Preparation and Characterization of a Novel Triple Composite Scaffold Containing Silk Fiborin, Chitosan, and Alginate for 3D Culture of Colonic Carcinoma Cells In Vitro

Abstract

Background Three-dimensional (3D) cell-culture scaffolds are ideal in vitro models to bridge the gap between two-dimensional cell culture in vitro and in vivo cancer models. Construction of 3D scaffolds using two kinds of biomaterials has been reported, but there are still many defects. To improve the performance of the scaffolds for 3D cell culture of colonic carcinoma (CC) cells in vitro , we attempted to construct triple composite scaffolds using silk fibroin (SF), chitosan (Cs), and alginate (Alg). Material/Methods We explored the suitability of triple composite scaffolds of SF/Cs/Alg at ratios of 1: 1: 0.5, 1: 1: 1, and 1: 1: 2 for 3D culture of CC cells, and used the dual composite scaffold of SF/Cs (1: 1) as a control group. We analyzed the physicochemical characteristics of these scaffolds and studied cell adhesion, cell proliferation, migration, colony-forming ability, microstructure and ultrastructure, and spheroid-forming capacity of the commercially available CC cell line HCT-116 on the prepared scaffolds. Results Our results show that SF/Cs/Alg (1: 1: 1) scaffolds demonstrated the best profile, the highest uniform porosity and connectivity, and excellent hydroscopicity, and also exhibited appropriate and controlled swelling and degradation characteristics. The adhesion, proliferation, colony-forming, and wound-healing assays, green fluorescent protein-labeled HCT116 cell imaging, 4′,6-diamidino-2-phenylindole and DY-554-phalloidin staining, scanning electron microscopy, and haematoxylin and eosin staining revealed that the triple composite scaffolds of SF/CS/Alg (1: 1: 1) supported cell adhesion, proliferation, migration, colony-forming ability, and spheroid formation far better than the dual composite scaffold of SF/CS (1: 1). Conclusions This study successfully demonstrated the potential of SF/Cs/Alg (1: 1: 1) scaffold as an alternative for the 3D in vitro culture of CC cells.

High-Throughput Dispensing of Viscous Solutions for Biomedical Applications

“…To demonstrate the ability of the scaffold dispenser to uniformly distribute scaffold materials into microplates, two exemplary polymer solutions were chosen. Chitosan and alginate are two widely used biocompatible materials that achieve excellent cell models when they are used as the substrate for cell culture [ 35 , 36 , 37 , 38 , 39 ]. Chitosan, a derivative of chitin, is a natural cationic polysaccharide derived from shrimp shells; alginate is a natural anionic polysaccharide derived from brown algae.…”

Evaluation of Anticancer and Cytotoxic Effects of Genistein on PC3 Prostate Cell Line under Three-Dimensional Culture Medium

“…Recent scientific reports have shown that GNT has anticancer effects on breast [27] , colon [28] , lung [29] , stomatch [30] , and prostate [31] tissues in a 2D culture medium. However, in many cases, the toxicological results of 2D cell culture are not very reliable for use in human experiments; therefore, 3D cell culture studies have been suggested to produce more accurate results and also to reduce the cost of research [32] . Based on the literature survey, the addition of GNT to LNCaP PCa cells at concentrations more than 10 µM suppressed PCa cell growth for three days [33] .…”

Development of a Composite Hydrogel Containing Statistically Optimized PDGF-Loaded Polymeric Nanospheres for Skin Regeneration: In Vitro Evaluation and Stem Cell Differentiation Studies

“…At specific time intervals, the scaffolds were removed from the PBS solution, and after drying their surface, their weight was measured and recorded with an analytical balance. The swelling ratio of the scaffolds was then obtained from the following equation. , swelling .25em ratio = W 2 − W 1 W 1 where W 1 and W 2 are the initial weight of the scaffolds and the weight of the scaffolds after swelling, respectively.…”