Abstract
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.
Highlights
The three-dimensional order (3D) microenvironment complexity contributes to regulate tumor growth in vivo [1,2,3,4]
The use of cell culture plates and wells is efficient, convenient, and lends itself to high-throughput processing, cell morphology and function can vary greatly based on the used extracellular matrix, which can potentially skew the results of growth, proliferation, differentiation, and chemo-/radiotherapy studies [17,18,19,20]
We developed a simple and green method utilizing substrates derived from butterfly wings with natural anisotropic microstructures
Summary
The three-dimensional order (3D) microenvironment complexity contributes to regulate tumor growth in vivo [1,2,3,4]. Biomimetics 2018, 3, 2 suited to growing hepatocytes into aggregates in order to mimic the in vivo liver microenvironment. As tumor formation and growth stems from specific gene expression profiles, a cancer cell line that may have undergone multiple passages may produce biased results. Hepatocytes cultured on two-dimensional (2D) monolayer models have some limitations as shown in a drug screening study where cultured hepatocytes resulted in a decrease of liver-specific functionality and gene expression, and viability [25,26,27]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
