CAJ | 학술논문

目的构建仿生肝脏三维空间血管网络,探讨这一组织工程实质脏器三维血管网络构建新方法的可行性.方法利用64排计算机断层扫描(CT)采集正常人肝脏血管并转换成4 cm×4 cm血管断层平面坐标图,根据血管不同断面平面坐标图进行激光打孔处理.扫描电子显微镜(SEM)检测打孔后,膜片变化.将打孔后的各膜片按次序通过基质胶粘贴在一起.以绿色荧光蛋白(GFP)-腺病毒标记骨髓干细胞诱导的内皮细胞,将标记的内皮细胞灌注于仿生血管网络中.放入37 ℃、5％CO2培养箱培养,在培养的第3周检测血管形成状况.结果 GFP-腺病毒按照感染复数(MOI) =0、5、25、50、100转染间充质干细胞(MSCs)1周后,MOI=100阳性标记率最高[(82.13±3.65)％],细胞增殖实验显示5组间细胞增殖率差异无统计学意义(P＞0.05).内皮细胞迁移实验6h后,5组细胞迁移数分别为(32.54±1.14)、(28.13±1.43)、(28.89±1.01)、(27.28 ±0.52)、(33.14±0.98)个/视野,差异无统计学意义(P＞0.05).激光打孔后,扫描电镜观察孔径边缘,孔径边缘光滑平整.经3周后免疫荧光检测,可见管壁形成内皮细胞层.结论针对组织工程脏器血管网络的构建,将为今后组织工程脏器构建奠定研究基础.
목적 구건방생간장삼유공간혈관망락,탐토저일조직공정실질장기삼유혈관망락구건신방법적가행성.방법 이용64배계산궤단층소묘(CT)채집정상인간장혈관병전환성4 cm×4 cm혈관단층평면좌표도,근거혈관불동단면평면좌표도진행격광타공처리.소묘전자현미경(SEM)검측타공후,막편변화.장타공후적각막편안차서통과기질효점첩재일기.이록색형광단백(GFP)-선병독표기골수간세포유도적내피세포,장표기적내피세포관주우방생혈관망락중.방입37 ℃、5％CO2배양상배양,재배양적제3주검측혈관형성상황.결과 GFP-선병독안조감염복수(MOI) =0、5、25、50、100전염간충질간세포(MSCs)1주후,MOI=100양성표기솔최고[(82.13±3.65)％],세포증식실험현시5조간세포증식솔차이무통계학의의(P＞0.05).내피세포천이실험6h후,5조세포천이수분별위(32.54±1.14)、(28.13±1.43)、(28.89±1.01)、(27.28 ±0.52)、(33.14±0.98)개/시야,차이무통계학의의(P＞0.05).격광타공후,소묘전경관찰공경변연,공경변연광활평정.경3주후면역형광검측,가견관벽형성내피세포층.결론 침대조직공정장기혈관망락적구건,장위금후조직공정장기구건전정연구기출.
Objective To construct biomimetic vascular network of tissue engineering liver scaffold.Methods Using 64 row CT to collect normal liver blood vessels image,then transform the CT images into 4 cm× 4 cm vascular fault plane coordinate chart,according to liver vessels different section plane coordinates,laser prepare biomimetic vascular network on the electrospun Poly (EC-CL) scaffolds.The scanning election microscopy (SEM) tested the bionic vascular network.each of the liver tissue engineering scaffolds will punch in the order attached together using the matrigel adhesive.Using green fluorescent protein (GFP)-adenovirus marked euro-collins solution (ECs),then implanted ECs into bionic blood vessels in the bionic network.Cultured the bionic liver vessel network at 37 ℃,5％ CO2 incubator,after 3 weeks culture,tested the formation of blood vessels with fluorescence microscope.Results After GFP-adenovirus transfected mesenchymal stem cells (MSCs) as MOI =0,5,25,50,100 a week,the group of MOI =100 has the highest transfected rate (82.13 ± 3.65) ％.Cells proliferation test shows no statistical significance in all 5 groups (P ＞ 0.05).Endothelial cells migration test still has no statistical significance (P＞0.05).the 5 groups migration date:(32.54 ± 1.14),(28.13±1.43),(28.89± 1.01),(27.28 ± 0.52) and (33.14 ± 0.98)/HP respectively.After laser drilling,scanning electron microscopy (sem) shows the bionic vascular edge smooth.After 3 weeks culture,immune fluorescent detected visible wall endothelial layer formation.Conclusion Helped to promote metabolism functional tissue engineering viscera research progress.