Exploring the Therapeutic Potential of T₃C₂ MXene Using an In-Ovo Model

    MXenes, a new class of 2D nanomaterials, holds promises for revolutionizing various biomedical treatments. Their unique combination of a large surface area and specific chemical properties makes them ideal candidates for various applications, including biomedicine. In particular, MXenes demonstrated significant potential as photosensitizers, paving the way for a powerful infrared laser photothermal therapy of cancer. However, further exploration is necessary to fully unlock the full therapeutic potential of MXenes.

    The in-ovo model is gaining attention as a more ethical and cost-efficient in-vivo alternative to the traditional animal based testing. This approach allows to study the impact of treatments, drugs, or specific conditions directly on developing embryos within a fertilized egg, offering a valuable and economic in-vivo method.

    Melanoma, a highly aggressive form of skin cancer, can spread rapidly throughout the body if left undetected. Relatively easy accessibility of melanoma makes it a promising candidate for treatment with photothermal therapy (PTT). This approach is based on the ability of the light of certain wavelengths to penetrate to some extent into the living tissues. Thus, given the superficial nature of melanoma, PTT can be a promising method for targeting of melanoma by low invasive treatment options. Therefore, in this project we aimed at investigating conditions for efficient tumor ablation of melanoma with MXene as the photosensitizer using an in-ovo chicken embryo model.

    B16F10 mouse melanoma cells were cultured under standard conditions in a DMEM/F12 medium supplemented with 10% FBS. The cells were loaded with Ti3C2 MXene at 6,25 µg/ml concentration. The fertilized chicken eggs were incubated for 10 days in a standard egg incubator under recommended conditions. Then, a window was created in the egg shell and 2-3 million cells suspended in 50 µl of medium with 5 mg/ml hyaluronic acid were grafted onto the chorioallantoic membrane (CAM). To enhance tumor formation efficiency, the CAM was subjected to a brief, 20-second irritation with filter paper just before cell engraftment. Then, 6-7 days post-inoculation, the eggs were irradiated with a 1064 nm pulsed laser for 80 sec at 3.1 J/cm2 power density, 200 msec pulses at 1 Hz.

    The melanoma tumor could be efficiently grown on chicken CAM. The tumor can reach up to 2.5x1.6x1 cm (width x height x thickness) in size and of up to 1.7 grams of weight. The immunohistochemistry analysis confirmed the melanoma nature of the xenograft tumor. Tumors were successfully grown also with the MXene loaded cells. Currently, the experiment is still in progress to figure out possible changes in the structure and dynamics of tumor development after irradiation.