Liu Yanbing, Fang Yan
Department of Spinal Surgery, The Third Hospital of Shijiazhuang, Hebei, China.
Department of Cardiac Surgery, The Second Hospital of Hebei Medical University, Hebei, China.
Turk J Biol. 2022 Sep 2;46(5):414-425. doi: 10.55730/1300-0152.2627. eCollection 2022.
BACKGROUND/AIM: Icariin (ICA)-loaded zein/PLGA nanofiber membrane combined with MSCs was prepared by coaxial electrospinning and bioelectrospraying.
SEM and TEM were used to evaluate the surface morphology and microstructure of the fiber membrane. Ultraviolet spectrophotometry was used to detect drug release. A LIVE/DEAD Viability/Cytotoxicity Kit and fluorescence staining were used to detect cell morphology and activity. Alkaline phosphatase and calcium mineralization deposition were used to evaluate the osteoinductive activity of the scaffold. Dynamic mechanical analysis was used to determine the Young's modulus, maximum load, and maximum elongation of the prepared scaffold. Western blot was used to detect the related protein expression in MSCs induced by drug-loaded scaffolds.
Good mechanical properties and stability were observed in the prepared drug-loaded scaffolds. SEM showed that there were a considerable number of MSCs dispersed in the scaffold. MSCs were evenly distributed, could grow evenly between fibers, and were arranged orderly along the fibers. Sustained release of ICA confers cell laden scaffold higher ALP activity and ECM mineral deposition through Runx2, OPN and OCN pathways.
Isotropic sustained release of ICA grant cell laden scaffolds rapid bone regeneration compacity which can provide a good osteogenic environment for loaded MSCs.
背景/目的:采用同轴静电纺丝和生物电喷雾法制备了负载淫羊藿苷(ICA)的玉米醇溶蛋白/聚乳酸-羟基乙酸共聚物(PLGA)纳米纤维膜并与间充质干细胞(MSCs)复合。
采用扫描电子显微镜(SEM)和透射电子显微镜(TEM)评估纤维膜的表面形态和微观结构。采用紫外分光光度法检测药物释放情况。使用活/死细胞活力/细胞毒性试剂盒和荧光染色检测细胞形态和活性。采用碱性磷酸酶和钙矿化沉积评估支架的骨诱导活性。采用动态力学分析测定所制备支架的杨氏模量、最大载荷和最大伸长率。采用蛋白质免疫印迹法检测载药支架诱导的MSCs中相关蛋白的表达。
所制备的载药支架具有良好的力学性能和稳定性。SEM显示支架中有大量分散的MSCs。MSCs分布均匀,可在纤维间均匀生长,并沿纤维有序排列。ICA的持续释放通过Runx2、骨桥蛋白(OPN)和骨钙素(OCN)途径赋予负载细胞的支架更高的碱性磷酸酶活性和细胞外基质矿化沉积。
ICA的各向同性持续释放赋予负载细胞的支架快速的骨再生能力,可为负载的MSCs提供良好的成骨环境。