Li Zhengqiang, Liu Peng, Yang Ting, Sun Ying, You Qi, Li Jiale, Wang Zilin, Han Bing
Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Jilin University, Changchun, China Jilin Provincial Key Laboratory of Tooth Development and Bone Remodeling, Changchun, China.
Department of Stomatology, School of Medicine, Yanbian University, Yanji, China.
J Biomater Appl. 2016 May;30(10):1552-65. doi: 10.1177/0885328216638587. Epub 2016 Apr 7.
Nanofibrous materials produced by electrospinning have attracted considerable attention from researchers in regenerative medicine. A combination of nanofibrous scaffold and chondrocytes is considered promising for repair of cartilage defect or damage. In the present study, we fabricated a poly(l-lactic-acid) (PLLA)/silk fibroin (SF) nanofibrous scaffold by electrospinning and evaluated its chondrogenic potential. The PLLA/SF nanofibers were characterized for diameter, surface wettability, swelling ratio, and tensile strength. Throughin vitroexperiments, PLLA/SF scaffold-chondrocyte interactions were investigated relative to the unmodified PLLA scaffold with regard to cellular adhesion, spreading, and proliferation by scanning electron microscopy and confocal laser scanning microscopy, and through analyses of DNA, sulfated glycosaminoglycan, and collagen. In addition, hematoxylin-eosin and Alcian blue-nuclear fast red staining were used to observe growth of chondrocytes, and secretion and distribution of cartilage-specific extracellular matrices in the scaffolds. Expressions of cartilage-related genes (collagen II, aggrecan, sox9, collagen I, and collagen X) were detected by real-time quantitative PCR. The PLLA/SF scaffold had better hydrophilicity, and could support chondrocytes adhesion and spreading more effectively than the unmodified PLLA scaffold. Chondrocytes secreted more cartilage-specific extracellular matrices and maintained their phenotype on the PLLA/SF scaffold. So it is concluded that the PLLA/SF scaffold is more conducive toin vitroformation of cartilage-like new tissues than the unmodified PLLA scaffold, and may be a promising material in cartilage tissue engineering.
通过静电纺丝制备的纳米纤维材料已引起再生医学领域研究人员的广泛关注。纳米纤维支架与软骨细胞的组合被认为在修复软骨缺损或损伤方面具有广阔前景。在本研究中,我们通过静电纺丝制备了聚(L-乳酸)(PLLA)/丝素蛋白(SF)纳米纤维支架,并评估了其软骨生成潜力。对PLLA/SF纳米纤维的直径、表面润湿性、溶胀率和拉伸强度进行了表征。通过体外实验,利用扫描电子显微镜和共聚焦激光扫描显微镜,以及通过对DNA、硫酸化糖胺聚糖和胶原蛋白的分析,研究了PLLA/SF支架与软骨细胞的相互作用,涉及细胞黏附、铺展和增殖,并与未改性的PLLA支架进行了比较。此外,采用苏木精-伊红染色和阿尔辛蓝-核固红染色观察软骨细胞的生长以及支架中软骨特异性细胞外基质的分泌和分布。通过实时定量PCR检测软骨相关基因(胶原蛋白II、聚集蛋白聚糖、sox9、胶原蛋白I和胶原蛋白X)的表达。PLLA/SF支架具有更好的亲水性,并且比未改性的PLLA支架更能有效地支持软骨细胞的黏附和铺展。软骨细胞在PLLA/SF支架上分泌更多的软骨特异性细胞外基质并维持其表型。因此得出结论,与未改性的PLLA支架相比,PLLA/SF支架更有利于体外形成软骨样新组织,可能是软骨组织工程中有前景的材料。