CAJ | 학술논문

目的探讨脉动式张应力环境下,在生物反应器内进行组织工程血管(TEBV)培养的可行性.方法从人脐动脉获取血管平滑肌细胞(VSMC),经体外培养扩增后接种到聚乙醇酸(PGA)支架并置于生物反应器内进行三维培养.实验组利用气动式左心循环辅助泵对VSMC施加模拟人体内血管环境(压力120 mm Hg,1 mm Hg=0.133 kPa,频率60次/min)的脉动式张应力作用.对照组无张力环境下进行三维培养.2周后收获TEBV分别行扫描电镜(SEM)检测、组织切片HE染色和Masson染色.结果实验组TEBV外观色泽鲜亮,具有一定厚度和弹性,能自我维持管腔形状.SEM显示实验组TEBV管壁表面光滑平整,横切面有丰富的细胞外基质(ECM),ECM分布均匀,把PGA完全包裹.HE染色管壁均匀致密,ECM及细胞的排列具有一定方向性,其中含有未降解的PGA碎片.Masson染色管壁胶原纤维丰富.对照组TEBV外观色泽暗淡,管壁薄,弹性差,从支架取下后管腔塌陷.SEM显示管壁表面欠光滑,横切面上ECM含量少,大部分PGA没有被包裹.组织切片染色管壁结构疏松,ECM和胶原纤维含量较少,层次感差.结论在生物反应器内的脉动式张应力环境下,可培养出具有良好形态结构的组织工程血管样组织.
목적 탐토맥동식장응력배경하,재생물반응기내진행조직공정혈관(TEBV)배양적가행성.방법 종인제동맥획취혈관평활기세포(VSMC),경체외배양확증후접충도취을순산(PGA)지가병치우생물반응기내진행삼유배양.실험조이용기동식좌심순배보조빙대VSMC시가모의인체내혈관배경(압력120 mm Hg,1 mm Hg=0.133 kPa,빈솔60차/min)적맥동식장응력작용.대조조무장력배경하진행삼유배양.2주후수획TEBV분별행소묘전경(SEM)검측、조직절편HE염색화Masson염색.결과 실험조TEBV외관색택선량,구유일정후도화탄성,능자아유지관강형상.SEM현시실험조TEBV관벽표면광활평정,횡절면유봉부적세포외기질(ECM),ECM분포균균,파PGA완전포과.HE염색관벽균균치밀,ECM급세포적배렬구유일정방향성,기중함유미강해적PGA쇄편.Masson염색관벽효원섬유봉부.대조조TEBV외관색택암담,관벽박,탄성차,종지가취하후관강탑함.SEM현시관벽표면흠광활,횡절면상ECM함량소,대부분PGA몰유피포과.조직절편염색관벽결구소송,ECM화효원섬유함량교소,층차감차.결론 재생물반응기내적맥동식장응력배경하,가배양출구유량호형태결구적조직공정혈관양조직.
Objective To explore the feasibility of incubation of tissue engineered blood vessels (TEBV) subjected to pulsatile stress in a bioreactor.Methods The vascular smooth muscle cells (VSMCs) isolated from human umbilical artery were,following in vitro incubation,seeded on polyglycolic acid (PGA) scaffold and placed into the bioreactor for three-dimensional culture.The pulsatile stress was simulated by using a gas-driven left ventricular assisted pump mimicking the human cardiovascular internal environment (pressure:120 mm Hg,1 mm Hg=0.133 kPa,frequency:60 beats/min).The control group was cultured without pulsatile stress.This was followed by harvesting of TEBVs for subsequent scanning electron microscopy (SEM),HE staining and Masson staining at week 2.Results The TEBVs in the experimental group appeared bright-colored,with considerable thickness and flexibility sufficient to maintain the lumen architecture.As revealed by SEM,the internal wall lining the TEBV was smooth,with abundant extracellular matrix (ECM) that was evenly distributed and totally surrounded the PGA in the cross-section.HE staining showed a dense uniform vascular wall with well-oriented ECM and cells juxtaposed against undegraded PGA fragments.An abundance of collagen fibers lining the vascular wall was evidenced by Masson staining.In the control group,the dim-colored TEBV was characterized by its thin wall with poor flexibility that collapsed instantaneously following removal from the scaffold.SEM revealed a rough surface with little ECM by which PGA was not mostly surrounded.Histopathology staining showed a loose architecture characterized by unremarkable ECM and collagen fibers lining the vascular wall.Conclusion TEBV-like tissue with an ideal morphology can be cultured under the pulsatile stress in a bioreactor.