Functionalized Graphene Oxide Thin Films for Anti-tumor Drug Delivery to Melanoma Cells

Livia E Sima; Gabriela Chiritoiu; Irina Negut; Valentina Grumezescu; Stefana Orobeti; Cristian V A Munteanu; Felix Sima; Emanuel Axente

doi:10.3389/fchem.2020.00184

Functionalized Graphene Oxide Thin Films for Anti-tumor Drug Delivery to Melanoma Cells

Front Chem. 2020 Mar 23:8:184. doi: 10.3389/fchem.2020.00184. eCollection 2020.

Authors

Livia E Sima¹, Gabriela Chiritoiu¹, Irina Negut², Valentina Grumezescu², Stefana Orobeti^{1

2}, Cristian V A Munteanu³, Felix Sima², Emanuel Axente²

Affiliations

¹ Department of Molecular Cell Biology, Institute of Biochemistry, Romanian Academy, Bucharest, Romania.
² Photonic Investigations Laboratory, Center for Advanced Laser Technologies, National Institute for Lasers, Plasma and Radiation Physics, Magurele, Romania.
³ Department of Bioinformatics and Structural Biochemistry, Institute of Biochemistry, Romanian Academy, Bucharest, Romania.

Abstract

Since Graphene discovery, their associated derivate nanomaterials, Graphene Oxide (GO) and reduced-GO were in the forefront of continuous developments in bio-nano-technology due to unique physical-chemical properties. Although GO nano-colloids (GON) were proposed as drug release matrix for targeting cancer cells, there is still a concern regarding its cytotoxicity issues. In this study, we report on the fabrication of functional GON bio-coatings by Matrix-Assisted Pulsed Laser Evaporation (MAPLE) to be used as drug carriers for targeting melanoma cells. We first performed a thorough in vitro cytotoxicity assay for comparison between GON and protein functionalized GON coatings. As functionalization protein, Bovine Serum Albumin (BSA) was non-covalently conjugated to GO surface. Safe concentration windows were identified in cytotoxicity tests by live/dead staining and MTS assays for five different human melanoma cell lines as well as for non-transformed melanocytes and human dermal fibroblasts. Hybrid GON-BSA nano-scaled thin coatings incorporating Dabrafenib (DAB) and Trichostatin A (TSA) inhibitors for cells bearing BRAF^V600E pathway activating mutation were assembled on solid substrates by MAPLE technique. We further demonstrated the successful immobilization for each drug-containing GON-BSA assembling systems by evaluating cellular BRAF activity inhibition and histone deacetylases activity blocking, respectively. DAB activity was proven by the decreased ERK phosphorylation in primary melanoma cells (SKmel28 BRAF^V600E cell line), while TSA effect was evidenced by acetylated histones accumulation in cell's nuclei (SKmel23 BRAF WT cell line). In addition, melanoma cells exposed to GON-BSA coatings with compositional gradient of inhibitors evidenced a dose-dependent effect on target activity. Such functional bio-platforms could present high potential for cell-biomaterial interface engineering to be applied in personalized cancer therapy studies.

Keywords: MAPLE; cytotoxicity; drug delivery; graphene oxide; melanoma; nanomaterials; thin films.