3D Silk Fibroin-Gelatin/Hyaluronic Acid/Heparan Sulfate Scaffold Enhances Expression of Stemness and EMT Markers in Cholangiocarcinoma

In Vivo. 2022 May-Jun;36(3):1155-1167. doi: 10.21873/invivo.12815.

Abstract

Background/aim: Cholangiocarcinoma (CCA) is a stem cell-based cancer. The in vivo tumor microenvironment is not present in two-dimensional (2D) cultures, which is one of the limitations in cancer stem cell (CSC) research. Thus, we aimed to establish three-dimensional (3D) culture mimicking extracellular matrix (ECM) that could serve as a niche for CSC enrichment in CCA.

Materials and methods: Silk fibroin-gelatin/hyaluronic acid/heparan sulfate (SF-GHHs) scaffolds were fabricated by lyophilization in various ratios and compared to silk fibroin (SF) scaffold. The physical and biological characteristics of the scaffolds were investigated.

Results: The SF-GHHs 1:2 scaffold with pore size of 350±102 μm harbored optimal porosity, good water uptake, and stable beta-sheet that supported the increase in KKU-213A cell proliferation and aggregation. The CSC and the epithelial-mesenchymal transition (EMT) markers were significantly upregulated in this scaffold compared to 2D. Moreover, drug sensitivity against cisplatin and gemcitabine in 3D culture was significantly higher than that in 2D culture.

Conclusion: The SF-GHHs 1:2 scaffold could simulate ECM that may serve as a CSC niche of CCA, and reinforce stemness and EMT properties, suggesting its suitability for 3D CCA model, which supports CSC and new targeting drug research in CCA.

Keywords: Silk fibroin-based scaffold; cancer stem cell; cholangiocarcinoma; epithelial-mesenchymal transition; three-dimensional culture.

MeSH terms

  • Cholangiocarcinoma*
  • Epithelial-Mesenchymal Transition
  • Fibroins* / metabolism
  • Fibroins* / pharmacology
  • Gelatin
  • Heparitin Sulfate
  • Humans
  • Hyaluronic Acid
  • Tissue Engineering / methods
  • Tissue Scaffolds
  • Tumor Microenvironment

Substances

  • Gelatin
  • Hyaluronic Acid
  • Fibroins
  • Heparitin Sulfate