仿生取向纤维的刚度变化对巨噬细胞极化特性的影响  

Effect of stiffness change of biomimetic aligned fibrous substrates on macrophage polarity

作　　者：沈勇 易兵成 沈炎冰[1] 唐寒 周璟 薛苏桐 张彦中[1] SHEN Yong;YI Bingcheng;SHEN Yanbing;TANG Han;ZHOU Jing;XUE Sutong;ZHANG Yanzhong(College of Chemistry,Chemical Engineering and Biotechnology,Donghua University,Shanghai 201620,China)

机构地区：[1]东华大学化学化工与生物工程学院,上海201620

出　　处：《东华大学学报（自然科学版）》2022年第2期1-9,17,共10页Journal of Donghua University(Natural Science)

基　　金：国家科技部重点研发专项子课题资助项目(2016YFC1100203);国家自然科学基金资助项目(32071345,31771050,31570969);中央高校基本科研业务费专项资金资助项目(2232019A3-09)。

摘　　要：为探索仿生取向纤维基质刚度变化对巨噬细胞响应行为的影响,采用同轴静电纺技术制备了以弹性聚(L-丙交酯-co-己内酯)(poly(lactide-co-caprolactone),PLCL)为壳层、刚性左旋聚乳酸(poly(L-lactic acid),PLLA)为芯层的不同刚度的PLCL/PLLA取向纤维。采用扫描电子显微镜、荧光染色以及cell counting kit-8等研究PLCL/PLLA取向纤维刚度变化对巨噬细胞RAW 264.7的形貌、增殖及迁移等行为的影响;利用实时定量聚合酶链反应、酶联免疫吸附检测及免疫荧光技术在基因和蛋白水平探究该巨噬细胞的极化状态变化。研究结果表明,与刚度较低的PLCL/PLLA取向纤维相比,培养在高刚度PLCL/PLLA取向纤维上的巨噬细胞具有较高的铺展、增殖和迁移能力,并且更倾向于朝促炎表型极化,巨噬细胞的表型对纤维刚度具有高度的依赖性。In order to explore the stiffness effect of biomimetic aligned fibrous substrates(AFSs) on the responsive behavior of macrophages, the elastic poly(L-lactide-co-caprolactone)(PLCL, as shell component) and rigid poly(L-lactic acid)(PLLA, as core component) were produced into aligned core-shell structured composite fibers of PLCL/PLLA by coaxial electrospinning. Then the effects of fiber stiffness on the morphology, proliferation and migration of RAW 264.7 cells were investigated by scanning electron microscope, fluorescence staining and cell counting kit-8 assay. Quantitative real time polymerase chain reaction, enzyme-linked immunosorbent assay and immunofluorescence were performed to identify the polarization states of macrophages at the gene and protein levels. The results show that compared with PLCL/PLLA aligned fibers with lower stiffness, PLCL/PLLA aligned fibers with higher stiffness promote the macrophages a higher capacity in spreading, proliferation and migration, and predispose to direct the cells into pro-inflammatory phenotype. The phenotypic function of macrophages is highly dependent on the fiber stiffness.

关 键 词：仿生取向纤维 刚度 巨噬细胞极性 静电纺丝 血管组织工程 

分 类 号：Q811.6[生物学—生物工程]