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Fertility preservation is a critical concern for cancer patients, especially for those diagnosed with malignancies that carry a high risk of ovarian metastasis, such as leukemia and Burkitt lymphoma. Current methods, including ovarian tissue cryopreservation and transplantation, restore ovarian function in over 90% of cases, with more than 200 live births reported globally. However, transplantation is not allowed for patients with tumors at high metastasis risk due to the potential for reintroducing malignant cells. This creates a need for alternative strategies to restore fertility in such patients. Our laboratory is working on developing a transplantable engineered ovary to restore fertility in these patients. A 3D matrix of the engineered ovary plays a crucial role in protecting follicles during transplantation, maintaining cell-cell and cell-matrix interactions, and preserving follicular architecture. Mimicking the ovarian extracellular matrix (ECM) is essential but remains challenging using conventional tissue engineering methods, which struggle to replicate the complexity of natural tissues, including their physical structure, mass transfer, and oxygen diffusion. To overcome these limitations, advanced bioprinting techniques were employed. Collaborating with Aspect Biosystems and utilizing their RX1-Microfluidic 3D Bioprinter, we developed a novel oxygen-generating ovarian scaffold. This bioprinting system enables the creation of core-shell filaments, where the shell consists of alginate, PEGylated fibrinogen, and ovarian stromal cells, and the core comprises alginate, liposomal catalase, and liposomal hydrogen peroxide (H₂O₂). Optimization of bioprinting parameters was conducted, achieving ideal conditions: 75 mbar for the shell, 200 mbar for the core, 1 mbar for the crosslinker, a printing speed of 14 mm/s, and a nozzle distance of 0.6 mm. These parameters allowed the successful fabrication of cubic structures with the desired composition. Hypoxia significantly impacts follicular survival and development post-transplantation. To mitigate this, an oxygen-releasing liposomal system was developed, incorporating catalase and H₂O₂ into liposomes. This system demonstrated effective decomposition of cytotoxic H₂O₂, enhanced oxygen release, and maintained cell viability under hypoxic conditions in both 2D culture and cell-encapsulated hydrogels. This project represents a promising step toward addressing fertility preservation for cancer patients at high risk of ovarian metastasis. By leveraging advanced bioprinting and oxygenation strategies, we aim to develop a functional transplantable engineered ovary capable of restoring fertility.
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Dadashzadeh, A., Moghassemi, S., & Andrade Amorim, C. (2023). Bioprinting of an ovarian tissue. Discovery Ecosystem Virtual Symposium, Canada (Online). https://hdl.handle.net/2078.5/242713