immunohistochemical detection: The histological examination of the engineered vessel showed that at 2 weeks, the vessels in both group contained mainly undegraded PGA fibers, while at 6 weeks, the PGA fibers were almost completely degraded in both groups and in the control group only fibrous-like tissue formed. Contrastly, in experimental group a typical vascular structure formed, Masson's trichrome stain, which stains collagen green, smooth muscle fibers red and cells purple, showed significant amounts of stainable collagen and smooth muscle fibers in the wall of the engineered vessel, furthermore,immunohistochemistry examination revealed that there were an endothelial cell layer formed in the inner surface of the engineered vessel which was confirmed by positive staining of yon Willebrand factor, meanwhile, the smooth muscle cells in the wall of the engineered vessel were confirmed by the positive staining of smooth muscle α-actin.CONCLUSION: The subcutaneous implantation of polyglicolide acid cocultured with vascular endothelial cells and smooth muscle cells may form vessels, which are similar to normal vascular histological structure.