Development of reduced graphene oxide-incorporated gelatin-polymannose hydrogel scaffolds as a 3D cell culture model for cancer research.

Authors

Hema Priya Manivannan, Vishnu Priya Veeraraghavan, Arul Prakash Francis

Published in

Journal of biomaterials science. Polymer edition. Pages 1-38. Jul 18, 2026. Epub Jul 18, 2026.

Abstract

In this study, we have created a 3D environment conducive to A549 cell growth by developing a novel gelatin-polymannose scaffold incorporated with reduced graphene oxide (Gel:OPM:rGO). The Gel:OPM:rGO scaffold was fabricated and characterised by FTIR, XRD and contact angle analysis, which confirmed the successful fabrication of hydrophilic scaffolds. Scanning electron microscopy (SEM) revealed a porous nanofibrous architecture with pore sizes ranging from 50 to 150 µm. Compression testing further demonstrated enhanced mechanical stability following rGO incorporation. A549 cells cultured on the Gel:OPM:rGO scaffold exhibited significantly higher proliferation than the Gel:OPM scaffold and were comparable to those observed in Matrigel (p < 0.01, n = 3). Live staining assays confirmed progressive cell migration and high cell viability. Confocal microscopy images further confirmed elevated ROS generation within the 3D scaffold, indicating the establishment of a hypoxia-like tumor microenvironment. Gene expression analysis showed elevated levels of HIF-1α, MMP2, β-catenin, and CDH1 in 3D cultures compared with 2D cultures. These findings were further supported by Western blot analysis, which showed increased expression of HIF-1α, MMP-2, β-catenin, and E-cadherin. Notably, the molecular expression profile of the Gel:OPM:rGO scaffold closely resembled that of Matrigel. Drug response was evaluated using doxorubicin (Dox) as a model anticancer drug. A549 cells cultured in the Gel:OPM:rGO scaffold and Matrigel exhibited higher inhibitory concentration of Dox when compared to the 2D culture. Our results suggest that the Gel:OPM:rGO scaffold is better suited for A549 cell culture and may serve as a preclinical 3D lung cancer model for drug testing.

PMID:
42471034
Bibliographic data and abstract were imported from PubMed on 19 Jul 2026.

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