CAJ | 학술논문

目的使用可降解材料聚乳酸和聚羟基乙酸二元共聚物[poly(lactic-co-glycolic acid),PLGA]作为组织工程人工血管支架,探讨种子细胞培养条件,为供临床使用的小口径人工血管研制提供参考.方法以PLGA作为细胞基质,新生儿脐带血管提取的平滑肌细胞和内皮细胞作种子细胞,体外培养4周后,将平滑肌细胞种植到PLGA上,4周后将内皮细胞种植到平滑肌细胞上,再4周后取出含此细胞的血管片段行细胞性质及活性检测,了解细胞在PLGA上的存活及生长状态.结果免疫组化检测人工血管中平滑肌肌动蛋白、内皮细胞Ⅷ因子及CD31、CD34均呈阳性表达,证实接种细胞为平滑肌细胞和内皮细胞；放免法检测PLGA细胞培养液,内皮素和6-酮-前列腺素含量明显高于单纯无血清培养液(P＜0.05),证明种植到PLGA内表面的细胞具有活性；光镜和电镜下接种细胞呈典型的平滑肌细胞、内皮细胞形态特征.结论新生儿脐带血管提取的内皮细胞及平滑肌细胞作为种子细胞,体外培养并种植到PLGA上,短期内仍具有生物活性.
목적 사용가강해재료취유산화취간기을산이원공취물[poly(lactic-co-glycolic acid),PLGA]작위조직공정인공혈관지가,탐토충자세포배양조건,위공림상사용적소구경인공혈관연제제공삼고.방법 이PLGA작위세포기질,신생인제대혈관제취적평활기세포화내피세포작충자세포,체외배양4주후,장평활기세포충식도PLGA상,4주후장내피세포충식도평활기세포상,재4주후취출함차세포적혈관편단행세포성질급활성검측,료해세포재PLGA상적존활급생장상태.결과 면역조화검측인공혈관중평활기기동단백、내피세포Ⅷ인자급CD31、CD34균정양성표체,증실접충세포위평활기세포화내피세포；방면법검측PLGA세포배양액,내피소화6-동-전렬선소함량명현고우단순무혈청배양액(P＜0.05),증명충식도PLGA내표면적세포구유활성；광경화전경하접충세포정전형적평활기세포、내피세포형태특정.결론 신생인제대혈관제취적내피세포급평활기세포작위충자세포,체외배양병충식도PLGA상,단기내잉구유생물활성.
Objective The degradable artificial material—poly(lactic-co-glycolic acid) (PLGA) was used as the scaffold of the tissue engineering blood vessel for cells implantation,and the better requirement for cell culture was studied.These results may be useful to the manufacture of small diameter tissue engineering blood vessel used in clinical in future.Methods The nonwoven mesh degradable PLGA was used as tissue engineering scaffold.The smooth muscular cell(SMC) and endothelial cell (EC) harvested from fetus umbilical blood vessel were collected as the seeding cells.The SMC and EC were cultivated for 4 weeks respectively in vitro.Then,the SMC was seeded on the surface of the PLGA 4 weeks later,the EC was seeded on the surface of the SMC layer and PLGA.After another 4 weeks,the artificial blood vessel segment with seeding cells was examined in the cellular quality and activity.Results By immunohistochemical method,all test expressions were positive in staining smooth musle actin,yon Willebrand 、CD31 and CD34.By radioimmunoassay,the levels of endothelin and 6-Keto-PGF1 a in PLGA cell culture solution were higher than in control soulture (P ＜ 0.05).These demonstrated that all cells seeded on PLGA still have cellular activity.All seeding cells showed the typical SMC or EC morphological appearances in light microscopic and scanning electric microscopic analyses.Conclusion The results show that the SMC and EC from umbilical vessel,after being cultivated in vitro,as the seeding cells seeded sequentially on the degradable blood vessel scaffold (PLGA),still possess the cellular activity.This may be useful in further studying the small diameter tissue engineering blood vessel.