基于不同仿生纤维密度的细胞响应行为研究  

Effect of Different Biomimetic Fiber Density on Cellular Responses

作　　者：周璟 唐寒 易兵成 沈炎冰[1] 沈勇 薛苏桐 张彦中[1] ZHOU Jing;TANG Han;YI Bingcheng;SHEN Yanbing;SHEN Yong;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：《中国细胞生物学学报》2021年第4期747-758,共12页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:32071345、31771050、31570969);国家重点研究发展计划(批准号:2016YFC1100203);东华大学中央高校基本科研业务费学科交叉(理工科)重点计划项目(批准号:2232019A3-09)资助的课题。

摘　　要：在组织再生中,电纺纤维具有仿生天然细胞外基质(ECM)纤维结构的优势,可为理解细胞对其生存微环境的动态重塑过程提供理想的研究平台,但常规电纺纤维膜的致密性带来的纤维间横纵交叉相互牵掣问题,最终导致其作用与表面呈纳米纤维状拓扑结构的实体材料类似,因而不能"精准"了解细胞对仿生纤维微环境的重塑作用。该研究通过控制电纺纤维的沉积时间,在经硅烷化处理的玻片上接收不同密度的明胶/聚己内酯(GT/PCL)纤维基质,然后通过CCK-8、骨架染色、SEM、qRT-PCR、ELISA、免疫荧光染色等方法,研究纤维密度变化对C3H/10T1/2细胞的形貌、增殖、ECM分泌及基质降解能力的影响。结果表明,细胞活力与纤维密度相关,合适的低密度微量纤维(103.0±13.2) μg/cm^(2)更利于实现细胞重塑纤维基质微环境。该研究为今后"精准"设计仿生纤维基质用于组织再生以及深入了解细胞–纤维间的相互作用提供了新的思路和研究基础。In the field of tissue regeneration,electrospun fibers with the characteristics of biomimicking fibrous structure of the native ECM (extracellular matrix) could provide an ideal platform for exploring the dynamic remodeling process of cells in their residing microenvironment.However,the compactness of conventional electrospun fibrous mats with the nanoscaled fibers crisscrossing and entangling with each other,eventually leads to a topological phenomenon similar to that of the nanofiber-like topography embossed on the surface of a solid matter.Therefore,it is impossible to accurately determine the remodeling effect of cells on the biomimicking fibrous microenvironment.In this study,electrospun fibrous matrices of GT/PCL (gelatin/polycaprolactone) with varied fiber densities were prepared by varying the collecting time of the GT/PCL fibers deposited on prior-silanized glass slides.Then,the effects of fiber-density on the cytomorphology,proliferation,ECM secretion,and matrix degradation of the C3H/10T1/2 cells were examined by performing CCK-8 assay,cytoskeleton staining,SEM imaging,qRT-PCR,ELISA,and immunofluorescence staining.The results showed that the cell viability was correlated with the fiber-density,and there existed a suitable fiber-density (103.0±13.2) μg/cm^(2) from which cells remodeled the fibrous microenvironment effectively.This study paved the way for the precise design of biomimetic fibrous matrix for tissue regeneration and laid a foundation for furthering our understanding of cell-fiber interactions.

关 键 词：电纺仿生纤维 纤维密度 细胞外基质 基质降解 重塑 

分 类 号：R318.08[医药卫生—生物医学工程]