Rapid preparation of high-precision hydrogel micropatterns and its induction of cell behavior

Zhang Weicai; Zheng Meiling; Dong Xianzi; Liu Jie; Jin Feng

doi:10.12086/oee.2022.210336

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Zhang W C, Zheng M L, Dong X Z, et al. Rapid preparation of high-precision hydrogel micropatterns and its induction of cell behavior[J]. Opto-Electron Eng, 2022, 49(2): 210336. doi: 10.12086/oee.2022.210336

Citation:

Zhang W C, Zheng M L, Dong X Z, et al. Rapid preparation of high-precision hydrogel micropatterns and its induction of cell behavior[J]. Opto-Electron Eng, 2022, 49(2): 210336. doi: 10.12086/oee.2022.210336

Rapid preparation of high-precision hydrogel micropatterns and its induction of cell behavior

1.
Laboratory of Organic NanoPhotonics and CAS Key Laboratory of Bio-Inspired Materials and Interfacial Science, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, Beijing 100190, China
2.
School of Future Technology, University of Chinese Academy of Sciences, Beijing 101407, China

Fund Project: National Natural Science Foundation of China (61975213, 61475164, 51901234), Cooperative R&D Projects between Austria, FFG and China, CAS (GJHZ1720), and International Partnership Program of Chinese Academy of Sciences (GJHZ2021130)

More Information

Corresponding author: zhengmeiling@mail.ipc.ac.cn

Received Date 21 October 2021

Revised Date 14 January 2022

Published Date 25 February 2022

Abstract

Abstract

In order to investigate the regulation effect of hydrogel micropatterns of different sizes on cell behaviors, large-area polygons and polygonal microstructures of different sizes were achieved by a femtosecond laser maskless optical projection lithography technique, in combination with a large-area splicing method. The optimum processing conditions and the wettability of the microstructure are studied in detail. The coculture experimental results of fibroblasts with patterned substrates showed that the spatial confinement in the microstructure can change the cell morphology and effectively regulate the cell growth behaviors. In particular, in the small polygonal and polygonal star microstructure, the nucleus tends to fall into the concave lacunae of the microstructure, and the spreading of cytoskeleton gradually tends to be consistent with the morphology of microstructures. This study indicates that the size of microstructural pattern units is very important for inducing cell behavior and function, which would provide a new technology and new method for cell research in vitro by using biocompatible hydrogel microstructures.
- maskless optical projection lithography /
- hydrogel /
- micropattern /
- cell behavior

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References

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Overview

Overview

Previous researches have indicated that pattern surfaces usually exhibit contact induction effects on cells. The microstructure can effectively regulate the adhesion and proliferation of cells. Cell morphology and cellular function are inseparable. For example, stem cells have a large degree of differentiation of different lineage depending on the adhesion form of the cell. Nanostructures affect cell adhesion behavior at the molecular level. Nanowires have been used to stimulate and record individual neuronal activities, and nanopores can be used for single-cell detection. Therefore, it will bring significant contributions to the fields of organizational engineering to create a variety of scales, different topological substrate in vitro to investigate cell-material, cell-environmental interaction mechanism.

Hydrogels are commonly used in tissue engineering since they have a similar component to the extracellular matrix and have advantages such as good biocompatibility, non-toxic and degradability. A large number of bionic cell scaffolds have been prepared by techniques such as microfluidic technology, 3D printing, soft printing, self-assembly, ultraviolet photolithography etc. This provides important guidance for cancer transfer, wound healing and inflammatory treatment. However, the sizes of these microstructures are usually large with a relatively low structure resolution, accounting for the limitation of hydrogel properties and manufacturing methods. Thus, it is still a challenge to rapidly fabricate large-area hydrogel micro/nano-structures with complex and arbitrary patterns.

In this paper, we have prepared the hydrogel by using PEGDA, PE-3A and the mixing photoinitiators (Irgure 369 and Benzil) with the weight ratios of 39.2: 59.2: 0.8: 0.8. A home-made maskless optical projection photolithography system is used to fabricate hydrogel patterns. A femtosecond laser, as the light source and high numerical aperture objective lens are used to improve structural resolution, and a splicing method is used to obtain large-area structures with high processing efficiency. Large-area polygons and polygonal stars with cavities in the center of itself are fabricated. First, the optimum processing conditions and the wettability of substrate with different patterns are studied. Then, Fibroblasts L929 are cultured on all kinds of pre-fabricated patterns. The cell behavior in micropattern with large cavity is similar to those on 2D flat substrates. Only the skeleton of cells close to the microstructure will produce deformation and interaction. For micropatterns with small sizes of cavities, the length of cells is significantly reduced because of the small space limit. In particular, the cell skeleton on the small-sized micropatterns exhibits a consistent distribution of the topography. The nuclear would fall into the center depression likes “bone trap” due to gravity. This study indicates that the size of microstructural pattern units is very important for inducing cell behavior and function, which would provide a novel method to prepare hydrogel micropatterns to study cell behavior in the field of organizational engineering.