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We present the development and characterization of Methylene-Blue-loaded nanoparticles (MB-NP) as well as their phototherapeutic application in an in vivo rat glioma model. The photo sensitizer, Methylene-Blue was covalently conjugated to the nanoparticle matrix to prevent the potential for leaching and non-specific staining of target and ancillary tissues. In vitro experiments demonstrate high efficiency in producing reactive oxygen species (ROS) and concomitant cytotoxicity. In vivo experiments were performed by first optimizing therapeutic parameters such as nanoparticle surface modification by attachment of F3-peptide targeting moieties, dose/concentration administered, incubation time, lightfluence and time of illumination. The quantitative growth patterns of the glioma were determined through visual observation of the tumorigenic response to various treatment parameters, using a cranial window model in vivo. The cranial window permitted non-invasive external illumination of the tumor and evaluation of loading of tumors with nanoparticles, the effects of particle targeting on retention in situ, and efficacy. The targeted MB-NP produced significant delay in tumor growth compared to control groups, demonstrating some potential advantages of nanoparticle-based PDT and its promise for the rapid eradication of local tumors. Furthermore, this approach holds promise as a potential surgical adjuvant palliation of any remaining tumor mass.

Towards Glioma Intra-operative Photodynamic Therapy: Targeted Photo-active Nanoparticles in a Brain Tumor Window Model