左旋聚乳酸/明胶复合纳米纤维支架在软骨组织工程中的应用

The application of composite poly(L-lactic-acid)/gelatin scaffold in cartilage tissue engineering

目的观察静电纺丝技术制备左旋聚乳酸/明胶(PLLA/Gel)复合纳米纤维支架在软骨组织工程中的应用潜力,为其作为软骨组织工程支架提供实验基础。方法通过静电纺丝技术制备PLLA/Gel复合纳米纤维支架,将其与软骨细胞共培养,通过倒置荧光显微镜、激光共聚焦显微镜(CLSM)、扫描电镜(SEM)等观察细胞、支架及细胞-支架复合物的形态结构;通过MTT实验、生物化学实验等定量检测软骨细胞在支架上的增殖及分泌细胞外基质(ECM)的能力;通过实时定量PCR检测软骨细胞在支架上表达软一型胶原、二型胶原及十型胶原基因的相对定量。结果静电纺丝PLLA/Gel复合纳米纤维支架为三维多孔结构,软骨细胞能够在此支架上很好的粘附、增殖并分泌软骨特异性的ECM,且能稳定的表达软骨相关基因。结论静电纺丝PLLA/Gel复合纳米纤维支架对软骨细胞有着良好的相容性,可以作为支架材料应用于软骨组织工程中。

Objective To evaluated the chondrogenic potential of composite nanofibrous PLLA/Gel scaffold fabricated by electrospinning,and provide the experimental basis for cartilage tissue engineering. Methods We fabricated the compositenanofibrous PLLA/Gel scaffold by electrospinning. Chondrocytes were seeded on the scaffolds and cultured together. The structures and morphology of scaffold-chondrocyte were investigated by inverted fluorescence microscope,scanning electron microscopy and confocal laser scanning microscopy,and detecting the abilities of the proliferation and secreting through analyses of MTT,biochemistry experiment,etc. Expressions of cartilage-related genes（collagen I,collagen II and collagen X） were detected by real-time quantitative PCR. Results The PLLA/Gel scaffold made by electrospinning had a three-dimensional porous structure,and could support chondrocytes adhesion and proliferation. Chondrocytes could secret cartilage-specific extracellular matrices and maintained their phenotype on the PLLA/Gel scaffold. Conclusion The PLLA/Gel scaffold had a good biocompatibility with chondrocytes,and may be a promising material in cartilage tissue engineering.