超细纤维
肝细胞
细胞外
组织工程
细胞生物学
细胞外基质
化学
微流控
材料科学
生物物理学
纳米技术
生物医学工程
体外
生物化学
工程类
生物
复合材料
作者
Rie Utoh,Sakiko Enomoto,Masao Yamada,Keigo Yamanaka,Yuya Yajima,Kazuya Furusawa,Minoru Seki
标识
DOI:10.1016/j.msec.2021.112417
摘要
Artificial biological scaffolds made of extracellular matrix (ECM) components, such as type I collagen, provide ideal physicochemical cues to various cell culture platforms. However, it remains a challenge to fabricate micrometer-sized ECM materials with precisely controlled morphologies that could reconstitute the 3-dimensional (3D) microenvironments surrounding cells. In the present study, we proposed a unique process to fabricate fragmented collagen microfibers using a microfluidic laminar-flow system. The continuous flow of an acidic collagen solution was neutralized to generate solid fibers, which were subsequently fragmented by applying a gentle shear stress in a polyanion-containing phosphate buffer. The morphology of the fiber fragment was controllable in a wide range by changing the type and/or concentration of the polyanion and by tuning the applied shear stress. The biological benefits of the fragmented fibers were investigated through the formation of multicellular spheroids composed of primary rat hepatocytes and microfibers on non-cell-adhesive micro-vessels. The microfibers enhanced the survival and functions of the hepatocytes and reproduced proper cell polarity, because the fibers facilitated the formation of cell−cell and cell−matrix interactions while modulating the close packing of cells. These results clearly indicated that the microengineered fragmented collagen fibers have great potential to reconstitute extracellular microenvironments for hepatocytes in 3D culture, which will be of significant benefit for cell-based drug testing and bottom-up tissue engineering. • Type I collagen-based microfibers were microfluidically engineered. • Polyanion solutions triggered controllable fragmentation of the collagen microfiber. • Primary hepatocytes formed composite spheroids with the fragmented microfibers. • The fragmented collagen microfibers remodeled the extracellular microenvironments.
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