复合碱性成纤维细胞生长因子聚乳酸/胶原支架修复兔尿道缺损  

Basic fibroblast growth factor combined with poly(lactic acid)/collagen scaffold for urethral defect in rabbits

作　　者：刘沛[1] 张冠英 余泉峰 李泽宇 韩广业[1] 吴春磊[1] Liu Pei;Zhang Guanying;Yu Quanfeng;Li Zeyu;Han Guangye;Wu Chunlei(Department of Urology Surgery,First Affiliated Hospital of Xinxiang Medical College,Xinxiang 453100,Henan Province,China)

机构地区：[1]新乡医学院第一附属医院泌尿外科,河南省新乡市453100

出　　处：《中国组织工程研究》2022年第34期5468-5474,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:碱性成纤维细胞生长因子作为有效的血管形成因子之一,不但可增加缺血组织的血液灌注量、加快组织微血管再生,还可提高肌肉组织的血管形成,增加肌肉的血流灌注量。目的:将碱性成纤维细胞生长因子复合于聚乳酸/胶原支架中,修复兔尿道缺损,提升组织工程支架诱导尿道再生的能力。方法:制备电纺聚乳酸/胶原尿道支架与负载碱性成纤维细胞生长因子的电纺聚乳酸/胶原支架,分别命名为对照支架与实验支架,检测实验支架的体外缓释性能。将第3代兔脂肪间充质干细胞接种于两种支架表面7 d,分别进行CCK-8实验、细胞黏附实验与细胞活死染色。取成年雄性新西兰大白兔24只,建立长约5 cm的尿道缺损模型,随机分2组治疗,每组12只,分别植入对照支架+脂肪间充质干细胞复合体与实验支架+脂肪间充质干细胞复合体,术后12,24周分别进行逆行尿道造影与尿道组织学观察。结果与结论:(1)体外缓释实验:在前6 d时,实验支架中的碱性成纤维细胞生长因子以平均2.5 ng/d的速率释放,在第7-14天时以平均1.52 ng/d的速率释放,此后以平均1.12 ng/d的速率释放,至第21天后释放速率逐步下降。(2)体外细胞实验显示,实验支架上的细胞增殖速率快于对照支架(P<0.05);脂肪间充质干细胞可黏附于两种支架表面,生长状态良好并伸出伪足,其中实验支架上的细胞增殖迅速;实验支架上的细胞存活率高于对照支架(P<0.05)。(3)体内实验结果:术后24周尿道造影下可见,实验支架组未见尿道狭窄,对照支架组可见轻度的尿道狭窄。术后24周组织学观察显示,实验支架组尿道组织可见较厚且较完整的上皮层,可见大量肌纤维束与血管形成;对照支架组尿道组织上皮层形成较薄,也可见少量的肌纤维束与血管形成。(4)结果表明,采用负载碱性成纤维细胞生长因子的电纺聚乳酸/胶原支架修复兔尿道缺损可BACKGROUND:As one of the effective angiogenic factors,basic fibroblast growth factor can not only increase the blood perfusion of ischemic tissues and accelerate the regeneration of tissue microvessels,but also improve the formation of blood vessels in muscle tissues and increase the blood perfusion of muscles.OBJECTIVE:Basic fibroblast growth factor was incorporated into poly(lactic acid)/collagen scaffold to repair rabbit urethral defects and to improve the ability of tissue engineered scaffolds to induce urethral regeneration.METHODS:The electrospun poly(lactic acid)/collagen urethral scaffold and the electrospun poly(lactic acid)/collagen scaffold loaded with basic fibroblast growth factor were prepared and named as control scaffold and experimental scaffold,respectively.The in vitro release properties of the scaffolds were tested.The third passage of rabbit adipose derived mesenchymal stem cells was seeded on the two scaffolds for 7 days.CCK-8 assay,cell adhesion test,and cell livedead staining were performed respectively.Twenty-four adult male New Zealand white rabbits were used to establish a 5 cm long urethral defect model and randomly divided into two groups(n=12).The control scaffold+adipose derived mesenchymal stem cell complex and the experimental scaffold+adipose derived mesenchymal stem cell complex were implanted,respectively.At 12 and 24 weeks after operation,retrograde urethrography and histology were performed.RESULTS AND CONCLUSION:(1)In vitro sustained release experiment:Basic fibroblast growth factor was released at an average rate of 2.5 ng/d in the first 6 days,1.52 ng/d from the seventh to fourteenth days,and 1.12 ng/d thereafter.The release rate gradually decreased after 21 days.(2)In vitro cell experiments showed that the cell proliferation rate on the experimental scaffold was faster than that on the control scaffold(P<0.05).Adipose-derived mesenchymal stem cells could adhere to the surfaces of the two scaffolds,grow well and protrude pseudopodia,among which the cells on the experimenta

关 键 词：碱性成纤维细胞生长因子 聚乳酸 胶原 支架 尿道缺损 尿道再生 组织工程 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]