Theranostic Potential of Adaptive Cold Atmospheric Plasma with Temozolomide to Checkmate Glioblastoma: An In Vitro Study

SIMPLE SUMMARY: Glioblastoma (GBM) is an aggressive form of brain cancer. Here, we present a combination therapy of cold atmospheric plasma (CAP) and temozolomide (TMZ) to treat GBM in vitro. We analyze the effects of the co-treatment in two GBM (TMZ-resistant and -sensitive) cell lines. The aim of this study is mainly to sensitize these cells using CAP so that they respond well to TMZ. We further found that the removal of cell culture media after CAP treatment does not affect the sensitivity of CAP to cancer cells but enhances the effects of TMZ. However, it was observed in our study that keeping the CAP-treated media for a shorter time did not significantly inhibit T98G cells. Interestingly, keeping the same plasma-treated media for a longer duration resulted in a decrease in cell viability. On the contrary, TMZ-sensitive cell A172 responded well to the co-treatment. This could be a potential reason for the sensitization of the combination therapy. ABSTRACT: Cold atmospheric plasma (CAP) has been used for the treatment of various cancers. The anti-cancer properties of CAP are mainly due to the reactive species generated from it. Here, we analyze the efficacy of CAP in combination with temozolomide (TMZ) in two different human glioblastoma cell lines, T98G and A172, in vitro using various conditions. We also establish an optimized dose of the co-treatment to study potential sensitization in TMZ-resistant cells. The removal of cell culture media after CAP treatment did not affect the sensitivity of CAP to cancer cells. However, keeping the CAP-treated media for a shorter time helped in the slight proliferation of T98G cells, while keeping the same media for longer durations resulted in a decrease in its survivability. This could be a potential reason for the sensitization of the cells in combination treatment. Co-treatment effectively increased the lactate dehydrogenase (LDH) activity, indicating cytotoxicity. Furthermore, apoptosis and caspase-3 activity also significantly increased in both cell lines, implying the anticancer nature of the combination. The microscopic analysis of the cells post-treatment indicated nuclear fragmentation, and caspase activity demonstrated apoptosis. Therefore, a combination treatment of CAP and TMZ may be a potent therapeutic modality to treat glioblastoma. This could also indicate that a pre-treatment with CAP causes the cells to be more sensitive to chemotherapy treatment.