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Hepatocyte Aggregate Formation on Chitin-Based Anisotropic Microstructures of Butterfly Wings

by Abdelrahman Elbaz 1,2, Bingbing Gao 1,2, Zhenzhu He 1,2 and Zhongze Gu 1,2,3,*
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China
National Demonstration Center for Experimental Biomedical Engineering Education, Southeast University, Nanjing 210096, China
Laboratory of Environment and Biosafety Research Institute of Southeast University in Suzhou, Suzhou 215123, China
Author to whom correspondence should be addressed.
Biomimetics 2018, 3(1), 2;
Received: 5 December 2017 / Revised: 9 January 2018 / Accepted: 13 January 2018 / Published: 18 January 2018
(This article belongs to the Special Issue Biomimetic Nanotechnology)
Scaffold nanotopography plays the most significant role in the mimicry of the in vivo microenvironment of the hepatocytes. Several attempts have been made to develop methods and substrates suited to growing hepatocytes into aggregates. Functional biomaterials, particularly biodegradable polymers, have been used in several studies aimed to develop improved scaffolds with ordered geometry and nanofibrous architecture for tissue engineering. However, there are still some limitation in their fabrication: it is not cost-efficient, is time-consuming, and exhibits some technological complications. The synthetic scaffolds are usually non-biodegradable and can be non-biocompatible compared to the naturally derived biomaterials. Here, we utilized a simple, cost-effective, and green method with two-step chemical treatment to get more selected hydrophilic butterfly wings from Morpho menelaus, Papilio ulysses telegonus, and Ornithoptera croesus lydius as a chitin-based natural scaffolds to growing hepatocyte aggregates. We established a three-dimensional (3D) in vitro model for culture of HepG2 cells and aggregate formation that maintained the hepatocytes function on these natural anisotropic microstructures. Cells cultured on these substrates show higher viability than those cultured on a two-dimensional (2D) culture plate. Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay results revealed excellent viability of HepG2 cells on P. u. telegonus wings (fibrous area). The results also demonstrated appropriate cell activity, cell retention, and stable and functional expression in terms of albumin secretion and urea synthesis activity compared to the 2D monolayer culture of hepatocytes on the culture dish surface. With a slightly different degree, the other substrates also shown similar results. We anticipate that these natural anisotropic, biodegradable, and biocompatible substrates can maintain long-term hepatic culture as an in vitro 3D model for potential therapeutic applications and regenerative tissue applications. The model presented here provides a feasible alternative to the synthetic scaffolds and is expected to be more reliable for 3D organotypic liver culture models based on such scaffolds. View Full-Text
Keywords: organotypic culture models; cell aggregate; HepG2; 3D scaffolds; anisotropic microstructure organotypic culture models; cell aggregate; HepG2; 3D scaffolds; anisotropic microstructure
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Elbaz, A.; Gao, B.; He, Z.; Gu, Z. Hepatocyte Aggregate Formation on Chitin-Based Anisotropic Microstructures of Butterfly Wings. Biomimetics 2018, 3, 2.

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