CAJ | 학술논문

背景：胆管损伤修复是腹部外科手术的难题，组织工程胆管是解决这一难题的理想方法，而构建性能优异的组织工程胆管支架是这一研究的关键。目的：观察猪骨髓间充质干细胞诱导分化的胆管内皮细胞与电纺纳米纤维的生物相容性。方法：将猪骨髓间充质干细胞于体外定向分化为肝干细胞，再进一步分化为胆管内皮细胞，并通过形态学及RT-PCR对分化完成的细胞加以鉴定。用静电纺丝方法制备聚乳酸-羟基乙酸纳米纤维膜，通过扫描电镜观察其形态，并测定其短期(2周)内体外降解率。将分化的胆管内皮细胞与纳米纤维复合培养，观察细胞在纳米纤维表面的黏附、增殖能力，荧光染色观察细胞形态及在材料表面的分布情况，扫描电镜观察细胞在纳米纤维表面生长形貌。结果与结论：骨髓间充质干细胞体外定向分化4周后，细胞呈现典型树枝状胆管内皮细胞形态学改变，并且有CK19的表达。扫描电镜照片显示电纺材料为连续的纳米纤维，纤维直径均分布在200-500 nm范围内，在2周内聚乳酸-羟基乙酸纳米纤维没有发生明显的降解。通过细胞黏附率计算、MTT 法检测、荧光染色及扫描电镜观察证明猪骨髓间充质干细胞诱导分化后的胆管内皮细胞与聚乳酸-羟基乙酸纳米纤维具有良好的生物相容性。
배경：담관손상수복시복부외과수술적난제，조직공정담관시해결저일난제적이상방법，이구건성능우이적조직공정담관지가시저일연구적관건。목적：관찰저골수간충질간세포유도분화적담관내피세포여전방납미섬유적생물상용성。방법：장저골수간충질간세포우체외정향분화위간간세포，재진일보분화위담관내피세포，병통과형태학급RT-PCR대분화완성적세포가이감정。용정전방사방법제비취유산-간기을산납미섬유막，통과소묘전경관찰기형태，병측정기단기(2주)내체외강해솔。장분화적담관내피세포여납미섬유복합배양，관찰세포재납미섬유표면적점부、증식능력，형광염색관찰세포형태급재재료표면적분포정황，소묘전경관찰세포재납미섬유표면생장형모。결과여결론：골수간충질간세포체외정향분화4주후，세포정현전형수지상담관내피세포형태학개변，병차유CK19적표체。소묘전경조편현시전방재료위련속적납미섬유，섬유직경균분포재200-500 nm범위내，재2주내취유산-간기을산납미섬유몰유발생명현적강해。통과세포점부솔계산、MTT 법검측、형광염색급소묘전경관찰증명저골수간충질간세포유도분화후적담관내피세포여취유산-간기을산납미섬유구유량호적생물상용성。
BACKGROUND:Repair of extrahepatic biliary tract injury is a difficult problem in the abdominal surgery. Tissue-engineered extrahepatic biliary tract is an ideal selection for this problem. Construction of tissue-engineered extrahepatic biliary tract with excelent performance is a key to related studies. OBJECTIVE:To investigate the biocompatibility of bile duct endothelial cels differentiated by porcine bone marrow mesenchymal stem cels with electrospun nanofibers. METHODS:Porcine bone marrow mesenchymal stem cels were induced toward biliary tract endothelial cels, which were then identified by morphology and RT-PCR. Polylactic-co-glycolic acid (PLGA) nanofiber membranes were prepared by electrospinning. The morphology was determined by scanning electron microscopy and the short-term (2-week)in vitro degradation rate was determined. Adhesion and proliferation of biliary tract endothelial cels on the nanofiber surface was analyzed by calculating the cel adhesion rate and MTT assay, respectively. Cel growth, morphology and distribution on the material surface were observed by fluorescence staining and scanning electron microscopy, respectively. RESULTS AND CONCLUSION: After 4 weeks of directed differentiation of bone marrow mesenchymal stem celsin vitro, cels showed typical morphology of dendritic bile duct endothelial cels and had the expression of CK19. Scanning electron micrographs showed that electrospun materials were continuous nanofibers with diameters between 200 and 500 nm. No significant degradation of the PLGA nanofibers was observed within 2 weeks. Based on the measured cel adhesion rate, MTT assay, fluorescence staining, and scanning electron microscopy, the differentiated cels possessed a good proliferative capacity on PLGA nanofibers. Bone marrow mesenchymal stem cels differentiated into bile duct endothelial cels in vitro. Materials prepared by the electrospinning method had a nanofiber structure, which did not significantly degrade within 2 weeks. Differentiated cels exhibit good biocompatibility with the nanofibers.