Abstract
Tissue engineering technology has made major advances with respect to the repair of injured tissues, for which scaffolds and cells are key factors. However, there are still some issues with respect to the relationship between scaffold and cell growth parameters, especially that between the pore size and cells. In this study, we prepared scaffolds with different pore sizes by melt electrowritten (MEW) and used bone marrow mensenchymal stem cells (BMSCs), chondrocytes (CCs), and tendon stem cells (TCs) to study the effect of the scaffold pore size on cell adhesion, proliferation, and differentiation. It was evident that different cells demonstrated different adhesion and proliferation rates on the scaffold. Furthermore, different cell types showed differential preferences for scaffold pore sizes, as evidenced by variations in cell viability. The pore size also affected the differentiation and gene expression pattern of cells. Among the tested cells, BMSCs exhibited the greatest viability on the 200-μm-pore-size scaffold, CCs on the 200- and 100-μm scaffold, and TCs on the 300-μm scaffold. The scaffolds with 100- and 200-μm pore sizes induced a significantly higher proliferation, chondrogenic gene expression, and cartilage-like matrix deposition after in vitro culture relative to the scaffolds with smaller or large pore sizes (especially 50 and 400 μm). Taken together, these results show that the architecture of 10 layers of MEW scaffolds for different tissues should be different and that the pore size is critical for the development of advanced tissue engineering strategies for tissue repair.
Highlights
In recent years, tissue engineering technology has made tremendous contributions to tissue regeneration (Guo et al, 2018; Cunniffe et al, 2019; Ruvinov et al, 2019)
We have developed many intricate methods to generate complex 2D models or prototypes with mechanical or chemical gradients, these culture conditions may not apply to many cell types (Karageorgiou and Kaplan, 2005; Tanaka et al, 2010; Panadero et al, 2016)
The following reagents were used in this study: PCL, a-DMEM, DMEM/F12 (HyClone, UT, United States), fetal bovine serum (FBS; Gibco, NY, United States), TRITC and FITC phalloidin (R415 and A12379, Invitrogen, CA, United States), 4,6-diamidino-2-phenylindole (DAPI, C0060, MaoKang, Shanghai, China), cell counting kit-8 (CCK-8; Dojindo, Kumamoto, Japan), primary antibodies anti-collagen type II (COL II; ab185430), anti-collagen type I (COL I; ab6308), anti-aggrecan (AGC; ab3778), SOX-9 (SOX9; ab76997; Abcam, Cambridge, United Kingdom), Alexa Fluor 555-conjugated anti-mouse antibody (A32727, Invitrogen, CA, United States), RNeasy mini kit (Qiagen, Hilden, Germany), SuperScriptTM III Reverse Transcriptase (Bimake, Shanghai, China), and Sirius red solution, Safranin, and fix-green solution (YifanBio, Shanghai, China)
Summary
Tissue engineering technology has made tremendous contributions to tissue regeneration (Guo et al, 2018; Cunniffe et al, 2019; Ruvinov et al, 2019). Tissue engineering technology simulates the regeneration of tissues and organs by combining elements such as biological materials, cells, and biologically active molecules to mimic the structure and function of native tissues and organs (Kang et al, 2018; Kumai et al, 2019). These studies reveal that tissue engineering scaffolds are vital components, whose composition and structure affect the proliferation, differentiation, and gene expression in cells (Mannoor et al, 2013; Li et al, 2017; Schon et al, 2017; Wang et al, 2018). The specific effects of the micro-architecture on cells, especially that of the scaffold pore size, are not completely clear
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