Abstract
Three-dimensional (3D) micronano structures have attracted much attention in tissue engineering since they can better simulate the microenvironment in vivo. Two-photon polymerization (TPP) technique provides a powerful tool for printing arbitrary 3D structures with high precision. Here, the desired 3D biocompatible hydrogel microscaffolds (3D microscaffold) with structure design referring to fibroblasts L929 have been fabricated by TPP technology, particularly considering the relative size of cell seed (cell suspension), spread cell, strut and strut spacing of scaffold. Modulation of the cell behavior has been studied by adjusting the porosity from 69.7% to 89.3%. The cell culture experiment results reveal that the obvious modulation of F-actin can be achieved by using the 3D microscaffold. Moreover, cells on 3D microscaffolds exhibit more lamellipodia than those on 2D substrates, and thus resulting in a more complicated 3D shape of single cell and increased cell surface. 3D distribution can be also achieved by employing the designed 3D microscaffold, which would effectively improve the efficiency of information exchange and material transfer. The proposed protocol enables us to better understand the cell behavior in vivo, which would provide high prospects for the further application in tissue engineering.
Highlights
Accepted: 3 September 2021Damage or defect of tissues and organs from traumas and tumors seriously threaten human health, the repair and reconstruction of these defects has emerged as one of the challenges in modern medicine
A large number of studies have focused on the influence of the porosity and pore size of the 3D microscaffold on cell behaviors
Hyclone Dulbecco’s Modified Eagle Medium (DMEM)/High Glucose culture medium was purchased from GE Healthcare Life Sciences (Logan, Utah, USA)
Summary
Accepted: 3 September 2021Damage or defect of tissues and organs from traumas and tumors seriously threaten human health, the repair and reconstruction of these defects has emerged as one of the challenges in modern medicine. Tissue engineering is based on a small number of cells and supported by biological materials for trauma-induced repair and in vitro tissue reconstruction [1,2]. With the continuous development of three-dimensional (3D) scaffolds that mimic the real microenvironment in vivo [5,6,7,8], investigations of scaffold-induced cell behavior for wound repair and tissue healing are becoming a new research focus [9,10,11]. Previous studies have considered the induction behavior of cells modulated by the parameters of 3D scaffolds such as pore size [12,13,14,15], pore shape [16,17,18], surface roughness [19,20,21], support strength [22,23], and wettability [21]. It is worth mentioning that the struts of these scaffolds mostly overlap in the vertical direction, forming a space similar to a “patio” [24,25], and the struts and pores are either much smaller than the cell seed (cell suspension) [26], or much larger than
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