Assessment of Safety and Anti-Cancer Potential of Chicken Feather Keratin Nanoparticles Encapsulated Doxorubicin in Chicken Chorioalantoic Membrane: An Alternate to Animal Model

Abstract

Background: Nanoencapsulation of chemotherapeutics can improve the therapeutic index of chemotherapeutics like doxorubicin (Dox) by reducing systematic toxicity and enhancing tumor targeting. Methods: This study reports on the synthesis, characterization and in vivo evaluation of a novel nanokeratin encapsulated doxorubicin (NKD). The formulation was tested against human breast cancer tumor cell line (MCF-7) using chorioallantoic membrane (CAM) of chicken embryo as an alternate model to animal experiments and real-time PCR was used for apoptosis related genes expression. Results: NKD exhibited desirable physicochemical properties, including a particle size of 150 nm and a negative zeta potential. The in vivo CAM model demonstrated that NKD was not toxic to the embryo, showing a 100% survival rate. This stands in contrast to free doxorubicin, which induced moderate toxicity and a significant decrease in embryo survival. Furthermore, quantitative real- time PCR findings showed a significant upregulation of pro-apoptotic gene BAX and p53 in NKD compared to Dox. Histopathological analysis showed reduced tumor cell density, increased apoptosis and signs of tumor architecture returning to normal compared to control group. Conclusion: These findings suggest that the nanokeratin based formulation is a safe and effective strategy for doxorubicin delivery, with significant potential for further development in breast cancer therapy.