Purpose: Glioblastoma Multiforme (GBM) is the most common and most aggressive form of brain cancer. There is currently no cure for GBM. Treatment is largely palliative and usually includes surgery followed by radiation therapy and/or chemotherapy. Hence, there is intensive research for new therapies. Nanoparticle-mediated therapy (NPRT) is an emerging option. The goal of this research is to advance NPRT for GBM using fluorescence guided cell morphometry.
Methods: Clinically relevant doses were delivered to two GBM cell lines (T98G and U87), along with several nanoparticles including carbon quantum dots, graphene quantum dots and CdSe(ZnS) quantum dots. Real time cell migration measurements were done using an electric cell impedance sensing device (ECIS). Clonogenic assays were performed using CytoSMART Omni, a cloud-based real-time cell imaging device. Fluorescence images were taken using both Hoechst and Calcein dyes. These fluorescent dyes stain nucleic acid and cytoplasm, respectively. Morphometric analysis was done using the ImageJ software and parameters such circularity, nuclear to cytoplasm ration, mitotic cell count, were extracted.
Results: The mitotic cell ratios between all conditions provided the most distinguishing criterion and holds promise as an early read-out of possible treatment outcomes. Preliminary results suggest that the use of carbon quantum dots in conjunction with radiotherapy significantly decreases the mitotic cell ratio within 24 hours of treatment. This trend also fits with the 14-day and 21-day survival fractions from clonogenic assays.
Conclusion: Fluorescence imaging enabled very early detection of parameters possibly predictive of treatment outcomes for various nanoparticles tested for their ability to enhance radiotherapy against glioblastoma cells. These findings may facilitate the development of effective NPRT against brain tumors.
Radioimmunotherapy, Radiation Therapy, Morphology